MICROORGANISMS AND METHODS FOR THE PRODUCTION OF FATTY ACIDS AND FATTY ACID DERIVED PRODUCTS

Abstract

This invention relates to metabolically engineered microorganism strains, such as bacterial strains, in which there is an increased utilization of malonyl-CoA for production of a fatty acid or fatty acid derived product, wherein the modified microorganism produces fatty acyl-CoA intermediates via a malonyl-CoA dependent but malonyl-ACP independent mechanism.

Claims

1-42. (canceled)

43. A genetically modified organism comprising: a heterologous nucleic acid encoding a first NphT7 polypeptide; and a heterologous nucleic acid encoding a second NphT7 polypeptide, the second NphT7 polypeptide comprising one or more amino acid substitutions at a position selected from the group consisting of Ser84, Thr85, Gln90, Val114, Tyr144, Ile147, Val157, Phe159, Ile194, Val196, Phe217, Gly288, and Gly318, wherein said microorganism produces a fatty acid or fatty acid-derived product having a carbon chain length of C6 or greater.

44. The genetically modified organism of claim 43 wherein the second NphT7 polypeptide comprises amino acid substitutions at Ile147 and Phe217.

45. The genetically modified organism of claim 43 wherein the second NphT7 polypeptide comprises amino acid substitutions of I147S and F217V.

46. The genetically modified organism of claim 43, wherein the fatty acid or fatty acid-derived product has a carbon chain length of C8 or greater.

47. The genetically modified organism of claim 43, wherein the fatty acid or fatty acid-derived product has a carbon chain length of C10 or greater.

48. The genetically modified organism of claim 43, wherein the microorganism further comprises: a heterologous nucleic acid encoding a ketoacyl-CoA reductase; a heterologous nucleic acid encoding a hydroxyacyl-CoA dehydratase; and a heterologous nucleic acid encoding an enoyl-CoA reductase.

49. The genetically modified organism of claim 48, wherein: the ketoacyl-CoA reductase is selected from the group consisting of SEQ ID NO: 183 and SEQ ID NO: 137; the hydroxyacyl-CoA dehydratase is selected from the group consisting of SEQ ID NO: 183, SEQ ID NO: 273, and SEQ ID NO: 274; and the enoyl-CoA reductase is SEQ ID NO: 275.

50. The genetically modified organism of claim 43 further comprising a heterologous nucleic acid encoding a polypeptide selected from the group consisting of SEQ ID NO: 3-120.

51. The genetically modified organism of claim 43 further comprising a heterologous nucleic acid sequence encoding a termination enzyme that catalyzes production of the fatty acid-derived product selected from the group comprising a fatty alcohol, a fatty aldehyde, a fatty alkene, a fatty amide, a fatty ester, a fatty alkane, and a fatty diacid.

52. The genetically modified organism of claim 51, wherein: the termination enzyme comprises a wax ester synthase; and the fatty acid-derived product is a fatty acid ester.

53. The genetically modified organism of claim 52, wherein the wax ester synthase comprises an amino acid sequence of at least 70% homology to any one of SEQ ID NO 289, SEQ ID NO 290, SEQ ID NO 291, and SEQ ID NO 292.

54. The genetically modified organism of claim 52 wherein the second NphT7 polypeptide comprises amino acid substitutions at Ile147 and Phe217.

55. The genetically modified organism of claim 52 wherein the second NphT7 polypeptide comprises amino acid substitutions of I147S and F217V.

56. The genetically modified organism of claim 52, wherein the fatty acid ester has a carbon chain length of C8 or greater.

57. The genetically modified organism of claim 52, wherein the fatty acid ester has a carbon chain length of C10 or greater.

58. The genetically modified organism of claim 52, wherein the microorganism further comprises: a heterologous nucleic acid encoding a ketoacyl-CoA reductase; a heterologous nucleic acid encoding a hydroxyacyl-CoA dehydratase; and a heterologous nucleic acid encoding an enoyl-CoA reductase.

59. The genetically modified organism of claim 58, wherein: the ketoacyl-CoA reductase is selected from the group consisting of SEQ ID NO: 183 and SEQ ID NO: 137; the hydroxyacyl-CoA dehydratase is selected from the group consisting of SEQ ID NO: 183, SEQ ID NO: 273, and SEQ ID NO: 274; and the enoyl-CoA reductase is SEQ ID NO: 275.

60. The genetically modified organism of claim 52 further comprising a heterologous nucleic acid encoding a polypeptide selected from the group consisting of SEQ ID NO: 3-120.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0047] FIG. 1 is a chart showing the carbon chain length distribution in oleochemical feedstocks used to make fatty acids and fatty acid derivatives.

[0048] FIG. 2 is a diagram that illustrates various complete bioproduction pathways of the present invention, and provides a representative example of the conversion of various carbon sources to a C10 fatty acid or C10 fatty ester.

[0049] FIG. 3 is a diagram showing production of even chain fatty acids using acetyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule.

[0050] FIG. 4 is a diagram showing production of even chain fatty acid esters using acetyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule.

[0051] FIG. 5 is a diagram showing production of odd chain fatty acids using propionyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule.

[0052] FIG. 6 is a diagram showing production of odd chain fatty acid esters using propionyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule.

[0053] FIG. 7-FIG. 14 are a series of various reaction pathways in accordance with the present invention.

[0054] FIG. 15 is a bar chart showing the formation of acyl-CoA products produced with NphT7 variants and NphT7 mutants acting (100 g, 30 min. assays) on Malonyl-CoA and C4-, C6-, or C10-CoA to generate the corresponding C6-, C8- and C12-hydroxyacyl-CoA in the presence of PaFabG hydroxyacyl-CoA reductase.

[0055] FIG. 16 is a bar chart showing activity of thioesterases on acyl-CoA substrates of different carbon chain lengths.

[0056] FIG. 17 is a bar chart showing production rates of C8-C18 free fatty acids produced in different bacteria strains.

[0057] FIG. 18 is a bar chart showing titers of C6-C18 free fatty acids produced in different bacteria strains.

[0058] FIG. 19 is a pie chart showing the chain length specificity distribution of free fatty acids produced in strain sample 3.

[0059] FIG. 20 is a bar chart showing the chain length specificity preference of different 3HDh.

[0060] FIG. 21 is a graph showing the fatty acid carbon chain length distribution produced by various microorganisms in accordance with the present invention.

[0061] FIG. 22 includes a series of pie charts showing the fatty acid carbon chain length distribution produced by various microorganisms in accordance with the present invention.

[0062] FIG. 23 is a bar chart showing the amounts of C4, C6, and C8 free fatty acid produced by various genetically modified microorganisms in accordance with the present invention.

[0063] FIG. 24 is a bar chart showing the amounts of total fatty acids (C4-C18) produced by various genetically modified microorganisms in accordance with the present invention.

[0064] FIG. 25 includes a series of pie charts showing the distribution of free fatty acids produced by various genetically modified microorganisms in accordance with the present invention.

[0065] FIG. 26 shows a fatty acid pathway that comprises four steps which utilizes a pathway that is similar to the type II fatty acid synthesis (FAS) system utilized by bacteria. Both fatty acid syntheses are shown in FIG. 26. A. In step 1, 3-ketoacyl-CoA synthase catalyzes the condensation of acyl-CoA (or acetyl-CoA at initial step of chain elongation) with malonyl-CoA to yield -ketoacyl-CoA. In the subsequent steps, -ketoacyl-CoA undergoes reduction by -ketoacyl-CoA reductase (step 2), dehydration by -hydroxyacyl-CoA dehydratase (step 3), and a final reduction by enoyl-CoA reductase (step 4). Reactions are repeated and each cycle adds two carbons to the acyl-CoA chain. (B) Type II FAS System. Fatty acid synthesis is initiated by (3-ketoacyl-ACP synthase (KASIII) (step 1a) which catalyzes the condensation of acetyl-CoA with malonyl-ACP to yield -ketoacyl-ACP. In the subsequent steps, -ketoacyl-ACP undergoes reduction (step 2), dehydration (step 3), and reduction (step 4) similar to CoA specific pathway. Further elongation steps are initiated by KASI or KASII (step 1b) which catalyzes the condensation of acyl-ACP with malonyl-ACP. FIG. 27 depicts the novel CoA dependent fatty acid pathway and the key enzymes associated therewith.

[0066] FIG. 27 shows the novel CoA dependent fatty acid pathway and the key enzymes associated therewith.

DESCRIPTION OF EMBODIMENTS

[0067] The details of one or more inventive embodiments are set forth in the accompanying drawings, the claims, and the description herein. Other features, objects, and advantages of the inventive embodiments disclosed and contemplated herein can be combined with any other embodiment unless explicitly excluded.

[0068] The present invention relates generally to various production methods and/or genetically modified microorganisms that have utility for fermentative production of various chemical products, to methods of making such chemical products that utilize populations of these microorganisms in vessels, and to systems for chemical production that employ these microorganisms and methods. Among the benefits of the present invention is increased specific productivity when such microorganisms produce a chemical product during a fermentation event or cycle.

[0069] The present invention provides production techniques and/or genetically modified microorganisms to produce a chemical product of interest, such as a fatty acid or fatty acid derived product. The invention provides for one or more means for modulating conversion of malonyl-CoA to fatty acyl molecules, wherein the production pathway comprises a malonyl-CoA dependent pathway that includes an enzymatic conversion step that uses malonyl-CoA as a substrate. In accordance with certain embodiments, the malonyl-CoA dependent pathway is also a malonyl-ACP independent pathway, and is used in combination with the inhibition of a microorganism's native malonyl-ACP dependent fatty acid synthase pathway. In accordance with certain other embodiments of the present invention, fatty acid or fatty acid derived products are produced in a manner dependent on both a malonyl-CoA dependent pathway and a malonyl-ACP dependent pathway.

[0070] The genetically modified microorganisms of the invention are metabolically engineered to increase utilization of malonyl-CoA for production of a fatty acid or fatty acid derived product, through a metabolic pathway that is at least in part malonyl-CoA dependent. The fatty acid derived products may include esters, aldehydes, alcohols, alkanes, alkenes, and diacids, with various degrees of desaturation and chain branching, and further downstream products made from such chemical products. Also, genetic modifications may be made to provide one or more chemical products.

[0071] The present invention also relates to genetically engineered microorganisms having encoded therein unique enzymes and combinations of enzymes that function within the malonyl-CoA dependent pathway to produce fatty acids or fatty acid derived products of specific chain lengths. The microorganisms and methods provide a cost-competitive means of producing relatively high concentrations of fatty acids or fatty acid derived products of specific chain lengths or products having a relatively narrow carbon chain length distribution (i.e., 2, 3, 4 or less than 5 different carbon chain lengths with in the fatty acid product, e.g. C8/C10 or C8/C10/C12).

I. Definitions/Nomenclature

[0072] As used herein unless otherwise indicated, open terms such as contain, containing, include, including, and the like mean comprising.

[0073] Some embodiments herein contemplate numerical ranges. When a numerical range is provided, the range includes the range endpoints. Numerical ranges include all values and subranges therein as if explicitly written out.

[0074] As used herein, the article a means one or more unless explicitly stated otherwise.

[0075] As used herein, herein unless otherwise indicated, the one letter abbreviations A, R, N, C, D, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y and V, represent the twenty amino acids commonly found in peptides synthesized in nature.

[0076] As used herein, unless otherwise indicated, the convention Letter1NumberLetter2, when applied to polypeptides, means that the amino acid having the one letter code Letter 1, at Number position in the polypeptide, is substituted with the amino acid having the one letter code Letter2. For example, I147T means that the amino acid I, found at position 147 in the peptide, is substituted with the amino acid T.

[0077] As used herein, unless otherwise indicated, the symbol CNumber means a carbon backbone chain length having the indicated number of carbon atoms. For example, C20 means a chemical backbone having a 20 carbon chain length. Note that the number of carbons included in the carbon backbone does not include carbon contained in functional units attached to the backbone (e.g., a functional unit in a fatty acid derived product).

[0078] As used herein, reduced enzymatic activity, reducing enzymatic activity, decreased enzymatic activity, decreasing enzymatic activity, and the like is meant to indicate that a microorganism cell's, or an isolated enzyme, exhibits a lower level of activity than that measured in a comparable cell of the same species or its native enzyme. That is, enzymatic conversion of the indicated substrate(s) to indicated product(s) under known standard conditions for that enzyme is at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 percent less than the enzymatic activity for the same biochemical conversion by a native (non-modified) enzyme under a standard specified condition. These terms also can include elimination of that enzymatic activity. A decrease in enzymatic activity may be achieved in variety a ways known to those skilled in the art, including for example, a gene disruption or a gene deletion. A decrease in enzymatic activity may be temporal, be controlled through the expression of various genetic elements, or decrease in response to the cultivation conditions of the cell. A cell having reduced enzymatic activity of an enzyme can be identified using any method known in the art. For example, enzyme activity assays can be used to identify cells having reduced enzyme activity. See, for example, Enzyme Nomenclature, Academic Press, Inc., New York 2007.

[0079] As used herein, increase enzymatic activity, increasing enzymatic activity, and the like is meant to indicate that a microorganism cell's, or an isolated enzyme, exhibits a higher level of activity than that measured in a comparable cell of the same species or its native enzyme. That is, enzymatic conversion of the indicated substrate(s) to indicated product(s) under known standard conditions for that enzyme is at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 percent greater than the enzymatic activity for the same biochemical conversion by a native (non-modified) enzyme under a standard specified condition. These terms also can include addition of an exogenous enzymatic activity. An increase in enzymatic activity may be temporal, be controlled through the expression of various genetic elements, or increase in response to the cultivation conditions of the cell. A cell having increased enzymatic activity of an enzyme can be identified using any method known in the art, including the enzyme activity assays noted above used to identify cells having reduced enzyme activity.

[0080] As used herein, the term gene disruption, or grammatical equivalents thereof (and including to disrupt enzymatic function, disruption of enzymatic function, and the like), is intended to mean a genetic modification to a microorganism that renders the encoded gene product as having a reduced polypeptide activity compared with polypeptide activity in or from a microorganism cell not so modified. The genetic modification can be, for example, deletion of the entire gene, deletion or other modification of a regulatory sequence required for transcription or translation, deletion of a portion of the gene which results in a truncated gene product (e.g., enzyme) or by any of various mutation strategies that reduces activity (including to no detectable activity level) of the encoded gene product. A disruption may broadly include a deletion of all or part of the nucleic acid sequence encoding the enzyme, and also includes, but is not limited to other types of genetic modifications, e.g., introduction of stop codons, frame shift mutations, introduction or removal of portions of the gene, and introduction of a degradation signal, those genetic modifications affecting mRNA transcription levels and/or stability, and altering the promoter or repressor upstream of the gene encoding the enzyme.

[0081] In various contexts, a gene disruption is taken to mean any genetic modification to the DNA, mRNA encoded from the DNA, and the corresponding amino acid sequence that results in reduced polypeptide activity. Many different methods can be used to make a cell having reduced polypeptide activity. For example, a cell can be engineered to have a disrupted regulatory sequence or polypeptide-encoding sequence using common mutagenesis or knock-out technology. See, e.g., Methods in Yeast Genetics (1997 edition), Adams et al., Cold Spring Harbor Press (1998). One particularly useful method of gene disruption is complete gene deletion because it reduces or eliminates the occurrence of genetic reversions in the genetically modified microorganisms of the invention. Accordingly, a disruption of a gene whose product is an enzyme thereby disrupts enzymatic function. Alternatively, antisense technology can be used to reduce the activity of a particular polypeptide. For example, a cell can be engineered to contain a cDNA that encodes an antisense molecule that prevents a polypeptide from being translated. Further, gene silencing can be used to reduce the activity of a particular polypeptide.

[0082] The term heterologous is intended to include the term exogenous as the latter term is generally used in the art. Heterologous can refer to polypeptides and/or nucleic acids which are not ordinarily produced by the host cell. Such heterologous polypeptides and/or nucleic acid thus may comprise polypeptides which either do not have substantial amino acid sequence homology with those proteins produced by the host cell or may comprise polypeptides with substantial but incomplete homology to proteins produced by the host cell or the cell line from which the host cell is derived.

[0083] The term heterologous DNA, heterologous nucleic acid sequence, and the like as used herein refers to a nucleic acid sequence wherein at least one of the following is true: (a) the sequence of nucleic acids is foreign to (i.e., not naturally found in) a given host microorganism; (b) the sequence may be naturally found in a given host microorganism, but in an unnatural amount (e.g., greater than expected) or position; or (c) the sequence of nucleic acids comprises two or more subsequences that are not found in the same relationship to each other in nature. For example, regarding instance (c), a heterologous nucleic acid sequence that is recombinantly produced will have two or more sequences from unrelated genes arranged to make a new functional nucleic acid.

[0084] The term antisense molecule as used herein encompasses any nucleic acid molecule or nucleic acid analog (e.g., peptide nucleic acids) that contains a sequence that corresponds to the coding strand of an endogenous polypeptide. An antisense molecule also can have flanking sequences (e.g., regulatory sequences). Thus, antisense molecules can be ribozymes or antisense oligonucleotides.

[0085] As used herein, a ribozyme can have any general structure including, without limitation, hairpin, hammerhead, or axhead structures, provided the molecule cleaves RNA.

[0086] Bio-production, as used herein, may be aerobic, microaerobic, or anaerobic.

[0087] As used herein, the language sufficiently homologous refers to proteins or portions thereof that have amino acid sequences that include a minimum number of identical or equivalent amino acid residues when compared to an amino acid sequence of the amino acid sequences provided in this application (including the SEQ ID Nos./sequence listings) such that the protein or portion thereof is able to achieve the respective enzymatic reaction and/or other function. To determine whether a particular protein or portion thereof is sufficiently homologous may be determined by an assay of enzymatic activity, such as those commonly known in the art.

[0088] Descriptions and methods for sequence identity and homology are intended to be exemplary and it is recognized that these concepts are well-understood in the art. Further, it is appreciated that nucleic acid sequences may be varied and still encode an enzyme or other polypeptide exhibiting a desired functionality, and such variations are within the scope of the present invention.

[0089] Further to nucleic acid sequences, hybridization refers to the process in which two single-stranded polynucleotides bind non-covalently to form a stable double-stranded polynucleotide. The term hybridization may also refer to triple-stranded hybridization. The resulting (usually) double-stranded polynucleotide is a hybrid or duplex. Hybridization conditions will typically include salt concentrations of less than about 1M, more usually less than about 500 mM and less than about 200 mM. Hybridization temperatures can be as low as 5 C., but are typically greater than 22 C., more typically greater than about 30 C., and often are in excess of about 37 C. Hybridizations are usually performed under stringent conditions, i.e. conditions under which a probe will hybridize to its target subsequence. Stringent conditions are sequence-dependent and are different in different circumstances. Longer fragments may require higher hybridization temperatures for specific hybridization. As other factors may affect the stringency of hybridization, including base composition and length of the complementary strands, presence of organic solvents and extent of base mismatching, the combination of parameters is more important than the absolute measure of any one alone. Generally, stringent conditions are selected to be about 5 C. lower than the T.sub.m (temperature at which half the DNA is present in a single-stranded (denatured) form) for the specific sequence at a defined ionic strength and pH. Exemplary stringent conditions include salt concentration of at least 0.01 M to no more than 1 M Na ion concentration (or other salts) at a pH 7.0 to 8.3 and a temperature of at least 25 C. For example, conditions of 5SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and a temperature of 25-30 C. are suitable for allele-specific probe hybridizations. For stringent conditions, see for example, Sambrook and Russell and Anderson Nucleic Acid Hybridization 1.sup.st Ed., BIOS Scientific Publishers Limited (1999), which is hereby incorporated by reference for hybridization protocols. Hybridizing specifically to or specifically hybridizing to or like expressions refer to the binding, duplexing, or hybridizing of a molecule substantially to or only to a particular nucleotide sequence or sequences under stringent conditions when that sequence is present in a complex mixture (e.g., total cellular) DNA or RNA.

[0090] The use of the phrase segment of interest is meant to include both a gene and any other nucleic acid sequence segment of interest. One example of a method used to obtain a segment of interest is to acquire a culture of a microorganism, where that microorganism's genome includes the gene or nucleic acid sequence segment of interest.

[0091] When the genetic modification of a gene product, i.e., an enzyme, is referred to herein, including the claims, it is understood that the genetic modification is of a nucleic acid sequence, such as or including the gene, that normally encodes the stated gene product, i.e., the enzyme.

[0092] In some embodiments a truncated respective polypeptide has at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99% of the full length of a polypeptide encoded by a nucleic acid sequence encoding the respective native enzyme, and more particularly at least 95% of the full length of a polypeptide encoded by a nucleic acid sequence encoding the respective native enzyme. By a polypeptide having an amino acid sequence at least, for example, 95% identical to a reference amino acid sequence of a polypeptide is intended that the amino acid sequence of the claimed polypeptide is identical to the reference sequence except that the claimed polypeptide sequence can include up to five amino acid alterations per each 100 amino acids of the reference amino acid of the polypeptide. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a reference amino acid sequence, up to 5% of the amino acid residues in the reference sequence can be deleted or substituted with another amino acid, or a number of amino acids up to 5% of the total amino acid residues in the reference sequence can be inserted into the reference sequence. These alterations of the reference sequence can occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence. In other embodiments truncation may be more substantial, as described elsewhere herein.

[0093] Species and other phylogenic identifications are according to the classification known to a person skilled in the art of microbiology.

[0094] Where methods and steps described herein indicate certain events occurring in certain order, those of ordinary skill in the art will recognize that the ordering of certain steps may be modified and that such modifications are in accordance with the variations of the invention. Additionally, certain steps may be performed concurrently in a parallel process when possible, as well as performed sequentially.

[0095] The meaning of abbreviations is as follows: C means Celsius or degrees Celsius, as is clear from its usage, DCW means dry cell weight, s means second(s), min means minute(s), h, hr, or hrs means hour(s), psi means pounds per square inch, nm means nanometers, d means day(s), L or uL or ul means microliter(s), mL means milliliter(s), L means liter(s), mm means millimeter(s), nm means nanometers, mM means millimolar, M or uM means micromolar, M means molar, mmol means millimole(s), mol or uMol means micromole(s), g means gram(s), g or ug means microgram(s) and ng means nanogram(s), PCR means polymerase chain reaction, OD means optical density, OD.sub.600 means the optical density measured at a photon wavelength of 600 nm, kDa means kilodaltons, g means the gravitation constant, bp means base pair(s), kbp means kilobase pair(s), % w/v means weight/volume percent, % v/v means volume/volume percent, IPTG means isopropyl--D-thiogalactopyranoiside, RBS means ribosome binding site, rpm means revolutions per minute, HPLC means high performance liquid chromatography, UPLC means ultra performance liquid chromatography, and GC means gas chromatography.

[0096] By means for modulating is meant any one of the following: 1) providing in a microorganism cell at least one polynucleotide that encodes at least one polypeptide having certain enzymatic activity, wherein such enzymatic activity of the polypeptide so encoded is either: (a) exogenous, (b) native but is lower or higher than the enzymatic activity of its native form (such as by mutation and/or promoter substitution, etc.), or (c) modulated to have a reduced or increased enzymatic activity at any point during a fermentation process (such as by temperature sensitivity, inducible promoter, etc.); or 2) providing to a vessel comprising a microorganism cell or population an inhibitor that inhibits enzymatic activity or a supplement that increases enzymatic activity. These means may be provided in combination with one another.

[0097] As used herein, references to synthase III, synthase IV, synthase V, and synthase VI (except in the context of the name of a specific enzyme sequence included in a FASTA header in one of the Tables) shall refer to the third, fourth, fifth and sixth synthase, respectively, that is included in among a group of synthases. Synthase III, synthase IV, synthase V, and synthase VI may be any 3-ketoacyl-CoA synthase disclosed herein. For example, a reference herein to a genetically modified organism comprising a heterologous nucleic acid sequence encoding a 3-ketoacyl-CoA synthase selected from the group consisting of synthase III and synthase IV means either: (1) a genetically modified organism comprising a heterologous nucleic acid sequence encoding at least three 3-ketoacyl-CoA synthases wherein at least one of such 3-ketoacyl-CoA synthases is a 3-ketoacyl-CoA synthase disclosed herein; or (2) a genetically modified organism comprising a heterologous nucleic acid sequence encoding at least four 3-ketoacyl-CoA synthases wherein at least one of such 3-ketoacyl-CoA synthases is a 3-ketoacyl-CoA synthase disclosed herein.

II. The Bioproduction Pathways of the Present Invention

[0098] A. CoA Dependent Pathways

[0099] The present invention relates to a fatty acid pathway that comprises four steps which utilizes a pathway that is similar to the type II fatty acid synthesis (FAS) system utilized by bacteria. Both fatty acid syntheses are shown below in Scheme 1. As illustrated in FIG. 26 in Scheme 1, both pathways are cyclical processes that involve: 1) condensation of acyl chain, 2) reduction of the condensation product, 3) dehydration, and 4) reduction to produce an acyl chain that is two carbon atoms longer and the process is repeated with each cycle adding two additional carbons. Given the similarities between the two processes, most enzymes utilized for the type II FAS system can also function in the propose fatty acid pathway. However, a key step involving the chain elongation of acyl moiety is quite different. In accordance with the present invention, a condensation step of the proposed fatty acid pathway employs, inter alia, a ketoacyl-CoA synthase that catalyzes the condensation of acyl-CoA with malonyl-CoA, while type II FAS system utilizes ketoacyl-ACP synthases that catalyzes the condensation of acyl-ACP with malonyl-ACP. This type of CoA dependent pathway has been previously known for elongation of longer fatty acid chain lengths (e.g., elongation to C14 to C16 or higher). In accordance with the present invention, however, applicants have discovered novel genetically modified microorganisms capable of producing fatty acids through the elongation pathway illustrated in Scheme 1A and which is capable of elongation of lower carbon chain lengths through this pathway (e.g., elongation of C4 to C6, C6 to C8, C8 to C10, C10 to C12, and C12 to C14). (Note that -Ketoacyl and 3-ketoacyl are synonymous.)

[0100] The novel CoA dependent fatty acid pathway and the key enzymes associated therewith are illustrated in FIG. 27 for the production of a C6 fatty acid. One skilled in the art will appreciate the cyclic nature of this pathway, wherein a malonyl-CoA is added during each cycle until the desired carbon chain length is reached. The cyclic nature of this novel pathway is further illustrated in FIG. 3 to FIG. 6. FIG. 3 illustrates the production of even chain fatty acids using acetyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule. FIG. 4 illustrates the production of even chain fatty acid esters using acetyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule. FIG. 5 illustrates the production of odd chain fatty acids using propionyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule. FIG. 6 illustrates the production of odd chain fatty acid esters using propionyl-CoA as a primer and malonyl-CoA (MCA) as the extender molecule.

[0101] In accordance with the present invention, fatty acid or fatty acid derived products are produced in a manner dependent at least in part on a malonyl-CoA dependent pathway. In accordance with certain embodiments, the malonyl-CoA dependent pathway is also a malonyl-ACP independent fatty acid production pathway, and may be used in combination with the inhibition of a microorganism's malonyl-ACP dependent fatty acid synthase pathway. In accordance with certain other embodiments of the present invention, fatty acid or fatty acid derived products are produced through a microorganism pathway that is partially malonyl-CoA dependent and partially malonyl-ACP dependent. In accordance with certain other embodiments of the present invention, fatty acid or fatty acid derived products are produced through a microorganism pathway that is initiated through the reaction of malonyl-CoA and acetyl-CoA via a CoA dependent pathway.

[0102] Referring to FIG. 7 to FIG. 14, examples of various malonyl-CoA dependent pathways are illustrated. The pathways illustrated are examples of the production of fatty acids or esters having carbon chain lengths of 4, 6, 8, or 10. One skilled in the art would appreciate that in view of cyclic nature of the pathways, the pathways could be extended to depict higher carbon chain lengths. In accordance with the present invention, genetically modified microorganisms are provided that include various combinations of enzymes that determine (1) the carbon chain lengths produced by the organism, and (2) the extent to which the pathway is CoA-dependent or both CoA- and ACP-dependent. In addition, if the acetyl-CoA precursor that initiates the pathways shown in FIG. 7 to FIG. 14 is changed to propionyl-CoA, then fatty acids and esters having a carbon chain length that is an odd number (i.e., 5, 7, 9, or 11) will be made through the pathways.

III. Genetic Modifications to Microorganisms

[0103] The present invention herein provides genetically modified microorganisms that are modified to enable and/or improve a microorganism's ability to produce fatty acids and/or fatty acid derivatives at least in part through a malonyl-CoA dependent pathway. The malonyl-CoA dependent pathway may be independent of a malonyl-ACP pathway or may be in combination with a malonyl-ACP pathway.

[0104] In general, the genetically modified organism herein can be Clostridium, Zymomonas, Escherichia, Salmonella, Rhodococcus, Pseudomonas, Streptomyces, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klebsiella, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Pichia, Candida, Hansenula, Thraustochytrids, Bacteriophage, Saccharomyces; can be a prokaryotic cell; can be a eukaryotic cell; and/or can be a bacteria, yeast, fungi, microalgae or algae cell. Preferably the genetically modified organism is Escherichia coli.

[0105] The genetic modifications contemplated by the present invention include enhancing the organism's function in three phases of a CoA-dependent fatty acid pathway contemplated herein: (1) initiation of the fatty acid pathway; (2) chain length extension (or elongation); and (3) termination of the process once a desired chain length is achieved. These three phases are exemplified in FIG. 7 to FIG. 14.

[0106] A. Genetic Modifications to Drive Phase OneReaction Initiation

[0107] The first phase of the malonyl-CoA dependent pathway is reaction initiation. The reaction to produce even chain fatty acid products is initiated through the conversion of acetyl-CoA+malonyl-CoA to 3-ketobutyryl-CoA. This conversion requires a synthasea ketobutyryl-CoA synthase. As illustrated in FIG. 7, the reaction initiation phase is completed by the conversion of ketobutyryl-CoA to butyryl-CoA by three enzymes: a ketoacyl-CoA reductase (KCR), a hydroxyacyl-CoA dehydratase (3HDh), and an enoyl-CoA reductase (EnCr). The reaction to produce odd chain fatty acid products is initiated through the conversion of propionyl-CoA+malonyl-CoA to 3-ketovaleryl-CoA with subsequent reduction and dehydration reactions catalyzed by KCR, 3HDh, and EnCr. Accordingly, a genetically modified microorganism of the present invention includes native or exogenous enzymes encoded therein that provide these functions.

[0108] (1) Phase One (Reaction Initiation)Synthases

[0109] In accordance with one aspect of the present invention, NphT7, a 3-ketoacyl-CoA synthase from Streptomyces sp. Strain CL190 acts as the ketobutyryl-CoA synthase that initiates fatty acid synthesis by catalyzing the condensation of acetyl-CoA with malonyl-CoA to 3-ketobutyryl-CoA and with reduction.fwdarw.dehydration.fwdarw.reduction to butyryl-CoA (C.sub.4-CoA). In accordance with one aspect of the present invention, NphT7 acts as the 3-ketovaleryl-CoA synthase that initiates fatty acid synthesis by catalyzing the condensation of propionyl-CoA with malonyl-CoA to 3-ketovaleryl-CoA and with reduction.fwdarw.dehydration.fwdarw.reduction to valeryl-CoA (C.sub.5CoA). The protein sequence for NphT7 (BAJ10048.1 GI:299758082) and its nucleotide sequence (AB540131.1 GI:299758081) are provided below (SEQ ID NO:1; SEQ ID NO:2).

TABLE-US-00001 SEQIDNO:1 MTDVRFRIIGTGAYVPERIVSNDEVGAPAGVDDDWITRKTGIRQRRWAAD DQATSDLATAAGRAALKAAGITPEQLTVIAVATSTPDRPQPPTAAYVQHH LGATGTAAFDVNAVCSGTVFALSSVAGTLVYRGGYALVIGADLYSRILNP ADRKTVVLFGDGAGAMVLGPTSTGTGPIVRRVALHTFGGLTDLIRVPAGG SRQPLDTDGLDAGLQYFAMDGREVRRFVTEHLPQLIKGFLHEAGVDAADI SHFVPHQANGVMLDEVFGELHLPRATMHRTVETYGNTGAASIPITMDAAV RAGSFRPGELVLLAGFGGGMAASFALIEW SEQIDNO:2 1 cctgcaggccgtcgagggcgcctggaaggactacgcggag caggacggccggtcgctgga 61 ggagttcgcggcgttcgtctaccaccagccgttcacgaag atggcctacaaggcgcaccg 121 ccacctgctgaacttcaacggctacgacaccgacaaggac gccatcgagggcgccctcgg 181 ccagacgacggcgtacaacaacgtcatcggcaacagctac accgcgtcggtgtacctggg 241 cctggccgccctgctcgaccaggcggacgacctgacgggc cgttccatcggcttcctgag 301 ctacggctcgggcagcgtcgccgagttcttctcgggcacc gtcgtcgccgggtaccgcga 361 gcgtctgcgcaccgaggcgaaccaggaggcgatcgcccgg cgcaagagcgtcgactacgc 421 cacctaccgcgagctgcacgagtacacgctcccgtccgac ggcggcgaccacgccacccc 481 ggtgcagaccaccggccccttccggctggccgggatcaac gaccacaagcgcatctacga 541 ggcgcgctagcgacacccctcggcaacggggtgcgccact gttcggcgcaccccgtgccg 601 ggctttcgcacagctattcacgaccatttgaggggcgggc agccgcatgaccgacgtccg 661 attccgcattatcggtacgggtgcctacgtaccggaacgg atcgtctccaacgatgaagt 721 cggcgcgccggccggggtggacgacgactggatcacccgc aagaccggtatccggcagcg 781 tcgctgggccgccgacgaccaggccacctcggacctggcc acggccgcggggcgggcagc 841 gctgaaagcggcgggcatcacgcccgagcagctgaccgtg atcgcggtcgccacctccac 901 gccggaccggccgcagccgcccacggcggcctatgtccag caccacctcggtgcgaccgg 961 cactgcggcgttcgacgtcaacgcggtctgctccggcacc gtgttcgcgctgtcctcggt 1021 ggcgggcaccctcgtgtaccggggcggttacgcgctggtc atcggcgcggacctgtactc 1081 gcgcatcctcaacccggccgaccgcaagacggtcgtgctg ttcggggacggcgccggcgc 1141 aatggtcctcgggccgacctcgaccggcacgggccccatc gtccggcgcgtcgccctgca 1201 caccttcggcggcctcaccgacctgatccgtgtgcccgcg ggcggcagccgccagccgct 1261 ggacacggatggcctcgacgcgggactgcagtacttcgcg atggacgggcgtgaggtgcg 1321 ccgcttcgtcacggagcacctgccgcagctgatcaagggc ttcctgcacgaggccggggt 1381 cgacgccgccgacatcagccacttcgtgccgcatcaggcc aacggtgtcatgctcgacga 1441 ggtcttcggcgagctgcatctgccgcgggcgaccatgcac cggacggtcgagacctacgg 1501 caacacgggagcggcctccatcccgatcaccatggacgcg gccgtgcgcgccggttcctt 1561 ccggccgggcgagctggtcctgctggccgggttcggcggc ggcatggccgcgagcttcgc 1621 cctgatcgagtggtagtcgcccgtaccaccacagcggtcc ggcgccacctgttccctgcg 1681 ccgggccgccctcggggcctttaggccccacaccgcccca gccgacggattcagtcgcgg 1741 cagtacctcagatgtccgctgcgacggcgtcccggagagc ccgggcgagatcgcgggccc 1801 ccttctgctcgtccccggcccctcccgcgagcaccacccg cggcggacggccgccgtcct 1861 ccgcgatacgccgggcgaggtcgcaggcgagcacgccgga cccggagaagccccccagca 1921 ccagcgaccggccgactccgtgcgcggccagggcaggctg cgcgccgtcgacgtcggtga 1981 gcagcaccaggagctcctgcggcccggcgtagaggtcggc cagccggtcgtagcaggtcg 2041 cgggcgcgcccggcggcgggatcagacagatcgtgcccgc ccgctcgtgcctcgccgccc 2101 gcagcgtgaccagcggaatg tcccgcccagctccgga

[0110] In some embodiments, the 3-ketobutyryl-CoA synthase of the present invention is a homolog to a synthase comprising a protein sequence of any one of SEQ ID NOs. 1-120, as shown in Table 1 below. In some embodiments, the 3-ketobutyryl-CoA synthase of the present invention is a 3-ketoacyl-CoA synthase that comprises an amino acid sequence having at least about 70%, about 75%, about 80%, about 85%, about 90%, about 92%, about 94%, about 96%, about 98%, or about 99%, but less than 100% or about 100% homology to any one of SEQ ID NOs. 1-120. In some embodiments, the method herein comprises selecting at least two of 3-ketoacyl-CoA synthases, wherein each synthase occupies a different branch of a phylogenetic tree. In one aspect, the present invention provides a library of NphT7 homologs herein selected by a method herein.

[0111] In some embodiments, the 3-ketovaleryl-CoA synthase of the present invention is a homolog to a synthase comprising a protein sequence of any one of SEQ ID NOs. 1-120, as shown in Table 1 below. In some embodiments, the 3-ketovaleryl-CoA synthase of the present invention is a 3-ketoacyl-CoA synthase that comprises an amino acid sequence having at least about 70%, about 75%, about 80%, about 85%, about 90%, about 92%, about 94%, about 96%, about 98%, or about 99%, but less than 100% or about 100% homology to any one of SEQ ID NOs. 1-120. In some embodiments, the method herein comprises selecting at least two of 3-ketoacyl-CoA synthases, wherein each synthase occupies a different branch of a phylogenetic tree. In one aspect, the present invention provides a library of NphT7 homologs herein selected by a method herein.

TABLE-US-00002 TABLE1 SynthaseSequences SEQIDNO FASTAHeader Proteinsequence SEQIDNO:3 >gi|18310050|ref| MKNAKMIGFGLYTPKNLVENERLQEFLETSDEWIRTRTGIERRYI NP_561984.1|/1- SLDENTSDLAVEASKKALSQARLSAEEIDLIIVATVTPDNFTPSTA 3243-oxoacyl- CIVQDKLGAKNAWAFDINAACTGFIYALKLGRSLIRSGEANNALI ACPsynthase IGAETLSKALNWEDRGSCVLFGDGAGATVLTSTEEDCGIKCVNV [Clostridium KSDGSKGDSLVIQGLPLNSPFKDGREVSENYINMNGREIFKFATK perfringensstr. VMEESIVEILEKENIKIEDIAAIIPHQANLRIIDYVVKRLGIPREKFIT 13] NLQNYGNTSGASIPIALCESIDEGNLKKGDNIIMVGFGGGLTWGA ALIKL SEQIDNO:4 >gi|21224866|ref| MHQGSRITAVGHYQPARILTNEDLAGMVDTSDEWIRSRVGIRTR NP_630645.1|/1- RIAGPDEPVDELAGHAAAKALASAGLTPADVDLVVVATSTAIDR 3163-oxoacyl- SPNTAARVAARLGIPGPAALDLNVVCAGFTHALATADHAVRAGS ACPsynthase ASRALVVGADKMSEVVDWTDRTTCVLVGDGAGAAVVEACAPG [Streptomyces EEPGIGPVLWGSVPEMGNAVRIEGTPPRFAQEGQSVYRWATTRL coelicolorA3(2)] PAIARQACERSGLEPADLAAVVLHQANLRIVEPLAAKIGAVNAV VARDVVESGNTSAASIPLALSKLAERGEITTGDPALLFGFGGNLS YAGQVVRCP SEQIDNO:5 >gi|23014672|ref| MIVRSQIIGCGSYLPSRLVTNAELAAKVDTTDEWIVERSGIRQRHI ZP_00054477.1|/1- AAEGETTSDLATNAALRALEAAGIAGSAVDLVIVATATPDNTFPA 324COG0332: TATKVQSRIGMKHGFAFDVQAVCSGFVYALSVADNFIKSGQVQT 3-oxoacyl-[acyl- ALVIGAETFSRILDWNDRTTCVLFGDGAGAVVLRANRGKGSSAD carrier-protein] RGILSTHLHSDGSHYDLLYVDGGPSSTQTVGHVHMEGREVFRHA synthaseIII VINLASVVGEALSANDLKASDIDWVVPHQANRRIIEGTAKKLGFP [Magnetospirillum LDKMVMTVDRHANTSAASIPLALTEAVSDGRIKPGQLVLLEAMG magnetotacticum GGFTWGSALVRM MS-1] SEQIDNO:6 >gi|28898830|ref| MYSKILGTGSYLPSQVRTNADLEKMVDTSDEWIVARTGIKERRIA NP_798435.1|/1- AEDETVADMAFYAAENAIDMAGIDKNDIDLIIVATTSSSHTFPSSA 3163-oxoacyl- CQVQAKLGIKGCPAFDLAAACSGFVYALSVADQHIKSGMCKNV ACPsynthase LVIGADALSKTCDPTDRSTIILFGDGAGAVVVGASQEPGIISTHIY [Vibrio ADGQFGDLLSLPVPERGKDVDKWLHMAGNEVFKVAVTQLSKLV parahaemolyticus KDTLEANDMHKSELDWLVPHQANYRIISATAKKLSMSLDQVVV RIMD2210633] TLDRHGNTSAATVPTALDEAVRDGRIKRGQTLLLEAFGGGFTWG SALVKF SEQIDNO:7 >gi|56419339|ref| MGAGIIGVGRYVPEKVLTNFDLEKMMDTSDEWIRTRTGIEERRIA YP_146657.1|/1- ADDIDTSDMAYFAAKRALQDAGMEAKDIDLILVATVTPDRPFPS 3103-oxoacyl- VACMLQERLGAVNAAALDISAACAGFMYGMVTAAQFIDTGAY ACPsynthase KYILVVGADKLSKITDWTDRNTAVLFGDGAGAVVMGPVSPGRGI [Geobacillus LSFELGADGTGGKHLYKDEYIVMNGREVFKFAVRQMGESSVRV kaustophilus LEKAGLTKDDVDFLIPHQANIRIVEAARQRLELPEEKISTTIRRYG HTA426] NTSAASIPISLVEELEAGKIHDDDLIIMVGFGGGLTWGAIALRWG R SEQIDNO:8 >gi|65318552|ref| MGILGIGRYVPEKVVTNHDLEKIMDTSDEWIRTRTGIAERRIADD ZP_00391511.1|/1- TIDTSYMAVEASKKALEDAGISGEDIDLILVATVTPDRAFPAVAC 308COG0332: VIQEAIGAKHAAAMDLSAACAGFMYGMITAQQFIQTGTYKNVL 3-oxoacyl-[acyl- VVGSDKLSKIVDWNDRNTAVLFGDGAGAIVMGAVSEGKGVLSF carrier-protein] ELGADGSGGKHLYQDEYVMMNGREVFKFAVRQLGDSCLRVLD synthaseIII KAGLTKEDVDFLVPHQANIRIMESARERLNLPQEKMSMTIEKFG [Bacillusanthracis NTSASSIPIAMVEELQNGRIQDGDLIILVGFGGGLTWGAVALRWG str.A2012] K SEQIDNO:9 >gi|86159172|ref| MRSLIAGTGSYAPEKVVTNADLEKLVDTNDQWIVERTGIRERHV YP_465957.1|/1- VADDQATSDLALEASRRALDAAGLDAKDVEMIVVGTVTPDYPF 3263-oxoacyl- PSVGAVLQGKLGNKKAFAFDVSAACAGSLYALSVADRFVASGA ACPsynthase VKNALVVGADALTRITDWTDRNTCILFGDGAGAMVLKPTDDPQ [Anaeromyxobacter RGIRAVRLHADGSLVPILLQPGGGSRDPISEKVVREKSHYVKMN dehalogenans GREVFKVAVRSLEESCREVLADEKLTPGDVTWVIAHQANKRILD 2CP-C] ATLHRLEIPESKCWMNLEKYGNTSAASVPMTLDEANRAGWLKP GDTVLMMAIGGGMAWGASVVRW SEQIDNO:10 >gi|93006238|ref| MTTCITGTGLYIPPFSISNEELVESFNQYVEKYNTKHAADIEAGTL YP_580675.1|/1- TALQPSSAAFIEKVSGIKSRYVMEKDGILNPDIMAPVIAYRNLGEE 3813-oxoacyl- LSIMAEMGVAALNDALADAGLEANDLDGIILACSNFQRTYPAVSI ACPsynthase EIQNAIGMVGGFAYDMNVACSAATFGLSQAHGSIASGLAKRVAV [Psychrobacter VNVEITSAHLNWRNRDSHFIFGDVATACIVEELDTPKGYEILNSK cryohalolentisK5] LFTQFSTNIKNEYGFMDRSEFLAAQTEMYPDIKEPVTDKLFLQNG RKVFREVCPKVSEVITEHLQENNIATSDVKMMWLHQANANMLD LILRTVIGKEADKAIVPSVIAEFANTSSASPMIVFHRYKDDLASGD LGVICSFGAGYSIGSVIVRKV SEQIDNO:11 >gi|109899602|ref| MTNSVVISGSGLWNPPHSISNEELVDAYNAYAQQFNEQNADEIES YP_662857.1|/1- GAITAKPFSSAEFIQKASGIRSRYCYMKDGVLDINRMRPIIPERGE 3743-oxoacyl- EELSDQAEMAINAAKLALEAANKTAEDIDVVIVSCAYTQRSYPA ACPsynthase LAIEVQGALGIKGFGFDMLVACSAATFALHRAYEMISAGTAKGV [Pseudoalteromonas LVINPELTSPQVNYCDRDSHFIFGDVATAMVVEHADTATSEHVF atlanticaT6c] DILSTKAITQYSNNIRSNFGYVSRANDVDPYGADKLFHQEGRKVF KEVCPMAAEHISEHLERHQLTSADVKRWWLHQANINMNTLISKR LLGREATVEEAPIVLDRYANTASAGSIIAFNLHHKDLQAGDYGLL CSFGAGYSIGSLLVRKR SEQIDNO:12 >gi|114047960|ref| MHTKILGTGSYLPVQVRSNQDLEKMVETSDQWIVERTGISERRIA YP_738510.1|/1- AQDETVSTMGYQAALKALEMAGIEASELDMIICGTTSAANAFPA 3193-oxoacyl- AACEIQAMLGVHTIPAFDIAAACSGFVYALSVADQFVKNGTAKK (acylcarrier VLVIGADVLSRLCEPEDRTTIILFGDGAGAAVIGASDEPGIISTHIY protein)synthase ADGRQGDLLKCAFPPRQGETSEAVGFMTMKGNDVFKVAVTQLS III[Shewanella HVVTETLRLNNIDKSEIDWLVPHQANFRIINATAKKLDMSLDKV sp.MR-7] VLTLAKHGNTSAASVPIALDEAVRDGRIQRGQLLLLEAFGAGFA WGSALVRF SEQIDNO:13 >gi|121533809|ref| MKANDIGVGILGLGCYVPEKVLTNHDLEKMVDTSDEWIVERTGI ZP_01665636.1|/ RERRIADPDVATSDLATRAAERALSNAGISADELDLIIVATATPD 1-3383-oxoacyl- MFFPSVACLVQDNLKATRAAAFDLVAGCSGFVYGLTVGAQFIKT (acyl-carrier- GLYKKVLVIGAETLSKILDWTDRNTCVLFGDGAGAAVLSETEPG protein)synthase YGLIGFHLGADGSGGDLLKLPAGGSRLPPSVETVTQRLHFVHMN III[Thermosinus GNEVFKFAVRVMGEAAVKALENAGLGHQDVDCLIPHQANIRIIQ carboxydivorans SAAKRLKLPMDKVIVNVDKYGNTSAASIPIALEEAVRNGRVKKG Nor1] DVVVLVGFGAGLTWASCVIKWCKEDNTIA SEQIDNO:14 >gi|146293464|ref| MKQVVISGSGLFTPPYSISNEALVESFNAYVDIFNLENAGLIEQGH YP_001183888.1|/ VAALSYSSSEFIEKASGIKHRYVMVKEGILDPEIMMPLIPERSSDE 1-3733-oxoacyl- LSMQAEIGVEAALMALNNANLKAEQIDLVIVACAYTQRAYPAM ACPsynthase AIEIQRALGTRGYGYDMQVACSSATFAIVAAANAIATGSASRVLV [Shewanella INPEICSAQVNYRDRDSHFIFGDVATALVLEEQSLVEPNKGFTILS putrefaciensCN- SRCFTDYSNNIRSNFGFLNRCDPSSAHQADKLFHQQGRKVFKELL 32] PMIYQHLDEHLAEQASTPQSFKRLWLHQANINMNQFVVRKMLG DEVSPEQAPVVLDEYANTASAGSVIAFHKYSSDFKAGDLGLLSSF GAGYSIGSVILQKR SEQIDNO:15 >gi|160900704|ref| MRRYARITGTGSYLPPRRLTNHDLAAELAQRGIETSDEWIVERTG YP_001566286.1|/ IHARHFAAPDVASSDLALEASKKALEAAGCQPQDIDLIIVATSTPD 1-3253-oxoacyl- MVFPSTACILQNKLGANGCAAFDVQAVCSGFVYALTVADAMIQ ACPsynthase SGAASRALVVGSEVFSRILDFNDRTTCVLFGDGAGAVVLEASEQ [Delftia PGILASDLHADGKHVGILCVPGNVSGGQVLGDPLLKMDGQAVF acidovoransSPH- KLAVGVLEKAARATLDKAGLTDADIDWLIPHQANIRIMQSTARK 1] LKLSMDKVVVTVDQHGNTSAASIPLALDHGVRNGQVKPGQTVL LEGVGGGFTWGAVLLKM SEQIDNO:16 >gi|166364688|ref| MNGFGAAVVITGCGSATPAQFLSNEELSQIVETSDEWIKSRTGIG YP_001656961.1|/ KRHLADRSVSLSQLAAQAAIKALEMAQVSPRDIDLILLATSTPDD 1-3333-oxoacyl- LFGSAAQVQSQIGANRAIAFDLTAACSGFLVGLVTATQFIRTGTY ACPsynthase RNVLVIGADVLSRWVDWNDRATCVLFGDGAGAVVCQANDTKD [Microcystis NILGFELHSDGSQNGSLNLAYEGEELPLKQGIRVQKGTYKPLRM aeruginosaNIES- NGREVYRFAVAKVPEVIEKALYRANLTTSDIDWLVLHQANQRIM 843] DAVSERLKLPPEKVISNLSEYGNTSAASIPLALDEAVRSGKVKKG DIIASSGFGAGLTWGGIIFRWGD SEQIDNO:17 >gi|169633183|ref| MGIRITGTGLFHPTEIISNEELADSLNAYVEQYNQENAEKIAAGEL YP_001706919.1|/ EELRGSSAEFIEKASGIKRRYVIEKSGILDPTRLRPRLSERSNDELSI 1-3683-oxoacyl- QAEWGVIAAKQAMENAGVTAEDIDVVILACSNMQRAYPAVAIEI ACPsynthase QSALGIQGYAYDMNVACSAATFGLKQAADAIRSGARRVLLVNV [Acinetobacter EITSGHLDYRNRDCHFIFGDVATASIIEETTTKTGFEILDIHLFTQFS baumanniiSDF] NNIRNNFGFLNRSEDAVVDDKLFRQDGRKVFKDVCPLVAKIINA QLEKMQLTANDIKRFWLHQANANMNELILKYVAGKDADLSRTP IILDEFANTSSAGVIIALHRTGHEVDDGEYGVISSFGAGYSVGSIV VQKHVA SEQIDNO:18 >gi|170781992|ref| MVERFTRIWGLGAARGELDVPNDDLVGPIDSSDEWIRQRTGIITR YP_001710324.1|/ KRAGADVDAVDLATTASLEAIAKAGIRPEQIGIVLVSTVSNTVQT 1-3243-oxoacyl- PSMAALLADRIGANPAPAYDISAACAGYTYGIAQADSFIRSGLAE ACPsynthase YVLVVGAEKLSDIVDPTDRSISFLLGDGAGAAIVGPSDTPGISPTV [Clavibacter WGSDGSNWDAVGMTGTLKSMRDGSAWPTLRQDGQKVFRWAV michiganensis WEMVKVAKEALDRAGVAPEQLAAFIPHQANMRIVDEFAKQLGL subsp. PESVAIARDIATTGNTSAASIPLATHRLLEEDPSLSGGLALQIGFGA sepedonicus] GLVFGAQVVVLP SEQIDNO:19 >gi|197104835|ref| MNDAVIAATGLYTPPLSLSNAELVETFNAYVERFNAANAEAIAR YP_002130212.1|/ GEVQPLQPSSVEFIEKASGIKSRFVVDKTGLVDPEIMRPIIPERPND 1-3703-oxoacyl- QLSILAEIAVEAAKDAIARWGKPVSEIDAVICAASNMQRAYPAM ACPsynthase AIEVQQALGIDGFAFDMNVACSSATFGIKTAADFVAGGAKAVLM [Phenylobacterium VNPEICSGHLNFRDRDSHFIFGDVATAVIVERADQATDGWDILGT zucineum RLKTQFSNNIRNNFGFLNRADPEGVGKPDKLFVQEGRKVFREVV HLK1] PMVSEMIVDHAADLGIDPTGLKRLWLHQANINMNEMIGRKVLG RDPAPGENVIILDEYANTSSAGSIIAFHKANDDFQTGDTGLICSFG AGYSAGTVFVRKR SEQIDNO:20 >gi|219849850|ref| MYDRKVARVSRERYAAVIGWGMAVPNRVVTNDDLAQRIDTSD YP_002464283.1|/ EWIRTRTGIRERRVAGPGESTSTFATAAGREALEMAGVSPATIDT 1-3423-oxoacyl- VIVATCTPDRPFPATACTVQANLQIPRATAFDLAAACSGFVYGLT (acyl-carrier- VATSLIKSGVSRRLLLIGADIFTHYINWNDRNTCVLFGDGAGAVV protein)synthase LEATDEPLGLIASNLSADGNLEDLMAVDAGGTRMPLTAELLAEG III[Chloroflexus RQYVYMNGREIFKHAVREMSESALHVVQAAGLTIDDIALVIPHQ aggregansDSM ANVRIIDAVARRLELPPERVMINLDRYGNTSAASIPIALYEAAQQE 9485] RIKAGDYVLMTAFGGGLTWGSGIVRWGRPSR SEQIDNO:21 >gi|227523050|ref| MKFENFKILATASQVPTRVVDNDELSTMMDTSDDWIVQRTGIRR ZP_03953099.1|/ RHIAVDETTSSLCTSVAKQLLEKTGLKPSEIDLIIVATMSPDYLTPS 1-3273-oxoacyl- VSAMVQGNLGADHAVAMDIDAACSGFVYGLNMVKQLLIAETPK (acylcarrier NAILIGGEMLSKLIDWQDRSTAVLFGDGAGGVLLKNTPKAEGAFI protein)synthase SENLKTLGKLGRYLTAGKTGAPTPFMEKKDEFSPFFQMNGRRVY III[Lactobacillus RFAVNNVPESINQALAEASLTTDDIDHFVLHQANSRIVEKIAETLG hilgardiiATCC VSMDKFPINIDEYGNTAAASEPILLDQLVTNGTIKRGDVVLLSGF 8290] GGGLTVGTMILKY SEQIDNO:22 >gi|238623523|emb| MRMSDLGILGTGAYVPDRVVSNDDVGAAAGVDDAWIRRKTAIR CAX48662.1|/1- ERRWAAPGQATSDLAAAAGRAALRSAGITADQLSVIVVATSTPD 327putative3- RPQPPTAAYVQHGLGAAGAAAFDVNAVCSGSVFALAVAEGLLA oxoacyl-[acyl- GRGGHALVIGADLYSRILNPADRRTVVLFGDGAGALVLGPAAQG carrier-protein] PRVRHLALHTFGELAGLIEVPAGGSRLPGDRAALEAGLQYFAMD synthase GREVRRFVAEQLPRLTKQFLHEAGVVPDDIGHFVPHQANGVLLD [Streptomyces AVTADLGLPRAASHRTLAHYGNTGAASIPITLDTAARAGAFRPG anulatus] DLILLAGFGGGMSAGLALVEW SEQIDNO:23 >gi|239623103|ref| MTTRIIGTGSYVPEQIVTNNDLAQIVETNDEWIRSRTGIGERRIATT ZP_04666134.1|/ ESTSYMAANAAMRALEQSGVKPEEIDLILLGTSSPDYCFPNGACE 1-3203-oxoacyl- VQGMIGAVNAACYDISAACTGFVYALNTAHAFISSGIYKTALVIG [acyl-carrier- SDVLSKLIDWTDRGTCVLFGDGAGAVVVKADETGILGINMHSDG protein]synthase TKGNVLTCGSRTNGNFLLGKKPELGYMTMDGQEVFKFAVRKVP III[Clostridiales ECIKQVLDDAGVAAAEVRYFVIHQANYRIIESIAKRLKVSVDCFP bacterium VNMEHYGNTSGASVPLLLDEINRKGMLESGDKIVFSGFGAGLTW 1_7_47_FAA] GATLLEW SEQIDNO:24 >gi|240850683|ref| MIRSIIRGVGSALPKRSLSNDEIAKFVETSDSWIVQRTGIRQRYIAS YP_002972083.1|/ ENETTVSLGVEAAQAALTNAGLTIKDIDCIILATSTPNRTFPASAV 1-3243-oxoacyl- EIQCALGMSHGFAFDIQAVCSGFIFALTTGDSYLRCGAAKRILVIG (acylcarrier SDTFSRILDWEDRTTCVLFGDGAGAAILEAQEIEGGIAFERGILSA protein)synthase KLRSNGAYIDKLYVDGGPSTTQTTGYLRMEGREVFKYAVGMITD III[Bartonella VVDDCFAAAGMDSSQLDWFVPHQANKRIIEASAKKLGISLDKVV grahamiias4aup] ITVDQHGNTSAASVPLALTTAVCDGKIKEGDLIMLEAMGGGFTW GAILIRW SEQIDNO:25 >gi|253681256|ref| MYNVKIISTGKYIPDNVVTNDDMSKFVDTNDKWISERTGIKERRI ZP_04862054.1|/ STGENTSHMAVKAALAALEKSSVKATDLDLIIIATCTPDSFVPSTA 1-3243-oxoacyl- CIVQDKLGATKATCFDISAACTGFIYALGVASQFIKTGQVKNALV [acyl-carrier- IGAETLSKILNWEDRSTCILFADGAGAAIIERSEEVGLISQYTGSDG protein]synthase TGGKALKCEALPVRNPYCKVDDKFKDTLSMEGREVFKFAVNAM 3[Clostridium IESINKVLENTEYTLDDIDYIVPHQANIRIIEFVSKKLGISQDKFYV botulinumDstr. NLHKYGNTSGASIPIALDEMNKKGMFKKGDNIILVGFGGGLTFG 1873] AHLIQWN SEQIDNO:26 >gi|254286853|ref| MYSKILGTGSYLPSQVRTNADLEKMVETSDEWIVARTGIRERRIA ZP_04961806.1|/ ADNETVADMAFFAAQNAIDMAGIDKHDIDMIIVATTSASHTFPSA 1-3123-oxoacyl- ACQVQGKLGIKGCPAFDLAAACSGFMYALSIADQHVKSGMCKH (acyl-carrier- VLVIGADALSKTCDPTDRSTIILFGDGAGAVVVGASNEPGILSTHI protein)synthase HADGEFGDLLSLEVPVRGGDSDKWLHMAGNEVFKVAVTQLSKL III[Vibrio VVDTLKANNMHKSELDWLVPHQANYRIISATAKKLSMSLDQVVI choleraeAM- TLDRHGNTSAATVPTALDEAVRDGRIQRGQMLLLEAFGGGFTW 19226] GSA SEQIDNO:27 >gi|254477647|ref| MTRRAVIAGIGHYLPERIVENAEFEATLDTSDEWIRSRSGIERRHF ZP_05091033.1|/ AAEGETTSNMATKAAQNALADAGMTADDIDAIVVATSTADLTF 1-3233-oxoacyl- PSAATMVQAQLGMTKGFAFDVQAVCAGFVYALSNANALVASG (acyl-carrier- QADKVLVIGAETFSKIMDWTDRSTCVLFGDGAGALVLEAQEGA protein)synthase GTSDDRGILATDLNSDGRFKDLLYVDGGVSTQNTGHLRMQGNQ III[Ruegeriasp. VFRHAVEKLASTAHTSLERAGLGADDVDWIVPHQANIRIIQGTA R11] KKMGLPMDKVVVTVQDHGNTSAASIPLALSVGKARGQIKQGDLI VTEAIGGGLAWGSVVLRW SEQIDNO:28 >gi|262375396|ref| MGIRITGTGLFHPEHVITNEELVESLNAYVELFNHENADKIAAGE ZP_06068629.1|/ VEARRGSSADFIEKASGVQRRYVVEKSGILDPKRLRPNLRERADD 1-3693-Oxoacyl- EISLQAEWGVIAAKQAMENAGVTAEDIDIVILSCSNLQRAYPAVA [acyl-carrier- IEIQTALGIKGYAYDMNVACSAATFGLKQAYDAIKAGARRVLLV protein(ACP)] NVEITSAHTDFRSRDCHFIFGDVATASIIENTDSKTGFEILDSELFT synthaseIII QFSNNIRNNFGFLNTSENADIDDKRFRQDGRKVFKEVCPLVAKMI familyprotein TAQLEKNQIEPTGVKRFWLHQANASMNELILKLVVGKENAKPGL [Acinetobacter VPIILNEFANTSSAGVIIALHRTAHEVEDGEYGVLCSFGAGYSVGS lwoffiiSH145] ILVQKRVA SEQIDNO:29 >gi|282854072|ref| MTAIKTRPVHGYSKFLSTGSARGSRVVTNEEMCTLIDSTPEWIEQ ZP_06263409.1|/ RTGITERRWATSSETVASMGTTAARTALERSGLEASQIDAIIVATV 1-3323-oxoacyl- SHHRPSPSLAAYIARELGLGDAAAFDLNGACAGFCYSTALADSM [acyl-carrier- IRTGSANYVLVIGVEKLSEMTNLDDRSTAFLFSDGAGAAIISASDE protein]synthase PGIGPVVWGSRSDQLKTIELEDWPTASADPNKIHPLIRMEGRAVF 3 KWAMTDVAKRAAEAVAEAGITPADLDVFIPHQANDRITDVVSR [Propionibacterium HLKLPESVTVCHDIADMGNTSAASVPIAIDRMLQRGQAHSGDLA acnesJ139] LIIGFGAGLVYAGQVIRLP SEQIDNO:30 >gi|291439887|ref| MAKIKPSKGAPYARILGVGGYRPTRVVPNEVILETIDSSDEWIRSR ZP_06579277.1|/ SGIETRHWASPEETVAAMSVEASGKAIADAGIDAAQIGAVVVST 1-3333-oxoacyl- VSHFAQTPAIATEIADRLGTDRAAAFDISAGCAGFGYGLTLAKG (acylcarrier MVVEGSAEYVLVIGVERLSDLTDLEDRATAFLFGDGAGAVVVGP protein)synthase SQEPAIGPTVWGSEGDKSETIKQTVPWTDYRDGTVEKFPAITQEG III[Streptomyces QAVFRWAVFEMAKVAQQALDAAGITADDLDVFIPHQANVRIIDS ghanaensisATCC MVKTLKLPEHVTVARDIRTTGNTSAASIPLAMERLLATGEAKSG 14672] DTALVIGFGAGLVYAASVVTLP SEQIDNO:31 >gi|294791665|ref| MTMMNKPVGIIGTGSFLPDNVVTNFDLEKMVDTNDQWIRERTGI ZP_06756813.1|/ EERRIAPEGMNTSYMATEAAKKAMQMANVTAEEIDMIIFATLTP 1-3313-oxoacyl- DMIIPSAACVLQANLGAKNAAAYDLQAACSGFVYGLITAASYISS (acyl-carrier- GIYKKVLVVGAEILSRRVNWNDRGTCILFGDGAGAAVVSEVPEG protein)synthase YGIKGIDMGADGTGGSALCIPAGGTAVVANDQRVEEGLTFIHMD III[Veillonellasp. GPEVYKFAVKTMGRTVLKSLERASMELNELDYFIPHQANIRIIDS 6_1_27] AAKRLHLPMEKVFVNLHKYGNTSAASVAIALDEANREGRFKRG DNVAFAGFGAGLTWASLVLKWY SEQIDNO:32 >gi|296388215|ref| MHKAVISGTGLYTPPYSISNDELVESFNTFVRQYNDQHAEAIAKG ZP_06877690.1|/ ELEALAESSSAFIEKASGIKSRFVMNKEGILDPQRMVPYLPERSND 1-3733-oxoacyl- EWSILCEMAVAAAREALQRAGRSAADIDGVIVACSNLQRAYPAI (acylcarrier AVEVQAALGIQGYGYDMNVACSSATFGIQAATTAIQTGQARAIL protein)synthase MVNPEICTGHLNFRDRDSHFIFGDACTAVIVERADLAVSKHQFDI III[Pseudomonas VSTRLLTQFSNNIRNNFGFLNRADESGIGKRDKLFVQEGRKVFKD aeruginosaPAb1] VCPMVAELIGEHLAANEIQVAEVKRFTVLHQANLNMNLLITRKLL GRDAEAHEAPVILDSYANTSSAGSVIALHKHQDDLPSGAIGVLSS FGAGYSIGSVILRKH SEQIDNO:33 >gi|302539498|ref| MTAIGILGTGSYLPADTVSNRVVGERAGVTEDWILQKTGIRERRY ZP_07291840.1|/ AAEYEATSDLAVEAARSALDAAGISAEQLSWIVVATSTPDSPQPA 1-3433-oxoacyl- TACLVQHRIGAVNAAAFDVNSVCSGFVFGLVAAARMLPGQDGG [acyl-carrier- VRGHALVIGADVYSRIIDREDRRTAVLFGDGAGAVVLGPVRSGY protein]synthase GVLGSYLASRGDQAELIRVEAGGSRLPASEKTVAEGLHHFRMNG III[Streptomyces RGVRDFVAAELPRAVGEVLDRHGLERSEVDHFVPHQANGVMLG sp.C] ETVPRLGLPRARTHLTVAEHGNTSAASIPLALDEAYRSGAFRDRD VVLLAGFGGGMSLGTVLVRWDEEAAPAPRKDSAA SEQIDNO:34 >gi|307083025|ref| MTEIATTSGARSVGLLSVGAYRPERVVTNDEICQHIDSSDEWIYT ZP_07492138.1|/ RTGIKTRRFAADDESAASMATEACRRALSNAGLSAADIDGVIVTT 1-3133-oxoacyl- NTHFLQTPPAAPMVAASLGAKGILGFDLSAGCAGFGYALGAAAD [acyl-carrier- MIRGGGAATMLVVGTEKLSPTIDMYDRGNCFIFADGAAAVVVG protein]synthase ETPFQGIGPTVAGSDGEQADAIRQDIDWITFAQNPSGPRPFVRLEG IIIfabH,partial PAVFRWAAFKMGDVGRRAMDAAGVRPDQIDVFVPHQANSRINE [Mycobacterium LLVKNLQLRPDAVVANDIEHTGNTSAASIPLAMAELLTTGAAKP tuberculosis GDL SUMu012] SEQIDNO:35 >gi|311113478|ref| MTTLKQYENNRYSRILGYGASRGEVIVHNNDIVEAINSSDEWIKQ YP_003984700.1|/ RTGISTRHRASENQTVNDLAIAAAHDALANSHVTGEQIDAVIISTI 1-3413-oxoacyl- SHPYATPSLAVLVADAIGSRCPAYDISAACAGFCYGIAQADAMV (acyl-carrier- RSGMAQNVLVIGVEKLSDFIDNTERSISFLLGDGAGAAVVSVSDE protein)synthase PGIAPTIWGSDGSRWGTVGMTHSLLDIRNRDFVVNPVQEDEKIW III[Rothia PTLRQDGPSVFRWAVWEMAKVAQQALESAGITPDELGALIPHQA dentocariosa NARIIDQMAKTLKLPENVAIARDIADAGNTSAASVPLAAHRLLQE ATCC17931] QPELSGKFALQIGFGAGLAYAAQVVVLP SEQIDNO:36 >gi|312793335|ref| MKQNVKILSTGRFVPEKILSNYDLEKMVETSDEWITQRTGIKERR YP_004026258.1|/ IVDGRTSTTDLAVQAARNAMQKAGISPDEIDLVIVATVTPEMFFP 1-3283-oxoacyl- STACLVQKELKLKNAFAFDISAACSGFIYGMAVATQFIQNGFCKT (acyl-carrier- ALVIGAEALSKITNWSDRSTCVLFGDGAGAAILTASSEEGILGFEL protein)synthase GSDGENGLLLYCHAFGLSDLSYSQFKDMPNFRKIYMDGNEVYKF iii AVKIMPYAVEKVLEKVGLSSSDIDVFIPHQANIRIIESAAKRLKIP [Caldicellulosiruptor MEKVFVNLHKYGNTSAASIPIALDEAIEEGRIKKGDRIVLVGFGG kristjanssonii GLTWASCAVKWI 177R1B] SEQIDNO:37 >gi|318080591|ref| MDNSELCATVASTPEWIETRSGIRARGFAAPDETLRFMGRAAAE ZP_07987923.1|/ KALARAGVLPDGIDLVLVASMSRLEQTPPLAVLLAEDLGARAAA 1-3073-oxoacyl- GLDVSGACAGFCHALALASDAVRAGSARHVLVVGTERMTDLVE (acyl-carrier- RADRTVSVLFADGAGAAVVGPSARPGISPPARGAAGRYAGALR protein)synthase MDRGWDAFAADPSLGRPWMRMDGRRVFRWAMDEVTPRAAEL III[Streptomyces LRESGIEPEALDAFVPHQANLRMIELMAERLGLPERTAVARDVV sp.SA3_actF] RAGNTSAASVPLALEALLDSGEVGSGDRALLVGFGAGLNYAAQ VVELP SEQIDNO:38 >gi|320116117|ref| MCEKIAAGILGTGSYVPEKVLTNFDLEKMVDTSDEWITTRTGIKE YP_004186276.1|/ RRIADPSQATSDLATEAAKKALEDAKVDPSEIDMIIVATVTPDMN 1-3313-oxoacyl- FPSTACIVQANLGAANAAAFDISVGCSGFIYGLAIAQQFVETGMY (acyl-carrier- NKILVIGAETLSKIINWKDRNTCVLFGDGAGAVVVGRVESGYGIL protein)synthase SSYLGADGTGGKHLYMPAGGSRMPASEETVKKNLHTIFMEGQE III VFKFAVKVMDSATIEALNRCGLKPEDIDMLIPHQANTRIIEAARK [Thermoanaerobacter RLKLSNDKVYINLDKYGNTSAASVAIALDEAYRKGLIKKGDVILT brockiisubsp. VAFGAGLTWASSVIRWSK finniiAko-1] SEQIDNO:39 >gi|320449672|ref| MSGILALGAYAPERVMKNEEFEAYLDTSDEWIVTRTGIRERRIAA YP_004201768.1|/ EDEYTSDLAFKAVEDLLGRHPGALEGVDGVIVATNTPDALFPDT 1-3223-oxoacyl- AALVQARFGIQGFAYDLLAGCPGWLYALAQAHAMVEAGLARK ACPsynthase VLVVGAEALSKIVDWNDRATAVLFGDAGGAAVVGKVSKGFGFR [Thermus SFVLGADGTGAKELYHACVAPRLPDGTSMRNRLYMNGREVFKF scotoductusSA- AVRVMNTATLEAIEKAGLTPEDIKVFVPHQANLRIIDAARERLGL 01] PWERVVVNVDRYGNTSTASIPLALKEAVDEGRIREGDHVLLVSF GAGLTWAAAVITWGGA SEQIDNO:40 >gi|322421910|ref| MIRAEILGTGGFVPARVVPNAHFNYLVDDADQWIHSRTGIRERRF YP_004201133.1|/ ASAEEATSDLATNAALLALENGDVDPLEIDCIIVSTSTPDMILPAT 1-3263-oxoacyl- ACMVQKNIGAAKAFAFDMNAVCSSFIYGMEVADNLIRSGKYRK (acyl-carrier- VLLIGADTYSKILDFDDKGSAPLFGDGAGAVILGAGLSGKGILQS protein)synthase VMHSDGNGWELIQVPSSGSRKPVTAESIAAKENTFKMAGKSVFT III[Geobactersp. FATDVIPRIISDLAERGGIRAEDIDHIIPHQANVRIIDFISRKTGIPKE M18] KFLLNLDRYGNTAAASVGLALDENRRNGVIKSGELVLMMGFGG GLSWGGVLLKA SEQIDNO:41 >gi|322513545|ref| MYSKILATGSYLPAQIRTNADLEKMVDTTDEWIFTRSGMKERRIA ZP_08066645.1|/ AADETVATMGAQAAKKALEMAKIDHNEIDLIVVGTTTNSHAYPS 1-3163-oxoacyl- AACQIQGMLEIKDAIAFDVAAACTGFVYALSVADQFVRTGKVKK (acyl-carrier- ALVIGSDLNSRALDETDRSTVVLFGDGAGAVILEASEEQGIISTHL protein)synthase HSSSDSEYMLALPAQKRGNEKSGFIQMQGNATFKLAVGQLSSVV III[Actinobacillus EETLEANNLQKSDLDWLVPHQANIRIIAATAKKLEMDMSQVVLT ureaeATCC VEKYGNNSAATVPVALDEAVRDGRIQRGQLLLLEAFGGGWTWG 25976] SALVRF SEQIDNO:42 >gi|325677042|ref| MPAPIATATPAAHAALLGLGVYRPRRVVPNSEIVDRIDSSDEWIR ZP_08156713.1|/ TRSGITARGWAEPDETIVSMSVAAARDALAAAGLVAEQIDAVVL 1-3453-oxoacyl- ATSSQMVLGPSAGAVVATELGMQDTAAFDISAGCAGFCYALGN (acyl-carrier- AASLVRAGQARHVLVIGVERLSDLLDPTDRTCAFIFADGAGAVV protein)synthase VGPSDSEGIGPVAWGSDGSQTKAIKQDKDFMQYFAEVAAAEAA III[Rhodococcus GGSTERPYIRMDGQAVFRWAITFLEKACRDALEKAGVTADDLD equiATCC AFVPHQANSRITDALIRTLGLPDSVAVARDIAESGNTSAASIPMA 33707] MEQLLRSGEARPGDTALLLGFGAGLAYAGQVVQLPAIS SEQIDNO:43 >gi|325917371|ref| MSKRIYSRIAGTGSYLPEKVLTNDDMSKIVDTSDEWIFSRTGIRER ZP_08179586.1|/ HIVADDQTTSDLAYFASLKAMEAAGVTADEIDLIVIGTTTPDLIFP 1-3253-oxoacyl- STACLLQARLGNVGCGAMDVNAACSGFVYALSVADKFVRSGD (acyl-carrier- AKTVLVVGAETLTRIVDWTDRTTCVLFGDGAGAVILKADEETGI protein)synthase LSTHLHADGSKKELLWDPVGVSVGFGEGKNGGGALLMKGNDV III[Xanthomonas FKYAVKALDSVVDETLAANGYDKHDLDWLIPHQANLRIIEATAK vesicatoriaATCC RLDLPMEQVVVTVDRHGNTSSASVPLALDEAVRSGRVQRGQLL 35937] LLEAFGGGFTWGSALLRY SEQIDNO:44 >gi|326203621|ref| MIKSTKSVGIIGTGSFVPEKVLTNNDLEKMVDTSDEWIIKRTGISE ZP_08193485.1|/ RRILDHDTPNYTMGIEAANRALEDAGLKAEDIDLLILSTEAPDYM 1-3323-oxoacyl- SPSMSCIIQGAIGAVNAIAFDLNAACTGFIYSLSVARQFIANGVYR (acyl-carrier- NALVIGCEGLSKIVDWKDRNTCILFGDASGAVVLGEVDEGYGIL protein)synthase DSFLGSNGAEGMNITIPNLYLSEEEKAKRVNEKYNTLWMDGKEV III[Clostridium FKFAVKAMSSATMHVLDNLNMDIKELDFIFPHQANTRIIDGAIKK papyrosolvens LGITDDKIHYIINKYGNISSASIPVAMDEAKRDGKLKKGDNMVLV DSM2782] AFGGGLTWGSMAVKWSK SEQIDNO:45 >gi|332670773ref| MTRPTLTQATGPAHSRILGIGGVRGERVVPNDDLVGPIDSSDEWI YP_004453781.1|/ RQRTGIVTRRRAGEGTDVLDLAEGAARAAIENAGLTGADIDAVIL 1-3343-oxoacyl- STVTYFHQTPAGAAIIADRIGATPAAAYDISAACAGYCYGIGQAD (acyl-carrier- ALVRAGAARHVLVIGAEKMSEFVDPTDRSISFLLGDGAGAVVIGP protein)synthase SDTPGIGPTVWGSDGAQAQAIRQTHSWLATRDEGAGWPTLRQE III[Cellulomonas GQSVFKWAVWQMAPVAQKALDAAGVTADQIDAFVPHQANMRI fimiATCC484] IDQMIKQLKLPETVVVGRDIADTGNTSAASIPLATERLLREGQVSS GALALQIGFGAGLVYAAQVVVLP SEQIDNO:46 >gi|339488784|ref| MISGTGLYTPAQSISNEELVASFNTWSQQFNEDNAAAIERGEVEA YP_004703312.1|/ APLSDAAFIEKASGIKSRFVMDKAGILDPQRMKPRLPERSNDEPS 1-3693-oxoacyl- VLCEMAVAAARQALERAGRTAADVDGVIVACSNLQRPYPAIAIE ACPsynthase VQQALGIQGFAFDMNVACSSATFGIQTAANSVALGQARAVLMV [Pseudomonas NPEVCTGHLNFRDRDSHFIFGDAATAVLLERADKATSAHQFDIVS putidaS16] SKLWTEFSNNIRNNFGFLNRAAEEGEGAADKLFIQEGRKVFREVC PKVAELIGEHLQENGLQPSDVKRFWLHQANLSMNHLIVKKLLGR EVAEEDAPVILDRYANTSSAGSVIAFHLYQDDLAKGSLGVLSSFG AGYSIGSVVLRKR SEQIDNO:47 >gi|339494943|ref| MYNVVISGTGLYTPASSISNDELVESFNTYVHRFNSENAAAIEAG YP_004715236.1|/ EVQPLAESSSAFIEKASGIKSRYVTDKAGILDPERMVPRIPERSND 1-3733-oxoacyl- EWSILCEMSVKAAEEALARAGKTAADIDGVIVACSNLQRAYPAI (acylcarrier AIEVQAALGIKGFGFDMNVACSSATFGIQNAVNSIKLGQARAILM protein)synthase VNPEICTGHMNFRDRDSHFIFGDACTAVVIEREDLATSAHQWEV III[Pseudomonas LSTKLVTEFSNNIRNNFGFLNRTAEEYMSNPDKLFIQEGRKVFKE stutzeriATCC VCPMVAELIGEHLSENGIAVESVKRFWLHQANLNMNHLIVRKLL 17588= LMG GRDATEEEAPVILDTYANTSSAGSVIAFHKHQDDLPSGSLGVLSS 11199] FGAGYSIGSVILRKR SEQIDNO:48 >gi|340361349|ref| MQYAKILGTGSYLPANRVSNDDLAKKVDTSDEWITTRTGIKFRHI ZP_08683778.1|/ ADEGEKTSDLAAEASRRALVAAGVTADEIDLIIVATATPDMQFPS 1-3203-oxoacyl- TATIVQQKLGIANGCPAFDVQAVCAGFMYALSTANAYIKSGMA [acyl-carrier- KKALVIGAETFSRIVDWNDRTTCVLFGDGAGAVVLGASDEAGII protein]synthase HSKLKADGNYLDLLNVPGQIANGQVCGSPYITMDGPGVFKFAVK III[Neisseria MLAKIADEVISEAGYTPDQIDWLVPHQANKRIIDSTAKHLGLDME macacaeATCC KVILTVQEHGNTSAASIPLALDVGIQNGQIKRGQNLLLEGIGGGF 33926] AWGAVLVKY SEQIDNO:49 >gi|344206308|ref| MSKRIYSRIAGTGSYLPEKVLTNADLEKMVETSDEWIQSRTGIRE YP_004791449.1|/ RHIAAEGETTSDLGYNAALRALEAAGIDASQLDMIVVGTTTPDLI 1-3253-oxoacyl- FPSTACLIQAKLGVAGCPAFDVNAACSGFVFALGVADKFIRSGDC ACPsynthase KHVLVIGTETLTRMVDWNDRTTCVLFGDGAGAVVLKADEETGI [Stenotrophomonas LSTHLHADGSKKELLWNPVGVSSGFKDGANGGGTINMKGNDVF maltophiliaJV3] KYAVKALDSVVDETLAANGLDKSDLDWLIPHQANLRHEATAKR LDMSMDQVVVTVDKHGNTSSGSVPLALDAAVRSGRVERGQLLL LEAFGGGFTWGSALLRY SEQIDNO:50 >gi|345304635|ref| MPYAAITAVGHFLPEDRLTNADLEKMVDTSDEWIRTRTGIRERRI YP_004826537.1|/ LRDPNKATSYMATEAARECLRKRGMDPEDVELIIVATVTPDMFF 1-3463-oxoacyl- PATACLVQANLGARNAWGFDLSAACSGFLFALSTAARFIESGKH ACPsynthaseIII KRVMVIGADKMSTITDYTDRKNCILFGDAAAAVLLEPDPECGVI [Rhodothermus DSVEHCDGNNWELLCMLGGGSLNPPTHETVDRKMHYLHQEGR marinus AVFKLAVEGMAQVAVEIMERNNLTADDVRYLVPHQANLRIIDA SG0.5JP17-172] TARRMGLSPDKVMVNIDRYGNTTAATIPLCLYDWERQLRRGDN LILAAFGGGFTWGAIYLKWAYDGDKVAAAAEATAETSTENA SEQIDNO:51 >gi|349685677|ref| MTAKRSLLSGFGGYLPERIVTNDELASRLDTSDEWIRGRTGIGQR ZP_08896819.1|/ HIAGENDTAVSMAAQAARRALDYAGAAPDDVDAIIVATSTPDQ 1-3233-oxoacyl- AFPSTAVRVQAELGMTSGFGFDLAAACSGFIYALSMADSLIRSGQ [acyl-carrier- ARSALVIGSEVYSRILDWSDRGTCVLFGDGAGAAFLTAAGPDDG protein]synthase DAGILSTHLHSDGQYGDLLYVDGATGQHDRPAHLRMQGRDVFR III HAVGKLSASVDEALAANNLSHADVNWLVPHQANLRIIDGVARK [Gluconacetobacter LALPAERVVVTVDRHANTSAASIPLALNEAVRDGRIRKGDLVLM oboediens EALGGGLTWGSALVRL 174Bp2] SEQIDNO:52 >gi|352106212|ref| MTHVVITGTGLYTPEHAIDNAALVAAFNAWVDGENEQHAEAIER ZP_08961263.1|/ GEREPLANSSSEFIEKASGIKSRYVLDASGILDPQRMRPKLPQRSN 1-3733-oxoacyl- DEPSLQCEMATEAAHQALAAAQVDAADIELVIVACSNLERAYPA (acylcarrier VAVEVQQTLGTSGYGFDMNVACSSATFALETAANAIASGSVNRA protein)synthase LVVNPEICSAHLNFRDRDSHFIFGDACTAVVLENSAVAVADEQFE III[Halomonas ILGTRLVTKFSNAIRNNAGFLNRVTDSDPMALDKLFVQEGRRVF sp.HAL1] KEVCPMVAKLITDHLASLELNGSDLKRMWLHQANRHMNDLIAR KVLGYDPSETQAPIILDRYANTSSAGSIIAFHLHREQFNQGDIGVIC SFGAGYSAGSVVIRRV SEQIDNO:53 >gi|358061230|ref| MNVGIKGFGAYAPENIIDNAYFEQFLETSDEWISKMTGIKERHWA ZP_09147893.1|/ DEDQDTSDLAYNASVKAIEDAGIKPEDIDMIIVATATGDMPFPSV 1-3133-oxoacyl- ANILQERLGTGKVASMDQLAACSGFMYSMITAKQYIQSGDYHNI (acylcarrier LVVGADKLSKITDLTDRSTAVLFGDGAGAVIIGEVSEGRGIISYEM protein)synthase GSDGSGGKYLYLDKETGKLKMNGREVFKFAVRIMGDASTRVVE III KANLTSDDIDLFIPHQANIRIMESARERLGISKDKMSVSVDKYGN [Staphylococcus TSAASIPLSINQELQNGKLKDDDTIVLVGFGGGLTWGAMTIKWG simiaeCCM K 7213] SEQIDNO:54 >gi|373112342|ref| MKSVGIKGLSSYVPERIMTNFEFEKIIDTSDEWIRTRTGIEERRFAS ZP_09526574.1|/ PEQATSDLCYEATQKLLATMKMDPQEIDFIMVCTCTPDYPVPSTA 1-3283-oxoacyl- CVLQSKLNLLGVPAVDINAACSGFMYGLAMATSMVQTGLYKNV [acyl-carrier- LVIGAETLSRIMDMQDRNTCVLFGDGAAAAIIGEVEEGSGILATH protein]synthase LGAEGEDEGILQIPGGGSRYPSTLESVHTKKQFVQMKGQNVYKF 3[Fusobacterium AVHALPEATLAALKKAKVEASQVARFFPHQANLRIIEAAAKRMN necrophorum VSLDKFHVNLHKVGNTSAASVGLALADALEKGMVKKGDYIALT subsp. GFGAGLTYGSVVMKWAY funduliforme 1_1_365] SEQIDNO:55 >gi|374851360|dbj| MGTTLTGIGYYLPPKVLTNFDLEKMVDTSDDWITTRTGIKERRIA BAL54322.1|/1- DNENVTQMAYMASLEALESANIQPEDIDLIILATLTPELKFPSTAC 3073-oxoacyl- LLQAKLGAKRAYAFDISAACSGFIYGLELADAYIKSGKAKKILLV [acyl-carrier- GAERLSEIVNWQDRSTCVLFGDGAGAVIISEGDGEVLSSKMLSDG protein]synthase ELWEILYAPKCGYINMKGKELFKLAVRSMEEVCRYVLESAGISIE III[uncultured DVSIMIPHQANIRIMEALAEKLGMPKEKVYSNIHKYGNTSAASIPI Aquificae AMYEAYKEGKLRRGDIVMLTAMGGGLTWGAALLRF bacterium] SEQIDNO:56 >gi|375098553|ref| MSTQDARGVAVLAGLGGWLPPRVVDNDELSRRLDTSDEWIRTR ZP_09744816.1|/ TGIAKRHVVHTGLSTVDMAVEAGRRALESAGPYGENVDAVVLA 1-3403-oxoacyl- TSTPDHVCPASAPQVAAELGLSGAAAFDVNAVCSGFVYALATAS (acyl-carrier- GLISGGVAKRVLLVGADAFTTLLDPDDRTTVPIFGDGAGAVVLR protein)synthase EGSADELGAVGPFDLHSDGELAELLIVPAGGSRRKKSENASDHFL III KMQGPAVFRHATARMASSSRAVLEKAGWTTSDVDRFVGHQAN [Saccharomonospora VRILTATAKNLGLPADSLVVNIGHTGNTSAASIPLAMVDAAVDG cyaneaNA- MLQPGDRVLVTAFGAGLTWGSTVLRWPELACAPLP 134] SEQIDNO:57 >gi|381164912|ref| MTRPTLTLAQGAKASRVLGVGSTQPDRVVTNDELSQHMDTSDQ ZP_09874142.1|/ WIRDRVGIIERRFAGEDERLVDMAVTAGAKALADAGVAPSEVDT 1-3263-oxoacyl- VIVPNCTMPAPIPNAAAQVADRIGVKAAGAFDLNAACAGFCYGL (acyl-carrier- GVASDLVRAGSAKKVLVIGAEKLTDVVDPTDRSTAIIFADGAGA protein)synthase ALVGPSDEPGIGPVAWGSAGDLVDVIYMRDNRYIFQEGQPVFRW III ATTQIAPVAMRAVELAGLELSDIDVLIPHQANLRIVEAIAKRLRA [Saccharomonospora KGARDDMVVADDIRYSGNTSSASIPMALDHMRAAGTVKPGDVV azureaNA- LTVGFGAGLSYAGQVLICP 128] SEQIDNO:58 >gi|383771442|ref| MTQIRSVVLGCGSYLPEQVVTNAQLAARIDTSDEWIVQRTGIRER YP_005450507.1|/ HIAAEGEFTSHLAIKAAQAALTDAGLDAQSIDLIVLATSTPDNTFP 1-3263-oxoacyl- ATAVAVQHGLGINHGAAFDLQAVCSGFVFALATADNFLRTGAF ACPsynthase KRALVIGAETFSRILDWNDRGTCVLFGDGAGAVVLEAQEQPGNA [Bradyrhizobium ATDRGVVTTHLRSDGRHKAKLFVDGGPSSTQTVGHLRMEGREV sp.S23321] FKHAVGMITDVIVDAFEATGLNADSIDWFVPHQANKRIIDASAH KLHIAPEKVVLTVDRHGNTSAASIPLALSVARRDGRIKRGDAVL MEAMGGGFTWGSALVRW SEQIDNO:59 >gi|384154990|ref| MIYAAFRSIGAYIPPKIMSNADFEKIIDTSDEWITKRTGIKERRIAN YP_005537805.1|/ EGEASSDLGARAGELAIERAGISKEEIDLVICATVTPDFLCMPSTA 1-3333-oxoacyl- CLIAAKLGLPNVMAFDVSAACTGFVYALNVAKAFIESGMKKNV ACPsynthase LIVGAEKYSAILDYTDRTTCFLFGDGAGAAIISATNDKNESIIDINC [Arcobacter SSDGNYEDLIKTPGGGSKNPCSQEVLENKMACIKMKGNETFKLA butzleriED-1] VKTLTSDVKTMLEKHNLTNEDINHFIPHQANYRIIKAVGEALDLS DEKTVVTVDKYGNTSAASIPMAMNYAFEQGKIKAGDTILFDAFG GGLTWGSALFKFAPIKR SEQIDNO:60 >gi|384450582|ref| MCVKKTRKASIWATGSYLPEKILSNSDLEQMVDTSDEWIVTRTGI YP_005663182.1|/ KERRIAAANEYTSIMGAKAAERAIQKAGLTKDQIECIIFSTSAPDY 1-3353-oxoacyl- IFPSSAALAQAYLGIKDIPAFDCMAACTGYLYGLSVAKAYVESG ACPsynthase MYNNVLLIAADKLSSFVNYKDRNTCVLFGDGGAACIIGESRPGA [Chlamydophila LEITNVNLGADGSVADLLSLPAGGSRVPASQETLEAGKHFISMEG psittaci6BC] KEVFKHAVRRMESAAKTCIAGAGIEESDIDWLVPHQANERIIDAI AKRFEIDEGKVFKTLCKYGNTAASSVCIALDELLQSHTIHSGEYL LLVAFGGGLSWGAVVLQQVES SEQIDNO:61 >gi|385331603|ref| MIKAVISGTGLYTPPATISNDELVEAFNQYVELFNAENADAIASG YP_005885554.1|/ DVTPLQPSSSSFIEKASGIKRRHVIDKDGILDPNRMKPYIPDRSNEE 1-3733-oxoacyl- PSVQCDMAVTACREALEQAGKSAEDVDAVIVACSNLQRAYPAV ACPsynthase SIEVQEALGIDGFAYDMNVACSSATFGLQAAVNSVENGSARAVL [Marinobacter VVSPEICSGHLNFRDRDSHFIFGDACTAILVEREEDTREGQGFEIL adhaerensHP15] GTSLKTKFSNNIRNNFGFLNRADESGVGKPDKLFVQQGRKVFKE VSPLVAETIQKQLQSLSLAPDDLRRMWLHQANLNMNQLIARKVL GRDATEEEAPVILDEYANTSSAGSIIAFHKNKDDLVSGDLGVICSF GAGYSIGSVVVRRR SEQIDNO:62 >gi|386265484|ref| MNSRILSTGSYLPSHIRTNADLEKMVDTSDEWIVTRSGIRERRIAA YP_005828976.1|/ ADETVATMGFEAAKNAIEAAQINPQDIELIIVATTSHSHAYPSAAC 1-316Beta- QVQGLLNIDDAISFDLAAACTGFVYALSVADQFIRAGKVKKALVI ketoacyl-ACP GSDLNSRKLDETDRSTVVLFGDGAGAVILEASEQEGIISTHLHAS synthaseIII ADKNNALVLAQPERGIEKSGYIEMQGNETFKLAVRELSNVVEET [Haemophilus LSANNLDKKDLDWLVPHQANLRIITATAKKLEMDMSQVVVTLD influenzaeR2846] KYANNSAATVPVALDEAIRDGRIQRGQLLLLEAFGGGWTWGSA LVRF SEQIDNO:63 >gi|386335197|ref| MHDVVISGTGLWVAPEVITNEELVASFNAYARHYNEANATAIAA YP_006031367.1|/ GTLAAVAESSVEFIEKASGIRQRYVIDKAGVLDPARMRPRLAPRG 1-3733-oxoacyl- DDALSLQAEIGVAAAREALAAAGRDAGDIDMLICSAANMQRPYP ACPsynthase AMGIEIQNALGADGYAFDMNVACSSATFGLEQAINAVRTGSARV [Ralstonia ALMVNPEITSGHLAWKDRDCHFIFGDVCTAVVVERADDARAPD solanacearum QWQVLGTRMATRFSNSIRNNAGFLSRSEDRDPDDRDQLFRQEGR Po82] KVFKEVCPMAAEHIAGHLQSLGHAPADVRRFWLHQANLGMNQ LIGKRLLGRDASADEAPVILDEFANTASAGSIIAFHRHRADLQPGD LGLICSFGAGYSIGSVAVRKR SEQIDNO:64 >gi|390454110|ref| MNKLRPVGIIGTGKYVPEKILTNKDLEAIVETSDEWIVSRTGIQER ZP_10239638.1|/ HIAAPEQATSDLAYEAAIKALKSAGMTAEDLDLIIVATVTPDMAF 1-3293-oxoacyl- PSTACILQDKLGAKGAAAFDLSAACSGFVYGLATATSFIKTGIYN (acyl-carrier- NALIIGADCLSRITDYTDRNTCVLFGDGAGAVVIGEVSEGRGFQS protein)synthase FDLGAEGAGGSLLNLAAGGSRLPASADTLENKQHYIYMNGREVF III[Paenibacillus KFAVRVMGTATVDVLEKAGLTKDDIDLFVPHQANIRIIQSAMQR peoriaeKCTC LDLPEEKVVINVNKYANTSAASIPLALVEAAEEGRMKEGDRVLM 3763] VGFGGGLTWGASVLVW SEQIDNO:65 >gi|392946737|ref| MLGLGVYRPARVVTNDEIAQRVETSDAWIQSRTGIATRRIADEEE ZP_10312379.1|/ TTVAMGAAAAEKALAAAGLTADTIDLVIGATCTSPSQIPGAGPQI 1-3073-oxoacyl- AHRIGADQAGAFDINGACAGFSYAVSTAADMVRAGSVRHVLVV (acyl-carrier- ATERLSDYTDWDDRSTCILLADGAGATVIGAAETDEIGPAVWGH protein)synthase DGSRPEAIRVPGYGDNMFRMEGQAVFRWAISLVPTVRQICERAG III[Frankiasp. VAPDELAGIVPHQANLRIVEALATGIGATNAAVARDVVDSGNTS QA3] AASIPLGLARLLDAGEIRRGDPVLLFGFGAGLTYCGQVVRCP SEQIDNO:66 >gi|397172008|ref| MQQVVISGSGLFTPQHIISNDELVVSFNQYVDQFNTEHAAQIAAG ZP_10495404.1|/ ELAALEYSSSEFIEKASGIKARHVLYKDGILDPKVMHPVFRKRGE 1-3723-oxoacyl- DELPEMVEMAVQAATQALAQANKTAADIDLIICAASNMQRPYP (acylcarrier ALSVELQQALGAGGYAFDMNVACSSATFAISNAVNAIRGGSAKV protein)synthase VLVVNPEFASPQVDYRSRDSHFIFGDVCTATIIEAESSCTSSQAFRI III[Alishewanella LGMRLKTTFSNNIRCDIGYTEHCFSEQDPKAPFFKQQGRKVFKEL aestuariiB11] LPIVAEVILDEMAAQQVTADDLKRLWLHQANINMNIFAAKKILG RDPLPEEAPLVLDTYANTASAGSIIAFHKYQQGLQSGDKAILCSF GAGYSVGCLVLEKC SEQIDNO:67 >gi|398305630|ref| MKAGILGVGRYIPEKVLTNHDLEKMVETSDEWIRTRTGIEERRIA ZP_10509216.1|/ ADDVYSSHMAVAAAKKALEQAEVAAEDLDMILVATVTPDQSFP 1-3123-oxoacyl- TVSCMIQEELGAKKACAMDISAACAGFMYGVVTGKQFIESGTYK (acylcarrier HVLVVGVEKLSSITDWEDRNTAVLFGDGAGAAVVGPVSDDRGIL protein)synthase SFELGADGTGGQHLYLNEKGHTIMNGREVFKFAVRQMGESCVN III[Bacillus VIEKAGLSKEDVDFLIPHQAMRIMEAARERLELPVEKMSKTVHK vallismortisDV1- YGNTSAASIPISLVEELEAGKIKDGDVVVMVGFGGGLTWGAIAIR F-3] WGR SEQIDNO:68 >gi|398884037|ref| MHNVVISGTGLYTPANSISNEELVQSFNAYVAQFNADNADAIAR ZP_10638982.1|/ GEVEALTESSAAFIEKASGIKSRFVMDKDGILDPQRMAPRLPERS 1-3733-oxoacyl- NDEWSVLCQMAIGAAEQALQRAGKTAADIDGVIVACSNLQRAY (acyl-carrier- PAIAIEVQEALGIQGFGFDMNVACSSATFGIQAAANSVQLGQARA protein)synthase VLMVNPEVCTGHLNFRDRDSHFIFGDAATAVIIERADLATSKYQF III[Pseudomonas DVVSTKLLTKFSNNIRNNFGFLNRAAEEGIGAKDKLFVQEGRKVF sp.GM60] KEVCPMVAELIGAHLEENQLNVGDVKRFWLHQANLSMNHLIVR KLLGREATEAEAPVILDTYANTSSAGSVIAFHKNQDDLAAGSLA VLSSFGAGYSIGSVILRKR SEQIDNO:69 >gi|399047091|ref| MRQMDKKRSVGILATGSYTPDRVLSNFDLEKMVETTDEWIVSRT ZP_10739223.1|/ GIRERRICSAEQASSDLAYEAAKKALERANISAEQLDMIIVATVTP 1-3423-oxoacyl- DMMFPSTACILQEKLGAKRAAALDVSAACTGFLYGITTAAQFIA (acyl-carrier- NGLYKYVLVVGVETLSKITNYKDRNTCVLFGDGAGAAVIGEVRE protein)synthase GFGFQSFELGADGAGGELLCLPAGGSRIPASSESVENNLHYLSMA III[Brevibacillus GGEVFKFAVRVMNSATEAVLSKAGVERENIDLLVPHQANKRIID sp.CF112] SAVQRFGLSEDKVAINLDRYGNMSSASIPVALDEAIAAGRVKEG DNVILVGFGGGLTWGATLLKWSTTPAEGSGQ SEQIDNO:70 >gi|400755130|ref| MFTPAITGTGVFTPSQTITNAELVAAFNAYADKTNAENAKAIAAG YP_006563498.1|/ EMEPLAHSSEEFILKASGIEQRYVMDKSGVLDPEVMHPLLRQRG 1-3743-oxoacyl- DDEPSIMAEMALDAAKKALAQAGKTAADVDTVICAASNMERAY [acyl-carrier- PALAIEIQDLLGIKGFAFDMNVACSSATFGIQAAADMVRSGSIRS protein]synthase ALVVNPEICSGHLEWRDRDCHFIFGDVATATLIERSEDATGAYFEI 3[Phaeobacter LSTRCATSFSNNIRNNNGYLRRSRPDGVEDRRDMQFMQNGRKVF gallaeciensis2.10] KEVLPMVSQHIAEHMEAEGVSNTDLKRLWLHQANKTMNDFIGK KVLGRTPEAGEQPNILQDYANTSSAGSIIAFSKYSDDLSAGDLGLI CSFGAGYSVGSVILRRVA SEQIDNO:71 >gi|400756529|ref| MMRARIVGTGSAVPSKVLTNFDLEKMVDTSDEWVTTRTGIKERR NP_952652.2|/1- IAVDGEYTSTFATLAAERALEMAGVKASDLDLLIVATITPDFPFPA 3263-oxoacyl- TACVVQSNLKATKAAAYDISAACSGFIYALAQASNAIRSGSARK ACPsynthase ALVIGAEVLSRIIDWTDRNTCLLFGDGAGAVVLEACDDGHGVLS [Geobacter THLHSDGSYWELLYQPGCGNRNPAVQKTLDDRRIYLMMQGNEV sulfurreducens FKLAVRAMEDAALEALDANGLTPADISLFIPHQANRRIIDAIGKRL PCA] GLPGEKVYVNLDRFGNTSAASIPLALDEANRSGRIKPNDVVVFD AFGGGLTWGSALVRW SEQIDNO:72 >gi|401563713|ref| MPKISAGILGTGYYVPERVLTNFDLEKMVQTNDAWIVERTGIHE ZP_10804658.1|/ RRIAADGEPVSVLAQRAAEMALADAGVDAADLDLIIMATLTSDR 1-334beta- IIPSTACVLQDRLGAKHAAAFDLSAACSGFVYAASIAAQFIESGV ketoacyl-acyl- YRHVLVIGGETLSKVVDWEDRNTCILFGDGAGAAVFGPVEDGY carrier-protein GIRAFDLGSDGSGGDALDIPSSGSLCPVTPETIEQRLNFVHMDGK synthaseIII AVFRFATKVMGRTVETSLERAGMQREDLDYLVPHQANIRIIQAA [Selenomonassp. AKRLDMPMDKVIINIHRYGNMSAASIPVALAEAAHAQQFKKGD FOBRC6] NIALAGFGAGLTWASCIMKWAKEENG SEQIDNO:73 >gi|402823152|ref| MIRSVLIGTGSALPRNAVSNAELAERVDTSDEWIVERTGISNRHIA ZP_10872590.1|/ EADETTSSLATEAGRKAIEAAGIDAESIDLIVLATATPDQTFPASA 1-3233-oxoacyl- TIVQSRLGCRAGGIAFDVAAVCSGFLYAVGVADSMLRTGMARR (acylcarrier ALVIGAETFSRILDWEDRTTCVLFGDGAGAVVLEAQEQVGETPR protein)synthase GILATRLHADGAHNQLLFVDGGPSTTGTVGKLRMKGREVFRHA III VVNLAEVLREVIEEAGLSTSDIDWLVPHQANARILDATAKKLSLP [Sphingomonas PEKVVMTVGQHANTSAASVPLALDVAVRDGRIKQGDLVMLEA sp.LH128] MGGGFTWGASLIRI SEQIDNO:74 >gi|407684813|ref| MSQQVVISGVGVWHPKDSITNEELVDSYNAYVDAFNEENKAQIE YP_006799987.1|/ SGDVAAMPYSSAEFIEKASGIKSRYIYQKEGALDITRMKPKIAPR 1-3743-oxoacyl- ADDELSHQAEIAVEAAKLALASANVTADEIDAVIVSCAYTQRAY ACPsynthase PAIAIEVQEALNIEGFGFDMLVACSAATFGMHRAYEMLSAKNAT [Alteromonas RVLVINPELVSPQINYADRDSHFIFGDVATATVLELAETAKSEHV macleodiistr. YDVLSTKALTKFSNNIRSNFGYMTRAEDVDPYGPDKLFHQAGRK EnglishChannel VFKEVCPLAAAHIEAHLASHDITPEGVKRWWLHQANINMNTLIC 673] KRLLGRDADRTEAPIVLDEYANTASAGSVIAFGLNHEDLVAGDV GVLCSFGAGYSIGSLVIRKR SEQIDNO:75 >gi|410479651|ref| MTPTMLNRSIILGTGSFAPANVLTNEDISRKVETSDLWIRERTGIR YP_006767288.1|/ ERRIASSGESTSDLALEAGRNALRNAALSPADLDGIIVATATPDLT 1-3413-oxoacyl- FPSTACLVQARLGIPGTFAFDVNAVCSGFMYALKIADSMIRSGQC (acyl-carrier- ETLLVIGAEVMSRFVDWSDRSTCILFGDGAGAVVLGKSGSPQTG protein)synthase GVGTVTLHADGRYWDLIHVPGGGSRSPVETEKPPGNACTIRMKG III SETFRMAVRSLEESVREVLKEEGIGVNELDWVVPHQAMRILEAL [Leptospirillum SERLGIPLGHFVVNIDRYGNTSAASIPMALDEAVQDKRIQPGHRIL ferriphilumML- LTAFGSGVTWGSGLVHWTQKAGGDR 04] SEQIDNO:76 >gi|410617776|ref| MNSRIIGTGSYYPSEVRTNADLSLMVDTSDEWITDRTGIKERRIIG ZP_11328741.1|/ ADETAASMGVEASKKALEAAGIDAKSLDMIVCATTSGRYALPST 1-3193-oxoacyl- ACEIQKALDIDGIPAFDVAAACAGYCYALSVADQYIKSGMAKRIL [acyl-carrier- VVGTDCLSRMISPEDRTMVILFGDAAGATIIEASEEPGILSTHIHAA protein]synthase GSYGDLLAIGNPTRGDEASIHENWGSMKGNEVFRVAVTKLSEVV 3protein1 EETLAANNMQKSDLDWLVPHQANFRIIKATAKKLNMSLDQVVL [Glaciecola TLERYGNTSAATVPTALDEAIRDGRIKRGQNLLLEAFGGGFAWA polarisLMG SALVRY 21857] SEQIDNO:77 >gi|411009303|ref| MHSKILGTGSYLPHSVRTNADLEQMVETSDEWIVERTGIRERRIA ZP_11385632.1|/ GADETVATLSHQAALRALEAAGLTAADLDMIVLATTSAENAFPA 1-3193-oxoacyl- AACELQGLLGVQGIPAFDVAAACAGFTYALSIADQFVKSGAARH (acylcarrier VLVVGADVLSRMCDPEDRGTIILFGDGAGAVVIGASDTPGILSTH protein)synthase LHADGRYGELLKLPQPRRGMPGAELEAYMYMKGNDVFKVAVT III[Aeromonas RLSEIVTETLAAAGIEPSELDWLVPHQANFRIISATAKKLGMGLD aquariorum KVVLTLDKHGNTSAASVPIAFDEGVRDGRIKPGQLVLLEAFGGG AAK1] FAWGSALVRL SEQIDNO:78 >gi|415794657|ref| YTKIIGTGSYLPEQVRTNADLEKMVDTSDEWIVTRTGIRERHIAA ZP_11496472.1|/ PNETVSTMGFEAATRAIEMAGIEKDQIGLIVVATTSATHAFPSAA 1-3163-oxoacyl- CQIQSMLGIKGCPAFDVAAACAGFTYALSVADQYVKSGAVKYA (acyl-carrier- LVVGSDVLARTCDPTDRGTIIIFGDGAGAAVLAASEEPGIISTHLH protein)synthase ADGSYGELLTLPNADRVNPENSIHLTMAGNEVFKVAVTELAHIV IIIfamilyprotein, DETLAANNLDRSQLDWLVPHQANLRIISATAKKLGMSMDNVVV partial TLDRHGNTSAASVPCALDEAVRDGRIKPGQLVLLEAFGGGFTWG [Escherichiacoli SALVRF E128010] SEQIDNO:79 >gi|417318270|ref| MDTSDEWIRTRTGIEERRIARDDEYTHDLAYEAAKVAIKNAGLTP ZP_12104859.1|/ DDIDLFIVATVTQEATFPSVANIIQDRLGAKNAAGMDVEAACAGF 1-2873-oxoacyl- TFGVVTAAQFIKTGAYKNIVVVGADKLSKITNWDDRTTAVLFGD (acylcarrier GAGAVVMGPVSDDHGLLSFDLGSDGSGGKYLNLDENKKIYMNG protein)synthase REVFRFAVRQMGEASLRVLERAGLEKEDLDLLIPHQAMRIMEAS III[Listeria RERLNLPEEKLMKTVHKYGNTSSSSIALALVDAVEEGRIKDNDN monocytogenes VLLVGFGGGLTWGALIIRWGK J1-220] SEQIDNO:80 >gi|417334430|ref| MLGIKGCPAFDVAAACAGFTYALSIADQYVKSGAVKHALVVGS ZP_12117640.1|/ DVLARTCDPGDRGTIIIFGDGAGAAVLSASEEPGIISTHLHADGRY 1-2213-oxoacyl- GELLTLPNADRVNPDNPIYLTMAGNEVFKVAVTELAHIVDETLA acyl-carrier- ANNLDRSELDWLVPHQANLRIISATAKKLGMSMDNVVVTLDRH proteinsynthase GNTSAASVPCALDEAVRDGRIKAGQLVLLEAFGGGFTWGSALIR KAS3 F [Salmonella entericasubsp. entericaserovar Alachuastr.R6- 377] SEQIDNO:81 >gi|417747984|ref| MKQIAATSGPTNIGLLSVGSYRPQRVVTNDELCQNIDSSDEWIYS ZP_12396438.1|/ RTGIKTRRFAARDESTASMATEAGREAIAKAGLEASDIDCVVVAT 1-3353-oxoacyl- STHFLQTPACGPAVAAALGATGVPAFDISAGCAGFGYALGVAAD (acyl-carrier- MVRGGTAGKVLVLGSEKLSPTVDMTDRSNCFIFADGAAGVVVG protein)synthase ETPTQGIGPTVWGSDGTQATAIRQDIDWMDYLDRPTGPRPFLRLE III GSAVFRWAAFEMGKVGQQAMDAAGVRPDEIDVFLPHQANSRIN [Mycobacterium EILAKSLELRPDAVIANDIEHTGNTSAASIPLAMAEVLATGAAKA aviumsubsp. GDLALLIGYGAGLSYAAQVVRLPPG paratuberculosis S397] SEQIDNO:82 >gi|420680190|ref| MLGIKDAASFDLAAACAGFTYALSVADQYVKSGAVKHAIVIGSD ZP_15164698.1|/ VLSRALDPEDRGTIILFGDGAGAVVLGASEQPGIMSTHLHADGRY 1-2203-oxoacyl- GELLALPYPDRQQDQPAYVTMAGNEVFKVAVTELAHIVDETLQ [acyl-carrier- ANNLDRTALDWLVPHQANLRIISATAKKLGMGMDKVVITLDRH protein]synthase GNTSAASVPSAFDEAVRDGRIQRGQLVLLEAFGGGFTWGSALVR 3[Yersiniapestis F PY-47] SEQIDNO:83 >gi|421612789|ref| MIETSSNVTANDLAAKSVNEESSAESTAVPTEAVSAVMPGNATT ZP_16053888.1|/ RGRMGNLKGVRIAGTGSYVPERIVTNEDLAALGCDSDWIVRRTG 1-3923-oxoacyl- ILQRRHAEPGQATSDLCYEAALRCLENANVSVDEIDLILVATITPD (acyl-carrier- HPTPSTACHLQRRLGAVAPAMDIGAACAGFMYALVTGAQFVSN protein)synthase GNARNVLVIGADLMSRTVDPEDKKTYPLFGDAAGAALLVPSTQ III DECQSTECNGSAADSTSQTDGLLAYQLGSEGCGGEMLCIPAGGS [Rhodopirellula RTPITTDGEDSASRYLQMDGRGVFKWAVRVFDESAKDVLRAAN balticaSH28] VSSDQLSLVVLHQANQRIIDSAVSDLNVPPEKVFVNLDKYGNTSG ASIPLALDEAARAGRLKEGDLVLLCGFGAGLAWGTALLRW SEQIDNO:84 >gi|421888767|ref| MPRCRFPPPLRPPTPHKGSAPGHPIPTPHMTRYARIIGTGSYLPPK ZP_16319848.1|/ RVTNHELAAQLAEKGIETSDEWIVTRSGIRARHYAEPDVTCSDLA 1-3553-oxoacyl- VKAAERAIEAAGIDRAEIDMILVATSTPDFVFPSAACLVQQKLGL (acyl-carrier- SNHCAAFDLQAVCSGFVYALATADKFIRAGGCRNVLVIGAEVFS protein)synthase RILDFNDRTTCVLFGDGAGAVVLQASDEPGILSTALHADGSHADI III(Beta-ketoacyl- LCVPGNVAAGAIKGSAFLYMDGQAVFKLAVNVLDKVAREALGL ACPsynthaseIII) ANVEASQIDWLIPHQANIRIMQGTAKKLGLPNERMVVTVDEHGN (KASIII) TSAASIPLALDAAVRDGRIRKGHHVLLEGVGGGFTWGAALLRF [Ralstonia solanacearum K60-1] SEQIDNO:85 >gi|422338672|ref| MQSIGIKGIGYYVPENVFTNFDFEKIIDTSDEWIRTRTGIVERRFAS ZP_16419632.1|/ KDQATSDLAREAALKAIENAKIKKEDVDMIILATTTPDYIAQGAA 1-3283-oxoacyl- CIVQNKLGLTSIPCFDLNAACTGFIYGLEVAYSLVKSGLYKNVLVI (acyl-carrier- GAETLSRIIDMQNRNTCVLFGDGAAAAIVGQVEEGYGFLGLSIGA protein)synthase EGEDDMILKVPAGGSKKPNDEETIKNRENFVIMKGQDVFKFAVS III TLPKVTLDALEKAKLDVNDLSMVFPHQANLRIIESAAKRMKFPL [Fusobacterium EKFYMNLSRYGNTSSASVGIALGEAVEKGLVKKGDNIALTGFGG nucleatumsubsp. GLTYGSAIIKWAY polymorphum F0401] SEQIDNO:86 >gi|423074933|ref| MVSVGIVGTGSYVPDKVLTNFDLEQMVDTNDQWIVSRTGIKERH ZP_17063653.1|/ IAEPETPVSELCYQAAVRALEDAKLPPEELDLVIVATITPDFVFPA 1-3313-oxoacyl- TACLVAERLGAKKAAGFDLQAACTGFLYGVATAAQFIATGIYKN (acylcarrier ALVIGGETLSKILNWEDRGTCILFGDGAGAAVLQPVEEGYGFLG protein)synthase YDLGMDGAGGSLLTMPGGGSMHPASAETVAKKMHTIQMAGSE III VFKFAVRIMGETALKALDKAGLGIGDVDCLIPHQANTRIVDAAV [Desulfitobacterium KRLGIDAKKVVVNLDRYGNMSAASIPVALDEAARSGRLNYGDI hafnienseDP7] MVMVGFGGGLTWGAAVVKWSKRGV SEQIDNO:87 >gi|423197564|ref| MTSIVISGSGLYTPPFAVSNEALVAAFNQYVDLYNEENASAIDAG ZP_17184147.1|/ QLPAKQHSSSEFIEKASGIKSRYLVSKEGVLDPDIMQPLLAERPDD 1-373 KPSIMVEMAVAAAEQALIAAGREPGEIDLVIVAASNMPRPYPALS hypothetical IELQHYLGASGMAFDMNVACSSATFGIKTAADMLAAGSARLAL protein VVNPEICSGHLNFRDRDSHFIFGDACTAVLLEREADCQVANPWQ HMPREF1171_ LVASKLVTQYSNNIRNNFGFLNRLSPRTRYGDDKLFRQQGRKVF 02179[Aeromonas KEVLPLVCDQIAGQLDEQGWAADSLSRLWLHQANLTMNQFIAR hydrophilaSSU] KLLGHDASQQEAPVILDSYGNTSSAGSIIAFHLYNRDLPAGARGV LCSFGAGYSIGSLLLRRL SEQIDNO:88 >gi|424068956|ref| MHNVVISGTGLFTPANSISNEELVQSFNAYVAQFNSDNAAAIERG ZP_17806404.1|/ DVQALSESSAAFIEKASGIKSRFVMDKEGILDPQRMKPNLPERSN 1-3733-oxoacyl- DEWSILCEMGVAAATQALQRAGKTAADIDGVIVACSNLQRAYP ACPsynthase AISIEIQQALGVAGYGFDMNVACSSATFGIQAACNSVQLGQARAL [Pseudomonas LVISPEICTAHLNFRDRDSHFIFGDGATAVVVERADLATSPYQFDI syringaepv. VSTRLLTQFSNNIRNNFGFLNRTSDEGQSAPDKLFVQEGRKVFRE avellanaestr. VCPMVAELVAAHLQDNGINITDVKRFWLHQANLSMNHLIVKKL ISPaVe013] LGRDASVEEAPVILDTYGNTSSAGSVIAFHTYQDDLPQGALAVLS SFGAGYSIGSVILRKR SEQIDNO:89 >gi|424853848|ref| MGKQIATVAGGRQSALLGLGVYRPERVVTNDEICELIDSNDEWI ZP_18278206.1|/ QSRSGIRNRRFAAEDENVVTMSIAAGRKAIEASGIDPEQIGCVIVA 1-3393-oxoacyl- TSTYLLLTPPAAAVVADALGTNGPGAFDLGGGCAGFCTALTVAS [acyl-carrier- DLVRGGSVDYALVVGVEKMSITTDPTDRSTRFIFGDGAGAVVVG protein]synthase KSDVAGIGPVEWGSDGAQADAIVQDLDWYEYITTPGATRPYIKM [Rhodococcus AGTAVFRWAAFEMGKVALRAVEKAGMSVDDLDAFVPHQANSR opacusPD630] ITEVIARSMKLPENVPVSDDIAESGNTSAASVPLAMEEMLQSGAT KPGDTALLLAFGAGLSYAAQVVTMPVLAKD SEQIDNO:90 >gi|427825838|ref| MMEKAMKYAKIAGSGGYLPERVVTNDDLAAELATRQISTSDEW ZP_18992900.1|/ IVERTGIRQRHLAERGVTTSQLATEAARRAMDDAGVQPDEIDMII 1-3293-oxoacyl- VATSTPDYVFPSTACLVQANLGAKGGAAFDVQAVCSGFVYAMT [acyl-carrier- TADSFIRAGRARCALVIGAEVFSRILDWNDRGTCVLFGDGAGAV protein]synthase VLKAADEPGILAAHLHADGSQTKILCAAGNVAYGDVTGDPFLR III[Bordetella MDGQAVFKQAVTVLDRSARDVCAEAGVEVDDIDWLIPHQANVR bronchiseptica ILNFLARKLRVPTERVVITVDQHANTSAASVPLALDVARRDGRV Bbr77] KPGQLVLMQGVGGGFTWGSVLARM SEQIDNO:91 >gi|441509582|ref| MSVIAANTGHQNVAMLGIGAYRPQRLVSNDEVCEVLDSSDEWIF ZP_20991498.1|/ ERSGVRNRRWISGDESARSMAAAAAERAIENSGIAKEKIGALILA 1-3563-oxoacyl- TNSWKTKIPHGGPIVAYDIGLNGIPAYDIAAGCGGFGYALGVAA [acyl-carrier- DTVRAGSAEYVLVVGVETMSVVMEPTDRNTAFIFGDGAGAVVV protein]synthase GPSEANGISPTVWGSDGENAEAIGQNYDIPEYMDRAQEYQHKDP III[Gordonia ETDPVGRMVVTMQGPRVFRWAAITLPKALTSVIERSGISADDIEV aichiensisNBRC FVPHQANARINELMKKNLGFPDDMPMANDIENTGNTSAASIPLA 108223] MEEMLATGKAKGGQTALLLGFGAGLSYAGAVVTLPPAPKVSSF DDLG SEQIDNO:92 >gi|443293257|ref| MTGSRIVSMGHYQPSRVVTNDDIAKLVDTNDEWIRDRVGIVSRRI ZP_21032351.1|/ ADGETVADMAAAAAGKALANSGLSASDIDLVVVATCSSIDRSPN 1-3143-oxoacyl- VACRVAAKLGIAAPGAFDVNTACSGFAYALGTVDHAVRAGASR (acyl-carrier- NALVIGAEKLSDFTDWTDRSTCIIFGDGAGAAVVTATADDEPAGI protein)synthase GPVVWGSVPEKSDAVRIEGWRPYIQQEGQSVFRWATTAIAPLAL III;acetylCoA QACERAGVDPSELAAFVPHQANARIIDGIAKRLNIPDAIIAKDIVE ACPtransacylase SGNTSAASVPLALSKLVERREVPSGAPVLLFGFGGGLTYAGQVV [Micromonospora RCP lupinistr.Lupac 08] SEQIDNO:93 >gi|443491493|ref| MEHRPECCCGCALAQMPSPPEESVPLPPTVGILGTAAFVPPRVVT YP_007369640.1|/ NNQAGASAGIDDAWIFARTGIRTRRWADPEQATSDLAVQAAEQ 1-3623-oxoacyl- ALANTAINAGQLGAIIVSTSTPDQPQPPTAAFVQNALHANSAYAF [acyl-carrier- DTNAVCSGFLFAINTAHALAQRDSIHVLVIGADVYSRILDPTDRK protein]synthase TVCLFGDGAGAVVVGPTTASSRHLRIVDTELHTFTQHINLIGVPG III,FabH_1 GGSRQPLTTATLDAGQHYFHMDGRGVRDFVTTTVPEQVRKFLA [Mycobacterium RHHLAVEDIDHVVMHQANGRMLDEIYSLLDLRNATCHQTIDRFG liflandii128FXT] NTGSASIPITLHHAYPELHGNILCIGFGGGMAAGITLLAAASGSAG DVGAHK SEQIDNO:94 >gi|444307652|ref| MSVPTLKQAPIQEHTRILGLGAYRPDVIVTNEDVCQWIDSSDEWI ZP_21143377.1|/ RQRTGIVTRHRAKADVSVIDMAEGAAREAMEKAGIEASELGAVI 1-3533-oxoacyl- VSTVTHPYATPSAAASLADRLGATPAPAFDISAACAGYCYGIAQ (acylcarrier GDALVRSGTAKYVLVVGAEKLSDVIDNRERTISFLLGDGAGAVV protein)synthase IGPSETPGIAPSVWGSDGSKWDAIGMTRSMLDVRDLGLAARQSD III[Arthrobacter STGDLALLEEAQELYPTLRQDGQTVFRWAVWEMAKVAQQALE sp.SJCon] AAGVEAEDLVAFIPHQANMRIIDEMVKKLKLPETVTVARDIADA GNTSAASIPLATHRLLQENPELSGGLALQIGFGAGLVFGAQVVVL P SEQIDNO:95 >gi|459055350|ref| MAVIADTTGIKNIGMLGIGAYRPERVVTNEEICQHIDSSDEWIYTR ZP_23152864.1|/ TGIKTRRFARRDESVMEMAVNAGRKAIANALLHGSDIDAVILAT 1-3383-oxoacyl- NTHLLLTPAGATKVATELGANGVPAFDVTVGCAGFGYGMALAS [acyl-carrier- DMIRGGSATHVLVIGAEQLSVTLDMTDRTNCFIFGDGAGAVVVG protein]synthase PTEEQELGPVVWGSDGSQFNAIRQDLDWVTFLDSDRKQRPYLRL III[Gordonia EGTAVFRWAAFEMGKVAHRALEAAKIGAEDLDVFVPHQANARI paraffinivorans NELLARSLKLREDAVVANDIEYTGNTSAASIPLAMEDLLSTGKAQ NBRC108238] PGQTALLLGFGAGLSYASQVVKLPPVPFE SEQIDNO:96 >gi|474659331|emb| MHRVIISGLGVEIPEPSITNEELVASFNAWVDTENVRRQASGEAPL CCV14840.1|/1- AKSDSAFIVHASGVQTRHVIEREGILDPTRMAPRIPARPDDALSLQ 373Beta- AEFGIASARKALDHAGLKPSDIDLVICSSSHQQRPYPAIAIEMQEA ketoacyl-acyl- LGTKGAGFDMGLGCSSAAAALHMAVNLVRSGAHKRVLVTTPEII carrier-protein TGHLNFRDRQTHFIFGDASVSMIVEGLAKGDKRPGRFEVLDTRIW synthaseI TQMSNNIRTNLGYHTRTAQDDPYMINLEGNLIKQVGNKVFKEVT [Mesorhizobium VAGHKFIVEFLAEHGLTPEAIRRFWLHQANARMNAMILKLSFGH sp.STM4661] EVGHDRAPMVLERLGNTAGAGAIIALSENHADMKPGDFGLLCAF GAGYSIGGALLRML SEQIDNO:97 >gi|478769383|gb| MPYARIIGTGSYLPEKALTNKDMEKMVDTTDQWIRERTGIERRHI ENO13968.1|/1- AAEGETTVDLAEQASLKAIEAAGIDVQDIDLIVFATSTPDKIFPSC 3223-oxoacyl- ACILQARLGIQGCPAFDIQAVCSGFVYALSTADKFIKTGASKKAL ACPsynthase VIGSEVFSRIVNWEDRGTCVLFGDGAGAVVLEANEETGILSTHIH [Marinobacter ADGQYEDLLHVPCGISDDFERVKAGQAFIEMKGNEVFKVAVNTL nanhaiticusD15- GKIVDETLEYNQMQKSDIDWLVPHQANLRHAATAKKLNMSMD 8W] QVVVTVNEHGNTSAASIPLALDVAVRDGRIKRNEVLLLEAFGGG FTWGSALLRY SEQIDNO:98 >gi|479875377|gb| MGIRITGTGLFHPTESISNEELVESLNAYVEQFNQENAEQIAAGEI ENU26638.1|/1- EALRGSSPEFIEKASGIQRRYVVEKSGILDPKRLRPRLQERSNDEL 368hypothetical SLQAEWGVIAAKQAMENAGVTAEDIDVVILACSNMQRAYPAVA protein IEIQSALGIQGYAYDMNVACSAATFGLKQAYDAVKCGARRVLLL F992_02187 NVEITSGHLDYRTRDAHFIFGDVATASIIEETETKSGYEILDIHLFT [Acinetobactersp. QFSNNIRNNFGFLNRSEDAVVDDKLFRQDGRKVFKEVCPLVAKII NIPH236] TAQLEKLELTPEQVKRFWLHQANANMNELILKLVVGKEADLER APIILDEFANTSSAGVIIAMHRTGEQVNNGEYAVISSFGAGYSVGS IIVQKHIA SEQIDNO:99 >gi|345301988|ref| MLPEQSLTTPLPATTTAAPARRAAVLGVGAALPAHREPSAETERR YP_004823890.1|/ LGLPPGWIARRTGIRERPLVGPDEATSDLAVRAGAAALAQAELSP 3-oxoacyl-ACP ERIGLLLLATSTPDHLLPPTAPVVAHRLGLKHAGAIDLAGACSGF synthaseIII LYALALADGYVRLQRTCVLVIGANVLSRRTNPDDPKTSALFADG [Rhodothermus AGAVVLGPSEGSRGIVACWLGADGSCWDDLYIPAGGSRRPLTPE marinus RVARGEHLMYMKDGRALFRRAATGMAEAGRRVLQQAGLDLDD SG0.5JP17-172] VAWWIPHQANLRLIEEARRQLGMPEARTVNLVDRIGNSSAATIPL ALALEAHRFAPGDLLLLTAVGAGLLSAAVLIQW SEQIDNO:100 >gi|471324089|ref| MTAPTAVLAGLGSALPPRVVTNHDLTARMDTSDEWIRTRTGIAE YP_007523119.1|/ RRIVDPGGATSDLAIEAGRRALDSAGGPDVGAVVVATATPDHPC 3-oxoacyl-[acyl- PATGPTVAAGLGLGTVPAFDVGAVCSGFLYALATGAGLIAASVA carrier-protein] DSVLVVGADAFTTIVDPYDRNTAPIFADGAGAVVLRAGRADEPG synthase3protein ALRRTELASDGMQADLIRVAAGGSRQRSHHSAALREDQYLTMR 3[Streptomyces GGEVFKNAVLRMTEASRTVLDRTGWSTAEVDLLVGHQANVRIL davawensisJCM HAVAEQLGIGQERAYVNIGHTGNTAAASIPLALDDAHGEGRLRA GDKVLLTAFGAGTTWGAITLTWPEGLQYRGAAGSAAA SEQIDNO:101 >gi|330444499|ref| MDKIKKAAILATGSYLPEKILSNADLEKMVDTSDEWIVTRTGIKE YP_004377485.1|/ RRIASDNEYTSDMGAKAAEKAlRASGLSKDLIDCIVFATSAPDYIF 3-oxoacyl-ACP PSSGALAQAYLGIKEVPAFDCLAACTGFLYGLSIAKAYVESGTYN synthaseIII HVLLIAADKLSSFVNYQDRNTCVLFGDGGAACIVGRSRPGALEIN [Chlamydophila QVCLGADGALGDLLSLPAGGSRNPATEATLKEGRHYISMEGKEV pecorumE58] FKHAVRRMEAASKASIAVAGIQEEQVGWLVPHQANERIIDAIAK RFNISEAKVFKSLYKYGNTAASSLGIALDELLNTETVLPHEYLLLT AFGGGLSWGSVVLEHV SEQIDNO:102 >gi|459068159|ref| MNSLYSVGITGIGSYVPEKVITNYDLCEIVDTSNEWIVERTGIQER ZP_23165498.1|/ RIVDQSLSTSDIGTIAANKALEDSNTNPKEIDLIIVATATPDMAFPS 3-oxoacyl-(acyl- TACIVQKNIQAINAAAFDISAGCSGFIYGLSIGFNFIKAGTYRKVL carrier-protein) VIGGETLSKIVNWEDRNTCVLFGDGAGACILERCEEGFGFLTFDL synthaseIII GSDGNNGHLLIQPAGGSRLPASYETVSNRLHTIKMDGREVFKFA [Clostridium VRIIEKSSKEVLRKANIPLEQIDLLIPHQANMRIIQSAIKKLQLEEN ultunenseEsp] KVYINLDKYGNMSSASIPVALDEAYKKEFFSKGDIVLLVAFGAGL TWGATLLRWNK SEQIDNO:103 >gi|383454618|ref| MARTHIIGTGSYAPTQVLTNQDLERLVETSDAWIRERTGIQERRQ YP_005368607.1|/ AAPDEATSDLAVNAARNALEMAGVAPGDLDLIVVGTVTADMP 3-oxoacyl-(acyl- MPSCAALVQSKLGAKRAFAFDVSAACAGGLYALSVADQFVRSG carrier-protein) QVKRALVVGADLLTRAVDWTDRNTCVLFGDGAGALVLGAEQD synthaseIII ADEDAMAPRGILSTHLRTDGDLANLLCIPAGGSRTPVTADNVDA [Corallococcus NLHKLKMNGKEVFRFAVRALVESTQASLGAHGMDTTQVDHVIA coralloidesDSM HQANLRILEAVMERLEIPKEKCWLNLHKYGNTSSASLPMSLDEA 2259] QRAGRLKRGDVIAMMAIGAGMAWGSAVVRW SEQIDNO:104 >gi|333371191|ref| MRIMGSVGIIGTGAYLPEKVLTNADLEKMVDTNDEWIVSRTGIRE ZP_08463153.1|/ RRIAADDQASSDLAVEAGRRALESAGIEAKDLDLIIVATVTPDMA 3-oxoacyl-[acyl- FPATACLVQDRLGAEKAATFDLSAACTGFLYGISVASQFISNGMY carrier-protein] RHALVIGVDCLSKITDFTDRNTCVLFGDGAGAAVLGPVEEGKGF synthaseIII LSFELGGDGSGGHLLKQPAGGSRIPASGKSVEDRLHFISMNGREV [Desmosporasp. FKFAVRVLGSSAEEALRKAGMTKEDVDFLIPHQANTRIIDTAVQR 8437] LGLSRDKVVVNLDRYGNMSSASIPVALDEAVQRGKIKKDDTLVL VGFGGGMTWGASVMKWTMETK SEQIDNO:105 >gi|390454110|ref| MNKLRPVGIIGTGKYVPEKILTNKDLEAIVETSDEWIVSRTGIQER ZP_10239638.1|/ HIAAPEQATSDLAYEAAIKALKSAGMTAEDLDLIIVATVTPDMAF 3-oxoacyl-(acyl- PSTACILQDKLGAKGAAAFDLSAACSGFVYGLATATSFIKTGIYN carrier-protein) NALIIGADCLSRITDYTDRNTCVLFGDGAGAVVIGEVSEGRGFQS synthaseIII FDLGAEGAGGSLLNLAAGGSRLPASADTLENKQHYIYMNGREVF [Paenibacillus KFAVRVMGTATVDVLEKAGLTKDDIDLFVPHQANIRIIQSAMQR peoriaeKCTC LDLPEEKVVINVNKYANTSAASIPLALVEAAEEGRMKEGDRVLM 3763] VGFGGGLTWGASVLVW SEQIDNO:106 >gi|392959403|ref| MNKKCVGIIGLGSYVPQRIMTNKDLEERMDTSDQWIVERTGIHE ZP_10324886.1|/ RRVAAENESTSDLAAKAGQKALEDAKISPAEIDLIIVATASPDMV 3-oxoacyl-(acyl- FPATACVVQENIKAVNAAAFDISAVCSGFLYAMITGSQFIKAGTY carrier-protein) RKVLVIGAETLSRFTDWSDRNTGMLFGDGAGAAVLGETPEGYGI synthase3 LGVDLGADGGGAELLKIPAGGSRHPATMETILQKQHFIYMNGNE [Pelosinus VFKFAVKVMGETTLKALKNANLTASDITYLVPHQANIRIIQSAAK fermentansDSM RLGIPMEKVVVNINKYGNTSAASIPIALDEAVKSGAIKSGDIVALA 17108] GFGGGLTWASSIMKWCK SEQIDNO:107 >gi|116626090|ref| MPKAKISALGCYTPPRVLTNQDLEKLVDTNDQWIMERTGIRERHI YP_828246.113- AAPEMATSDMAIEAARCALLQRGIDACEIDAIILCTVTPDHLFPST oxoacyl-ACP ACLVQNAIGAKGAWGFDLIAACSGFLYGLTTGAHFVMAGTHKK synthase VLVIGSDTMSRIIDYTDRATCVLFGDGAGAMLIEATDEADDGTGF [Candidatus IDFLGEIDGSGGEFLRMPAGGSRRPASHETVDQRMHYVHQEGSQ Solibacterusitatus VFKYASRKMYEVCRDLLERNHFKVEDVGLMIPHQANKRIIKAAG Ellin6076] DRLGIAPERVMINIERYGNTTAGTLPLATRDAISEGRLKKGDLVL FAAVGAGYTVGASLWRWAF SEQIDNO:108 >gi|323702691|ref| MSSNLVQAGIIGVGSYVPERILTNKDLEKMVDTSDEWITSRTGIK ZP_08114352.1|/ ERRIADPEESTSELAVKAARRALAHAGVKPEELDLIILATCTKDM 3-oxoacyl-(acyl- PFPASACLVQDQLGAVNAGAFDIEAGCTGFVYALTVGSQFVATG carrier-protein) SMKRVLVIGADNLSKVTNWEDRNTCVLFGDGAGAVVLGPVAPG synthaseIII EGILASKLAAEGAGWKYLSMPAGGSRMPASPLTVEKKLHYIHM [Desulfotomaculum QGREVFRYAVKVMEEEAANIVKAAGLALSDIDLLIPHQANIRIIEH nigrificans AAKKLKLSMDKVVVNVDRYGNTSTASIPLALDEAVKSGRVKAG DSM574] DNIVMVAFGAGLTSGAIVLKWSLGEGKE SEQIDNO:109 >gi|384566084|ref| MSTGILGAAGYLPPRVIDNDQVGAWVDRDPDWILERTGIKERHY ZP_10013188.1|/ AAPEVSTSDMACLAVEKLYASCPEKRASVGAVILGTSTPDHNFPS 3-oxoacyl-(cyl- TAAIVQGRMGLGRAFAFDLSAACSGYLFSFVTAHSLLSANPALEE carrier-protein) VLVIGADTISKVLYQSDRKTVTVFGDGAAATRVGRVPDGYGLLT synthaseIII HTLITDGCHADYVGQPAGGSRRPLDATTVNARERYMVMHGRKV [Saccharomonospora REYFEEVVPKLIHEVVEQAGVSLDDIDHFVFHQANPQMLADCIN glaucaK62] AMGIDPAKCPVPGVLSGNTGAASIPLVLSELRAERGDLVVMAAI GSGMTAGAAVLRWY SEQIDNO:110 >gi|298162138|gb| MNQGGVFPLPFKIAGLGRYVPADVVLSSDLEKKYDLPPGWCVE ADI59524.1| KQGIRERRWVKDETASFMGAEAAKEAVRDAGLKLEDIDLIINAS CorB GSPEQAVPDGGPLVQRELGLGRSGVPSITVNASCLSFFVALDVAA [Corallococcus NYLNMRRYKRILIVSSDISSVALDFRKPENFTLFGDAAAAAVVTL coralloides] PEPGEKSCIHASQVRTYGYGAEFSMVPGGGSRRHPNGKNTTPED NYLHMNGAELLKIGFEYLPRFNEALWKQCPDITIKDCRYVIPHQP SRVVLDYLSLTYPDDKLVRIIDRFANCIGASMPMALYEAVKVGG LRRGERGVLTGTGSGVSFVGMVFTY SEQIDNO:111 >gi|148359775|ref| MNFFRCEKPIYIKGPFVALPERVMSNQDVLNWMNSTQNPAVIGF YP_001250982.1|/ STGIKNRHWVNEDQACSDLAVRAAEHLFMEKPREKHKVNQVIL 3-oxoacyl-(acyl ATISGDYPSPPSSPLVQYRLGLQNAGAFDIGAACAGFVVGLHTSA carrierprotein) ALAQTNDGSVLLIASEIRSKFLNKNNFATSVLFGDGAAACCVSQD synthaseIIIFabH KEEADFRFIASALFADGEVYDAVSTPAGGSRLPAAVCNDNEQFYI [Legionella TIKESTALFVKAVHGMADSAKDFLKELNLTISDIQWLVPHQGNK pneumophilastr. NLVLSVAKQLGFPEEKTIKTVEETGNTSGSSVGIALDRLRSDGKIK Corby] SGEKVLLVAAGGGGIAACSLLEVI SEQIDNO:112 >gi|15824218|dbj| MTNEHLARRLDTDDAWIRTRTGIRRRHAVDPGQATSDLAVEAG BAB69376.1|3- RRALVCAATASVDAVVVATTTPDHSCPATAPAVAARLGLTGAA oxoacyl-(acyl AFDISAVCTGFVYGLASAAGLIAAGVAERVLLIGADTYSTIVDPL carrierprotein) DRANAIIFGDGAGAVVLRAGHPDEPGAVGHFDLGSDGAHEDLIM synthase VAAGGSRQRSRPGEPSRQDRHFGMRGKEVYRHAVTRMAESARA [Streptomyces TLSRAGWKTDDVDHFVPHQANLRILHSVADDLGLPRERCVTHVE ayermitilis] SVGNTGAASIPLALADAAAGQTLRPGDRVLLTAFGGGLTWGSCL LTWPTLPAPAPPYDPHAQGERTTS SEQIDNO:113 >gi|330468931|ref| MALSSHVEYESTTRTAVIAGLGAYVPDQVVKNEEIAARLGVTTD YP_004406674.1|/ WIRDRTGIEQRFVLNPEGATSDLAVEAARRALDSCGNPDIDFLIL 3-oxoacyl-(acyl ATCTPDHLFPSTAPSVASRLGFKGIAAFDLNAACSGFVYALSVST carrierprotein) GMLATGAYRTGLVIGADAISSILNHDDEITGPIFGDGGGAVVVRA synthaseIII GHLGETGSVSVQQLGSDGDLLDIMKTPGGGSRQRAAGVPVDIDS [Verrucosispora SYFTMSGRAVYKHAINRMSTVSRSVLERLGWTPDDVDWLIAHQ marisAB-18-032] ANRRILTATAEEIGIAPERAVINVDRVANTSAASIPLAMVDAVESG ALTAGDKVLLAAFGGGATWAAAGLTWPELTLAPTQTVR SEQIDNO:114 >gi|32444698|emb| MIETSSNVTANDLAAKSVNEESSAESTAVPTEAVSAVMPGNATT CAD74700.1|3- RGRMGNLKGVRIAGTGSYVPERIVTNEDLAALGCDSDWIVRRTG oxoacyl-(acyl- ILQRRHAEPGQATSDLCYEAALRCLENANVSVDEIDLILVATITPD carrierprotein) HPTPSTACHLQRRLGAVAPAMDIGAACAGFMYALVTGAQFVSN synthase GNARNVLVIGADLMSRTVDPEDKKTYPLFGDAAGAALLVPSTQ [Rhodopirellula DECQSTECNGSAADSTIQTDGLLAYQLGSEGCGGEMLCIPAGGSR balticaSH1] TPITTDGEDSASRYLQMDGRGVFKWAVRVFDESAKDVLRAANV SSDQLSLVVLHQANQRIIDSAVSDLNVPPEKVFVNLDKYGNTSGA SIPLALDEAARAGRLKEGDLVLLCGFGAGLAWGTALFRW SEQIDNO:115 >gi|392374495|ref| MYGSRIAGTGASVPDRVLTNAELEQMVSTSDEWIVTRTGISERRI YP_003206328.1|/ ASDDQATSDLAEGAARQALEASGVDPHDLDLILVNTVTPDMFFP 3-oxoacyl-[acyl- STACVLQERLGASRAAAFDLMAACAGFVYGLSVADAYLRAGV carrier-protein] MRNILVIGADTLSKVVDWSDRGTCVLFGDGAGAVVVQRTTADP synthaseIII(Beta- AILSTHLYSDGSKGRQLIIPGGGSRQPASQKVIDEKLVTIRMPNGN ketoacyl-ACP EVFKTAVRSMEEAAIAALKANGAEVSDVDLFISHQANARIIYAVA synthaseIII)(KAS ERLDLPRERIYMNIDRYGNTSAASIPIAMDEAVRAGRLKRGDLLL III)[Candidatus LTAFGGGFTWGSALIRW Methylomirabilis oxyfera]] SEQIDNO:116 >gi|317121784|ref| MVAAVRGVTIAGIGGCVPPAVVTNDDLAQVVETDDEWIRTRTGI YP_004101787.1|/ RQRRVADPGTATSDLAEVAARRALEEAGVRPDQVDLIIVATVTP 3-oxoacyl-(acyl- DMPFPSTACLLQDRLGATRAAGFDLEAACSGFVYALAAGAQFIA carrier-protein) AGLYDTVLVVGAETLSKIIDWSDRRTCVLLGDGAGAAVLRPAAP synthaseIII GEGILGLYLGADGSGGDLLKQPAGGSRLPASPETVARGLHFVQM [Thermaerobacter NGREVFKFAVKTMGDAAQAALAQAGLTFDDVDLYVPHQANFRI marianensisDSM IESSARRFDLPLERVVVNIDRYGNTSAASIPVALDEALSTGRIRAG 12885] QTVLLVAFGGGLTWGAAVVRWGYDRPAPRPLEMPGQEPRYGLP EWIREQAARGRARAGEPAQGEPAAAASEATAPAALAVPRAALD PAAVTAASPGSEGRPAWGGGGTR SEQIDNO:117 >gi|383787841|ref| MKVGVLGLGSYIPEKVVTNHDLEKFLDTSDEWIRTRTGIVERRIA YP_005472409.1|/ NENEATSDLASIAAKRALEDANLKPEDIDLIIVGTNSPDMLYPAT 3-oxoacyl-ACP ACLVQEKIGASGKCAAFDLQAGCPGFIYATVVGSQFVKSGAYKH synthase VLVIGAEVITRMMDPTDRGTYVLFGDGAGAVVLGEVEDNRGIV [Caldisericum DFELYADGSIAEHLTLPAGGSRKPFSEEVLKERSYFTKMNGGEVF exileAZM16c01] KFSVREISRISKKLLDKTGTKLEDIDWFIPHQANLRIIQAGAEKLGI PMEKVVVTIDKFGNSSAASIPVSLDTIRKEGKLKRGDLVLMVSFG AGMTSGAILMRW SEQIDNO:118 >gi|404450648|ref| MKKTRAVITGVQGWVPEYVLTNRELETMVDTNDEWITTRTGIKE ZP_11015628.1|/ RRILKGENQGTSVIGINAVKGLLEKTNTKAEDIDLIICATVTPDMP 3-oxoacyl-(acyl FPATANHADGVGAKNSYSYDISAACSGFLYALTIGSQFIETGMHK carrierprotein) KVIIVGADKMSSIIDYQDRATCHFGDGGGAVLLEPTQEKVGIMDS synthaseIII LLHADGSGAPFLHMKAGGSRKPASLETIAAREHFAFQEGSTVFKF [Indibacter AVTNMAEVSARIMERNNLASEDIAWLVPHQANKRIIDATANRM alkaliphilusLW1] GVGPDKVMLNIEKYGNTTAGTLPLCLWDYESQLKKGDNIILAAF GGGFTWGSIYLKWGYDPK SEQIDNO:119 >gi|189502112|ref| MRTAIRASITGVHGYVPEYILTNEKLEKMVDTNDEWITTRTGIKE YP_001957829.1|/ RRILEGTNQGTSVLGIPAVRGLLEKTNTDPREIDLLICATITPDMIT 3-oxoacyl-(acyl PATANIIAHAVGATNAFSYDLQAACSGFLYALITGVQFIETGKYK carrierprotein) KVVVVGADKMSSIVNYEDRNSCILFGDGAGAVLLEPNSQGYGII synthaseIII DSILKGDGNGEQYLHQKAGGSRRPPSAETIAAKEHYVYQEGRAV [Candidatus YRFAVEKMAEVVLEIMKKNNLHHEDIKFLVPHQANKRILDAVA Amoebophilus QRAGIKEEQVMITIQEFGNTTGATIPLCLWRYESQLQPGDKLIITT asiaticus5a2] FGGGFTWGAAYLTWAYK SEQIDNO:120 >gi|395801183|ref| MSAVITAIGGYVPSSILTNKKISETVDTSEEWIIKRTGIRERRIADD ZP_10480443.1|/ DTATSDLAAAAIENLIENYNVDREEIEALLVATATPDHILAPTASI 3-oxoacyl-ACP VCDKSGLTNAFGIDMNAACSGFLYALEMGANMIESGRYKKLIIV synthase GADKMSSIVDYEDRNTCILFGDGAGAILLEKSESDAGLMKTILKT [Flavobacterium DGSGVSSLAVPAGGSRNPTSMQSLLHRTHYLKQDGAFVFKRAV sp.F52] AAMSQVSQDALAKNELEADQIDWVVPHQANLRIITAVGESLGID FEKVKVNIDRYGNTTSATVPLCLWDFKDDFKEGQNVLITTFGAG FSWGATCLKWGVMRERKSAETITATTKAEAVLVEH

[0112] (2) Phase One (Reaction Initiation)Ketoacyl-CoA Reductase, 3-Hydroxyacyl-CoA Dehydratase and Enoyl-CoA Reductase

[0113] As noted above, the reaction initiation phase for even chain fatty acid products is completed by the conversion of 3-ketobutyryl-CoA to butyryl-CoA by three enzymes: a ketoacyl-CoA reductase, a hydroxyacyl-CoA dehydratase, and an enoyl-CoA reductase. For this phase, the ketoacyl-CoA reductase may be selected from the group consisting of 3-ketobutyryl-CoA reductase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and 3-hydroxybutyryl-CoA dehydrogenase (e.g., hbdSEQ ID NO 271); the hydroxyacyl-CoA dehydratase may be selected from the group consisting of 3-hydroxybutyryl-CoA dehydratase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and enoyl-CoA hydratase (e.g., crtSEQ ID NO 272); and the enoyl-CoA reductase may be trans-2-enoyl-reductase (e.g., terSEQ ID NO 275). Preferably, the bifunctional fadB is both the ketoacyl-CoA reductase and the hydroxyacyl-CoA dehydratase, or the ketoacyl-CoA reductase is hbd and the hydroxyacyl-CoA dehydratase is crt.

[0114] As noted above, the reaction initiation phase for odd chain fatty acid products is completed by the conversion of 3-ketovaleryl-CoA to valeryl-CoA by three enzymes: a ketoacyl-CoA reductase, a hydroxyacyl-CoA dehydratase, and an enoyl-CoA reductase. For this phase, the ketoacyl-CoA reductase may be selected from the group consisting of 3-ketovaleryl-CoA reductase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and 3-hydroxyvaleryl-CoA dehydrogenase (e.g., hbdSEQ ID NO 271); the hydroxyacyl-CoA dehydratase may be selected from the group consisting of 3-hydroxyvaleryl-CoA dehydratase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and enoyl-CoA hydratase (e.g., crtSEQ ID NO 272); and the enoyl-CoA reductase may be trans-2-enoyl-reductase (e.g., terSEQ ID NO 275). Preferably, the bifunctional fadB is both the ketoacyl-CoA reductase and the hydroxyacyl-CoA dehydratase, or the ketoacyl-CoA reductase is hbd and the hydroxyacyl-CoA dehydratase is crt.

[0115] (3) Phase One (Reaction Initiation)Malonyl-ACP Pathway

[0116] In accordance with an alternative embodiment, as shown in FIG. 12 and FIG. 13, the initiation phase may be achieved through a malonyl-ACP dependent pathway with at least a portion of one or more subsequent phases (i.e., elongation phase and/or termination phase) relying upon the malonyl-CoA dependent pathway. In accordance with this embodiment, the reaction to produce even chain fatty acid products is initiated through conversion of acetyl-CoA+malonyl-ACP to 3-ketobutyryl-ACP. In accordance with this embodiment, the reaction to produce even chain fatty acid products is initiated through conversion of propionyl-CoA+malonyl-ACP to 3-ketovaleryl-ACP. In accordance with this embodiment, a genetically modified microorganism is provided having encoded therein one or more enzymes described herein that catalyze reactions along the malonyl-CoA dependent pathway, and wherein native enzymes facilitate the initiation phase through the native malonyl-ACP pathway.

[0117] (4) Phase OneCoA/ACP or ACP/CoA Transition to Elongation

[0118] If a CoA-dependent elongation phase immediately follows an ACP-dependent initiation phase (see for example FIG. 13), the microorganism must also encode for a transacylase, such as butyryl-ACP:CoA transacylase, which will convert butyryl-ACP to butyryl-CoA or valeryl-ACP:CoA transacylase, which will convert valeryl-ACP to valeryl-CoA. Similarly, if an ACP-dependent elongation phase immediately follows an CoA-dependent initiation phase (see for example FIG. 11), the microorganism must also encode for a transacylase, such as butyryl-CoA:ACP transacylase, which will convert butyryl-CoA to butyryl-ACP or valeryl-CoA:ACP transacylase, which will convert valeryl-CoA to valeryl-ACP. Suitable butyryl-CoA:ACP transacylase include fabH, preferably from E. Coli, FASN, preferably from Homo sapiens, and FAS1, preferably from Saccharomyces cerevisiae. Additional transacylases include enzymes of the class 2.3.1.38, such as from Brassica juncea, Euglena gracilis, and ACT from Streptomyces collinus.

[0119] Alternatively, a genetically modified microorganism may be encoded for a gene that transitions a fatty acid production pathway from an ACP-dependent pathway to a CoA-dependent pathway, or conversely from a CoA-dependent pathway to an ACP-dependent pathway, by converting any ACP intermediate to its corresponding CoA intermediate, or vice versa. For example, the genetically modified microorganism may be encoded for phaG, preferably from Pseudomonas putida KT2440, which converts 3-hydroxyacyl-ACP to 3-hydroxyacyl-CoA.

[0120] B. Genetic Modifications to Drive Phase TwoChain Length Extensions (Elongation)

[0121] The second phase of the malonyl-CoA dependent pathway involves a cyclic process wherein the length of the carbon chain is extended by two carbons with each cycle. As illustrated in FIG. 7, this phase requires a ketoacyl-CoA synthase, a ketoacyl-CoA reductase, a hydroxyacyl-CoA dehydratase, and an enoyl-CoA reductase. Accordingly, a genetically modified microorganism of the present invention includes native or exogenous enzymes encoded therein that provide these functions.

[0122] (1) Phase Two (Elongation)Ketoacyl-CoA Synthase

[0123] NphT7 exhibits significant specificty for acetyl-CoA and propionyl-CoA as primers in the initiation phase, and it shows minimal activity with larger acyl-CoA chains during the elongation phase. Most 3-ketoacyl-CoA synthases that are capable of catalyzing the condensation of longer acyl-CoA chains are found in plants, mammals, yeast and other lower eukaryotes. Without a 3-ketoacyl-CoA synthase that has specificity for longer acyl-CoA, there will be no elongation of the acyl-CoA chain greater than C.sub.4-CoA or C.sub.5-CoA, and therefore 3-ketoacyl-CoA synthases that have specificity for longer acyl-CoA may be required.

[0124] In one aspect, the present invention provides a modified NphT7 polypeptide that functions as the ketoacyl-CoA synthase during the elongation phase in the malonyl-CoA dependent pathway. The modified NphT7 comprises an amino acid sequence having at least 70% but less than 100% or about 100% homology to SEQ ID NO:1 and one or more amino acid substitutions, deletions, or insertions, wherein the modified NphT7 polypeptide is capable of accepting an acyl-CoA substrate having a carbon chain length of C4 or greater, for example C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21 or C22. In some embodiments, the modified NphT7 polypeptide is capable of catalyzing a condensation reaction to condense an acyl-CoA substrate with a malonyl-CoA to produce a 3-ketoacyl-CoA having a carbon chain length of C6 or greater, for example C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21 or C22. In some embodiments, the modified NphT7 polypeptide comprises one or more amino acid substitutions selected from the group consisting of I147T, F217V, Y144L, V157F, G3095, G288S, a PDRP to HFLQ substitution for amino acids 86-89, I147F, I147M, I147Q, 1147S, I147C, 1147E, I147N, I147W, I147D, I147R, I147P, I147L, V196G, I147G, I147H, I147K, I147V, I147A, I147Y, F217G, F217A, F217L, F217I, F217M, F217T, F217P, F2175, F217E, F217L, F217W, and any combination thereof. In some embodiments, the modified NphT7 polypeptide comprises one amino acid substitution selected from the group consisting of I147V, I147F, I147M, I147Q, I147S, I147C, 1147E, I147N, I147W, I147D, I147R, I147P, I147L, I147G, I147H, I147K, I147A, I147Y, and F217V. In some embodiments, the modified NphT7 polypeptide comprises two amino acid substitutions selected from the group consisting of I147T and F217V, I147T and Y144L, I147T and V196G, I147F and F217V, I147M and F217V, I147S and F217V, I147T and HFLQ, I147T and V157F, I147T and F217G, I147T and F217A, I147T and F217L, I147T and F217I, I147T and F217M, I147T and F217P, I147T and F217S, I147T and F217E, I147S and F217G, I147S and F217A, I147S and F217L, I147S and F217I, I147S and F217M, I147S and F217W, I147S and F217S, I147S and F217E, I147S and F217K, I147F and F217A, I147F and F217L, I147F and F217I, I147F and F217M, I147F and F217P, I147F and F217E, I147M and F217G, I147M and F217A, I147M and F217L, I147M and F217I, I147M and F217M, I147M and F217P, I147M and F217S, I147M and F217E, and I147M and F217K. In some embodiments, the modified NphT7 polypeptide of any embodiment, comprising three amino acid substitutions selected from the group consisting of (Y144L, I147T, and F217V), (I147T, F217V, and HFLQ), (I147T, V147F, and F217V), and (Y144L, I147T, and V157F). In some embodiments, the modified NphT7 polypeptide comprises one or more amino acid substitutions at a position selected from the group consisting of Ser84, Val114, Gly288, Ile194, Gly318, Thr85, Gln90, Val196, Tyr144, Phe159, Ile147, Phe217, and any combination thereof. In some embodiments, the modified NphT7 polypeptide comprises an I147T amino acid substitution. In some embodiments, the modified NphT7 polypeptide comprises an F217V amino acid substitution. In some embodiments, the modified NphT7 polypeptide comprises two or more amino acid substitutions, deletions, or insertions, such as an I147T amino acid substitution and an F217V amino acid substitution. In some embodiments, the modified polypeptide is isolated and purified.

[0125] In one aspect, the present invention provides an isolated and purified polynucleotide encoding a modified NphT7 polypeptide. In some embodiments, the isolated and purified polynucleotide comprises a nucleic acid sequence having at least 70% but less than 100% or about 100% homology or complementarity to SEQ ID NO:2, wherein the polynucleotide encodes a modified NphT7 polypeptide of SEQ ID NO:1 having one or more amino acid substitutions, deletions, or insertions, wherein the modified NphT7 polypeptide is capable of accepting an acyl-CoA substrate having a carbon chain length of C4 or greater, for example C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21 or C22. In some embodiments, the isolated and purified polynucleotide encodes a modified NphT7 polypeptide capable of catalyzing a condensation reaction to condense an acyl-CoA substrate with a malonyl-CoA to produce a 3-ketoacyl-CoA having a carbon chain length of C6 or greater, for example C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21 or C22. The isolated and purified polynucleotide of any embodiment that encodes a modified NphT7 polypeptide comprising one or more amino acid substitutions selected from the group consisting of I147T, F217V, Y144L, V157F, G309S, G288S, a PDRP to HFLQ substitution for amino acids 86-89, I147F, I147M, I147Q, I147S, I147C, I147E, I147N, I147W, I147D, I147R, I147P, I147L, V196G, I147G, I147H, I147K, I147V, I147A, I147Y, F217G, F217A, F217L, F217I, F217M, F217T, F217P, F217S, F217E, F217L, F217W, and any combination thereof. In some embodiments, the isolated and purified polynucleotide encodes a modified NphT7 polypeptide comprising one amino acid substitution selected from the group consisting of I147V, I147F, I147M, I147Q, I147S, I147C, I147E, I147N, I147W, I147D, I147R, I147P, I147L, I147G, I147H, I147K, I147A, I147Y, and F217V. In some embodiments, the isolated and purified polynucleotide encodes a modified NphT7 polypeptide comprising two amino acid substitutions selected from the group consisting of I147T and F217V, I147T and Y144L, I147T and V196G, I147F and F217V, I147M and F217V, I147S and F217V, I147T and HFLQ, I147T and V157F, I147T and F217G, I147T and F217A, I147T and F217L, I147T and F217I, I147T and F217M, I147T and F217P, I147T and F217S, I147T and F217E, I147S and F217G, I147S and F217A, I147S and F217L, I147S and F217I, I147S and F217M, I147S and F217W, I147S and F217S, I147S and F217E, I147S and F217K, I147F and F217A, I147F and F217L, I147F and F217I, I147F and F217M, I147F and F217P, I147F and F217E, I147M and F217G, I147M and F217A, I147M and F217L, I147M and F217I, I147M and F217M, I147M and F217P, I147M and F217S, I147M and F217E, and I147M and F217K. In some embodiments, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide comprising three amino acid substitutions selected from the group consisting of (Y144L, I147T, and F217V), (I147T, F217V, and HFLQ), (I147T, V147F, and F217V), and (Y144L, I147T, and V157F). In some embodiments, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide comprising one or more amino acid substitutions at a position selected from the group consisting of Ser84, Val114, Gly288, Ile194, Gly318, Thr85, Gln90, Val196, Tyr144, Phe159, Ile147, Phe217, and any combination thereof. In some embodiments, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide comprising an I147T amino acid substitution. In some embodiments, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide comprising an F217V amino acid substitution. In some aspects, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide comprising two or more amino acid substitutions, deletions, or insertions. In some aspects, the isolated and purified polynucleotide herein encodes a modified NphT7 polypeptide, comprising an I147T amino acid substitution and an F217V amino acid substitution. In some embodiments, the isolated and purified polynucleotide herein is a RNA, mRNA, DNA, or cDNA. In some embodiments, the isolated and purified polynucleotide herein is a synthetic polynucleotide. In some embodiments, the isolated and purified polynucleotide herein is synthetic: RNA, mRNA, DNA, or cDNA.

[0126] In one aspect, the present invention provides for a ketoacyl-CoA synthase that is active during the elongation phase in the malonyl-CoA dependent pathway, wherein the ketoacyl-CoA synthase is selected from the group consisting of npht7 F217V, npht7 I147T, synthase III, synthase IV, synthase V, and synthase VI; wherein npht7 F217V and/or npht7 I147T catalyzes a reaction adding the 5.sup.th and/or 6.sup.th carbon in the elongation, npht7 F217V, npht7 I147T, and/or synthase III catalyzes a reaction adding the 7th and/or 8.sup.th carbon in the elongation, npht7 F217V, npht7 I147T, synthase III, synthase IV, and/or synthase V catalyzes a reaction adding the 9.sup.th and/or 10.sup.th carbon in the elongation, or wherein npht7 F217V, npht7 I147T, synthase III, synthase IV, synthase V, and/or synthase VI catalyzes a reaction adding the 9.sup.th and/or higher number carbon in the elongation.

[0127] (2) Phase Two (Elongation)Ketoacyl-CoA Reductase, 3-Hydroxyacyl-CoA Dehydratase and Enoyl-CoA Reductase

[0128] Referring again to FIG. 7, in addition to a 3-ketoacyl-CoA synthase, each cycle of the malonyl-CoA dependent elongation phase requires a 3-ketoacyl-CoA reductase (KCR), a hydroxyacyl-CoA dehydratase (3HDh), and an enoyl-CoA reductase (EnCr).

[0129] For the elongation phase, the ketoacyl-CoA reductase may be selected from the group consisting of 3-ketoacyl-CoA reductase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and 3-hydroxyacyl-CoA dehydrogenase (e.g., hbdSEQ ID NO 271); the hydroxyacyl-CoA dehydratase may be selected from the group consisting of 3-hydroxyacyl-CoA dehydratase (e.g., fadB, a bifunctional enzymeSEQ ID NO 183) and enoyl-CoA hydratase (e.g., crtSEQ ID NO 272); and the enoyl-CoA reductase may be trans-2-enoyl-reductase (e.g., terSEQ ID NO 275). Preferably, the bifunctional fadB is both the ketoacyl-CoA reductase and the hydroxyacyl-CoA dehydratase, or the ketoacyl-CoA reductase is hbd and the hydroxyacyl-CoA dehydratase is crt.

[0130] C. Genetic Modifications to Drive Phase ThreeChain Length Termination

[0131] The elongation phase ends with a termination step once the desired chain length is achieved. In one aspect of the invention, the genetically modified microorganism encodes an enzyme capable of terminating an acyl elongation cycle substantially at a desired chain length or substantially within a relatively narrow distribution of chain lengths (i.e., a distribution of 2-4 carbonse.g., C8-C10, C8-C12, C10-C12, C10-C14, etc.). In another aspect of the invention, the termination enzyme is a thioesterase such as an acyl-CoA esterase, and the microorganism produces a fatty acid. Suitable thioesterases include tesASEQ ID NO 277, tesASEQ ID NO 278, tesBSEQ ID NO 279, yciASEQ ID NO 280, ybgCSEQ ID NO 281, ybfFSEQ ID NO 282, fadMSEQ ID NO 283, AtTESEQ ID NO 284, CpTESEQ ID NO 285, CperfTESEQ ID NO 286, LpTESEQ ID NO 287, and PA2801TESEQ ID NO 288, and combinations thereof. Alternatively, the termination enzyme is a wax ester synthase and the microorganism produces a fatty ester. Suitable wax ester synthase include Maq1SEQ ID NO 289, Pcry1SEQ ID NO 290, Rjos1SEQ ID NO 291, and Abork1SEQ ID NO 292. Alternatively, it is within the skill of the art to add other known termination enzyme(s) that will enable the genetically modified microorganism to produce alternative fatty acid derivatives such as, for example, a fatty alcohol, a fatty aldehyde, a fatty alkene, a fatty amide, a fatty alkane, or a fatty diacid. By way of example, the termination enzyme may be a fatty acid or acyl-CoA reductase that catalyzes the production of a fatty alcohol or fatty aldehyde, or an aldehyde decarbonylase that catalyzes the production of fatty aldehyde, or an aldehyde decarbonylase together with an acyl-ACP reductase or an acyl-CoA reductase that catalyzes the production of an alkane.

[0132] D. Genetic Modifications Associated with Specific Chain Lengths

[0133] In accordance with one aspect of the invention, the genetically modified microorganism is engineered to produce a fatty acid or fatty acid derivative product having substantially a specific chain length or having a substantially narrow distribution of chain lengths (i.e., 2-4 carbons). Preferably, at least 50%, 60%, 70%, 80%, or 90% of the fatty acids or fatty acid derivative produced by the genetically modified microorganism of the present invention is of a desired chain length or within a desired narrow distribution of chain lengths. Applicants have determined that such specificity may be achieved through engineering a microorganism to encode for various combinations of genes that will lead to the production of fatty acids and fatty acid derivatives having specific chain lengths. Table 2 below sets forth certain unique combinations of genes that lead to the production of products having the carbon chain lengths indicated in Table 2.

TABLE-US-00003 TABLE 2 Chain length specificity of fatty Acid products by the enzyme combinations in the fatty acid pathways. Product pathway synthase KCR 3HDh EnCR Thioesterase C4 A nphT7 SEQ ID NO 1 Hbd SEQ ID NO 271 Crt SEQ ID NO 272 Ter SEQ ID NO 275 yciA SEQ ID NO 280 B npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 yciA SEQ ID NO 280 C6 A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 yciA SEQ ID NO 280 npht7 I147T, F217V B npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 PA2801TE SEQ ID NO 288 npht7 I147T, F217V C8 A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 yciA SEQ ID NO 280 B npht7 SEQ ID NO 1 fabG SEQ ID NO 270 Ech SEQ ID NO 273 ter SEQ ID NO 275 yciA SEQ ID NO 280 npht7 I147T, F217V synthase III C npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 PA2801TE SEQ ID NO 288 npht7 I147T, F217V synthase III D npht7 SEQ ID NO 1 fabG SEQ ID NO 270 ech SEQ ID NO 273 ter SEQ ID NO 275 PA2801TE SEQ ID NO 288 npht7 I147T, F217V synthase III C10 A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 AtTE SEQ ID NO 284 npht7 I147T, F217V synthase III, IV, V B npht7 SEQ ID NO 1 fabG SEQ ID NO 270 ech SEQ ID NO 273 ter SEQ ID NO 275 AtTE SEQ ID NO 284 npht7 I147T, F217V synthase III, IV, V C npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 ybgC SEQ ID NO 281 npht7 I147T, F217V synthase III, IV, V D npht7 SEQ ID NO 1 fabG SEQ ID NO 270 ech SEQ ID NO 273 ter SEQ ID NO 275 ybgC SEQ ID NO 281 npht7 I147T, F217V synthase III, IV, V C12 A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 tesA SEQ ID NO 278 npht7 I147T, F217V synthase III, IV, V, VI B npht7 SEQ ID NO 1 fabG SEQ ID NO 270 ech SEQ ID NO 273 ter SEQ ID NO 275 ybgC SEQ ID NO 281 npht7 I147T, F217V synthase III, IV, V, VI C npht7 SEQ ID NO 1 fadJ SEQ ID NO 185 fadJ SEQ ID NO 185 ter SEQ ID NO 275 ybfF SEQ ID NO 282 npht7 I147T, F217V synthase III, IV, V, VI C14-16 A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 tesA SEQ ID NO 278 npht7 I147T, F217V synthase III, IV, V, VI B npht7 SEQ ID NO 1 fadJ SEQ ID NO 185 fadJ SEQ ID NO 185 ter SEQ ID NO 275 fadM SEQ ID NO 283 npht7 I147T, F217Vsynthase III, IV, V, VI A npht7 SEQ ID NO 1 fadB SEQ ID NO 183 fadB SEQ ID NO 183 ter SEQ ID NO 275 tesA SEQ ID NO 278 npht7 I147T, F217V fadE SEQ ID NO 180 synthase III, IV, V, VI ydiO SEQ ID NO 186 B npht7 SEQ ID NO 1 fadJ SEQ ID NO 185 fadJ SEQ ID NO 185 ter fadE fadM SEQ ID NO 283 npht7 I147T, F217V SEQ ID NO 180 synthase III, IV, V, VI ydiO SEQ ID NO 186

[0134] In accordance with one aspect of the invention, there is provided a genetically modified microorganism having encoded therein the genes included in Table 2 for a given pathway, wherein such microorganism is capable of producing a fatty acid or fatty acid derivative having a carbon chain length indicated in Table 2 for such pathway. There is also provided a genetically modified microorganism having encoded therein the genes included in Table 2 above for a combination of pathways, wherein the microorganism is capable of producing fatty acids or fatty acid derivatives within a substantially narrow distribution of chain lengths corresponding to the carbon chain lengths indicated in Table 2 for such combination of pathways. For example, there is provided a genetically modified microorganism comprising NphT7, fadB, ter, AtTE, and tesA (C10 pathway A and C12 pathway A), wherein said microorganism is capable of producing a fatty acid composition comprising C10 and C12 fatty acids.

[0135] In another aspect, applicants have discovered that chain length specificity can be controlled by utilizing certain enzymes from certain specific species of organisms. Accordingly, the present invention provides one or more isolated and purified polynucleotides comprising exogenous nucleic acid molecules encoding proteins comprising a 3-oxoacyl-(acyl carrier protein) synthase III from a species selected from the group consisting of Alishewanella aestuarii B11 (SEQ ID NO 236), Arcobacter butzleri ED-1 (SEQ ID NO 262), Clostridiales bacterium 1_7_47_FAA (SEQ ID NO 248), Gluconacetobacter oboediens 174Bp2 (SEQ ID NO 259), Gordonia aichiensis NBRC 108223 (SEQ ID NO 267), Mesorhizobium sp. STM 4661 (SEQ ID NO 246), Pelosinus fermentans DSM 17108 (SEQ ID NO 106), Phaeobacter gallaeciensis 2.10 (SEQ ID NO 70), Ralstonia solanacearum Po82 (SEQ ID NO 63), Saccharomonospora azurea NA-128 (SEQ ID NO 57), Saccharomonospora glauca K62 (SEQ ID NO 109), and Verrucosispora maxis AB-18-032 (SEQ ID NO 113), wherein the proteins encoded by the polynucleotides are capable of producing a fatty acid.

[0136] In some embodiments, the 3-oxoacyl-(acyl carrier protein) synthase III is from a species selected from the group consisting of Pelosinus fermentans DSM 17108 (SEQ ID NO 106), Saccharomonospora glauca K62 (SEQ ID NO), Verrucosispora maxis AB-18-032 (SEQ ID NO 113), and Clostridiales bacterium 1_7_47_FAA (SEQ ID NO 248), and wherein the proteins encoded by the polynucleotides are capable of producing an acetyl-CoA. In some embodiments, the 3-oxoacyl-(acyl carrier protein) synthase III is from a species selected from the group consisting of Saccharomonospora glauca K62 (SEQ ID NO 109), Saccharomonospora azurea NA-128 (SEQ ID NO 57), Mesorhizobium sp. STM 4661 (SEQ ID NO 246), and Clostridiales bacterium 1_7_47_FAA (SEQ ID NO 248), and wherein the proteins encoded by the polynucleotides are capable of producing a four carbon fatty acid. In some embodiments, the 3-oxoacyl-(acyl carrier protein) synthase III is from a species selected from the group consisting of Gordonia aichiensis NBRC 108223 (SEQ ID NO 267), Arcobacter butzleri ED-1 (SEQ ID NO 262), Clostridiales bacterium 1_7_47_FAA (SEQ ID NO 248), Saccharomonospora glauca K62 (SEQ ID NO 109), and Ralstonia solanacearum Po82 (SEQ ID NO 63), and wherein the proteins encoded by the polynucleotides are capable of producing a six carbon fatty acid. In some embodiments, the 3-oxoacyl-(acyl carrier protein) synthase III is from a species selected from the group consisting of Gordonia aichiensis NBRC 108223 (SEQ ID NO 267), Gluconacetobacter oboediens 174Bp2 (SEQ ID NO 259), Arcobacter butzleri ED-1 (SEQ ID NO 262), Ralstonia solanacearum Po82 (SEQ ID NO 63), and Phaeobacter gallaeciensis 2.10 (SEQ ID NO 70), and wherein the proteins encoded by the polynucleotides are capable of producing an eight carbon fatty acid. In some embodiments, the 3-oxoacyl-(acyl carrier protein) synthase III is from Alishewanella aestuarii B11 (SEQ ID NO 236), and wherein the proteins encoded by the polynucleotides are capable of producing a ten carbon fatty acid.

[0137] E. Genetic Modifications to Redirect Malonyl-CoA from Native Malonyl-ACP Dependent Fatty Acid Synthesis to Malonyl-CoA Dependent Fatty Acid Synthesis

[0138] As discussed above, certain aspects the present invention relate to microorganisms that are genetically modified to produce fatty acids and fatty acid derivatives through a malonyl-CoA dependent pathway that is also a malonyl-ACP independent pathway. This aspect of the invention may be used in combination with the inhibition of a microorganism's malonyl-ACP dependent fatty acid synthase pathway through one or more genetic modifications to reduce the activity of enzymes encoded by one or more of the microorganism's malonyl-ACP dependent fatty acid synthase system genes. The compositions may be used in the methods and systems of the present invention.

[0139] In many microorganism cells the fatty acid synthase system comprises polypeptides that have the following enzymatic activities: malonyl-CoA-acyl carrier protein (ACP) transacylase; 3-ketoacyl-ACP synthase; 3-keto acyl-ACP reductase; 3-hydroxyacyl-ACP dehydratase; 3-hydroxyacyl-ACP dehydratase; and enoyl-ACP reductase. In various embodiments nucleic acid sequences that encode temperature-sensitive forms of these polypeptides may be introduced in place of the native enzymes, and when such genetically modified microorganisms are cultured at elevated temperatures (at which these thermolabile polypeptides become inactivated, partially or completely, due to alterations in protein structure or complete denaturation), there is observed an increase in flux through the malonyl-CoA dependent pathway and a decrease in flux through the malonyl-ACP dependent pathway.

[0140] In E. coli, these temperature-sensitive mutant genes could include fabI.sup.ts(S241F)(SEQ ID NO 141), fabB.sup.ts(A329V) or fabD.sup.ts(W257Q). In other embodiments other types of genetic modifications may be made to otherwise modulate, such as lower, enzymatic activities of one or more of these polypeptides. In various embodiments, a result of such genetic modifications is to shift malonyl-CoA utilization so that there is a reduced conversion of malonyl-CoA to fatty acids via the native pathway, overall biomass, and proportionally greater conversion of carbon source to a chemical product including a fatty acid or fatty acid derived product via a malonyl-CoA dependent, and in some cases a malonyl-ACP independent route. In various embodiments, the specific productivity for the microbially produced chemical product is unexpectedly high. Also, additional genetic modifications, such as to increase malonyl-CoA production, may be made for certain embodiments.

[0141] One enzyme, enoyl-acyl carrier protein reductase (EC No. 1.3.1.9, also referred to as enoyl-ACP reductase) is a key enzyme for fatty acid biosynthesis from malonyl-CoA. In Escherichia coli this enzyme, FabI (SEQ ID NO 132), is encoded by the gene fabI (See Enoyl-Acyl Carrier Protein (fabI) Plays a Determinant Role in Completing Cycles of Fatty Acid Elongation in Escherichia coli, Richard J. Heath and Charles O. Rock, J. Biol. Chem. 270:44, pp. 26538-26543 (1995), incorporated by reference for its discussion of fabI and the fatty acid synthase system).

[0142] The present invention may utilize a microorganism that is provided with a nucleic acid sequence (polynucleotide) that encodes a polypeptide having enoyl-ACP reductase enzymatic activity that may be modulated during a fermentation event. For example, a nucleic acid sequence encoding a temperature-sensitive enoyl-ACP reductase may be provided in place of the native enoyl-ACP reductase, so that an elevated culture temperature results in reduced enzymatic activity, which then results in a shifting utilization of malonyl-CoA to production of a desired chemical product. One such sequence is a mutant temperature-sensitive fabI (fabI.sup.TS) of E. coli or the fabI(S241F) (SEQ ID NO 141). This enzyme may exhibit reduced enzymatic activity at temperatures above 30 C. but normal enzymatic activity at 30 C., so that elevating the culture temperature to, for example to 34 C., 35 C., 36 C., 37 C. or even 42 C., reduces enzymatic activity of enoyl-ACP reductase. In such case, more malonyl-CoA is converted to a fatty acid or fatty acid derived product or another chemical product through the non-native pathway under the current invention than at 30 C., where conversion of malonyl-CoA to fatty acids through its native fatty acid pathway is not impeded by a less effective enoyl-ACP reductase.

[0143] It is appreciated that nucleic acid and amino acid sequences for enoyl-ACP reductase in species other than E. coli are readily obtained by conducting homology searches in known genomics databases, such as BLASTN and BLASTP. Approaches to obtaining homologues in other species and functional equivalent sequences are described herein. Accordingly, it is appreciated that the present invention may be practiced by one skilled in the art for many microorganism species of commercial interest.

[0144] Approaches other than a temperature-sensitive enoyl-ACP reductase may be employed as known to those skilled in the art, such as, but not limited to, replacing a native enoyl-ACP or enoyl-CoA reductase with a nucleic acid sequence that includes an inducible promoter for this enzyme, so that an initial induction may be followed by no induction, thereby decreasing enoyl-ACP or enoyl-CoA reductase enzymatic activity after a selected cell density is attained. For example, a genetic modification may be made to reduce the enzymatic activity of the enoyl-ACP reductase gene (e.g., fabI in E. coli). In such example the promoter may be induced (such as with isopropyl--D-thiogalactopyranoiside (IPTG)) during a first phase of a method herein, and after the IPTG is exhausted, removed or diluted out the second step, of reducing enoyl-ACP reductase enzymatic activity, may begin. Other approaches may be applied to control enzyme expression and activity such as are described herein and/or known to those skilled in the art. For example promoters that are turned on in response to phosphate depletion may be used to controllably express desired genes. Such promoters could include the yibD (SEQ ID NO 170) or pstS (SEQ ID NO 171) gene promoters in E. coli.

[0145] Without being bound to a particular theory, it is believed that reducing the enzymatic activity of enoyl-ACP reductase (and/or of other enzymes of the fatty acid synthase system) in a microorganism leads to an accumulation and/or shunting of malonyl-CoA, a metabolic intermediate upstream of the enzyme, and such malonyl-CoA may then be converted to a chemical product for which the microorganism cell comprises a metabolic pathway that utilizes malonyl-CoA. In certain compositions, methods and systems of the present invention the reduction of enzymatic activity of enoyl-ACP reductase (or, more generally, of the fatty acid synthase system) is made to occur after a sufficient cell density of a genetically modified microorganism is attained. This bi-phasic culture approach balances a desired quantity of biocatalyst, in the cell biomass which supports a particular production rate, with yield, which may be partly attributed to having less carbon be directed to cell mass after the enoyl-ACP reductase activity (and/or activity of other enzymes of the fatty acid synthase system) is/are reduced. This results in a shifting net utilization of malonyl-CoA, thus providing for greater carbon flux to a desired chemical product.

[0146] Once the modulation is in effect to decrease the noted enzymatic activity(ies), each respective enzymatic activity so modulated may be reduced by at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 percent compared with the activity of the native, non-modulated enzymatic activity (such as in a cell or isolated). Similarly, the conversion of malonyl-CoA to fatty acyl-ACP or molecules may be reduced by at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 percent compared with such conversion in a non-modulated cell or other system. Likewise, the conversion of malonyl-CoA to fatty acid molecules through its native pathway may be reduced by at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 percent compared with such conversion in a non-modulated cell or other system.

[0147] F. Additional Genetic Modifications

[0148] The genetic modifications described hereinabove may be combined with various additional genetic modifications to further enhance production of a desired chemical product. Such additional genetic modifications may result in a variety of beneficial attributes, such as increasing glucose uptake, decreasing consumption of key intermediates by alternative reaction pathways leading to undesirable by-products, and driving carbon flux to malonyl-CoA. Certain of these additional genetic modifications are set forth in Table 3.

TABLE-US-00004 TABLE 3 Genetic Modifications E.C. Gene Name Enzyme Function Classification in E. coli Modifications Glucose transporter N/A GalP Increase function (SEQ ID NO 177) Pyruvate dehydrogenase E1p 1.2.4.1 AceE Increase function (SEQ ID NO 151) lipoate acetyltransferase/ 2.3.1.12 AceF Increase function dihydrolipoamide (SEQ ID acetyltransferase NO 152) Pyruvate dehydrogenase E3 1.8.1.4 Lpd Increase function or alter such (lipoamide dehydrogenase) (SEQ ID as by mutation to increase NO 153) resistance to NADH inhibition, Lactate dehydrogenase 1.1.1.28 LdhA Decrease function, including (SEQ ID by mutation NO 124) Pyruvate formate lyase (B 2.3.1. PflB Decrease function, including inactive) (SEQ ID by mutation NO 125) Pyruvate oxidase 1.2.2.2 PoxB Decrease function, including (SEQ ID by mutation NO 127) Phosphate acetyltransferase 2.3.1.8 Pta Decrease function, including (SEQ ID by mutation NO 128) acetate kinase 2.7.2.15 AckA Decrease function, including 2.7.2.1 (SEQ ID by mutation NO 129) methylglyoxal synthase 4.2.3.3 MgsA Decrease function, including (SEQ ID by mutation NO 126) Heat stable, histidyl N/A ptsH Decrease function, including phosphorylatable protein (of (HPr) by mutation PTS) Phosphoryl transfer protein (of N/A ptsI Decrease function, including PTS) by mutation Polypeptide chain (of PTS) N/A Crr Decrease function, including by mutation 3-oxoacyl-ACP synthase I 2.3.1.179 FabF Decrease or increase function, 3-oxoacyl-ACP synthase II 2.3.1.41 (SEQ ID including by mutation monomer NO 136) 3-ketoacyl-ACP synthase I, 3- 2.3.1.41 fabB Decrease or increase function, oxoacyl-ACP-synthase I 2.3.1. (SEQ ID including by mutation NO 133) Malonyl-CoA-ACP 2.3.1.39 fabD Decrease or increase function, transacylase (SEQ ID including by mutation NO 135) enoyl acyl carrier protein 1.3.1.9, fabI Decrease or increase function, reductase 1.3.1.10 (SEQ ID including by mutation NO 132) 3-ketoacyl-acyl carrier protein 2.3.1.180 fabH Decrease or increase function, synthase III (SEQ ID including by mutation NO 134) Carboxyl transferase subunit 6.4.1.2 accA Increase function subunit (SEQ ID NO 147) Biotin carboxyl carrier protein 6.4.1.2 accB Increase function (SEQ ID NO 148) Biotin carboxylase subunit 6.3.4.14 accC Increase function (SEQ ID NO 149) Carboxyl transferase subunit 6.4.1.2 accD Increase function subunit (SEQ ID NO 150) long chain fatty acyl 3.1.2.2, tesA Decrease or increase function thioesterase I 3.1.1.5 (SEQ ID as well as alter by mutation to NO 277) express in cytoplasm. acyl-CoA synthase 2.3.1.86 fadD Decrease via deletion or (SEQ ID mutation NO 181) acetate CoA-transferase 2.8.3.8 atoD Decrease via deletion or (SEQ ID mutation NO 190) acetate CoA-transferase 2.8.3.8 atoA Decrease via deletion or (SEQ ID mutation NO 191) Transporter atoE Decrease via deletion or (SEQ ID mutation NO 192) acetyl-CoA acetyltransferase 2.3.1.9 atoB Decrease via deletion or (SEQ ID mutation NO 193) pantothenate kinase 2.7.1.33 coaA Increase function (SEQ ID NO 173) lactose repressor lacI Decrease via deletion or mutation -glutamyl-- 1.2.1. puuC Decrease via deletion or aminobutyraldehyde mutation dehydrogenase malate synthase A 2.3.3.9 AceB Decrease via deletion or (SEQ ID mutation NO 157) isocitrate lyase 4.1.3.1 AceA Decrease via deletion or (SEQ ID mutation NO 156) isocitrate dehydrogenase 3.1.3.2.7.11.5. AceK Decrease via deletion or phosphatase/isocitrate (SEQ ID mutation dehydrogenase kinase NO 158) pyruvate formate-lyase 1.2.1.10 adhE Decrease via deletion or deactivase 1.1.1.1 (SEQ ID mutation NO 130) aldehyde dehydrogenase A, 1.2.1.21 aldA Decrease via deletion or NAD-linked 1.2.1.22 mutation acetaldehyde dehydrogenase 1.2.1.4 aldB Decrease via deletion or mutation Lambda phage DE3 lysogen DE3 Increase function T7 mRNA polymerase T7pol Increase function trigger factor 5.2.1.8 Tig Decrease via deletion or (SEQ ID mutation NO 189) 3-ketoacyl-CoA thiolase 2.3.1.16 FadA Increase or decrease function (SEQ ID NO 182) dodecenoyl-CoA - 5.3.3.8 fadB Increase or decrease function isomerase, enoyl-CoA 1.1.1.35 (SEQ ID hydratase, 3-hydroxybutyryl- 5.1.2.3 NO 183) CoA epimerase, 3- 4.2.1.17 hydroxyacyl-CoA dehydrogenase Sucrose permease cscB Increase function (SEQ ID NO 175) Invertase 3.2.1.26 CscA Increase function (SEQ ID NO 174) fructokinase 2.7.1.4 cscK Increase function (SEQ ID NO 1173) carbonic anhydrase 4.2.1.1 cynT Increase function (SEQ ID NO 168) carbonic anhydrase 4.2.1.1 Can Increase function (SEQ ID NO 167) pyridine nucleotide 1.6.1.2 PntAB Increase function transhydrogenase (SEQ ID NOS 145-146) pyridine nucleotide 1.6.1.1 udhA Increase function transhydrogenase (SEQ ID NO 144) acyl-CoA thioesterase 3.1.2.20 yciA Increase or decrease function 3.1.2.2 (SEQ ID NO 280) thioesterase II 3.1.2.20 tesB Increase or decrease function 3.1.2.2 (SEQ ID NO 279) thioesterase III 3.1.2. fadM Increase or decrease function (SEQ ID NO 283) hydroxyphenylacetyl-CoA paaI Increase or decrease function thioesterase esterase/thioesterase 3.1.2.28 ybgC Increase or decrease function (SEQ ID NO 281) proofreading thioesterase in entH Increase or decrease function enterobactin biosynthesis acetoacetyl-CoA synthase 2.3.1.194 npth07 Increase function (SEQ ID NO 1) 3-ketoacyl-CoA synthase/ 2.3.1 Elo1 Increase function elongase 3-ketoacyl-CoA synthase/ 2.3.1 Elo2 Increase function elongase 3-ketoacyl-CoA synthase/ 2.3.1 Elo3 Increase function elongase 3-Hydroxybutyryl-CoA 1.1.1.157 hbd Increase function dehydrogenase (SEQ ID NO 271) 3-oxoacyl-CoA reductase 1.1.1.100 fabG Increase function (SEQ ID NO 270) enoyl-CoA hydratase 4.2.1.17 crt Increase function (SEQ ID NO 272) enoyl-CoA hydratase 4.2.1.17 ech2 Increase function (SEQ ID NO 274) Trans-2-enoyl-reductase 1.3.1.9 ter Increase function (SEQ ID NO 275)

[0149] In addition to the above-described genetic modifications, in various embodiments genetic modifications also are provided to increase the pool and availability of the cofactor NADPH, and/or, consequently, the NADPH/NADP.sup.+ ratio. For example, in various embodiments for E. coli, this may be done by increasing activity, such as by genetic modification, of one or more of the following genes: pgi (in a mutated form), pntAB, overexpressed, gapA:gapN substitution/replacement, and disrupting or modifying a soluble transhydrogenase such as sthA, and/or genetic modifications of one or more of zwf, gnd, and edd.

[0150] Any of the genetic modifications described herein may be provided to species not having such functionality, or having a less than desired level of such functionality.

[0151] More generally, and depending on the particular metabolic pathways of a microorganism selected for genetic modification, any subgroup of genetic modifications may be made to decrease cellular production of fermentation product(s) selected from the group consisting of acetate, acetoin, acetone, acrylic, malate, fatty acid ethyl esters, glycerolipids, lipids, isoprenoids, glycerol, ethylene glycol, ethylene, propylene, butylene, isobutylene, ethyl acetate, vinyl acetate, other acetates, 1,4-butanediol, 2,3-butanediol, butanol, isobutanol, sec-butanol, butyrate, isobutyrate, 2-OH-isobutryate, 3-OH-butyrate, ethanol, isopropanol, D-lactate, L-lactate, pyruvate, itaconate, levulinate, glucarate, glutarate, caprolactam, adipic acid, propanol, isopropanol, fusel alcohols, and 1,2-propanediol, 1,3-propanediol, formate, fumaric acid, propionic acid, succinic acid, valeric acid, and maleic acid. Gene deletions may be made as disclosed generally herein, and other approaches may also be used to achieve a desired decreased cellular production of selected fermentation products.

[0152] In some embodiments, additional genetic modification is associated with a genotype or an enzyme selected from the group listed in Table 4-Table 13 below. The amino acid sequences of these enzymes are shown in Table 14.

TABLE-US-00005 TABLE 4 Base strain E. coli K12 BW25113 (Datsenko, K. A., and Wanner, B. L., Proc. Natl., Acad. Sci. USA 97: 6640-6645, 2000) Genotype Function EC # Organism Gene Gene ID Comments F- Mating negative E. coli araD b0061, araA ECK0062 araB b0062, ECK0063 b0063, ECK0064 (araD-araB)567 utilization of E. coli lacZ b0344, deletion of araB, arabinose ECK0341 araA, araD lacZ4787(::rrnB-3) utilization of lactose disruption of lacZ LAM- lambda phage E. coli pyrE b3642, lysogen deletion ECK3632 rph-1 1-bp deletion in pyrE E. coli rhaD b3902, rhaA ECK3895 rhaB b3903, ECK3896 b3904, ECK3897 (rhaD-rhaB)568 utilization of E. coli araD b0061, deletion of araB, rhamnose araA ECK0062 araA, araD araB b0062, ECK0063 b0063, ECK0064 hsdR514 restriction 3.1.21.3 E. coli lacZ b0344, endonuclease R ECK0341

TABLE-US-00006 TABLE 5 Host modifications for yield increase/byproduct elimination Enzyme Function Reaction EC # Organism Gene Gene ID Comments ldhA (SEQ lactate pyruvate + 1.1.1.28 E. coli ldhA bl380, deletion ID NO 124) dehydrogenase NADH = lactate + NAD ECK1377, NC_000913.2 pflB(SEQ pyruvate formate pyruvate + 2.3.1.54 E. coli pflB b0903, deletion ID NO 125) lyase CoASH = ECK0894 acetyl-CoA + formate mgsA(SEQ methylglyoxal dihydroxyacetone 4.2.3.3 E. coli mgsA b0963, deletion ID NO 126) synthase phosphate = ECK0954 methylglyoxal + Pi poxB(SEQ pyruvate oxidase pyruvate + an 1.2.2.2 E. coli poxB b9871, deletion ID NO 127) ubiquinone = ECK0862, CO2 + an NP_415392 ubiquinol + acetate pta(SEQ phosphotransacetylase acetyl-CoA + 2.3.1.8 E. coli pta b2296, deletion ID NO 128) Pi = acetyl-P + CoASH ECK2291, ack(SEQ acetate kinase acetyl-P + ADP = 2.7.2.1 E. coli ackA b2296, deletion ID NO 129) acetate + ATP ECK2290, NP 416799 adhE(SEQ bifunctional acetyl-CoA acetyl-CoA + 1.1.1.1, E. coli adhE b1241, deletion ID NO 130) reductase/alcohol NAD = 1.2.1.10 ECK1235, dehydrogenase acetaldehyde + NADH MG4323 acetaldehyde + NAD = ethanol + NADH

TABLE-US-00007 TABLE 6 Fatty acid synthesis (including temperature sensitive alleles used for increased malonyl-CoA availability) Enzyme Function Reaction EC # Organism Gene Gene ID Comments ACP (SEQ acyl carrier none none E. coli acpP b1094, ID NO 131) protein ECK1080, MG4178 fabI(SEQ enoyl-ACP a trans-enoyl- 1.3.1.9 E. coli fabI b1288, TS allele ID NO 132) reductase acyl-ACP + ECK1283, used: NADH = an NP_415804 S241F acyl-ACP + NAD fabB(SEQ 3-keto-acyl- an acyl-ACP + 2.3.1.41 E. coli fabB b2323, TS allele ID NO 133) ACP synthase malonyl-ACP = ECK2317 used: KASI a 3-keto- A329V acyl-ACP + CO2 + ACP-SH fabH(SEQ 3-keto-acyl- acetyl-CoA + 2.3.1.180 E. coli fabH b1091, ID NO 134) ACP synthase malonyl-ACP = ECK1077 KASIII acetoacetyl- CoA + CO2 + ACP-SH fabD(SEQ malonyl- ACP-SH + 2.3.1.29 E. coli fabD b1092, TS allele ID NO 135) CoA: ACP malonyl-CoA = 2.3.1.85, 86 ECK1078, used: transacylase malonyl- AP_002424 W257Q ACP + CoASH fabF(SEQ 3-keto-acyl- malonyl-ACP + 2.3.1.41, E. coli fabF b1095, ID NO 136) ACP synthase acetyl-ACP = 2.3.1.179 ECK1081 KASII acetoacetyl- ACP + ACP + CO2 fabG(SEQ 3-keto-acyl- 3-keto-acyl- 1.1.1.100 E. coli fabG b1093, ID NO 137) ACP reductase ACP + ECK1079 (NADPH-dep) NADPH = 3- OH-acyl-ACP + NADP fabA(SEQ 3-keto- 3-hydroxy- 4.2.1.60 E. coli fabA b0954, ID NO 138) hydroxyl-acyl- acyl-ACP = ECK0945 ACP dehydrase 3-enoyl-acyl- ACP + H2O fabZ(SEQ 3-keto- 3-hydroxy- 4.2.1. E. coli fabZ b0180, ID NO 139) hydroxyl-acyl- acyl-ACP = ECK0179, ACP dehydrase 3-enoyl-acyl- NP 414722 ACP + H2O fabR(SEQ transcriptional none none E. coli fabR b3963, ID NO 140) repressor NP_418398

TABLE-US-00008 TABLE 7 Malonyl-CoA synthesis and other genes related to optimizing flux Enzyme Function Reaction EC # Organism Gene Gene ID Comments udhA(SEQ NADP/NAD NAD+ + NADPH = 1.6.1.1 E. coli udhA = sthA b3962, ID NO 144) transhydrogenase NADP+ + NADH ECK3954 (soluble) pntAB(SEQ NADP/NAD NADP+ + NADH = 1.6.1.2 E. coli pntA, pntB b1603, ID NOS transhydrogenase NADPH + NAD+ ECK1598 145-146) (membrane, complex) b1602, ECK1597 PDH(SEQ Pyruvate pyruvate + NAD + 1.2.4.1 E. coli aceE b0114, ID NO 151) dehydrogenase, CoASH = acetyl- NP_414656 subunit E1 CoA + NADH + CO2 PDH(SEQ Pyruvate pyruvate + NAD + 1.2.4.1 E. coli aceF b0115, ID NO 152) dehydrogenase, CoASH = acetyl- 2.3.1.12 NP_414657 subunit E2 CoA + NADH + CO2 PDH(SEQ Lipoamide pyruvate + NAD + 1.2.4.1 E. coli lpd b0116, lpd* = ID NO 153) dehydrogenase CoASH = acetyl- 2.3.1.12, ECK0115 NADH- of Pyruvate CoA + NADH + CO2 1.8.1.4 resistant dehydrogenase mutant complex E354K coaA(SEQ pantothenate pantothenate + 2.7.1.33 E. coli coaA (panK) b3974, coaA* = ID NO 154) kinase ATP = ECK3966 feedback- phosphopantothenate + resistant ADP variant R106A panD(SEQ aspartate-1- aspartate = beta- 4.1.1.11 E. coli panD b0131, ID NO 155) decarboxylase alanine + CO2 ECK0130 (proenzyme) aceA(SEQ isocitrate lyase isocitrate = 4.1.3.1 H. elongata aceA b4015, ID NO 156) glyoxylate + ECK4007 succinate aceB(SEQ malate acetyl-CoA + 2.3.3.9 H. elongata aceB b4014, ID NO 157) synthase glyoxylate + H2O = ECK4006 Malate + CoASH + H+ aceK(SEQ isocitrate phosphorylated 3.1.3. H. elongata aceK b4016, ID NO 158) dehydrogenase isocitrate ECK4008 kinase/phosphatase dehydrogenase = isodictrate dehydrogenase + Pi GAPDH(SEQ glyceraldehyde glyceraldehyde-3-P + 1.2.1.12 H. elongata gapA b1779, ID NO 159) 3-P NAD+ + ECK1777 dehydrogenase Pi = 1,3-bisPi- glycerate + NADH + H+ pyk(SEQ pyruvate pyruvate + ATP = 2.7.1.10 E. coli pykA b1854, ID NO 160) kinase ADP + P-enolpyruvate ECK1855 pyk(SEQ pyruvate pyruvate + ATP = 2.4.1.40 E. coli pykF b1676, ID NO 161) kinase ADP + P-enolpyruvate ECK1672 gltA(SEQ citrate oxaloacetate + 2.3.3.1 E. coli gltA b0720, ID NO 162) synthase acetyl-CoA = ECK0709 citrate + CoASH CS citrate oxaloacetate + 2.3.3.1 E. coli Arthrobacter AAC45662 SKG loop synthase acetyl-CoA = strain DS2-3R insertion, citrate + CoASH K313L, A10E bicA(SEQ bicarbonate bicarbonate (out) = none E. coli Synechococcus sp. ABG46427 ID NO 163) transporter bicarbonate (in) PCC7942 GOGAT(SEQ glutamate glutamine + 2- 1.4.1.13 E. coli gltB, gltD b3212, ID NO 164) synthase oxoglutarate + ECK3202 complex NADPH = 2 b3213, (transaminating) glutamate + NADP ECK3203 GOGAT(SEQ glutamate glutamate + 1.4.1.4 E. coli gltB, gltD b3212, ID NO 165) synthase NADP = 2- ECK3202 complex oxoglutarate + b3213, (deaminating) NH3 + NADPH ECK3203 gdh(SEQ glutamate glutamate + 11.4.1.4 E. coli gdhA b1761, ID NO 166) dehydrogenase NADP = 2- ECK1759 oxoglutarate + NH3 + NADPH can(SEQ carbonic CO2 + H20 = 4.2.1.1 E. coli can b0126, ID NO 167) anhydrase bicarbonate + H+ ECK0125 cynT(SEQ carbonic CO2 + H20 = 4.2.1.1 E. coli cynT b0339, ID NO 168) anhydrase bicarbonate + H+ ECK0336 cynS(SEQ cyanase cyanate + 4.2.1.104 E. coli cynS b0340, ID NO 169) bicarbonate = ECK0337 carbamate + CO2 yibD(SEQ predicted none none E. coli yibD b3615, P-regulated ID NO 170) glycosyltransferase ECK3605 gene pstS(SEQ Phosphate Pi (out) + ATP = 3.6.3.27 Arthrobacter pstS b3729, P-regulated ID NO 171) ABC Pi (in) + ADP (Antarctic ECK3721 gene transporter, Pi bacterium) binding protein strain DS2-3R

TABLE-US-00009 TABLE 8 Sugar transport and utilization Enzyme Function Reaction EC # Organism Gene Gene ID Comments cscA(SEQ sucrose sucrose = 3.2.1.48 E. coli cscA CAA57219 ID NO 174) hydrolase glucose + fructose cscB(SEQ sucrose sucrose (out) = none E. coli cscB CAA57217 N234D, ID NO 175) transporter sucrose (in) I312V cscK(SEQ fructokinase fructose + ATP = 2.7.1.3 E. coli cscK CAA57218 ID NO 175) fructose-P + ADP galP(SEQ galactose galactose (out) = none E. coli galP b2943, ID NO 177) transporter galactose (in) ECK2938 galK(SEQ galactokinase galactose + ATP = 2.7.1.6 E. coli galK b0757, ID NO 178) galactose-P + ADP ECK0746

TABLE-US-00010 TABLE 9 Host modifications for fatty acid product Enzyme Function Reaction EC # Organism Gene Gene ID Comments fadE(SEQ acyl-CoA a saturated acyl- 1.3.8. E. coli fadE b0221, ID NO 180) dehydrogenase CoA + oxidized ECK0222 flavoprotein = a trans-enoyl-acyl- CoA + a reduced flavoprotein fadD(SEQ fatty acyl-CoA a saturated fatty 6.2.1.3 E. coli fadD b1805, deletion ID NO 181) synthetase acid + ATP + ECK1803 CoASH = acyl- CoA + AMP + PPi fadA(SEQ 3-keto-acyl-CoA acyl-CoA + 2.3.1.16 E. coli fadA b3845, ID NO 182) thiolase acetyl-CoA = 3- ECK3847 ketoacyl-CoA + CoASH fadB(SEQ fatty acid 3-ketoacyl-CoA 5.1.2.3, E. coli fadB b3846, ID NO 183) oxidation complex .fwdarw. 3- 1.1.1.35, ECK3838 hydroxyacyl-CoA 4.2.1.17, .fwdarw. enoyl-CoA 5.3.3.8 fadI(SEQ 3-keto-acyl-CoA acyl-CoA + 2.3.1.16 E. coli fadI b2342, ID NO 184) thiolase acetyl-CoA = 3- ECK2336 (anaerobic) ketoacyl-CoA + CoASH fadJ(SEQ fatty acid 3-ketoacyl-CoA 5.1.2.3, E. coli fadJ b2341, ID NO 185) oxidation complex .fwdarw. 3- 1.1.1.35, ECK2335 (anaerobic) hydroxyacyl-CoA 4.2.1.17, .fwdarw. enoyl-CoA 5.3.3.8 ydiO(SEQ predicted enoyl- enoyl-CoA + 1.3.8. E. coli ydiO b1695, ID NO 186) CoA reductase reduced ECK1963 flavoprotein = acyl-CoA + oxidized flavoprotein paaJ(SEQ 3-ketoacyl-CoA acyl-CoA + 2.3.1. paaJ b1397, ID NO 187) thiolase acetyl-CoA = 3- ECK1394 keto-acyl-CoA + CoASH yqeF(SEQ predicted E. coli yqeF b2844, ID NO 188) acyltransferase ECK2842 tig(SEQ molecular none none E. coli tig b0436, deletion ID NO 189) chaperone ECK0430 atoD(SEQ Predicted acetate- acetoacetate + 2.8.3. E. coli atoD b2221, deletion ID NO 190) CoA transferase, acetyl-CoA = ECK2214 alpha subunit acetoacetyl-CoA + acetate atoA(SEQ Predicted acetate- acetoacetate + 2.8.3. E. coli atoA b2222, deletion ID NO 191) CoA transferase, acetyl-CoA = ECK2215 beta subunit acetoacetyl-CoA + acetate atoE(SEQ Predicted fatty none E. coli atoE b2223, deletion ID NO 192) acid transporter ECK2216 atoB(SEQ acetyl-CoA 2 acetyl-CoA = 2.3.1.9 E. coli atoB b2224, deletion ID NO 193) acetyltransferase acetoacetyl-CoA + ECK2217 CoASH

TABLE-US-00011 TABLE 10 Fatty acid pathway 3-keto-acyl-CoA synthases Enzyme Function Reaction EC # Organism Gene ID NphT7(SEQ acetoacetyl-CoA acetyl-CoA + 2.3.1. Streptomyces Sp AB540131 ID NO 1) synthase malonyl-CoA = CL190 NphT7 acetoacetyl-CoA + CoASH + CO2 SaFabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Staphylococcus GI:75765832 ID NO 194) synthase malonyl-CoA = a 3- aureus MW2 ketoacyl-CoA + PRK09352 CoASH + CO2 BsFabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Bacillus subtilis YP_004207150 ID NO 195) synthase malonyl-CoA = a 3- 168 fabH1 ketoacyl-CoA + CoASH + CO2 PaFabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Pseudomonas NP_251976 ID NO 196) synthase malonyl-CoA = a 3- aeruginosa PAO1 ketoacyl-CoA + PRK07515 CoASH + CO2 MtFabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Mycobacterium CAB08984 ID NO 197) synthase malonyl-CoA = a 3- tuberculosis H37Rv ketoacyl-CoA + fabH CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rhodothermus gi|345301988 ID NO 198) synthase malonyl-CoA = a 3- marinus ketoacyl-CoA + SG0.5JP17-172 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces gi|471324089 ID NO 199) synthase malonyl-CoA = a 3- davawensis ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Chlamydophila gi|330444499 ID NO 200) synthase malonyl-CoA = a 3- pecorum E58 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Clostridium gi|459068159 ID NO 201) synthase malonyl-CoA = a 3- ultunense Esp ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Corallococcus gi|383454618 ID NO 202) synthase malonyl-CoA = a 3- coralloides DSM ketoacyl-CoA + 2259 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Desmospora sp. gi|333371191 ID NO 203) synthase malonyl-CoA = a 3- 8437 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Paenibacillus gi|390454110 ID NO 204) synthase malonyl-CoA = a 3- peoriae KCTC 3763 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Pelosinus gi|392959403 ID NO 205) synthase malonyl-CoA = a 3- fermentans DSM ketoacyl-CoA + 17108 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Candidatus gi|116626090 ID NO 206) synthase malonyl-CoA = a 3- Solibacter usitatus ketoacyl-CoA + Ellin6076 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Desulfotomaculum gi|323702691 ID NO 207) synthase malonyl-CoA = a 3- nigrificans DSM ketoacyl-CoA + 574 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Saccharomonospora gi|384566084 ID NO 208) synthase malonyl-CoA = a 3- glauca K62 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Corallococcus gi|298162138 ID NO 209) synthase malonyl-CoA = a 3- coralloides ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Legionella gi|148359775 ID NO 210) synthase malonyl-CoA = a 3- pneumophila str. ketoacyl-CoA + Corby CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces gi|15824218 ID NO 211) synthase malonyl-CoA = a 3- avermitilis ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Verrucosispora gi|330468931 ID NO 212) synthase malonyl-CoA = a 3- maris AB-18-032 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rhodopirellula gi|32444698 ID NO 213) synthase malonyl-CoA = a 3- baltica SH 1 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Candidatus gi|392374495 ID NO 214) synthase malonyl-CoA = a 3- methylomirabilis ketoacyl-CoA + oxyfera CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Thermaerobacter gi|317121784 ID NO 215) synthase malonyl-CoA = a 3- marianensis ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Caldisericum exile gi|383787841 ID NO 216) synthase malonyl-CoA = a 3- AZM16c01 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Indibacter gi|404450648 ID NO 217) synthase malonyl-CoA = a 3- alkaliphilus LW1 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Candidatus gi|189502112 ID NO 218) synthase malonyl-CoA = a 3- amoebophilus ketoacyl-CoA + asiaticus 5a2 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Flavobacterium sp. gi|395801183 ID NO 219) synthase malonyl-CoA = a 3- F52 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Anaeromyxobacter gi|86159172 ID NO 220) synthase malonyl-CoA = a 3- dehalogenans 2CP-C ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Microcystis gi|166364688 ID NO 221) synthase malonyl-CoA = a 3- aeruginosa NIES- ketoacyl-CoA + 843 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Chloroflexus gi|219849850 ID NO 222) synthase malonyl-CoA = a 3- aggregans DSM ketoacyl-CoA + 9485 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Lactobacillus gi|227523050 ID NO 223) synthase malonyl-CoA = a 3- hilgardii ATCC ketoacyl-CoA + 8290 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Bartonella grahamii gi|240850683 ID NO 224) synthase malonyl-CoA = a 3- as4aup ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Clostridium gi|253681256 ID NO 225) synthase malonyl-CoA = a 3- botulinum D str. ketoacyl-CoA + 1873 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Vibrio cholerae gi|254286853 ID NO 226) synthase malonyl-CoA = a 3- AM-19226 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Propionibacterium gi|282854072 ID NO 227) synthase malonyl-CoA = a 3- acnes J139 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces gi|291439887 ID NO 228) synthase malonyl-CoA = a 3- ghanaensis ATCC ketoacyl-CoA + 14672 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Veillonella sp. gi|294791665 ID NO 229) synthase malonyl-CoA = a 3- 6_1_27 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces sp. C gi|302539498 ID NO 230) synthase malonyl-CoA = a 3- ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces sp. gi|318080591 ID NO 231) synthase malonyl-CoA = a 3- SA3_actF ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. uncultured gi|374851360 ID NO 232) synthase malonyl-CoA = a 3- Aquificae ketoacyl-CoA + bacterium CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Saccharomonospora gi|381164912 ID NO 233) synthase malonyl-CoA = a 3- azurea NA-128 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Ralstonia gi|386335197 ID NO 234) synthase malonyl-CoA = a 3- solanacearum Po82 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Frankia sp. QA3 gi|392946737 ID NO 235) synthase malonyl-CoA = a 3- ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Alishewanella gi|397172008 ID NO 236) synthase malonyl-CoA = a 3- aestuarii B11 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Brevibacillus sp. gi|399047091 ID NO 237) synthase malonyl-CoA = a 3- CF112 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Sphingomonas sp. gi|402823152 ID NO 238) synthase malonyl-CoA = a 3- LH128 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Alteromonas gi|407684813 ID NO 239) synthase malonyl-CoA = a 3- macleodii str. ketoacyl-CoA + English Channel CoASH + CO2 673 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Leptospirillum gi|410479651 ID NO 240) synthase malonyl-CoA = a 3- ferriphilum ML-04 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Glaciecola polaris gi|410617776 ID NO 241) synthase malonyl-CoA = a 3- LMG 21857 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Listeria gi|417318270 ID NO 242) synthase malonyl-CoA = a 3- monocytogenes J1- ketoacyl-CoA + 220 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Mycobacterium gi|417747984 ID NO 243) synthase malonyl-CoA = a 3- avium subsp. ketoacyl-CoA + paratuberculosis CoASH + CO2 S397 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Fusobacterium gi|422338672 ID NO 244) synthase malonyl-CoA = a 3- nucleatum subsp. ketoacyl-CoA + polymorphum CoASH + CO2 F0401 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Mycobacterium gi|443491493 ID NO 245) synthase malonyl-CoA = a 3- liflandii 128FXT ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Mesorhizobium sp. gi|474659331 ID NO 246) synthase malonyl-CoA = a 3- STM 4661 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Streptomyces gi|21224866 ID NO 247) synthase malonyl-CoA = a 3- coelicolor A3(2) ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Clostridiales gi|239623103 ID NO 248) synthase malonyl-CoA = a 3- bacterium ketoacyl-CoA + 1_7_47_FAA CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Ruegeria sp. R11 gi|254477647 ID NO 249) synthase malonyl-CoA = a 3- ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rothia dentocariosa gi|311113478 ID NO 250) synthase malonyl-CoA = a 3- ATCC 17931 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Caldicellulosiruptor gi|312793335 ID NO 251) synthase malonyl-CoA = a 3- kristjanssonii ketoacyl-CoA + 177R1B CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Thermus gi|320449672 ID NO 252) synthase malonyl-CoA = a 3- scotoductus SA-01 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Geobacter sp. M18 gi|322421910 ID NO 253) synthase malonyl-CoA = a 3- ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rhodococcus equi gi|325677042 ID NO 254) synthase malonyl-CoA = a 3- ATCC 33707 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Clostridium gi|326203621 ID NO 255) synthase malonyl-CoA = a 3- papyrosolvens ketoacyl-CoA + DSM 2782 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Cellulomonas fimi gi|332670773 ID NO 256) synthase malonyl-CoA = a 3- ATCC 484 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Neisseria macacae gi|340361349 ID NO 257) synthase malonyl-CoA = a 3- ATCC 33926 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rhodothermus gi|345304635 ID NO 258) synthase malonyl-CoA = a 3- marinus ketoacyl-CoA + SG0.5JP17-172 CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Gluconacetobacter gi|349685677 ID NO 259) synthase malonyl-CoA = a 3- oboediens 174Bp2 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Halomonas sp. gi|352106212 ID NO 260) synthase malonyl-CoA = a 3- HAL1 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Saccharomonospora gi|375098553 ID NO 261) synthase malonyl-CoA = a 3- cyanea NA-134 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Arcobacter butzleri gi|384154990 ID NO 262) synthase malonyl-CoA = a 3- ED-1 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Marinobacter gi|385331603 ID NO 263) synthase malonyl-CoA = a 3- adhaerens HP15 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Phaeobacter gi|400755130 ID NO 264) synthase malonyl-CoA = a 3- gallaeciensis 2.10 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Aeromonas gi|423197564 ID NO 265) synthase malonyl-CoA = a 3- hydrophila SSU ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Rhodococcus gi|424853848 ID NO 266) synthase malonyl-CoA = a 3- opacus PD630 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Gordonia aichiensis gi|441509582 ID NO 267) synthase malonyl-CoA = a 3- NBRC 108223 ketoacyl-CoA + CoASH + CO2 FabH(SEQ 3-ketoacyl-CoA a fatty acyl-CoA + 2.3.1. Acinetobacter sp. gi|479875377 ID NO 268) synthase malonyl-CoA = a 3- NIPH 236 ketoacyl-CoA + CoASH + CO2 PaFabG(SEQ 3-ketoacyl-CoA a 3-ketoacyl-CoA + 1.1.1.35 Pseudomonas NP_251657 ID NO 269) reductase NADH = a 3- aeruginosa PAO1 hydroxyacyl-CoA + NAD+ fabG(SEQ 3-ketoacyl-CoA a 3-ketoacyl-CoA + 1.1.1.35 Pseudomonas ABR85110 ID NO 270) reductase NADH = a 3- aeruginosa PA7 hydroxyacyl-CoA + NAD+ hbd(SEQ 3-hydroxybutyryl- acetoacetyl-CoA + 1.1.1.35 Clostridium AF494018_5 ID NO 271) CoA dehydrogenase NADH = 3- beijerinckii hydroxybutyryl-CoA + NAD+ crt(SEQ crotonase/enoyl- 3-hydroxybutyryl- 4.2.1.55 Clostridium AAA95967 ID NO 272) CoA hydratase CoA = crotonyl- acetobutylicum CoA + H2O ech(SEQ enoyl-CoA 3-hydroxybutyryl- 4.2.1.55 Pseudomonas ABA10805 ID NO 272) hydratase CoA = crotonyl- putida CoA + H2O ech2(SEQ bifunctional 3- a 3-ketoacyl-CoA + 1.1.1.35 Rattus norvegicus NP_077368 ID NO 274) hydroxyacyl-CoA NADH = a 3- 4.2.1.55 dehydrogenase/ hydroxyacyl-CoA + enoyl-CoA NAD+ hydratase 3-hydroxyacyl-CoA = enoyl-CoA + H2O ter(SEQ crotonase/enoyl- a enoyl-CoA + 1.1.1.36 Treponema WP_002681770 ID NO 275) CoA hydratase NADH = a denticola TDE0597 fattyacyl-CoA + NAD+ ccr(SEQ crotonase/enoyl- a enoyl-CoA + 1.1.1.36 Streptomyces GI:81309006, ID NO 276) CoA hydratase NADH = a collinus Q53865 fattyacyl-CoA + NAD+

TABLE-US-00012 TABLE 11 Thioesterases Enzyme Function Reaction EC # Organism Gene Gene ID Comments TesA acyl-CoA acyl-ACP + 3.1.2.14 E. coli tesA b0494, (SEQ ID thioesterase, H2O = fatty 3.1.2.2 ECK0488 NO 277) protease, acid + ACP phosphlipase acyl-CoA + (periplasmic) H2O = fatty acid + CoASH tesA acyl-CoA acyl-ACP + 3.1.2.14 E. coli tesA del2-24 (SEQ ID thioesterase H2O = fatty 3.1.2.2 (signal NO 278) (cytoplasmic) acid + ACP sequence) acyl-CoA + H2O = fatty acid + CoASH tesB thioesterase acyl-CoA + 3.1.2.20 E. coli tesB b0452, (SEQ ID H2O = fatty ECK0446 NO 279) acid + CoASH yciA acyl-CoA acyl-CoA + 3.1.2.20 E. coli yciA b1253, (SEQ ID thioesterase H2O = fatty ECK1247 NO 280) acid + CoASH ybgC acyl-CoA acyl-CoA + 3.1.2.28 E. coli ybgC b0736, (SEQ ID thioesterase H2O = fatty ECK0725 NO 281) acid + CoASH ybfF predicted acyl-CoA + 3.1.2.20 E. coli ybfF b0686, (SEQ ID thioesterase H2O = fatty ECK0674 NO 282) acid + CoASH fadM thioesterase acyl-CoA + 3.1.2. E. coli fadM b0443, (SEQ ID H2O = fatty ECK0437 NO 283) acid + CoASH AtTE thioesterase acyl-CoA + 3.1.2.20 Anaerococcus EEI82564 (SEQ ID H2O = fatty tetradius NO 284) acid + CoASH ATCC35098 CpTE thioesterase acyl-CoA + 3.1.2.20 Cuphea AAC49179 (SEQ ID H2O = fatty palustris NO 285) acid + CoASH CperfTE thioesterase acyl-CoA + 3.1.2.20 Clostridium ABG82470 (SEQ ID H2O = fatty perfringens NO 286) acid + CoASH ATCC13124 LpTE thioesterase acyl-CoA + 3.1.2.20 Lactobacillus CAD63310 (SEQ ID H2O = fatty plantarum NO 287) acid + CoASH ACFS1 fat PA2801TE thioesterase acyl-CoA + 3.1.2.20 Pseudomonas NP_251491 (SEQ ID H2O = fatty aeruginosa NO 288) acid + CoASH PAO1

TABLE-US-00013 TABLE 12 Wax ester synthases Enzyme Function Reaction EC # Organism Gene ID Maq1(SEQ ID Wax ester a fatty acyl-CoA + an 2.3.1.20 Marinobacter aquaeolei YP_957462 NO 289) synthase alcohol = a fatty acyl VT8 (ATCC700491) Ma1 methyl ester + CoASH Pcry1(SEQ ID Wax ester a fatty acyl-CoA + an 2.3.1.20 Psychrobacter cryohalolentis YP_579515 NO 290) synthase alcohol = a fatty acyl K5 Ps1 methyl ester + CoASH Rjos1(SEQ ID Wax ester a fatty acyl-CoA + an 2.3.1.20 Rhodococcus jostii YP_701572 NO 291) synthase alcohol = a fatty acyl RHA1 Rh1 methyl ester + CoASH Abork1(SEQ ID Wax ester a fatty acyl-CoA + an 2.3.1.20 Alcanivorax borkumensis YP_694462 NO 292) synthase alcohol = a fatty acyl strain SK2 atfA1 methyl ester + CoASH

TABLE-US-00014 TABLE 13 Miscellaneous Source/Genes E. coli Enzyme Function Reaction EC # unless noted Gene ID Comments prpE(SEQ propionyl-CoA propionate + CoASH + 6.2.1.17 Salmonella NP_454966 ID NO 293) synthetase ATP = propionyl-CoA + enterica AMP + PPi subsp typhimirium phaA(SEQ acetyl-CoA 2 acetyl-CoA = 2.3.1.9 Cupriavides YP_353824 ID NO 294) acetyltransferase/ acetoacetyl-CoA + necator thiolase CoASH (Rhodobacter sphaeroides 2.4.1) phaB(SEQ acetoacetyl- 3-ketoacyl-CoA + 1.1.1.35 Cupriavides YP_353825 ID NO 295) CoA reductase NAD(P)H = 3- necator hydroxyacyl-CoA + (Rhodobacter NAD(P)+ sphaeroides 2.4.1) phaC(SEQ PHA synthase hydroxyacyl-CoA + 2.3.1. Pseudomonas AAO59383 ID NO 296) [hydroxyalkanoate]n = stutzeri [hydroxyalkanoate]n + 1 + phaC1 CoASH phaC(SEQ PHA synthase hydroxyacyl-CoA + 2.3.1. Pseudomonas AAA25932 ID NO 297) [hydroxyalkanoate]n = oleovorans [hydroxyalkanoate]n + 1 + CoASH phaC(SEQ PHA synthase hydroxyacyl-CoA + 2.3.1. Pseudomonas AAG08441 ID NO 298) [hydroxyalkanoate]n = aeruginosa [hydroxyalkanoate]n + 1 + PAO1 CoASH THNS THN synthase 5 malonyl-CoA = THN unknown Streptomycs CAC01488 (tetrahydroxynaphthalene) coelicolor .fwdarw. flaviolin bcsA THNS THN synthase 5 malonyl-CoA = THN unknown Streptomycs CAC01488 C184S, variant C184S, (tetrahydroxynaphthalene) coelicolor del351-374 del25 .fwdarw. flaviolin bcsA

TABLE-US-00015 TABLE14 ProteinSequencesoftheenzymesinvolvedinthegeneticmodification. (Aminoacidslistedinboldandunderlinedrepresentmodificationsmadeinapplicants' alleles) SEQIDNO Enzyme FASTAHeader ProteinSequence-NCBIDatabase Hostmodificationsforyieldincrease/byproductelimination SEQIDNO:124 ldhA >gi|16129341|ref|NP_ MKLAVYSTKQYDKKYLQQVNESFGFELEFF 415898.11 DFLLTEKTAKTANGCEAVCIFVNDDGSRPVL fermentativeD-lactate EELKKHGVKYIALRCAGFNNVDLDAAKELG dehydrogenase,NAD- LKVVRVPAYDPEAVAEHAIGMMMTLNRRIH dependent RAYQRTRDANFSLEGLTGFTMYGKTAGVIG [Escherichiacolistr. TGKIGVAMLRILKGFGMRLLAFDPYPSAAAL K-12substr.MG1655] ELGVEYVDLPTLFSESDVISLHCPLTPENYHL LNEAAFEQMKNGVMIVNTSRGALIDSQAAIE ALKNQKIGSLGMDVYENERDLFFEDKSNDVI QDDVFRRLSACHNVLFTGHQAFLTAEALTSI SQTTLQNLSNLEKGETCPNELV SEQIDNO:125 pflB >gi|16128870|ref|NP_ MSELNEKLATAWEGFTKGDWQNEVNVRDFI 415423.1| pyruvate QKNYTPYEGDESFLAGATEATTTLWDKVME formatelyaseI GVKLENRTHAPVDFDTAVASTITSHDAGYIN [Escherichiacolistr. KQLEKIVGLQTEAPLKRALIPFGGIKMIEGSC K-12substr.MG1655] KAYNRELDPMIKKIFTEYRKTHNQGVFDVY TPDILRCRKSGVLTGLPDAYGRGRIIGDYRR VALYGIDYLMKDKLAQFTSLQADLENGVNL EQTIRLREEIAEQHRALGQMKEMAAKYGYD ISGPATNAQEAIQWTYFGYLAAVKSQNGAA MSFGRTSTFLDVYIERDLKAGKITEQEAQEM VDHLVMKLRMVRFLRTPEYDELFSGDPIWA TESIGGMGLDGRTLVTKNSFRFLNTLYTMGP SPEPNMTILWSEKLPLNFKKFAAKVSIDTSSL QYENDDLMRPDFNNDDYAIACCVSPMIVGK QMQFFGARANLAKTMLYAINGGVDEKLKM QVGPKSEPIKGDVLNYDEVMERMDHFMDW LAKQYITALNIIHYMHDKYSYEASLMALHD RDVIRTMACGIAGLSVAADSLSAIKYAKVKP IRDEDGLAIDFEIEGEYPQFGNNDPRVDDLA VDLVERFMKKIQKLHTYRDAIPTQSVLTITSN VVYGKKTGNTPDGRRAGAPFGPGANPMHG RDQKGAVASLTSVAKLPFAYAKDGISYTFSI VPNALGKDDEVRKTNLAGLMDGYFHHEASI EGGQHLNVNVMNREMLLDAMENPEKYPQL TIRVSGYAVRFNSLTKEQQQDVITRTFTQSM SEQIDNO:126 mgsA >gi|90111195|ref|NP_ MELTTRTLPARKHIALVAHDHCKQMLMSW 415483.21 VERHQPLLEQHVLYATGTTGNLISRATGMN methylglyoxal VNAMLSGPMGGDQQVGALISEGKIDVLIFF synthase[Escherichia WDPLNAVPHDPDVKALLRLATVWNIPVATN colistr.K-12substr. VATADFIIQSPHFNDAVDILIPDYQRYLADRL MG1655] K SEQIDNO:127 poxB >gi|16128839|ref|NP_ MKQTVAAYIAKTLESAGVKRIWGVTGDSLN 415392.1| pyruvate GLSDSLNRMGTIEWMSTRHEEVAAFAAGAE dehydrogenase AQLSGELAVCAGSCGPGNLHLINGLFDCHRN (pyruvateoxidase), HVPVLAIAAHIPSSEIGSGYFQETHPQELFREC thiamin-dependent, SHYCELVSSPEQIPQVLAIAMRKAVLNRGVS FAD-binding VVVLPGDVALKPAPEGATMHWYHAPQPVV [Escherichiacolistr. TPEEEELRKLAQLLRYSSNIALMCGSGCAGA K-12substr.MG1655] HKELVEFAGKIKAPIVHALRGKEHVEYDNPY DVGMTGLIGFSSGFHTMMNADTLVLLGTQF PYRAFYPTDAKIIQIDINPASIGAHSKVDMAL VGDIKSTLRALLPLVEEKADRKFLDKALEDY RDARKGLDDLAKPSEKAIHPQYLAQQISHFA ADDAIFTCDVGTPTVWAARYLKMNGKRRLL GSFNHGSMANAMPQALGAQATEPERQVVA MCGDGGFSMLMGDFLSVVQMKLPVKIVVF NNSVLGFVAMEMKAGGYLTDGTELHDTNF ARIAEACGITGIRVEKASEVDEALQRAFSIDG PVLVDVVVAKEELAIPPQIKLEQAKGFSLYM LRAIISGRGDEVIELAKTNWLR SEQIDNO:128 pta >gi|16130232|ref|NP_ MSRIIMLIPTGTSVGLTSVSLGVIRAMERKGV 416800.11phosphate RLSVFKPIAQPRTGGDAPDQTTTIVRANSSTT acetyltransferase TAAEPLKMSYVEGLLSSNQKDVLMEEIVAN [Escherichiacolistr. YHANTKDAEVVLVEGLVPTRKHQFAQSLNY K-12substr.MG1655] EIAKTLNAEIVFVMSQGTDTPEQLKERIELTR NSFGGAKNTNITGVIVNKLNAPVDEQGRTRP DLSEIFDDSSKAKVNNVDPAKLQESSPLPVL GAVPWSFDLIATRAIDMARHLNATIINEGDIN TRRVKSVTFCARSIPHMLEHFRAGSLLVTSA DRPDVLVAACLAAMNGVEIGALLLTGGYEM DARISKLCERAFATGLPVFMVNTNTWQTSLS LQSFNLEVPVDDHERIEKVQEYVANYINAD WIESLTATSERSRRLSPPAFRYQLTELARKAG KRIVLPEGDEPRTVKAAAICAERGIATCVLLG NPAEINRVAASQGVELGAGIEIVDPEVVRES YVGRLVELRKNKGMTETVAREQLEDNVVL GTLMLEQDEVDGLVSGAVHTTANTIRPPLQL IKTAPGSSLVSSVFFMLLPEQVYVYGDCAINP DPTAEQLAEIAIQSADSAAAFGIEPRVAMLSY STGTSGAGSDVEKVREATRLAQEKRPDLMID GPLQYDAAVMADVAKSKAPNSPVAGRATV FIFPDLNTGNTTYKAVQRSADLISIGPMLQG MRKPVNDLSRGALVDDIVYTIALTAIQSAQQ Q SEQIDNO:129 ack >gi|16130231|ref|NP_ MSSKLVLVLNCGSSSLKFAIIDAVNGEEYLS 416799.11acetate GLAECFHLPEARIKWKMDGNKQEAALGAG kinaseAand AAHSEALNFIVNTILAQKPELSAQLTAIGHRI propionatekinase2 VHGGEKYTSSVVIDESVIQGIKDAASFAPLH [Escherichiacolistr. NPAHLIGIEEALKSFPQLKDKNVAVFDTAFH K-12substr.MG1655] QTMPEESYLYALPYNLYKEHGIRRYGAHGT SHFYVTQEAAKMLNKPVEELNIITCHLGNGG SVSAIRNGKCVDTSMGLTPLEGLVMGTRSG DIDPAIIFHLHDTLGMSVDAINKLLTKESGLL GLTEVTSDCRYVEDNYATKEDAKRAMDVY CHRLAKYIGAYTALMDGRLDAVVFTGGIGE NAAMVRELSLGKLGVLGFENDHERNLAARF GKSGFINKEGTRPAVVIPTNEELVIAQDASRL TA SEQIDNO:130 adhE >gi|16129202|ref|NP_ MAVTNVAELNALVERVKKAQREYASFTQE 415757.11fused QVDKIFRAAALAAADARIPLAKMAVAESGM acetaldehyde-CoA GIVEDKVIKNHFASEYIYNAYKDEKTCGVLS dehydrogenase/iron- EDDTFGTITIAEPIGIICGIVPTTNPTSTAIFKSL dependentalcohol ISLKTRNAIIFSPHPRAKDATNKAADIVLQAA dehydrogenase/pyruva IAAGAPKDLIGWIDQPSVELSNALMHHPDIN te-formatelyase LILATGGPGMVKAAYSSGKPAIGVGAGNTP deactivase VVIDETADIKRAVASVLMSKTFMGVICASE [Escherichiacolistr. QSVVVVDSVYDAVRERFATHGGYLLQGKEL K-12substr.MG1655] KAVQDVILKNGALNAAIVGQPAYKIAELAGF SVPENTKILIGEVTVVDESEPFAHEKLSPTLA MYRAKDFEDAVEKAEKLVAMGGIGHTSCL YTDQDNQPARVSYFGQKMKTARILINTPASQ GGIGDLYNFKLAPSLTLGCGSWGGNSISENV GPKHLINKKTVAKRAENMLWHKLPKSIYFR RGSLPIALDEVITDGHKRALIVTDRFLFNNGY ADQITSVLKAAGVETEVFFEVEADPTLSIVRK GAELANSFKPDVIIALGGGSPMDAAKIMWV MYEHPETHFEELALRFMDIRKRIYKFPKMGV KAKMIAVTTTSGTGSEVTPFAVVTDDATGQ KYPLADYALTPDMAIVDANLVMDMPKSLC AFGGLDAVTHAMEAYVSVLASEFSDGQALQ ALKLLKEYLPASYHEGSKNPVARERVHSAA TIAGIAFANAFLGVCHSMAHKLGSQFHIPHG LANALLICNVIRYNANDNPTKQTAFSQYDRP QARRRYAEIADHLGLSAPGDRTAAKIEKLLA WLETLKAELGIPKSIREAGVQEADFLANVDK LSEDAFDDQCTGANPRYPLISELKQILLDTYY GRDYVEGETAAKKEAAPAKAEKKAKKSA Fattyacidsynthesis(includingtemperaturesensitiveallelesusedforincreasedmalonyl- CoAavailability) SEQIDNO:131 ACP >gi|16129057|ref|NP_ MSTIEERVKKIIGEQLGVKQEEVTNNASFVE 415612.1| acylcarrier DLGADSLDTVELVMALEEEFDTEIPDEEAEKI protein(ACP) TTVQAAIDYINGHQA [Escherichiacolistr. K-12substr.MG1655] SEQIDNO:132 fabI >gi|16129249|ref|NP_ MGFLSGKRILVTGVASKLSIAYGIAQAMHRE 415804.1| enoyl-[acyl- GAELAFTYQNDKLKGRVEEFAAQLGSDIVL carrier-protein] QCDVAEDASIDTMFAELGKVWPKFDGFVHS reductase,NADH- IGFAPGDQLDGDYVNAVTREGFKIAHDISSY dependent SFVAMAKACRSMLNPGSALLTLSYLGAERAI [Escherichiacolistr. PNYNVMGLAKASLEANVRYMANAMGPEGV K-12substr.MG1655] RVNAISAGPIRTLAASGIKDFRKMLAHCEAV TPIRRTVTIEDVGNSAAFLCSDLSAGISGEVV HVDGGFSIAAMNELELK SEQIDNO:133 fabB >gi|16130258|ref|NP_ MKRAVITGLGIVSSIGNNQQEVLASLREGRS 416826.1| 3-oxoacyl- GITFSQELKDSGMRSHVWGNVKLDTTGLIDR [acyl-carrier-protein] KVVRFMSDASIYAFLSMEQAIADAGLSPEAY synthaseI QNNPRVGLIAGSGGGSPRFQVFGADAMRGP [Escherichiacolistr. RGLKAVGPYVVTKAMASGVSACLATPFKIH K-12substr.MG1655] GVNYSISSACATSAHCIGNAVEQIQLGKQDIV FAGGGEELCWEMACEFDAMGALSTKYNDT PEKASRTYDAHRDGFVIAGGGGMVVVEELE HALARGAHIYAEIVGYGATSDGADMVAPSG EGAVRCMKMAMHGVDTPIDYLNSHGTSTPV GDVKELAAIREVFGDKSPAISATKAMTGHSL GAAGVQEAIYSLLMLEHGFIAPSINIEELDEQ AAGLNIVTETTDRELTTVMSNSFGFGGTNAT LVMRKLKD SEQIDNO:134 fabH >gi|16129054|ref|NP_ MYTKIIGTGSYLPEQVRTNADLEKMVDTSDE 415609.1| 3-oxoacyl- WIVTRTGIRERHIAAPNETVSTMGFEAATRAI [acyl-carrier-protein] EMAGIEKDQIGLIVVATTSATHAFPSAACQIQ synthaseIII SMLGIKGCPAFDVAAACAGFTYALSVADQY [Escherichiacolistr. VKSGAVKYALVVGSDVLARTCDPTDRGTIII K-12substr.MG1655] FGDGAGAAVLAASEEPGIISTHLHADGSYGE LLTLPNADRVNPENSIHLTMAGNEVFKVAVT ELAHIVDETLAANNLDRSQLDWLVPHQANL RIISATAKKLGMSMDNVVVTLDRHGNTSAA SVPCALDEAVRDGRIKPGQLVLLEAFGGGFT WGSALVRF SEQIDNO:135 fabD >gi|16129055|ref|NP_ MTQFAFVFPGQGSQTVGMLADMAASYPIVE 415610.1| malonyl- ETFAEASAALGYDLWALTQQGPAEELNKTW CoA-[acyl-carrier- QTQPALLTASVALYRVWQQQGGKAPAMMA protein]transacylase GHSLGEYSALVCAGVIDFADAVRLVEMRGK [Escherichiacolistr. FMQEAVPEGTGAMAAIIGLDDASIAKACEEA K-12substr.MG1655] AEGQVVSPVNFNSPGQVVIAGHKEAVERAG AACKAAGAKRALPLPVSVPSHCALMKPAAD KLAVELAKITFNAPTVPVVNNVDVKCETNG DAIRDALVRQLYNPVQWTKSVEYMAAQGV EHLYEVGPGKVLTGLTKRIVDTLTASALNEP SAMAAALEL SEQIDNO:136 fabF >gi|16129058|ref|NP_ MSKRRVVVTGLGMLSPVGNTVESTWKALL 415613.1| 3-oxoacyl- AGQSGISLIDHFDTSAYATKFAGLVKDFNCE [acyl-carrier-protein] DIISRKEQRKMDAFIQYGIVAGVQAMQDSGL synthaseII EITEENATRIGAAIGSGIGGLGLIEENHTSLMN [Escherichiacolistr. GGPRKISPFFVPSTIVNMVAGHLTIMYGLRGP K-12substr.MG1655] SISIATACTSGVHNIGHAARIIAYGDADVMV AGGAEKASTPLGVGGFGAARALSTRNDNPQ AASRPWDKERDGFVLGDGAGMLVLEEYEH AKKRGAKIYAELVGFGMSSDAYHMTSPPEN GAGAALAMANALRDAGIEASQIGYVNAHGT STPAGDKAEAQAVKTIFGEAASRVLVSSTKS MTGHLLGAAGAVESIYSILALRDQAVPPTIN LDNPDEGCDLDFVPHEARQVSGMEYTLCNS FGFGGTNGSLIFKKI SEQIDNO:137 fabG >gi|16129056|ref|NP_ MNFEGKIALVTGASRGIGRAIAETLAARGAK 415611.1| 3-oxoacyl- VIGTATSENGAQAISDYLGANGKGLMLNVT [acyl-carrier-protein] DPASIESVLEKIRAEFGEVDILVNNAGITRDN reductase[Escherichia LLMRMKDEEWNDIIETNLSSVFRLSKAVMR colistr.K-12substr. AMMKKRHGRIITIGSVVGTMGNGGQANYA MG1655] AAKAGLIGFSKSLAREVASRGITVNVVAPGFI ETDMTRALSDDQRAGILAQVPAGRLGGAQE IANAVAFLASDEAAYITGETLHVNGGMYMV SEQIDNO:138 fabA >gi|16128921|ref|NP_ MVDKRESYTKEDLLASGRGELFGAKGPQLP 415474.1| beta- APNMLMMDRVVKMTETGGNFDKGYVEAEL hydroxydecanoyl DINPDLWFFGCHFIGDPVMPGCLGLDAMWQ thioesterdehydrase LVGFYLGWLGGEGKGRALGVGEVKFTGQV [Escherichiacolistr. LPTAKKVTYRIHFKRIVNRRLIMGLADGEVL K-12substr.MG1655] VDGRLIYTASDLKVGLFQDTSAF SEQIDNO:139 fabZ >gi|16128173|ref|NP_ MTTNTHTLQIEEILELLPHRFPFLLVDRVLDF 414722.1| (3R)- EEGRFLRAVKNVSVNEPFFQGHFPGKPIFPG hydroxymyristolacyl VLILEAMAQATGILAFKSVGKLEPGELYYFA carrierprotein GIDEARFKRPVVPGDQMIMEVTFEKTRRGLT dehydratase RFKGVALVDGKVVCEATMMCARSREA [Escherichiacolistr. K-12substr.MG1655] SEQIDNO:140 fabR >gi|145698338|ref|NP_ MGVRAQQKEKTRRSLVEAAFSQLSAERSFA 418398.2| DNA- SLSLREVAREAGIAPTSFYRHFRDVDELGLT bindingtranscriptional MVDESGLMLRQLMRQARQRIAKGGSVIRTS repressor[Escherichia VSTFMEFIGNNPNAFRLLLRERSGTSAAFRA colistr.K-12substr. AVAREIQHFIAELADYLELENHMPRAFTEAQ MG1655] AEAMVTIVFSAGAEALDVGVEQRRQLEERL VLQLRMISKGAYYWYRREQEKTAIIPGNVK DE SEQIDNO:141 fabI enoyl-ACPreductase, MGFLSGKRILVTGVASKLSIAYGIAQAMHRE NADH-dependent, GAELAFTYQNDKLKGRVEEFAAQLGSDIVL temperature-sensitive QCDVAEDASIDTMFAELGKVWPKFDGFVHS applicantsallele IGFAPGDQLDGDYVNAVTREGFKIAHDISSY SFVAMAKACRSMLNPGSALLTLSYLGAERAI PNYNVMGLAKASLEANVRYMANAMGPEGV RVNAISAGPIRTLAASGIKDFRKMLAHCEAV TPIRRTVTIEDVGNSAAFLCSDLSAGIFGEVV HVDGGFSIAAMNELELK SEQIDNO:142 fabB 3-ketoacyl-ACP MKRAVITGLGIVSSIGNNQQEVLASLREGRS synthaseI, GITFSQELKDSGMRSHVWGNVKLDTTGLIDR temperaturesensitive KVVRFMSDASIYAFLSMEQAIADAGLSPEAY applicantsallele QNNPRVGLIAGSGGGSPRFQVFGADAMRGP RGLKAVGPYVVTKAMASGVSACLATPFKIH GVNYSISSACATSAHCIGNAVEQIQLGKQDIV FAGGGEELCWEMACEFDAMGALSTKYNDT PEKASRTYDAHRDGFVIAGGGGMVVVEELE HALARGAHIYAEIVGYGATSDGADMVAPSG EGAVRCMKMAMHGVDTPIDYLNSHGTSTPV GDVKELAAIREVFGDKSPAISATKVMTGHSL GAAGVQEAIYSLLMLEHGFIAPSINIEELDEQ AAGLNIVTETTDRELTTVMSNSFGFGGTNAT LVMRKLKD SEQIDNO:143 fabD malonyl-CoA:ACP MTQFAFVFPGQGSQTVGMLADMAASYPIVE transacylase, ETFAEASAALGYDLWALTQQGPAEELNKTW temperaturesensitive QTQPALLTASVALYRVWQQQGGKAPAMMA applicantsallele GHSLGEYSALVCAGVIDFADAVRLVEMRGK FMQEAVPEGTGAMAAIIGLDDASIAKACEEA AEGQVVSPVNFNSPGQVVIAGHKEAVERAG AACKAAGAKRALPLPVSVPSHCALMKPAAD KLAVELAKITFNAPTVPVVNNVDVKCETNG DAIRDALVRQLYNPVQQTKSVEYMAAQGV EHLYEVGPGKVLTGLTKRIVDTLTASALNEP SAMAAAL Malonyl-CoAsynthesisandothergenesrelatedtooptimizingflux SEQIDNO:144 udhA >gi|90111670|ref|NP_ MPHSYDYDAIVIGSGPGGEGAAMGLVKQGA 418397.2| pyridine RVAVIERYQNVGGGCTHWGTIPSKALRHAV nucleotide SRIIEFNQNPLYSDHSRLLRSSFADILNHADN transhydrogenase, VINQQTRMRQGFYERNHCEILQGNARFVDE soluble[Escherichia HTLALDCPDGSVETLTAEKFVIACGSRPYHP colistr.K-12substr. TDVDFTHPRIYDSDSILSMHHEPRHVLIYGAG MG1655] VIGCEYASIFRGMDVKVDLINTRDRLLAFLD QEMSDSLSYHFWNSGVVIRHNEEYEKIEGCD DGVIMHLKSGKKLKADCLLYANGRTGNTDS LALQNIGLETDSRGQLKVNSMYQTAQPHVY AVGDVIGYPSLASAAYDQGRIAAQALVKGE ATAHLIEDIPTGIYTIPEISSVGKTEQQLTAMK VPYEVGRAQFKHLARAQIVGMNVGTLKILF HRETKEILGIHCFGERAAEIIHIGQAIMEQKG GGNTIEYFVNTTFNYPTMAEAYRVAALNGL NRLF SEQIDNO:145 pntA >gi|1612956|ref|NP_ MRIGIPRERLTNETRVAATPKTVEQLLKLGFT 416120.1| pyridine VAVESGAGQLASFDDKAFVQAGAEIVEGNS nucleotide VWQSEIILKVNAPLDDEIALLNPGTTLVSFIW transhydrogenase, PAQNPELMQKLAERNVTVMAMDSVPRISRA alphasubunit QSLDALSSMANIAGYRAIVEAAHEFGRFFTG [Escherichiacolistr. QITAAGKVPPAKVMVIGAGVAGLAAIGAAN K-12substr.MG1655] SLGAIVRAFDTRPEVKEQVQSMGAEFLELDF KEEAGSGDGYAKVMSDAFIKAEMELFAAQA KEVDIIVTTALIPGKPAPKLITREMVDSMKAG SVIVDLAAQNGGNCEYTVPGEIFTTENGVKV IGYTDLPGRLPTQSSQLYGTNLVNLLKLLCK EKDGNITVDFDDVVIRGVTVIRAGEITWPAPP IQVSAQPQAAQKAAPEVKTEEKCTCSPWRK YALMALAIILFGWMASVAPKEFLGHFTVFAL ACVVGYYVVWNVSHALHTPLMSVTNAISGII VVGALLQIGQGGWVSFLSFIAVLIASINIFGG FTVTQRMLKMFRKN SEQIDNO:146 pntB >gi|16129560|ref|NP_ MSGGLVTAAYIVAAILFIFSLAGLSKHETSRQ 416119.1| pyridine GNNFGIAGMAIALIATIFGPDTGNVGWILLA nucleotide MVIGGAIGIRLAKKVEMTEMPELVAILHSFV transhydrogenase,beta GLAAVLVGFNSYLHHDAGMAPILVNIHLTE subunit[Escherichia VFLGIFIGAVTFTGSVVAFGKLCGKISSKPLM colistr.K-12substr. LPNRHKMNLAALVVSFLLLIVFVRTDSVGLQ MG1655] VLALLIMTAIALVFGWHLVASIGGADMPVV VSMLNSYSGWAAAAAGFMLSNDLLIVTGAL VGSSGAILSYIMCKAMNRSFISVIAGGFGTDG SSTGDDQEVGEHREITAEETAELLKNSHSVII TPGYGMAVAQAQYPVAEITEKLRARGINVR FGIHPVAGRLPGHMNVLLAEAKVPYDIVLE MDEINDDFADTDTVLVIGANDTVNPAAQDD PKSPIAGMPVLEVWKAQNVIVFKRSMNTGY AGVQNPLFFKENTHMLFGDAKASVDAILKA L SEQIDNO:147 ACCase >gi|16128178|ref|NP_ MSLNFLDFEQPIAELEAKIDSLTAVSRQDEKL 414727.1| acetyl-CoA DINIDEEVHRLREKSVELTRKIFADLGAWQIA carboxylase, QLARHPQRPYTLDYVRLAFDEFDELAGDRA carboxytransferase, YADDKAIVGGIARLDGRPVMIIGHQKGRETK alphasubunit EKIRRNFGMPAPEGYRKALRLMQMAERFKM [Escherichiacolistr. PIITFIDTPGAYPGVGAEERGQSEAIARNLRE K-12substr.MG1655] MSRLGVPVVCTVIGEGGSGGALAIGVGDKV NMLQYSTYSVISPEGCASILWKSADKAPLAA EAMGIIAPRLKELKLIDSIIPEPLGGAHRNPEA MAASLKAQLLADLADLDVLSTEDLKNRRYQ RLMSYGYA SEQIDNO:148 ACCase >gi|16131143|ref|NP_ MDIRKIKKLIELVEESGISELEISEGEESVRISR 417721.1| acetylCoA AAPAASFPVMQQAYAAPMMQQPAQSNAAA carboxylase,BCCP PATVPSMEAPAAAEISGHIVRSPMVGTFYRT subunit[Escherichia PSPDAKAFIEVGQKVNVGDTLCIVEAMKMM colistr.K-12substr. NQIEADKSGTVKAILVESGQPVEFDEPLVVIE MG1655] SEQIDNO:149 ACCase >gi|16131144|ref|NP_ MLDKIVIANRGEIALRILRACKELGIKTVAVH 417722.1| acetyl-CoA SSADRDLKHVLLADETVCIGPAPSVKSYLNIP carboxylase,biotin AIISAAEITGAVAIHPGYGFLSENANFAEQVE carboxylasesubunit RSGFIFIGPKAETIRLMGDKVSAIAAMKKAG [Escherichiacolistr. VPCVPGSDGPLGDDMDKNRAIAKRIGYPVII K-12substr.MG1655] KASGGGGGRGMRVVRGDAELAQSISMTRAE AKAAFSNDMVYMEKYLENPRHVEIQVLAD GQGNAIYLAERDCSMQRRHQKVVEEAPAPG ITPELRRYIGERCAKACVDIGYRGAGTFEFLF ENGEFYFIEMNTRIQVEHPVTEMITGVDLIKE QLRIAAGQPLSIKQEEVHVRGHAVECRINAE DPNTFLPSPGKITRFHAPGGFGVRWESHIYA GYTVPPYYDSMIGKLICYGENRDVAIARMK NALQELIIDGIKTNVDLQIRIMNDENFQHGGT NIHYLEKKLGLQEK SEQIDNO:150 ACCase >gi|16130251|ref|NP_ MSWIERIKSNITPTRKASIPEGVWTKCDSCGQ 416819.1| acetyl-CoA VLYRAELERNLEVCPKCDHHMRMTARNRL carboxylase,beta HSLLDEGSLVELGSELEPKDVLKFRDSKKYK (carboxyltransferase) DRLASAQKETGEKDALVVMKGTLYGMPVV subunit[Escherichia AAAFEFAFMGGSMGSVVGARFVRAVEQALE colistr.K-12substr. DNCPLICFSASGGARMQEALMSLMQMAKTS MG1655] AALAKMQERGLPYISVLTDPTMGGVSASFA MLGDLNIAEPKALIGFAGPRVIEQTVREKLPP GFQRSEFLIEKGAIDMIVRRPEMRLKLASILA KLMNLPAPNPEAPREGVVVPPVPDQEPEA SEQIDNO:151 PDH >gi|16128107|ref|NP_ MSERFPNDVDPIETRDWLQAIESVIREEGVER 414656.1| pyruvate AQYLIDQLLAEARKGGVNVAAGTGISNYINT dehydrogenase, IPVEEQPEYPGNLELERRIRSAIRWNAIMTVL decarboxylase RASKKDLELGGHMASFQSSATIYDVCFNHFF componentE1, RARNEQDGGDLVYFQGHISPGVYARAFLEG thiamin-binding RLTQEQLDNFRQEVHGNGLSSYPHPKLMPEF [Escherichiacolistr. WQFPTVSMGLGPIGAIYQAKFLKYLEHRGLK K-12substr.MG1655] DTSKQTVYAFLGDGEMDEPESKGAITIATRE KLDNLVFVINCNLQRLDGPVTGNGKIINELE GIFEGAGWNVIKVMWGSRWDELLRKDTSG KLIQLMNETVDGDYQTFKSKDGAYVREHFF GKYPETAALVADWTDEQIWALNRGGHDPK KIYAAFKKAQETKGKATVILAHTIKGYGMG DAAEGKNIAHQVKKMNMDGVRHIRDRFNV PVSDADIEKLPYITFPEGSEEHTYLHAQRQKL HGYLPSRQPNFTEKLELPSLQDFGALLEEQS KEISTTIAFVRALNVMLKNKSIKDRLVPIIAD EARTFGMEGLFRQIGIYSPNGQQYTPQDREQ VAYYKEDEKGQILQEGINELGAGCSWLAAA TSYSTNNLPMIPFYIYYSMFGFQRIGDLCWA AGDQQARGFLIGGTSGRTTLNGEGLQHEDG HSHIQSLTIPNCISYDPAYAYEVAVIMHDGLE RMYGEKQENVYYYITTLNENYHMPAMPEG AEEGIRKGIYKLETIEGSKGKVQLLGSGSILR HVREAAEILAKDYGVGSDVYSVTSFTELARD GQDCERWNMLHPLETPRVPYIAQVMNDAPA VASTDYMKLFAEQVRTYVPADDYRVLGTD GFGRSDSRENLRHHFEVDASYVVVAALGEL AKRGEIDKKVVADAIAKFNIDADKVNPRLA SEQIDNO:152 PDH >gi|16128108|ref|NP_ MAIEIKVPDIGADEVEITEILVKVGDKVEAEQ 414657.1| pyruvate SLITVEGDKASMEVPSPQAGIVKEIKVSVGD dehydrogenase, KTQTGALIMIFDSADGAADAAPAQAEEKKE dihydrolipoyltransacet AAPAAAPAAAAAKDVNVPDIGSDEVEVTEIL ylasecomponentE2 VKVGDKVEAEQSLITVEGDKASMEVPAPFA [Escherichiacolistr. GTVKEIKVNVGDKVSTGSLIMVFEVAGEAG K-12substr.MG1655] AAAPAAKQEAAPAAAPAPAAGVKEVNVPDI GGDEVEVTEVMVKVGDKVAAEQSLITVEGD KASMEVPAPFAGVVKELKVNVGDKVKTGSL IMIFENEGAAPAAAPAKQEAAAPAPAAKAE APAAAPAAKAEGKSEFAENDAYVHATPLIR RLAREFGVNLAKVKGTGRKGRILREDVQAY VKEAIKRAEAAPAATGGGIPGMLPWPKVDF SKFGEIEEVELGRIQKISGANLSRNWVMIPHV THFDKTDITELEAFRKQQNEEAAKRKLDVKI TPVVFIMKAVAAALEQMPRFNSSLSEDGQRL TLKKYINIGVAVDTPNGLVVPVFKDVNKKGI IELSRELMTISKKARDGKLTAGEMQGGCFTIS SIGGLGTTHFAPIVNAPEVAILGVSKSAMEPV WNGKEFVPRLMLPISLSFDHRVIDGADGARF ITIINNTLSDIRRLVM SEQIDNO:153 PDH >gi|16128109|ref|NP_ MSTEIKTQVVVLGAGPAGYSAAFRCADLGL 414658.1| lipoamide ETVIVERYNTLGGVCLNVGCIPSKALLHVAK dehydrogenase,E3 VIEEAKALAEHGIVFGEPKTDIDKIRTWKEK componentispartof VINQLTGGLAGMAKGRKVKVVNGLGKFTG threeenzyme ANTLEVEGENGKTVINFDNAIIAAGSRPIQLP complexes FIPHEDPRIWDSTDALELKEVPERLLVMGGGI [Escherichiacolistr. IGLEMGTVYHALGSQIDVVEMFDQVIPAAD K-12substr.MG1655] KDIVKVFTKRISKKFNLMLETKVTAVEAKED GIYVTMEGKKAPAEPQRYDAVLVAIGRVPN GKNLDAGKAGVEVDDRGFIRVDKQLRTNVP HIFAIGDIVGQPMLAHKGVHEGHVAAEVIAG KKHYFDPKVIPSIAYTEPEVAWVGLTEKEAK EKGISYETATFPWAASGRAIASDCADGMTKL IFDKESHRVIGGAIVGTNGGELLGEIGLAIEM GCDAEDIALTIHAHPTLHESVGLAAEVFEGSI TDLPNPKAKKK SEQIDNO:154 coaA >gi|16131808|ref|NP_ MSIKEQTLMTPYLQFDRNQWAALRDSVPMT 418405.1| LSEDEIARLKGINEDLSLEEVAEIYLPLSRLLN pantothenatekinase FYISSNLRRQAVLEQFLGTNGQRIPYIISIAGS [Escherichiacolistr. VAVGKSTTARVLQALLSRWPEHRRVELITTD K-12substr.MG1655] GFLHPNQVLKERGLMKKKGFPESYDMHRLV KFVSDLKSGVPNVTAPVYSHLIYDVIPDGDK TVVQPDILILEGLNVLQSGMDYPHDPHHVFV SDFVDFSIYVDAPEDLLQTWYINRFLKFREG AFTDPDSYFHNYAKLTKEEAIKTAMTLWKEI NWLNLKQNILPTRERASLILTKSANHAVEEV RLRK SEQIDNO:155 panD >gi|16128124|ref|NP_ MIRTMLQGKLHRVKVTHADLHYEGSCAIDQ 414673.1| aspartate1- DFLDAAGILENEAIDIWNVTNGKRFSTYAIA decarboxylase AERGSRIISVNGAAAHCASVGDIVIIASFVTM [Escherichiacolistr. PDEEARTWRPNVAYFEGDNEMKRTAKAIPV K-12substr.MG1655] QVA SEQIDNO:156 aceA >gi|16131841|ref|NP_ MKTRTQQIEELQKEWTQPRWEGITRPYSAED 418439.1| isocitrate VVKLRGSVNPECTLAQLGAAKMWRLLHGE lyase[Escherichiacoli SKKGYINSLGALTGGQALQQAKAGIEAVYLS str.K-12substr. GWQVAADANLAASMYPDQSLYPANSVPAV MG1655] VERINNTFRRADQIQWSAGIEPGDPRYVDYF LPIVADAEAGFGGVLNAFELMKAMIEAGAA AVHFEDQLASVKKCGHMGGKVLVPTQEAIQ KLVAARLAADVTGVPTLLVARTDADAADLI TSDCDPYDSEFITGERTSEGFFRTHAGIEQAIS RGLAYAPYADLVWCETSTPDLELARRFAQAI HAKYPGKLLAYNCSPSFNWQKNLDDKTIAS FQQQLSDMGYKFQFITLAGIHSMWFNMFDL ANAYAQGEGMKHYVEKVQQPEFAAAKDGY TFVSHQQEVGTGYFDKVTTIIQGGTSSVTAL TGSTEESQF SEQIDNO:157 aceB >gi|16131840|ref|NP_ MTEQATTTDELAFTRPYGEQEKQILTAEAVE 418438.1| malate FLTELVTHFTPQRNKLLAARIQQQQDIDNGT synthaseA LPDFISETASIRDADWKIRGIPADLEDRRVEIT [Escherichiacolistr. GPVERKMVINALNANVKVFMADFEDSLAPD K-12substr.MG1655] WNKVIDGQINLRDAVNGTISYTNEAGKIYQL KPNPAVLICRVRGLHLPEKHVTWRGEAIPGS LFDFALYFFHNYQALLAKGSGPYFYLPKTQS WQEAAWWSEVFSYAEDRFNLPRGTIKATLLI ETLPAVFQMDEILHALRDHIVGLNCGRWDYI FSYIKTLKNYPDRVLPDRQAVTMDKPFLNA YSRLLIKTCHKRGAFAMGGMAAFIPSKDEEH NNQVLNKVKADKSLEANNGHDGTWIAHPG LADTAMAVFNDILGSRKNQLEVMREQDAPI TADQLLAPCDGERTEEGMRANIRVAVQYIE AWISGNGCVPIYGLMEDAATAEISRTSIWQW IHHQKTLSNGKPVTKALFRQMLGEEMKVIAS ELGEERFSQGRFDDAARLMEQITTSDELIDFL TLPGYRLLA SEQIDNO:158 aceK >gi|16131842|ref|NP_ MPRGLELLIAQTILQGFDAQYGRFLEVTSGA 418440.1| isocitrate QQRFEQADWHAVQQAMKNRIHLYDHHVGL dehydrogenase VVEQLRCITNGQSTDAAFLLRVKEHYTRLLP kinase/phosphatase DYPRFEIAESFFNSVYCRLFDHRSLTPERLFIF [Escherichiacolistr. SSQPERRFRTIPRPLAKDFHPDHGWESLLMR K-12substr.MG1655] VISDLPLRLRWQNKSRDIHYIIRHLTETLGTD NLAESHLQVANELFYRNKAAWLVGKLITPS GTLPFLLPIHQTDDGELFIDTCLTTTAEASIVF GFARSYFMVYAPLPAALVEWLREILPGKTTA ELYMAIGCQKHAKTESYREYLVYLQGCNEQ FIEAPGIRGMVMLVFTLPGFDRVFKVIKDRF APQKEMSAAHVRACYQLVKEHDRVGRMAD TQEFENFVLEKRHISPALMELLLQEAAEKITD LGEQIVIRHLYIERRMVPLNIWLEQVEGQQL RDAIEEYGNAIRQLAAANIFPGDMLFKNFGV TRHGRVVFYDYDEICYMTEVNFRDIPPPRYP EDELASEPWYSVSPGDVFPEEFRHWLCADPR IGPLFEEMHADLFRADYWRALQNRIREGHV EDVYAYRRRQRFSVRYGEMLF SEQIDNO:159 GAPDH >gi|16129733|ref|NP_ MTIKVGINGFGRIGRIVFRAAQKRSDIEIVAIN 416293.1| DLLDADYMAYMLKYDSTHGRFDGTVEVKD glyceraldehyde-3- GHLIVNGKKIRVTAERDPANLKWDEVGVDV phosphate VAEATGLFLTDETARKHITAGAKKVVMTGP dehydrogenaseA SKDNTPMFVKGANFDKYAGQDIVSNASCTT [Escherichiacolistr. NCLAPLAKVINDNFGIIEGLMTTVHATTATQ K-12substr.MG1655] KTVDGPSHKDWRGGRGASQNIIPSSTGAAK AVGKVLPELNGKLTGMAFRVPTPNVSVVDL TVRLEKAATYEQIKAAVKAAAEGEMKGVL GYTEDDVVSTDFNGEVCTSVFDAKAGIALN DNFVKLVSWYDNETGYSNKVLDLIAHISK SEQIDNO:160 pyk >gi|16129807|ref|NP_ MSRRLRRTKIVTTLGPATDRDNNLEKVIAAG 416368.1| pyruvate ANVVRMNFSHGSPEDHKMRADKVREIAAKL kinaseII[Escherichia GRHVAILGDLQGPKIRVSTFKEGKVFLNIGD colistr.K-12substr. KFLLDANLGKGEGDKEKVGIDYKGLPADVV MG1655] PGDILLLDDGRVQLKVLEVQGMKVFTEVTV GGPLSNNKGINKLGGGLSAEALTEKDKADIK TAALIGVDYLAVSFPRCGEDLNYARRLARD AGCDAKIVAKVERAEAVCSQDAMDDIILAS DVVMVARGDLGVEIGDPELVGIQKALIRRAR QLNRAVITATQMMESMITNPMPTRAEVMDV ANAVLDGTDAVMLSAETAAGQYPSETVAA MARVCLGAEKIPSINVSKHRLDVQFDNVEEA IAMSAMYAANHLKGVTAIITMTESGRTALM TSRISSGLPIFAMSRHERTLNLTALYRGVTPV HIDSANDGVAAASEAVNLLRDKGYLMSGD LVIVTQGDVMSTVGSTNTTRILTVE SEQIDNO:161 pyk >gi|16129632|ref|NP_ MKKTKIVCTIGPKTESEEMLAKMLDAGMNV 416191.1| pyruvate MRLNFSHGDYAEHGQRIQNLRNVMSKTGKT kinaseI[Escherichia AAILLDTKGPEIRTMKLEGGNDVSLKAGQTF colistr.K-12substr. TFTTDKSVIGNSEMVAVTYEGFTTDLSVGNT MG1655] VLVDDGLIGMEVTAIEGNKVICKVLNNGDL GENKGVNLPGVSIALPALAEKDKQDLIFGCE QGVDFVAASFIRKRSDVIEIREHLKAHGGENI HIISKIENQEGLNNFDEILEASDGIMVARGDL GVEIPVEEVIFAQKMMIEKCIRARKVVITATQ MLDSMIKNPRPTRAEAGDVANAILDGTDAV MLSGESAKGKYPLEAVSIMATICERTDRVM NSRLEFNNDNRKLRITEAVCRGAVETAEKLD APLIVVATQGGKSARAVRKYFPDATILALTT NEKTAHQLVLSKGVVPQLVKEITSTDDFYRL GKELALQSGLAHKGDVVVMVSGALVPSGTT NTASVHVL SEQIDNO:162 gltA >gi|16128695|ref|NP_ MADTKAKLTLNGDTAVELDVLKGTLGQDVI 415248.1| citrate DIRTLGSKGVFTFDPGFTSTASCESKITFIDGD synthase[Escherichia EGILLHRGFPIDQLATDSNYLEVCYILLNGEK colistr.K-12substr. PTQEQYDEFKTTVTRHTMIHEQITRLFHAFR MG1655] RDSHPMAVMCGITGALAAFYHDSLDVNNPR HREIAAFRLLSKMPTMAAMCYKYSIGQPFV YPRNDLSYAGNFLNMMFSTPCEPYEVNPILE RAMDRILILHADHEQNASTSTVRTAGSSGAN PFACIAAGIASLWGPAHGGANEAALKMLEEI SSVKHIPEFVRRAKDKNDSFRLMGFGHRVY KNYDPRATVMRETCHEVLKELGTKDDLLEV AMELENIALNDPYFIEKKLYPNVDFYSGIILK AMGIPSSMFTVIFAMARTVGWIAHWSEMHS DGMKIARPRQLYTGYEKRDFKSDIKR SEQIDNO:163 bicA >gi|109820126|gb| MQITNKIHFRNIRGDIFGGLTAAVIALPMALA ABG46427.1| BicA FGVASGAGAEAGLWGAVLVGFFAALFGGTP [Synechococcussp. TLISEPTGPMTVVMTAVIAHFTASAATPEEGL PCC70021 AIAFTVVMMAGVFQIIFGSLKLGKYVTMMP YTVISGFMSGIGIILVILQLAPFLGQASPGGGV IGTLQNLPTLLSNIQPGETALALGTVAIIWFM PEKFKKVIPPQLVALVLGTVIAFIVFPPEVSD LRRIGEIRAGFPELVRPSFSPVEFQRMILDAA VLGMLGCIDALLTSVVADSLTRTEHNSNKEL IGQGLGNLFSGLFGGIAGAGATMGTVVNIQS GGRTALSGLVRAFVLLVVILGAASLTATIPLA VLAGIAFKVGVDIIDWSFLKRAHEISPKGALI MYGVILLTVLVDLIVAVGVGVFVANVLTIER MSNLQSEKVQTVSDADDNIRLTTTEKRWLD EGQGRVLLFQLSGPMIFGVAKAIAREHNAM GDCDALVFDIGEVPHMGVTASLALENAIEEA LDKERQVYIVGAAGQTRRRLEKLKLFKRVPP DKCLMSREEALKNAVLGIYPHLADGVTAPSS EMG SEQIDNO:164 GOGAT >gi|308209621|ref|NP MLYDKSLERDNCGFGLIAHIEGEPSHKVVRTAIHALA 417679.2| glutamate RMQHRGAILADGKTGDGCGLLLQKPDRFFRIVAQER synthase,largesubunit GWRLAKNYAVGMLFLNKDPELAAAARRIVEEELQRE [Escherichiacolistr. TLSIVGWRDVPTNEGVLGEIALSSLPRIEQIFVNAPAG K-12substr.MG1655] WRPRDMERRLFIARRRIEKRLEADKDFYVCSLSNLVNI YKGLCMPTDLPRFYLDLADLRLESAICLFHQRFSTNTV PRWPLAQPFRYLAHNGEINTITGNRQWARARTYKFQT PLIPDLHDAAPFVNETGSDSSSMDNMLELLLAGGMDII RAMRLLVPPAWQNNPDMDPELRAFFDFNSMHMEPW DGPAGIVMSDGRFAACNLDRNGLRPARYVITKDKLIT CASEVGIWDYQPDEVVEKGRVGPGELMVIDTRSGRIL HSAETDDDLKSRHPYKEWMEKNVRRLVPFEDLPDEE VGSRELDDDTLASYQKQFNYSAEELDSVIRVLGENGQ EAVGSMGDDTPFAVLSSQPRIIYDYFRQQFAQVTNPPI DPLREAHVMSLATSIGREMNVFCEAEGQAHRLSFKSPI LLYSDFKQLTTMKEEHYRADTLDITFDVTKTTLEATV KELCDKAEKMVRSGTVLLVLSDRNIAKDRLPVPAPM AVGAIQTRLVDQSLRCDANIIVETASARDPHHFAVLL GFGATAIYPYLAYETLGRLVDTHAIAKDYRTVMLNY RNGINKGLYKIMSKMGISTIASYRCSKLFEAVGLHDD VVGLCFQGAVSRIGGASFEDFQQDLLNLSKRAWLAR KPISQGGLLKYVHGGEYHAYNPDVVRTLQQAVQSGE YSDYQEYAKLVNERPATTLRDLLAITPGENAVNIADV EPASELFKRFDTAAMSIGALSPEAHEALAEAMNSIGG NSNSGEGGEDPARYGTNKVSRIKQVASGRFGVTPAYL VNADVIQIKVAQGAKPGEGGQLPGDKVTPYIAKLRYS VPGVTLISPPPHHDIYSIEDLAQLIFDLKQVNPKAMISV KLVSEPGVGTIATGVAKAYADLITIAGYDGGTGASPLS SVKYAGCPWELGLVETQQALVANGLRHKIRLQVDGG LKTGVDIIKAAILGAESFGFGTGPMVALGCKYLRICHL NNCATGVATQDDKLRKNHYHGLPFKVTNYFEFIARE TRELMAQLGVTRLVDLIGRTDLLKELDGFTAKQQKL ALSKLLETAEPHPGKALYCTENNPPFDNGLLNAQLLQ QAKPFVDERQSKTFWFDIRNTDRSVGASLSGYIAQTH GDQGLAADPIKAYFNGTAGQSFGVWNAGGVELYLTG DANDYVGKGMAGGLIAIRPPVGSAFRSHEASIIGNTCL YGATGGRLYAAGRAGERFGVRNSGAITVVEGIGDNG CEYMTGGIVCILGKTGVNFGAGMTGGFAYVLDESGD FRKRVNPELVEVLSVDALAIHEEHLRGLITEHVQHTGS QRGEEILANWSTFATKFALVKPKSSDVKALLGHRSRS AAELRVQAQ SEQIDNO:165 GOGAT >gi|16131103|ref|NP_ MSQNVYQFIDLQRVDPPKKPLKIRKIEFVEIY 417680.1| glutamate EPFSEGQAKAQADRCLSCGNPYCEWKCPVH synthase,4Fe-45 NYIPNWLKLANEGRIFEAAELSHQTNTLPEV protein,smallsubunit CGRVCPQDRLCEGSCTLNDEFGAVTIGNIER [Escherichiacolistr. YINDKAFEMGWRPDMSGVKQTGKKVAIIGA K-12substr.MG1655] GPAGLACADVLTRNGVKAVVFDRHPEIGGL LTFGIPAFKLEKEVMTRRREIFTGMGIEFKLN TEVGRDVQLDDLLSDYDAVFLGVGTYQSMR GGLENEDADGVYAALPFLIANTKQLMGFGE TRDEPFVSMEGKRVVVLGGGDTAMDCVRTS VRQGAKHVTCAYRRDEENMPGSRREVKNA REEGVEFKFNVQPLGIEVNGNGKVSGVKMV RTEMGEPDAKGRRRAEIVAGSEHIVPADAVI MAFGFRPHNMEWLAKHSVELDSQGRIIAPE GSDNAFQTSNPKIFAGGDIVRGSDLVVTAIAE GRKAADGIMNWLEV SEQIDNO:166 gdh >gi|16129715|ref|NP_ MDQTYSLESFLNHVQKRDPNQTEFAQAVRE 416275.1| glutamate VMTTLWPFLEQNPKYRQMSLLERLVEPERVI dehydrogenase, QIRVVWVDDRNQIQVNRAWRVQFSSAIGPY NADP-specific KGGMRFHPSVNLSILKFLGFEQTFKNALTTL [Escherichiacolistr. PMGGGKGGSDFDPKGKSEGEVMRFCQALM K-12substr.MG1655] TELYRHLGADTDVPAGDIGVGGREVGFMAG MMKKLSNNTACVFTGKGLSFGGSLIRPEATG YGLVYFTEAMLKRHGMGFEGMRVSVSGSG NVAQYAIEKAMEFGARVITASDSSGTVVDES GFTKEKLARLIEIKASRDGRVADYAKEFGLV YLEGQQPWSLPVDIALPCATQNELDVDAAH QLIANGVKAVAEGANMPTTIEATELFQQAG VLFAPGKAANAGGVATSGLEMAQNAARLG WKAEKVDARLHHIMLDIHHACVEHGGEGE QTNYVQGANIAGFVKVADAMLAQGVI SEQIDNO:167 can >gi|16128119|ref|NP_ MKDIDTLISNNALWSKMLVEEDPGFFEKLAQ 414668.1| carbonic AQKPRFLWIGCSDSRVPAERLTGLEPGELFV anhydrase HRNVANLVIHTDLNCLSVVQYAVDVLEVEH [Escherichiacolistr. IIICGHYGCGGVQAAVENPELGLINNWLLHIR K-12substr.MG1655] DIWFKHSSLLGEMPQERRLDTLCELNVMEQ VYNLGHSTIMQSAWKRGQKVTIHGWAYGIH DGLLRDLDVTATNRETLEQRYRHGISNLKLK HANHK SEQIDNO:168 cynT >gi|16128324|ref|NP_ MKEIIDGFLKFQREAFPKREALFKQLATQQSP 414873.1| carbonic RTLFISCSDSRLVPELVTQREPGDLFVIRNAG anhydrase NIVPSYGPEPGGVSASVEYAVAALRVSDIVIC [Escherichiacolistr. GHSNCGAMTAIASCQCMDHMPAVSHWLRY K-12substr.MG1655] ADSARVVNEARPHSDLPSKAAAMVRENVIA QLANLQTHPSVRLALEEGRIALHGWVYDIES GSIAAFDGATRQFVPLAANPRVCAIPLRQPT AA SEQIDNO:169 cynS >gi|16128325|ref|NP_ MIQSQINRNIRLDLADAILLSKAKKDLSFAEI 414874.1| cyanate ADGTGLAEAFVTAALLGQQALPADAARLVG aminohydrolase AKLDLDEDSILLLQMIPLRGCIDDRIPTDPTM [Escherichiacolistr. YRFYEMLQVYGTTLKALVHEKFGDGIISAIN K-12substr.MG1655] FKLDVKKVADPEGGERAVITLDGKYLPTKPF SEQIDNO:170 yibD >gi|16131486|ref|NP_ MMNSTNKLSVIIPLYNAGDDFRTCMESLITQ 418072.1| putative TWTALEIIIINDGSTDNSVEIAKYYAENYPHV glycosyltransferase RLLHQANAGASVARNRGIEVATGKYVAFVD [Escherichiacolistr. ADDEVYPTMYETLMTMALEDDLDVAQCNA K-12substr.MG1655] DWCFRETGETWQSIPTDRLRSTGVLTGPDW LRMGLSSRRWTHVVWMGVYRRDVIVKNNI KFIAGLHHQDIVWTTEFMFNALRARYTEQSL YKYYLHNTSVSRLHRQGNKNLNYQRHYIKI TRLLEKLNRNYADKIMIYPEFHQQITYEALR VCHAVRKEPDILTRQRMIAEIFTSGMYKRLIT NVRSVKVGYQALLWSFRLWQWRDKTRSHH RITRSAFNLR SEQIDNO:171 pstS >gi|16131597|ref|NP_ MCGIVGAIAQRDVAEILLEGLRRLEYRGYDS 418185.1| L- AGLAVVDAEGHMTRLRRLGKVQMLAQAAE glutamine:D-fructose- EHPLHGGTGIAHTRWATHGEPSEVNAHPHV 6-phosphate SEHIVVVHNGIIENHEPLREELKARGYTFVSE aminotransferase TDTEVIAHLVNWELKQGGTLREAVLRAIPQL [Escherichiacolistr. RGAYGTVIMDSRHPDTLLAARSGSPLVIGLG K-12substr.MG1655] MGENFIASDQLALLPVTRRFIFLEEGDIAEITR RSVNIFDKTGAEVKRQDIESNLQYDAGDKGI YRHYMQKEIYEQPNAIKNTLTGRISHGQVDL SELGPNADELLSKVEHIQILACGTSYNSGMV SRYWFESLAGIPCDVEIASEFRYRKSAVRRNS LMITLSQSGETADTLAGLRLSKELGYLGSLAI CNVPGSSLVRESDLALMTNAGTEIGVASTKA FTTQLTVLLMLVAKLSRLKGLDASIEHDIVH GLQALPSRIEQMLSQDKRIEALAEDFSDKHH ALFLGRGDQYPIALEGALKLKEISYIHAEAY AAGELKHGPLALIDADMPVIVVAPNNELLEK LKSNIEEVRARGGQLYVFADQDAGFVSSDN MHIIEMPHVEEVIAPIFYTVPLQLLAYHVALI KGTDVDQPRNLAKSVTVE SEQIDNO:172 PDH lipoamide MSTEIKTQVVVLGAGPAGYSAAFRCADLGL dehydrogenase, ETVIVERYNTLGGVCLNVGCIPSKALLHVAK NADH-inhibition VIEEAKALAEHGIVFGEPKTDIDKIRTWKEK resistant VINQLTGGLAGMAKGRKVKVVNGLGKFTG ANTLEVEGENGKTVINFDNAIIAAGSRPIQLP FIPHEDPRIWDSTDALELKEVPERLLVMGGGI IGLEMGTVYHALGSQIDVVEMFDQVIPAAD KDIVKVFTKRISKKFNLMLETKVTAVEAKED GIYVTMEGKKAPAEPQRYDAVLVAIGRVPN GKNLDAGKAGVEVDDRGFIRVDKQLRTNVP HIFAIGDIVGQPMLAHKGVHEGHVAAEVIAG KKHYFDPKVIPSIAYTEPEVAWVGLTEKEAK EKGISYETATFPWAASGRAIASDCADGMTKL IFDKESHRVIGGAIVGTNGGELLGEIGLAIEM GCDAEDIALTIHAHPTLHESVGLAAEVFEGSI TDLPNPKAKKK SEQIDNO:173 coaA pantothenatekinase, MSIKEQTLMTPYLQFDRNQWAALRDSVPMT feedback-resistant LSEDEIARLKGINEDLSLEEVAEIYLPLSRLLN FYISSNLRRQAVLEQFLGTNGQRIPYIISIAGS VAVGKSTTAAVLQALLSRWPEHRRVELITTD GFLHPNQVLKERGLMKKKGFPESYDMHRLV KFVSDLKSGVPNVTAPVYSHLIYDVIPDGDK TVVQPDILILEGLNVLQSGMDYPHDPHHVFV SDFVDFSIYVDAPEDLLQTWYINRFLKFREG AFTDPDSYFHNYAKLTKEEAIKTAMTLWKEI NWLNLKQNILPTRERASLILTKSANHAVEEV RLRK Sugartransportandutilization SEQIDNO:174 cscA >gi|608708|emb|CAA MTQSRLHAAQNALAKLHERRGNTFYPHFHL 57219.1| sucrose APPAGWMNDPNGLIWFNDRYHAFYQHHPM hydrolase[Escherichia SEHWGPMHWGHATSDDMIHWQHEPIALAP coli] GDENDKDGCFSGSAVDDNGVLSLIYTGHVW LDGAGNDDAIREVQCLATSRDGIHFEKQGVI LTPPEGIMHFRDPKVWREADTWWMVVGAK DPGNTGQILLYRGSSLREWTFDRVLAHADA GESYMWECPDFFSLGDQHYLMFSPQGMNAE GYSYRNRFQSGVIPGMWSPGRLFAQSGHFTE LDNGHDFYAPQSFVAKDGRRIVIGWMDMW ESPMPSKREGWAGCMTLARELSESNGKLLQ RPVHEAESLRQQHQSISPRTISNKYVLQENA QAVEIQLQWALKNSDAEHYGLQLGAGMRL YIDNQSERLVLWRYYPHENLDGYRSIPLPQG DMLALRIFIDTSSVEVFINDGEAVMSSRIYPQ PEERELSLYASHGVAVLQHGALWQLG SEQIDNO:175 cscB >gi|608706|emb|CAA MALNIPFRNAYYRFASSYSFLFFISWSLWWS 57217.1| sucrose LYAIWLKGHLGLTGTELGTLYSVNQFTSILF permease[Escherichia MMFYGIVQDKLGLKKPLIWCMSFILVLTGPF coli] MIYVYEPLLQSNFSVGLILGALFFGLGYLAG CGLLDSFTEKMARNFHFEYGTARAWGSFGY AIGAFFAGIFFSISPHINFWLVSLFGAVFMMIN MRFKDKDHQCIAADAGGVKKEDFIAVFKDR NFWVFVIFIVGTWSFYNIFDQQLFPVFYAGLF ESHDVGTRLYGYLNSFQVVLEALCMAIIPFF VNRVGPKNALLIGVVIMALRILSCALFVNPW IISLVKLLHAIEVPLCVISVFKYSVANFDKRLS STIFLIGFQIASSLGIVLLSTPTGILFDHAGYQT VFFAISGIVCLMLLFGIFFLSKKREQIVMETPV PSAI SEQIDNO:176 cscK >gi|20451632|emb|CA MSAKVWVLGDAVVDLLPESDGRLLPCPGGA A57218.2| D- PANVAVGIARLGGTSGFIGRVGDDPFGALM fructokinase QRTLLTEGVDITYLKQDEWHRTSTVLVDLN [Escherichiacoli] DQGERSFTFMVRPSADLFLETTDLPCWRHGE WLHLCSIALSAEPSRTSAFTAMTAIRHAGGF VSFDPNIREDLWQDEHLLRLCLRQALQLAD VVKLSEEEWRLISGKTQNDRDICALAKEYEI AMLLVTKGAEGVVVCYRGQVHHFAGMSVN CVDSTGAGDAFVAGLLTGLSSTGLSTDERE MRRIIDLAQRCGALAVTAKGAMTALPCRQE LESEK SEQIDNO:177 galP >gi|16130844|ref|NP_ MPDAKKQGRSNKAMTFFVCFLAALAGLLFG 417418.1| D-galactose LDIGVIAGALPFIADEFQITSHTQEWVVSSMM transporter FGAAVGAVGSGWLSFKLGRKKSLMIGAILF [Escherichiacolistr. VAGSLFSAAAPNVEVLILSRVLLGLAVGVAS K-12substr.MG1655] YTAPLYLSEIAPEKIRGSMISMYQLMITIGILG AYLSDTAFSYTGAWRWMLGVIIIPAILLLIGV FFLPDSPRWFAAKRRFVDAERVLLRLRDTSA EAKRELDEIRESLQVKQSGWALFKENSNFRR AVFLGVLLQVMQQFTGMNVIMYYAPKIFEL AGYTNTTEQMWGTVIVGLTNVLATFIAIGLV DRWGRKPTLTLGFLVMAAGMGVLGTMMHI GIHSPSAQYFAIAMLLMFIVGFAMSAGPLIW VLCSEIQPLKGRDFGITCSTATNWIANMIVGA TFLTMLNTLGNANTFWVYAALNVLFILLTL WLVPETKHVSLEHIERNLMKGRKLREIGAH D SEQIDNO:178 galK >gi|16128725|ref|NP_ MSLKEKTQSLFANAFGYPATHTIQAPGRVNL 415278.1| IGEHTDYNDGFVLPCAIDYQTVISCAPRDDR galactokinase KVRVMAADYENQLDEFSLDAPIVAHENYQ [Escherichiacolistr. WANYVRGVVKHLQLRNNSFGGVDMVISGN K-12substr.MG1655] VPQGAGLSSSASLEVAVGTVLQQLYHLPLD GAQIALNGQEAENQFVGCNCGIMDQLISALG KKDHALLIDCRSLGTKAVSMPKGVAVVIINS NFKRTLVGSEYNTRREQCETGARFFQQPALR DVTIEEFNAVAHELDPIVAKRVRHILTENART VEAASALEQGDLKRMGELMAESHASMRDD FEITVPQIDTLVEIVKAVIGDKGGVRMTGGG FGGCIVALIPEELVPAVQQAVAEQYEAKTGI KETFYVCKPSQGAGQC SEQIDNO:179 cscB sucrosepermease MALNIPFRNAYYRFASSYSFLFFISWSLWWS mutantwithincreased LYAIWLKGHLGLTGTELGTLYSVNQFTSILF activity MMFYGIVQDKLGLKKPLIWCMSFILVLTGPF MIYVYEPLLQSNFSVGLILGALFFGLGYLAG CGLLDSFTEKMARNFHFEYGTARAWGSFGY AIGAFFAGIFFSISPHINFWLVSLFGAVFMMIN MRFKDKDHQCIAADAGGVKKEDFIAVFKDR NFWVFVIFIVGTWSFYDIFDQQLFPVFYAGLF ESHDVGTRLYGYLNSFQVVLEALCMAIIPFF VNRVGPKNALLIGVVIMALRILSCALFVNPW VISLVKLLHAIEVPLCVISVFKYSVANFDKRL SSTIFLIGFQIASSLGIVLLSTPTGILFDHAGYQ TVFFAISGIVCLMLLFGIFFLSKKREQIVMETP VPSAI Hostmodificationsforfattyacidproduction SEQIDNO:180 fadE >gi|90111100|ref|NP_ MMILSILATVVLLGALFYHRVSLFISSLILLA 414756.2| acyl WTAALGVAGLWSAWVLVPLAIILVPFNFAP coenzymeA MRKSMISAPVFRGFRKVMPPMSRTEKEAIDA dehydrogenase GTTWWEGDLFQGKPDWKKLHNYPQPRLTA [Escherichiacolistr. EEQAFLDGPVEEACRMANDFQITHELADLPP K-12substr.MG1655] ELWAYLKEHRFFAMIIKKEYGGLEFSAYAQS RVLQKLSGVSGILAITVGVPNSLGPGELLQH YGTDEQKDHYLPRLARGQEIPCFALTSPEAG SDAGAIPDTGIVCMGEWQGQQVLGMRLTW NKRYITLAPIATVLGLAFKLSDPEKLLGGAE DLGITCALIPTTTPGVEIGRRHFPLNVPFQNGP TRGKDVFVPIDYIIGGPKMAGQGWRMLVEC LSVGRGITLPSNSTGGVKSVALATGAYAHIR RQFKISIGKMEGIEEPLARIAGNAYVMDAAA SLITYGIMLGEKPAVLSAIVKYHCTHRGQQSI IDAMDITGGKGIMLGQSNFLARAYQGAPIAI TVEGANILTRSMMIFGQGAIRCHPYVLEEME AAKNNDVNAFDKLLFKHIGHVGSNKVRSFW LGLTRGLTSSTPTGDATKRYYQHLNRLSANL ALLSDVSMAVLGGSLKRRERISARLGDILSQ LYLASAVLKRYDDEGRNEADLPLVHWGVQ DALYQAEQAMDDLLQNFPNRVVAGLLNVVI FPTGRHYLAPSDKLDHKVAKILQVPNATRSR IGRGQYLTPSEHNPVGLLEEALVDVIAADPIH QRICKELGKNLPFTRLDELAHNALVKGLIDK DEAAILVKAEESRLRSINVDDFDPEELATKPV KLPEKVRKVEAA SEQIDNO:181 fadD >gi|16129759|ref|NP_ MKKVWLNRYPADVPTEINPDRYQSLVDMFE 416319.1| acyl-CoA QSVARYADQPAFVNMGEVMTFRKLEERSRA synthetase(long- FAAYLQQGLGLKKGDRVALMMPNLLQYPV chain-fatty-acid-CoA ALFGILRAGMIVVNVNPLYTPRELEHQLNDS ligase)[Escherichia GASAIVIVSNFAHTLEKVVDKTAVQHVILTR colistr.K-12substr. MGDQLSTAKGTVVNFVVKYIKRLVPKYHLP MG1655] DAISFRSALHNGYRMQYVKPELVPEDLAFLQ YTGGTTGVAKGAMLTHRNMLANLEQVNAT YGPLLHPGKELVVTALPLYHIFALTINCLLFIE LGGQNLLITNPRDIPGLVKELAKYPFTAITGV NTLFNALLNNKEFQQLDFSSLHLSAGGGMP VQQVVAERWVKLTGQYLLEGYGLTECAPL VSVNPYDIDYHSGSIGLPVPSTEAKLVDDDD NEVPPGQPGELCVKGPQVMLGYWQRPDAT DEIIKNGWLHTGDIAVMDEEGFLRIVDRKKD MILVSGFNVYPNEIEDVVMQHPGVQEVAAV GYPSGSSGEAVKIFVVKKDPSLTEESLVTFCR RQLTGYKVPKLVEFRDELPKSNVGKILRREL RDEARGKVDNKA SEQIDNO:182 fadA >gi|49176430|ref|YP_ MEQVVIVDAIRTPMGRSKGGAFRNVRAEDL 026272.1| 3-ketoacyl- SAHLMRSLLARNPALEAAALDDIYWGCVQQ CoAthiolase(thiolase TLEQGFNIARNAALLAEVPHSVPAVTVNRLC I)[Escherichiacolistr. GSSMQALHDAARMIMTGDAQACLVGGVEH K-12substr.MG1655] MGHVPMSHGVDFHPGLSRNVAKAAGMMG LTAEMLARMHGISREMQDAFAARSHARAW AATQSAAFKNEIIPTGGHDADGVLKQFNYDE VIRPETTVEALATLRPAFDPVNGMVTAGTSS ALSDGAAAMLVMSESRAHELGLKPRARVRS MAVVGCDPSIMGYGPVPASKLALKKAGLSA SDIGVFEMNEAFAAQILPCIKDLGLIEQIDEKI NLNGGAIALGHPLGCSGARISTTLLNLMERK DVQFGLATMCIGLGQGIATVFERV SEQIDNO:183 fadB >gi|16131692|ref|NP_ MLYKGDTLYLDWLEDGIAELVFDAPGSVNK 418288.1| fused3- LDTATVASLGEAIGVLEQQSDLKGLLLRSNK hydroxybutyryl-CoA AAFIVGADITEFLSLFLVPEEQLSQWLHFANS epimerase/delta(3)-cis- VFNRLEDLPVPTIAAVNGYALGGGCECVLAT delta(2)-trans-enoyl- DYRLATPDLRIGLPETKLGIMPGFGGSVRMP CoAisomerase/enoyl- RMLGADSALEIIAAGKDVGADQALKIGLVD CoAhydratase/3- GVVKAEKLVEGAKAVLRQAINGDLDWKAK hydroxyacyl-CoA RQPKLEPLKLSKIEATMSFTIAKGMVAQTAG dehydrogenase KHYPAPITAVKTIEAAARFGREEALNLENKS [Escherichiacolistr. FVPLAHTNEARALVGIFLNDQYVKGKAKKL K-12substr.MG1655] TKDVETPKQAAVLGAGIMGGGIAYQSAWK GVPVVMKDINDKSLTLGMTEAAKLLNKQLE RGKIDGLKLAGVISTIHPTLDYAGFDRVDIVV EAVVENPKVKKAVLAETEQKVRQDTVLASN TSTIPISELANALERPENFCGMHFFNPVHRMP LVEIIRGEKSSDETIAKVVAWASKMGKTPIV VNDCPGFFVNRVLFPYFAGFSQLLRDGADFR KIDKVMEKQFGWPMGPAYLLDVVGIDTAH HAQAVMAAGFPQRMQKDYRDAIDALFDAN RFGQKNGLGFWRYKEDSKGKPKKEEDAAV EDLLAEVSQPKRDFSEEEIIARMMIPMVNEV VRCLEEGIIATPAEADMALVYGLGFPPFHGG AFRWLDTLGSAKYLDMAQQYQHLGPLYEV PEGLRNKARHNEPYYPPVEPARPVGDLKTA SEQIDNO:184 fadI >gi|16130275|ref|NP_ MGQVLPLVTRQGDRIAIVSGLRTPFARQATA 416844.1| beta- FHGIPAVDLGKMVVGELLARSEIPAEVIEQL ketoacyl-CoAthiolase, VFGQVVQMPEAPNIAREIVLGTGMNVHTDA anaerobic,subunit YSVSRACATSFQAVANVAESLMAGTIRAGIA [Escherichiacolistr. GGADSSSVLPIGVSKKLARVLVDVNKARTM K-12substr.MG1655] SQRLKLFSRLRLRDLMPVPPAVAEYSTGLRM GDTAEQMAKTYGITREQQDALAHRSHQRAA QAWSDGKLKEEVMTAFIPPYKQPLVEDNNIR GNSSLADYAKLRPAFDRKHGTVTAANSTPL TDGAAAVILMTESRAKELGLVPLGYLRSYAF TAIDVWQDMLLGPAWSTPLALERAGLTMSD LTLIDMHEAFAAQTLANIQLLGSERFAREAL GRAHATGEVDDSKFNVLGGSIAYGHPFAAT GARMITQTLHELRRRGGGFGLVTACAAGGL GAAMVLEAE SEQIDNO:185 fadJ >gi|16130274|ref|NP_ MEMTSAFTLNVRLDNIAVITIDVPGEKMNTL 416843.1| fusedenoyl- KAEFASQVRAIIKQLRENKELRGVVFVSAKP CoAhydrataseand DNFIAGADINMIGNCKTAQEAEALARQGQQ epimeraseand LMAEIHALPIQVIAAIHGACLGGGLELALAC isomerase/3- HGRVCTDDPKTVLGLPEVQLGLLPGSGGTQ hydroxyacyl-CoA RLPRLIGVSTALEMILTGKQLRAKQALKLGL dehydrogenase VDDVVPHSILLEAAVELAKKERPSSRPLPVR [Escherichiacolistr. ERILAGPLGRALLFKMVGKKTEHKTQGNYP K-12substr.MG1655] ATERILEVVETGLAQGTSSGYDAEARAFGEL AMTPQSQALRSIFFASTDVKKDPGSDAPPAP LNSVGILGGGLMGGGIAYVTACKAGIPVRIK DINPQGINHALKYSWDQLEGKVRRRHLKAS ERDKQLALISGTTDYRGFAHRDLIIEAVFENL ELKQQMVAEVEQNCAAHTIFASNTSSLPIGDI AAHATRPEQVIGLHFFSPVEKMPLVEIIPHAG TSAQTIATTVKLAKKQGKTPIVVRDKAGFYV NRILAPYINEAIRMLTQGERVEHIDAALVKFG FPVGPIQLLDEVGIDTGTKIIPVLEAAYGERFS APANVVSSILNDDRKGRKNGRGFYLYGQKG RKSKKQVDPAIYPLIGTQGQGRISAPQVAER CVMLMLNEAVRCVDEQVIRSVRDGDIGAVF GIGFPPFLGGPFRYIDSLGAGEVVAIMQRLAT QYGSRFTPCERLVEMGARGESFWKTTATDL Q SEQIDNO:186 ydiO >gi|90111318|ref|NP_ MDFSLTEEQELLLASIRELITTNFPEEYFRTCD 416210.4| putative QNGTYPREFMRALADNGISMLGVPEEFGGIP acyl-CoA ADYVTQMLALMEVSKCGAPAFLITNGQCIH dehydrogenase SMRRFGSAEQLRKTAESTLETGDPAYALALT [Escherichiacolistr. EPGAGSDNNSATTTYTRKNGKVYINGQKTFI K-12substr.MG1655] TGAKEYPYMLVLARDPQPKDPKKAFTLWW VDSSKPGIKINPLHKIGWHMLSTCEVYLDNV EVEESDMVGEEGMGFLNVMYNFEMERLINA ARSTGFAECAFEDAARYANQRIAFGKPIGHN QMIQEKLALMAIKIDNMRNMVLKVAWQAD QHQSLRTSAALAKLYCARTAMEVIDDAIQIM GGLGYTDEARVSRFWRDVRCERIGGGTDEI MIYVAGRQILKDYQNK SEQIDNO:187 paaJ >gi|16129358|ref|NP_ MREAFICDGIRTPIGRYGGALSSVRADDLAAI 415915.1| 3- PLRELLVRNPRLDAECIDDVILGCANQAGED oxoadipyl-CoA/3-oxo- NRNVARMATLLAGLPQSVSGTTINRLCGSGL 5,6-dehydrosuberyl- DALGFAARAIKAGDGDLLIAGGVESMSRAPF CoAthiolase VMGKAASAFSRQAEMFDTTIGWRFVNPLMA [Escherichiacolistr. QQFGTDSMPETAENVAELLKISREDQDSFAL K-12substr.MG1655] RSQQRTAKAQSSGILAEEIVPVVLKNKKGVV TEIQHDEHLRPETTLEQLRGLKAPPRANGVIT AGNASGVNDGAAALIIASEQMAAAQGLTPR ARIVAMATAGVEPRLMGLGPVPATRRVLER AGLSIHDMDVIELNEAFAAQALGVLRELGLP DDAPHVNPNGGAIALGHPLGMSGARLALAA SHELHRRNGRYALCTMCIGVGQGIAMILERV SEQIDNO:188 yqeF >gi|90111494|ref|NP_ MKDVVIVGALRTPIGCPRGALAGHSAVELGS 417321.2| putative LVVKALIERTGVPAYAVDEVILGQVLTAGA acyltransferase GQNPARQSAIKGGLPNSVSAITINDVCGSGL [Escherichiacolistr. KALHLATQAIQCGEADIVIAGGQENMSRAPH K-12substr.MG1655] VLTDSRTGAQLGNSQLVDSLVHDGLWDAFN DYHIGVTAENLAREYGISRQLQDAYALSSQQ KARAAIDAGRFKDEIVPVMTQSNGQTLVVD TDEQPRTDASAEGLARLNPSFDSLGSVTAGN ASSINDGAAAVMMMSEAKARALNLPVLARI RAFASVGVDPALMGIAPVYATRRCLERVGW QLAEVDLIEANEAFAAQALSVGKMLEWDER RVNVNGGAIALGHPIGASGCRILVSLVHEMV KRNARKGLATLCIGGGQGVALTIERDE SEQIDNO:189 tig >gi|16128421|ref|NP_ MQVSVETTQGLGRRVTITIAADSIETAVKSEL 414970.1| peptidyl- VNVAKKVRIDGFRKGKVPMNIVAQRYGASV prolylcis/trans RQDVLGDLMSRNFIDAIIKEKINPAGAPTYVP isomerase(trigger GEYKLGEDFTYSVEFEVYPEVELQGLEAIEV factor)[Escherichia EKPIVEVTDADVDGMLDTLRKQQATWKEK colistr.K-12substr. DGAVEAEDRVTIDFTGSVDGEEFEGGKASDF MG1655] VLAMGQGRMIPGFEDGIKGHKAGEEFTIDVT FPEEYHAENLKGKAAKFAINLKKVEERELPE LTAEFIKRFGVEDGSVEGLRAEVRKNMEREL KSAIRNRVKSQAIEGLVKANDIDVPAALIDSE IDVLRRQAAQRFGGNEKQALELPRELFEEQA KRRVVVGLLLGEVIRTNELKADEERVKGLIE EMASAYEDPKEVIEFYSKNKELMDNMRNVA LEEQAVEAVLAKAKVTEKETTFNELMNQQA SEQIDNO:190 atoD >gi|16130158|ref|NP_ MKTKLMTLQDATGFFRDGMTIMVGGFMGI 416725.1| acetyl- GTPSRLVEALLESGVRDLTLIANDTAFVDTGI CoA:acetoacetyl-CoA GPLIVNGRVRKVIASHIGTNPETGRRMISGEM transferase,alpha DVVLVPQGTLIEQIRCGGAGLGGFLTPTGVG subunit[Escherichia TVVEEGKQTLTLDGKTWLLERPLRADLALIR colistr.K-12substr. AHRCDTLGNLTYQLSARNFNPLIALAADITL MG1655] VEPDELVETGELQPDHIVTPGAVIDHIIVSQES K SEQIDNO:191 atoA >gi|16130159|ref|NP_ MDAKQRIARRVAQELRDGDIVNLGIGLPTM 416726.1| acetyl- VANYLPEGIHITLQSENGFLGLGPVTTAHPDL CoA:acetoacetyl-CoA VNAGGQPCGVLPGAAMFDSAMSFALIRGGH transferase,beta IDACVLGGLQVDEEANLANWVVPGKMVPG subunit[Escherichia MGGAMDLVTGSRKVIIAMEHCAKDGSAKIL colistr.K-12substr. RRCTMPLTAQHAVHMLVTELAVFRFIDGKM MG1655] WLTEIADGCDLATVRAKTEARFEVAADLNT QRGDL SEQIDNO:192 atoE >gi|16130160|ref|NP_ MIGRISRFMTRFVSRWLPDPLIFAMLLTLLTF 416727.1| shortchain VIALWLTPQTPISMVKMWGDGFWNLLAFG fattyacidtransporter MQMALIIVTGHALASSAPVKSLLRTAASAAK [Escherichiacolistr. TPVQGVMLVTFFGSVACVINWGFGLVVGA K-12substr.MG1655] MFAREVARRVPGSDYPLLIACAYIGFLTWGG GFSGSMPLLAATPGNPVEHIAGLIPVGDTLFS GFNIFITVALIVVMPFITRMMMPKPSDVVSID PKLLMEEADFQKQLPKDAPPSERLEESRILTL IIGALGIAYLAMYFSEHGFNITINTVNLMFMI AGLLLHKTPMAYMRAISAAARSTAGILVQFP FYAGIQLMMEHSGLGGLITEFFINVANKDTF PVMTFFSSALINFAVPSGGGHWVIQGPFVIPA AQALGADLGKSVMAIAYGEQWMNMAQPF WALPALAIAGLGVRDIMGYCITALLFSGVIF VIGLTLF SEQIDNO:193 atoB >gi|16130161|ref|NP_ MKNCVIVSAVRTAIGSFNGSLASTSAIDLGAT 416728.1| acetyl-CoA VIKAAIERAKIDSQHVDEVIMGNVLQAGLGQ acetyltransferase NPARQALLKSGLAETVCGFTVNKVCGSGLK [Escherichiacolistr. SVALAAQAIQAGQAQSIVAGGMENMSLAPY K-12substr.MG1655] LLDAKARSGYRLGDGQVYDVILRDGLMCAT HGYHMGITAENVAKEYGITREMQDELALHS QRKAAAAIESGAFTAEIVPVNVVTRKKTFVF SQDEFPKANSTAEALGALRPAPDKAGTVTA GNASGINDGAAALVIMEESAALAAGLTPLAR IKSYASGGVPPALMGMGPVPATQKALQLAG LQLADIDLIEANEAFAAQFLAVGKNLGFDSE KVNVNGGAIALGHPIGASGARILVTLLHAMQ ARDKTLGLATLCIGGGQGIAMVIERLN Fattyacidpathway3-keto-acyl-CoAsynthases SEQIDNO:1 NphT7 >gi|299758082|dbj|BA MTDVRPRIIGTGAYVPERIVSNDEVGAPAGV J10048.1| acetyl- DDDWITRKTGIRQRRWAADDQATSDLATAA CoA:malonyl-CoA GRAALKAAGITPEQLTVIAVATSTPDRPQPPT acyltransferase AAYVQHHLGATGTAAFDVNAVCSGTVFALS [Streptomycessp. SVAGTLVYRGGYALVIGADLYSRILNPADRK CL190] TVVLFGDGAGAMVLGPTSTGTGPIVRRVAL HTFGGLTDLIRVPAGGSRQPLDTDGLDAGLQ YFAMDGREVRRFVTEHLPQLIKGFLHEAGV DAADISHFVPHQANGVMLDEVFGELHLPRA TMHRTVETYGNTGAASIPITMDAAVRAGSFR PGELVLLAGFGGGMAASFALIEW SEQIDNO:194 SaFabH >gi|75765832|pdb|1Z MNVGIKGFGAYAPEKIIDNAYPEQFLDTSDE OW|AChainA, WISKMTGIKERHWADDDQDTSDLAYEASVK CrystalStructureOfS. AIADAGIQPEDIDMIIVATATGDMPFPTVAN AureusFabh,Beta- MLQERLGTGKVASMDQLAACSGFMYSMIT KetoacylCarrier AKQYVQSGDYHNILVVGADKLSKITDLTDR ProteinSynthaseIii STAVLFGDGAGAVIIGEVSEGRGIISYEMGSD GTGGKHLYLDKDTGKLKMNGREVFKFAVRI MGDASTRVVEKANLTSDDIDLFIPHQANIRI MESARERLGISKDKMSVSVNKYGNTSAASIP LSIDQELKNGKLKDDDTIVLVGFGGGLTWG AMTIKWGK SEQIDNO:195 BsFabH >gi|321314863|ref|YP_ MKAGILGVGRYIPEKVLTNHDLEKMVETSD 004207150.1| 3- EWIRTRTGIEERRIAADDVFSSHMAVAAAKN oxoacyl-(acylcarrier ALEQAEVAAEDLDMILVATVTPDQSFPTVSC protein)synthaseIII MIQEQLGAKKACAMDISAACAGFMYGVVT [Bacillussubtilis GKQFIESGTYKHVLVVGVEKLSSITDWEDRN BSn5] TAVLFGDGAGAAVVGPVSDDRGILSFELGA DGTGGQHLYLNEKRHTIMNGREVFKFAVRQ MGESCVNVIEKAGLSKEDVDFLIPHQANIRI MEAARERLELPVEKMSKTVHKYGNTSAASI PISLVEELEAGKIKDGDVVVMVGFGGGLTW GAIAIRWGR SEQIDNO:196 PaFabH >gi|15598482|ref|NP_ MESFNTFVRQYNDQHAEAIAKGELEALAESS 251976.1| 3-oxoacyl- SAFIEKASGIKSRFVMNKEGILDPQRMVPYLP ACPsynthase ERSNDEWSILCEMAVAAAREALQRAGRSAA [Pseudomonas DIDGVIVACSNLQRAYPAIAVEVQAALGIQG aeruginosaPAO1] YGYDMNVACSSATFGIQAATTAIQTGQARAI LMVNPEICTGHLNFRDRDSHFIFGDACTAVI VERADLAVSKHQFDIVSTRLLTQFSNNIRNN FGFLNRADESGIGKRDKLFVQEGRKVFKDV CPMVAELIGEHLAANEIQVAEVKRFWLHQA NLNMNLLITRKLLGRDAEAHEAPVILDSYAN TSSAGSVIALHKHQDDLPSGAIGVLSSFGAG YSIGSVILRKH SEQIDNO:197 MtFabH >gi|2113995|emb|CAB MTEIATTSGARSVGLLSVGAYRPERVVTNDE 08984.1| 3- ICQHIDSSDEWIYTRTGIKTRRFAADDESAAS OXOACYL-[ACYL- MATEACRRALSNAGLSAADIDGVIVTTNTHF CARRIER-PROTEIN] LQTPPAAPMVAASLGAKGILGFDLSAGCAGF SYNTHASEIII GYALGAAADMIRGGGAATMLVVGTEKLSPT FABH(BETA- IDMYDRGNCFIFADGAAAVVVGETPFQGIGP KETOACYL-ACP TVAGSDGEQADAIRQDIDWITFAQNPSGPRP SYNTHASEIII) FVRLEGPAVFRWAAFKMGDVGRRAMDAAG (KASIII) VRPDQIDVFVPHQANSRINELLVKNLQLRPD [Mycobacterium AVVANDIEHTGNTSAASIPLAMAELLTTGAA tuberculosisH37Rv] KPGDLALLIGYGAGLSYAAQVVRMPKG SEQIDNO:198 FabH >gi|345301988|ref|YP_ MLPEQSLTTPLPATTTAAPARRAAVLGVGA 004823890.1| 3- ALPAHREPSAETERRLGLPPGWIARRTGIRER oxoacyl-ACPsynthase PLVGPDEATSDLAVRAGAAALAQAELSPERI III[Rhodothermus GLLLLATSTPDHLLPPTAPVVAHRLGLKHAG marinusSG0.5JP17- AIDLAGACSGFLYALALADGYVRLQRTCVL 172] VIGANVLSRRTNPDDPKTSALFADGAGAVV LGPSEGSRGIVACWLGADGSCWDDLYIPAG GSRRPLTPERVARGEHLMYMKDGRALFRRA ATGMAEAGRRVLQQAGLDLDDVAWWIPHQ ANLRLIEEARRQLGMPEARTVNLVDRIGNSS AATIPLALALEAHRFAPGDLLLLTAVGAGLL SAAVLIQW SEQIDNO:199 FabH >gi|471324089|ref|YP_ MTAPTAVLAGLGSALPPRVVTNHDLTARMD 007523119.1| 3- TSDEWIRTRTGIAERRIVDPGGATSDLAIEAG oxoacyl-[acyl-carrier- RRALDSAGGPDVGAVVVATATPDHPCPATG protein]synthase3 PTVAAGLGLGTVPAFDVGAVCSGFLYALAT protein3 GAGLIAASVADSVLVVGADAFTTIVDPYDRN [Streptomyces TAPIFADGAGAVVLRAGRADEPGALRRTEL davawensisJCM ASDGMQADLIRVAAGGSRQRSHHSAALRED 4913] QYLTMRGGEVFKNAVLRMTEASRTVLDRTG WSTAEVDLLVGHQANVRILHAVAEQLGIGQ ERAYVNIGHTGNTAAASIPLALDDAHGEGRL RAGDKVLLTAFGAGTTWGAITLTWPEGLQY RGAAGSAAA SEQIDNO:200 FabH >gi|330444499|ref|YP_ MDKIKKAAILATGSYLPEKILSNADLEKMVD 004377485.1| 3- TSDEWIVTRTGIKERRIASDNEYTSDMGAKA oxoacyl-ACPsynthase AEKAIRASGLSKDLIDCIVFATSAPDYIFPSSG III[Chlamydophila ALAQAYLGIKEVPAFDCLAACTGFLYGLSIA pecorumE58] KAYVESGTYNHVLLIAADKLSSFVNYQDRN TCVLFGDGGAACIVGRSRPGALEINQVCLGA DGALGDLLSLPAGGSRNPATEATLKEGRHYI SMEGKEVFKHAVRRMEAASKASIAVAGIQE EQVGWLVPHQANERIIDAIAKRFNISEAKVF KSLYKYGNTAASSLGIALDELLNTETVLPHE YLLLTAFGGGLSWGSVVLEHV SEQIDNO:201 FabH >gi|459068159|ref|ZP_ MNSLYSVGITGIGSYVPEKVITNYDLCEIVDT 23165498.1| 3- SNEWIVERTGIQERRIVDQSLSTSDIGTIAAN oxoacyl-(acyl-carrier- KALEDSNTNPKEIDLIIVATATPDMAFPSTAC protein)synthaseIII IVQKNIQAINAAAFDISAGCSGFIYGLSIGFNF [Clostridiumultunense IKAGTYRKVLVIGGETLSKIVNWEDRNTCVL Esp] FGDGAGACILERCEEGFGFLTFDLGSDGNNG HLLIQPAGGSRLPASYETVSNRLHTIKMDGR EVFKFAVRIIEKSSKEVLRKANIPLEQIDLLIP HQANMRIIQSAIKKLQLEENKVYINLDKYGN MSSASIPVALDEAYKKEFFSKGDIVLLVAFG AGLTWGATLLRWNK SEQIDNO:202 FabH >gi|383454618|ref|YP_ MARTHIIGTGSYAPTQVLTNQDLERLVETSD 005368607.1| 3- AWIRERTGIQERRQAAPDEATSDLAVNAAR oxoacyl-(acyl-carrier- NALEMAGVAPGDLDLIVVGTVTADMPMPSC protein)synthaseIII AALVQSKLGAKRAFAFDVSAACAGGLYALS [Corallococcus VADQFVRSGQVKRALVVGADLLTRAVDWT coralloidesDSM DRNTCVLFGDGAGALVLGAEQDADEDAMA 2259] PRGILSTHLRTDGDLANLLCIPAGGSRTPVTA DNVDANLHKLKMNGKEVFRFAVRALVEST QASLGAHGMDTTQVDHVIAHQANLRILEAV MERLEIPKEKCWLNLHKYGNTSSASLPMSLD EAQRAGRLKRGDVIAMMAIGAGMAWGSAV VRW SEQIDNO:203 FabH >gi|333371191|ref|ZP_ MRIMGSVGIIGTGAYLPEKVLTNADLEKMV 08463153.113- DTNDEWIVSRTGIRERRIAADDQASSDLAVE oxoacyl-[acyl-carrier- AGRRALESAGIEAKDLDLIIVATVTPDMAFP protein]synthaseIII ATACLVQDRLGAEKAATFDLSAACTGFLYGI [Desmosporasp. SVASQFISNGMYRHALVIGVDCLSKITDFTD 8437] RNTCVLFGDGAGAAVLGPVEEGKGFLSFEL GGDGSGGHLLKQPAGGSRIPASGKSVEDRLH FISMNGREVFKFAVRVLGSSAEEALRKAGM TKEDVDFLIPHQANTRIIDTAVQRLGLSRDK VVVNLDRYGNMSSASIPVALDEAVQRGKIK KDDTLVLVGFGGGMTWGASVMKWTMETK SEQIDNO:204 FabH >gi|390454110|ref|ZP_ MNKLRPVGIIGTGKYVPEKILTNKDLEAIVET 10239638.1| 3- SDEWIVSRTGIQERHIAAPEQATSDLAYEAAI oxoacyl-(acyl-carrier- KALKSAGMTAEDLDLIIVATVTPDMAFPSTA protein)synthaseIII CILQDKLGAKGAAAFDLSAACSGFVYGLAT [Paenibacilluspeoriae ATSFIKTGIYNNALIIGADCLSRITDYTDRNTC KCTC3763] VLFGDGAGAVVIGEVSEGRGFQSFDLGAEG AGGSLLNLAAGGSRLPASADTLENKQHYIY MNGREVFKFAVRVMGTATVDVLEKAGLTK DDIDLFVPHQANIRIIQSAMQRLDLPEEKVVI NVNKYANTSAASIPLALVEAAEEGRMKEGD RVLMVGFGGGLTWGASVLVW SEQIDNO:205 FabH >gi|392959403|ref|ZP_ MNKKCVGIIGLGSYVPQRIMTNKDLEERMD 10324886.1| 3- TSDQWIVERTGIHERRVAAENESTSDLAAKA oxoacyl-(acyl-carrier- GQKALEDAKISPAEIDLIIVATASPDMVFPAT protein)synthase3 ACVVQENIKAVNAAAFDISAVCSGFLYAMIT [Pelosinusfermentans GSQFIKAGTYRKVLVIGAETLSRFTDWSDRN DSM17108] TGMLFGDGAGAAVLGETPEGYGILGVDLGA DGGGAELLKIPAGGSRHPATMETILQKQHFI YMNGNEVFKFAVKVMGETTLKALKNANLT ASDITYLVPHQANIRIIQSAAKRLGIPMEKVV VNINKYGNTSAASIPIALDEAVKSGAIKSGDI VALAGFGGGLTWASSIMKWCK SEQIDNO:206 FabH >gi|116626090|ref|YP_ MPKAKISALGCYTPPRVLTNQDLEKLVDTN 828246.1| 3-oxoacyl- DQWIMERTGIRERHIAAPEMATSDMAIEAAR ACPsynthase CALLQRGIDACEIDAIILCTVTPDHLFPSTACL [CandidatusSolibacter VQNAIGAKGAWGFDLIAACSGFLYGLTTGA usitatusEllin6076] HFVMAGTHKKVLVIGSDTMSRIIDYTDRATC VLFGDGAGAMLIEATDEADDGTGFIDFLGEI DGSGGEFLRMPAGGSRRPASHETVDQRMHY VHQEGSQVFKYASRKMYEVCRDLLERNHFK VEDVGLMIPHQANKRIIKAAGDRLGIAPERV MINIERYGNTTAGTLPLATRDAISEGRLKKG DLVLFAAVGAGYTVGASLWRWAF SEQIDNO:207 FabH >gi|323702691|ref|ZP_ MSSNLVQAGIIGVGSYVPERILTNKDLEKMV 08114352.1| 3- DTSDEWITSRTGIKERRIADPEESTSELAVKA oxoacyl-(acyl-carrier- ARRALAHAGVKPEELDLIILATCTKDMPFPA protein)synthaseIII SACLVQDQLGAVNAGAFDIEAGCTGFVYAL [Desulfotomaculum TVGSQFVATGSMKRVLVIGADNLSKVTNWE nigrificansDSM574] DRNTCVLFGDGAGAVVLGPVAPGEGILASK LAAEGAGWKYLSMPAGGSRMPASPLTVEK KLHYIHMQGREVFRYAVKVMEEEAANIVKA AGLALSDIDLLIPHQANIRIIEHAAKKLKLSM DKVVVNVDRYGNTSTASIPLALDEAVKSGR VKAGDNIVMVAFGAGLTSGAIVLKWSLGEG KE SEQIDNO:208 FabH >gi|384566084|ref|ZP_ MSTGILGAAGYLPPRVIDNDQVGAWVDRDP 10013188.1| 3- DWILERTGIKERHYAAPEVSTSDMACLAVEK oxoacyl-(acyl-carrier- LYASCPEKRASVGAVILGTSTPDHNFPSTAAI protein)synthaseIII VQGRMGLGRAFAFDLSAACSGYLFSFVTAH [Saccharomonospora SLLSANPALEEVLVIGADTISKVLYQSDRKTV glaucaK62] TVFGDGAAATRVGRVPDGYGLLTHTLITDG CHADYVGQPAGGSRRPLDATTVNARERYM VMHGRKVREYFEEVVPKLIHEVVEQAGVSL DDIDHFVFHQANPQMLADCINAMGIDPAKC PVPGVLSGNTGAASIPLVLSELRAERGDLVV MAAIGSGMTAGAAVLRWY SEQIDNO:209 FabH >gi|298162138|gb|ADI MNQGGVFPLPFKIAGLGRYVPADVVLSSDLE 59524.1| CorB KKYDLPPGWCVEKQGIRERRWVKDETASFM [Corallococcus GAEAAKEAVRDAGLKLEDIDLIINASGSPEQ coralloides] AVPDGGPLVQRELGLGRSGVPSITVNASCLS FFVALDVAANYLNMRRYKRILIVSSDISSVA LDFRKPENFTLFGDAAAAAVVTLPEPGEKSC IHASQVRTYGYGAEFSMVPGGGSRRHPNGK NTTPEDNYLHMNGAELLKIGFEYLPRFNEAL WKQCPDITIKDCRYVIPHQPSRVVLDYLSLT YPDDKLVRIIDRFANCIGASMPMALYEAVKV GGLRRGERGVLTGTGSGVSFVGMVFTY SEQIDNO:210 FabH >gi|148359775|ref|YP_ MNFFRCEKPIYIKGPFVALPERVMSNQDVLN 001250982.1| 3- WMNSTQNPAVIGFSTGIKNRHWVNEDQACS oxoacyl-(acylcarrier DLAVRAAEHLFMEKPREKHKVNQVILATISG protein)synthaseIII DYPSPPSSPLVQYRLGLQNAGAFDIGAACAG FabH[Legionella FVVGLHTSAALAQTNDGSVLLIASEIRSKFLN pneumophilastr. KNNFATSVLFGDGAAACCVSQDKEEADFRFI Corby] ASALFADGEVYDAVSTPAGGSRLPAAVCND NEQFYITIKESTALFVKAVHGMADSAKDFLK ELNLTISDIQWLVPHQGNKNLVLSVAKQLGF PEEKTIKTVEETGNTSGSSVGIALDRLRSDGK IKSGEKVLLVAAGGGGIAACSLLEVI SEQIDNO:211 FabH >gi|15824218|dbj|BA MTNEHLARRLDTDDAWIRTRTGIRRRHAVD B69376.1| 3-oxoacyl- PGQATSDLAVEAGRRALVCAATASVDAVVV (acylcarrierprotein) ATTTPDHSCPATAPAVAARLGLTGAAAFDIS synthase AVCTGFVYGLASAAGLIAAGVAERVLLIGA [Streptomyces DTYSTIVDPLDRANAIIFGDGAGAVVLRAGH avermitilis] PDEPGAVGHFDLGSDGAHEDLIMVAAGGSR QRSRPGEPSRQDRHFGMRGKEVYRHAVTRM AESARATLSRAGWKTDDVDHFVPHQANLRI LHSVADDLGLPRERCVTHVESVGNTGAASIP LALADAAAGQTLRPGDRVLLTAFGGGLTWG SCLLTWPTLPAPAPPYDPHAQGERTTS SEQIDNO:212 FabH >gi|330468931|ref|YP_ MALSSHVEYESTTRTAVIAGLGAYVPDQVV 004406674.1| 3- KNEEIAARLGVTTDWIRDRTGIEQRFVLNPE oxoacyl-(acylcarrier GATSDLAVEAARRALDSCGNPDIDFLILATC protein)synthaseIII TPDHLFPSTAPSVASRLGFKGIAAFDLNAACS [Verrucosisporamans GFVYALSVSTGMLATGAYRTGLVIGADAISS AB-18-032] ILNHDDEITGPIFGDGGGAVVVRAGHLGETG SVSVQQLGSDGDLLDIMKTPGGGSRQRAAG VPVDIDSSYFTMSGRAVYKHAINRMSTVSRS VLERLGWTPDDVDWLIAHQANRRILTATAE EIGIAPERAVINVDRVANTSAASIPLAMVDA VESGALTAGDKVLLAAFGGGATWAAAGLT WPELTLAPTQTVR SEQIDNO:213 FabH >gi|32444698|emb|CA MIETSSNVTANDLAAKSVNEESSAESTAVPT D74700.1| 3-oxoacyl- EAVSAVMPGNATTRGRMGNLKGVRIAGTGS (acyl-carrierprotein) YVPERIVTNEDLAALGCDSDWIVRRTGILQR synthase RHAEPGQATSDLCYEAALRCLENANVSVDEI [Rhodopirellula DLILVATITPDHPTPSTACHLQRRLGAVAPA balticaSH1] MDIGAACAGFMYALVTGAQFVSNGNARNV LVIGADLMSRTVDPEDKKTYPLFGDAAGAA LLVPSTQDECQSTECNGSAADSTIQTDGLLA YQLGSEGCGGEMLCIPAGGSRTPITTDGEDS ASRYLQMDGRGVFKWAVRVFDESAKDVLR AANVSSDQLSLVVLHQANQRIIDSAVSDLNV PPEKVFVNLDKYGNTSGASIPLALDEAARAG RLKEGDLVLLCGFGAGLAWGTALFRW SEQIDNO:214 FabH >gi|392374495|ref|YP_ MYGSRIAGTGASVPDRVLTNAELEQMVSTS 003206328.1| 3- DEWIVTRTGISERRIASDDQATSDLAEGAAR oxoacyl-[acyl-carrier- QALEASGVDPHDLDLILVNTVTPDMFFPSTA protein]synthaseIII CVLQERLGASRAAAFDLMAACAGFVYGLSV (Beta-ketoacyl-ACP ADAYLRAGVMRNILVIGADTLSKVVDWSDR synthaseIII)(KASIII) GTCVLFGDGAGAVVVQRTTADPAILSTHLYS [Candidatus DGSKGRQLIIPGGGSRQPASQKVIDEKLVTIR Methylomirabilis MPNGNEVFKTAVRSMEEAAIAALKANGAEV oxyfera] SDVDLFISHQANARIIYAVAERLDLPRERIYM NIDRYGNTSAASIPIAMDEAVRAGRLKRGDL LLLTAFGGGFTWGSALIRW SEQIDNO:215 FabH >gi|317121784|ref|YP_ MVAAVRGVTIAGIGGCVPPAVVTNDDLAQV 004101787.1| 3- VETDDEWIRTRTGIRQRRVADPGTATSDLAE oxoacyl-(acyl-carrier- VAARRALEEAGVRPDQVDLIIVATVTPDMPF protein)synthaseIII PSTACLLQDRLGATRAAGFDLEAACSGFVY [Thermaerobacter ALAAGAQFIAAGLYDTVLVVGAETLSKIIDW marianensisDSM SDRRTCVLLGDGAGAAVLRPAAPGEGILGL 12885] YLGADGSGGDLLKQPAGGSRLPASPETVAR GLHFVQMNGREVFKFAVKTMGDAAQAALA QAGLTFDDVDLYVPHQANFRIIESSARRFDLP LERVVVNIDRYGNTSAASIPVALDEALSTGRI RAGQTVLLVAFGGGLTWGAAVVRWGYDRP APRPLEMPGQEPRYGLPEWIREQAARGRAR AGEPAQGEPAAAASEATAPAALAVPRAALD PAAVTAASPGSEGRPAWGGGGTR SEQIDNO:216 FabH >gi|383787841|ref|YP_ MKVGVLGLGSYIPEKVVTNHDLEKFLDTSD 005472409.1| 3- EWIRTRTGIVERRIANENEATSDLASIAAKRA oxoacyl-ACPsynthase LEDANLKPEDIDLIIVGTNSPDMLYPATACLV [Caldisericumexile QEKIGASGKCAAFDLQAGCPGFIYATVVGSQ AZM16c01] FVKSGAYKHVLVIGAEVITRMMDPTDRGTY VLFGDGAGAVVLGEVEDNRGIVDFELYADG SIAEHLTLPAGGSRKPFSEEVLKERSYFTKMN GGEVFKFSVREISRISKKLLDKTGTKLEDIDW FIPHQANLRIIQAGAEKLGIPMEKVVVTIDKF GNSSAASIPVSLDTIRKEGKLKRGDLVLMVS FGAGMTSGAILMRW SEQIDNO:217 FabH >gi|404450648|ref|ZP_ MKKTRAVITGVQGWVPEYVLTNRELETMV 11015628.1| 3- DTNDEWITTRTGIKERRILKGENQGTSVIGIN oxoacyl-(acylcarrier AVKGLLEKTNTKAEDIDLIICATVTPDMPFPA protein)synthaseIII TANIIADGVGAKNSYSYDISAACSGFLYALTI [Indibacter GSQFIETGMHKKVIIVGADKMSSIIDYQDRAT alkaliphilusLW1] CIIFGDGGGAVLLEPTQEKVGIMDSLLHADG SGAPFLHMKAGGSRKPASLETIAAREHFAFQ EGSTVFKFAVTNMAEVSARIMERNNLASEDI AWLVPHQANKRIIDATANRMGVGPDKVML NIEKYGNTTAGTLPLCLWDYESQLKKGDNII LAAFGGGFTWGSIYLKWGYDPK SEQIDNO:218 FabH >gi|189502112|ref|YP_ MRTAIRASITGVHGYVPEYILTNEKLEKMVD 001957829.1| 3- TNDEWITTRTGIKERRILEGTNQGTSVLGIPA oxoacyl-(acylcarrier VRGLLEKTNTDPREIDLLICATITPDMITPATA protein)synthaseIII NIIAHAVGATNAFSYDLQAACSGFLYALITG [Candidatus VQFIETGKYKKVVVVGADKMSSIVNYEDRN Amoebophilus SCILFGDGAGAVLLEPNSQGYGIIDSILKGDG asiaticus5a2] NGEQYLHQKAGGSRRPPSAETIAAKEHYVY QEGRAVYRFAVEKMAEVVLEIMKKNNLHH EDIKFLVPHQANKRILDAVAQRAGIKEEQVM ITIQEFGNTTGATIPLCLWRYESQLQPGDKLII TTFGGGFTWGAAYLTWAYK SEQIDNO:219 FabH >gi|395801183|ref|ZP_ MSAVITAIGGYVPSSILTNKKISETVDTSEEWI 10480443.1| 3- IKRTGIRERRIADDDTATSDLAAAAIENLIEN oxoacyl-ACPsynthase YNVDREEIEALLVATATPDHILAPTASIVCDK [Flavobacteriumsp. SGLTNAFGIDMNAACSGFLYALEMGANMIE F52] SGRYKKLIIVGADKMSSIVDYEDRNTCILFGD GAGAILLEKSESDAGLMKTILKTDGSGVSSL AVPAGGSRNPTSMQSLLHRTHYLKQDGAFV FKRAVAAMSQVSQDALAKNELEADQIDWV VPHQANLRIITAVGESLGIDFEKVKVNIDRYG NTTSATVPLCLWDFKDDFKEGQNVLITTFGA GFSWGATCLKWGVMRERKSAETITATTKAE AVLVEH SEQIDNO:220 FabH >gi|86159172|ref|YP_ MRSLIAGTGSYAPEKVVTNADLEKLVDTND 465957.1| 3-oxoacyl- QWIVERTGIRERHVVADDQATSDLALEASRR ACPsynthase ALDAAGLDAKDVEMIVVGTVTPDYPFPSVG [Anaeromyxobacter AVLQGKLGNKKAFAFDVSAACAGSLYALSV dehalogenans2CP-C] ADRFVASGAVKNALVVGADALTRITDWTDR NTCILFGDGAGAMVLKPTDDPQRGIRAVRL HADGSLVPILLQPGGGSRDPISEKVVREKSH YVKMNGREVFKVAVRSLEESCREVLADEKL TPGDVTWVIAHQANKRILDATLHRLEIPESK CWMNLEKYGNTSAASVPMTLDEANRAGWL KPGDTVLMMAIGGGMAWGASVVRW SEQIDNO:221 FabH >gi|166364688|ref|YP_ MNGFGAAVVITGCGSATPAQFLSNEELSQIV 001656961.1| 3- ETSDEWIKSRTGIGKRHLADRSVSLSQLAAQ oxoacyl-ACPsynthase AAIKALEMAQVSPRDIDLILLATSTPDDLFGS [Microcystis AAQVQSQIGANRAIAFDLTAACSGFLVGLVT aeruginosaNIES-843] ATQFIRTGTYRNVLVIGADVLSRWVDWNDR ATCVLFGDGAGAVVCQANDTKDNILGFE,LH SDGSQNGSLNLAYEGEELPLKQGIRVQKGTY KPLRMNGREVYRFAVAKVPEVIEKALYRAN LTTSDIDWLVLHQANQRIMDAVSERLKLPPE KVISNLSEYGNTSAASIPLALDEAVRSGKVK KGDIIASSGFGAGLTWGGIIFRWGD SEQIDNO:222 FabH >gi|219849850|ref|YP_ MYDRKVARVSRERYAAVIGWGMAVPNRVV 002464283.1| 3- TNDDLAQRIDTSDEWIRTRTGIRERRVAGPG oxoacyl-(acyl-carrier- ESTSTFATAAGREALEMAGVSPATIDTVIVA protein)synthaseIII TCTPDRPFPATACTVQANLQIPRATAFDLAA [Chloroflexus ACSGFVYGLTVATSLIKSGVSRRLLLIGADIF aggregansDSM9485] THYINWNDRNTCVLFGDGAGAVVLEATDEP LGLIASNLSADGNLEDLMAVDAGGTRMPLT AELLAEGRQYVYMNGREIFKHAVREMSESA LHVVQAAGLTIDDIALVIPHQANVRIIDAVAR RLELPPERVMINLDRYGNTSAASIPIALYEAA QQERIKAGDYVLMTAFGGGLTWGSGIVRW GRPSR SEQIDNO:223 FabH >gi|227523050|ref|ZP_ MKFENFKILATASQVPTRVVDNDELSTMMD 03953099.1| 3- TSDDWIVQRTGIRRRHIAVDETTSSLCTSYAK oxoacyl-(acylcarrier QLLEKTGLKPSEIDLIIVATMSPDYLTPSVSA protein)synthaseIII MVQGNLGADHAVAMDIDAACSGFVYGLNM [Lactobacillus VKQLLIAETPKNAILIGGEMLSKLIDWQDRST hilgardiiATCC8290] AVLFGDGAGGVLLKNTPKAEGAFISENLKTL GKLGRYLTAGKTGAPTPFMEKKDEFSPFFQ MNGRRVYRFAVNNVPESINQALAEASLTTD DIDHFVLHQANSRIVEKIAETLGVSMDKFPIN IDEYGNTAAASEPILLDQLVTNGTIKRGDVV LLSGFGGGLTVGTMILKY SEQIDNO:224 FabH >gi|240850683|ref|YP_ MIRSIIRGVGSALPKRSLSNDEIAKFVETSDS 002972083.1| 3- WIVQRTGIRQRYIASENETTVSLGVEAAQAA oxoacyl-(acylcarrier LTNAGLTIKDIDCIILATSTPNRTFPASAVEIQ protein)synthaseIII CALGMSHGFAFDIQAVCSGFIFALTTGDSYL [Bartonellagrahamii RCGAAKRILVIGSDTFSRILDWEDRTTCVLFG as4aup] DGAGAAILEAQEIEGGIAFERGILSAKLRSNG AYIDKLYVDGGPSTTQTTGYLRMEGREVFK YAVGMITDVVDDCFAAAGMDSSQLDWFVP HQANKRIIEASAKKLGISLDKVVITVDQHGN TSAASVPLALTTAVCDGKIKEGDLIMLEAMG GGFTWGAILIRW SEQIDNO:225 FabH >gi|253681256|ref|ZP_ MYNVKIISTGKYIPDNVVTNDDMSKFVDTN 04862054.1| 3- DKWISERTGIKERRISTGENTSHMAVKAALA oxoacyl-[acyl-carrier- ALEKSSVKATDLDLIIIATCTPDSFVPSTACIV protein]synthase3 QDKLGATKATCFDISAACTGFIYALGVASQFI [Clostridium KTGQVKNALVIGAETLSKILNWEDRSTCILF botulinumDstr.1873] ADGAGAAIIERSEEVGLISQYTGSDGTGGKA LKCEALPVRNPYCKVDDKFKDTLSMEGREV FKFAVNAMIESINKVLENTEYTLDDIDYIVPH QANIRIIEFVSKKLGISQDKFYVNLHKYGNTS GASIPIALDEMNKKGMFKKGDNIILVGFGGG LTFGAHLIQWN SEQIDNO:226 FabH >gi|254286853|ref|ZP_ MYSKILGTGSYLPSQVRTNADLEKMVETSDE 04961806.1| 3- WIVARTGIRERRIAADNETVADMAFFAAQN oxoacyl-(acyl-carrier- AIDMAGIDKHDIDMIIVATTSASHTFPSAACQ protein)synthaseIII VQGKLGIKGCPAFDLAAACSGFMYALSIAD [VibriocholeraeAM- QHVKSGMCKHVLVIGADALSKTCDPTDRSTI 19226] ILFGDGAGAVVVGASNEPGILSTHIHADGEF GDLLSLEVPVRGGDSDKWLHMAGNEVFKV AVTQLSKLVVDTLKANNMHKSELDWLVPH QANYRIISATAKKLSMSLDQVVITLDRHGNT SAATVPTALDEAVRDGRIQRGQMLLLEAFG GGFTWGSA SEQIDNO:227 FabH >gi|282854072|ref|ZP_ MTAIKTRPVHGYSKFLSTGSARGSRVVTNEE 06263409.1| 3- MCTLIDSTPEWIEQRTGITERRWATSSETVAS oxoacyl-[acyl-carrier- MGTTAARTALERSGLEASQIDAIIVATVSHH protein]synthase3 RPSPSLAAYIARELGLGDAAAFDLNGACAGF [Propionibacterium CYSTALADSMIRTGSANYVLVIGVEKLSEMT acnesJ139] NLDDRSTAFLFSDGAGAAIISASDEPGIGPVV WGSRSDQLKTIELEDWPTASADPNKIHPLIR MEGRAVFKWAMTDVAKRAAEAVAEAGITP ADLDVFIPHQANDRITDVVSRHLKLPESVTV CHDIADMGNTSAASVPIAIDRMLQRGQAHS GDLALIIGFGAGLVYAGQVIRLP SEQIDNO:228 FabH >gi|291439887|ref|ZP_ MAKIKPSKGAPYARILGVGGYRPTRVVPNEV 06579277.1| 3- ILETIDSSDEWIRSRSGIETRHWASPEETVAA oxoacyl-(acylcarrier MSVEASGKAIADAGIDAAQIGAVVVSTVSHF protein)synthaseIII AQTPAIATEIADRLGTDRAAAFDISAGCAGF [Streptomyces GYGLTLAKGMVVEGSAEYVLVIGVERLSDL ghanaensisATCC TDLEDRATAFLFGDGAGAVVVGPSQEPAIGP 14672] TVWGSEGDKSETIKQTVPWTDYRDGTVEKF PAITQEGQAVFRWAVFEMAKVAQQALDAA GITADDLDVFIPHQANVRIIDSMVKTLKLPEH VTVARDIRTTGNTSAASIPLAMERLLATGEA KSGDTALVIGFGAGLVYAASVVTLP SEQIDNO:229 FabH >gi|294791665|ref|ZP_ MTMMNKPVGIIGTGSFLPDNVVTNFDLEKM 06756813.1| 3- VDTNDQWIRERTGIEERRIAPEGMNTSYMAT oxoacyl-(acyl-carrier- EAAKKAMQMANVTAEEIDMIIFATLTPDMII protein)synthaseIII PSAACVLQANLGAKNAAAYDLQAACSGFV [Veillonellasp. YGLITAASYISSGIYKKVLVVGAEILSRRVNW 6_1_27] NDRGTCILFGDGAGAAVVSEVPEGYGIKGID MGADGTGGSALCIPAGGTAVVANDQRVEEG LTFIHMDGPEVYKFAVKTMGRTVLKSLERA SMELNELDYFIPHQANIRIIDSAAKRLHLPME KVFVNLHKYGNTSAASVAIALDEANREGRF KRGDNVAFAGFGAGLTWASLVLKWY SEQIDNO:230 FabH >gi|302539498|ref|ZP_ MTAIGILGTGSYLPADTVSNRVVGERAGVTE 07291840.1| 3- DWILQKTGIRERRYAAEYEATSDLAVEAARS oxoacyl-[acyl-carrier- ALDAAGISAEQLSWIVVATSTPDSPQPATAC protein]synthaseIII LVQHRIGAVNAAAFDVNSVCSGFVFGLVAA [Streptomycessp.C] ARMLPGQDGGVRGHALVIGADVYSRIIDRE DRRTAVLFGDGAGAVVLGPVRSGYGVLGSY LASRGDQAELIRVEAGGSRLPASEKTVAEGL HHFRMNGRGVRDFVAAELPRAVGEVLDRH GLERSEVDHFVPHQANGVMLGETVPRLGLP RARTHLTVAEHGNTSAASIPLALDEAYRSGA PRDRDVVLLAGFGGGMSLGTVLVRWDEEA APAPRKDSAA SEQIDNO:231 FabH >gi|318080591|ref|ZP_ MDNSELCATVASTPEWIETRSGIRARGFAAP 07987923.1| 3- DETLRFMGRAAAEKALARAGVLPDGIDLVL oxoacyl-(acyl-carrier- VASMSRLEQTPPLAVLLAEDLGARAAAGLD protein)synthaseIII VSGACAGFCHALALASDAVRAGSARHVLVV [Streptomycessp. GTERMTDLVERADRTVSVLFADGAGAAVV 5A3_actF] GPSARPGISPPARGAAGRYAGALRMDRGWD AFAADPSLGRPWMRMDGRRVFRWAMDEVT PRAAELLRESGIEPEALDAFVPHQANLRMIEL MAERLGLPERTAVARDVVRAGNTSAASVPL ALEALLDSGEVGSGDRALLVGFGAGLNYAA QVVELP SEQIDNO:232 FabH >gi|374851360|dbj|BA MGTTLTGIGYYLPPKVLTNFDLEKMVDTSD L54322.1| 3-oxoacyl- DWITTRTGIKERRIADNENVTQMAYMASLE [acyl-carrier-protein] ALESANIQPEDIDLIILATLTPELKFPSTACLL synthaseIII QAKLGAKRAYAFDISAACSGFIYGLELADAY [unculturedAquificae IKSGKAKKILLVGAERLSEIVNWQDRSTCVL bacterium] FGDGAGAVIISEGDGEVLSSKMLSDGELWEI LYAPKCGYINMKGKELFKLAVRSMEEVCRY VLESAGISIEDVSIMIPHQANIRIMEALAEKLG MPKEKVYSNIHKYGNTSAASIPIAMYEAYKE GKLRRGDIVMLTAMGGGLTWGAALLRF SEQIDNO:233 FabH >gi|381164912|ref|ZP_ MTRPTLTLAQGAKASRVLGVGSTQPDRVVT 09874142.1| 3- NDELSQHMDTSDQWIRDRVGIIERRFAGEDE oxoacyl-(acyl-carrier- RLVDMAVTAGAKALADAGVAPSEVDTVIVP protein)synthaseIII NCTMPAPIPNAAAQVADRIGVKAAGAFDLN [Saccharomonospora AACAGFCYGLGVASDLVRAGSAKKVLVIGA azureaNA-128] EKLTDVVDPTDRSTAIIFADGAGAALVGPSD EPGIGPVAWGSAGDLVDVIYMRDNRYIFQE GQPVFRWATTQIAPVAMRAVELAGLELSDID VLIPHQANLRIVEAIAKRLRAKGARDDMVV ADDIRYSGNTSSASIPMALDHMRAAGTVKP GDVVLTVGFGAGLSYAGQVLICP SEQIDNO:234 FabH >gi|386335197|ref|YP_ MHDVVISGTGLWVAPEVITNEELVASFNAY 006031367.1| 3- ARHYNEANATAIAAGTLAAVAESSVEFIEKA oxoacyl-ACPsynthase SGIRQRYVIDKAGVLDPARMRPRLAPRGDD [Ralstonia ALSLQAEIGVAAAREALAAAGRDAGDIDMLI solanacearumP082] CSAANMQRPYPAMGIEIQNALGADGYAFDM NVACSSATFGLEQAINAVRTGSARVALMVN PEITSGHLAWKDRDCHFIFGDVCTAVVVERA DDARAPDQWQVLGTRMATRFSNSIRNNAGF LSRSEDRDPDDRDQLFRQEGRKVFKEVCPM AAEHIAGHLQSLGHAPADVRRFWLHQANLG MNQLIGKRLLGRDASADEAPVILDEFANTAS AGSIIAFHRHRADLQPGDLGLICSFGAGYSIG SVAVRKR SEQIDNO:235 FabH >gi|392946737|ref|ZP_ MLGLGVYRPARVVTNDEIAQRVETSDAWIQ 10312379.1| 3- SRTGIATRRIADEEETTVAMGAAAAEKALAA oxoacyl-(acyl-carrier- AGLTADTIDLVIGATCTSPSQIPGAGPQIAHRI protein)synthaseIII GADQAGAFDINGACAGFSYAVSTAADMVR [Frankiasp.QA3] AGSVRHVLVVATERLSDYTDWDDRSTCILL ADGAGATVIGAAETDEIGPAVWGHDGSRPE AIRVPGYGDNMFRMEGQAVFRWAISLVPTV RQICERAGVAPDELAGIVPHQANLRIVEALA TGIGATNAAVARDVVDSGNTSAASIPLGLAR LLDAGEIRRGDPVLLFGFGAGLTYCGQVVRC P SEQIDNO:236 FabH >gi|397172008|ref|ZP_ MQQVVISGSGLFTPQHIISNDELVVSFNQYV 10495404.1| 3- DQFNTEHAAQIAAGELAALEYSSSEFIEKASG oxoacyl-(acylcarrier IKARHVLYKDGILDPKVMHPVFRKRGEDELP protein)synthaseIII EMVEMAVQAATQALAQANKTAADIDLIICA [Alishewanella ASNMQRPYPALSVELQQALGAGGYAFDMN aestuariiB11] VACSSATFAISNAVNAIRGGSAKVVLVVNPE FASPQVDYRSRDSHFIFGDVCTATIIEAESSCT SSQAFRILGMRLKTTFSNNIRCDIGYTEHCFS EQDPKAPFFKQQGRKVFKELLPIVAEVILDE MAAQQVTADDLKRLWLHQANINMNIFAAK KILGRDPLPEEAPLVLDTYANTASAGSIIAFH KYQQGLQSGDKAILCSFGAGYSVGCLVLEK C SEQIDNO:237 FabH >gi|399047091|ref|ZP_ MRQMDKKRSVGILATGSYTPDRVLSNFDLE 10739223.1| 3- KMVETTDEWIVSRTGIRERRICSAEQASSDL oxoacyl-(acyl-carrier- AYEAAKKALERANISAEQLDMIIVATVTPDM protein)synthaseIII MFPSTACILQEKLGAKRAAALDVSAACTGFL [Brevibacillussp. YGITTAAQFIANGLYKYVLVVGVETLSKITN CF112] YKDRNTCVLFGDGAGAAVIGEVREGFGFQS FELGADGAGGELLCLPAGGSRIPASSESVEN NLHYLSMAGGEVFKFAVRVMNSATEAVLSK AGVERENIDLLVPHQANKRIIDSAVQRFGLSE DKVAINLDRYGNMSSASIPVALDEAIAAGRV KEGDNVILVGFGGGLTWGATLLKWSTTPAE GSGQ SEQIDNO:238 FabH >gi|402823152|ref|ZP_ MIRSVLIGTGSALPRNAVSNAELAERVDTSD 10872590.1| 3- EWIVERTGISNRHIAEADETTSSLATEAGRKA oxoacyl-(acylcarrier IEAAGIDAESIDLIVLATATPDQTFPASATIVQ protein)synthaseIII SRLGCRAGGIAFDVAAVCSGFLYAVGVADS [Sphingomonassp. MLRTGMARRALVIGAETFSRILDWEDRTTC LH128] VLFGDGAGAVVLEAQEQVGETPRGILATRL HADGAHNQLLFVDGGPSTTGTVGKLRMKG REVFRHAVVNLAEVLREVIEEAGLSTSDIDW LVPHQANARILDATAKKLSLPPEKVVMTVG QHANTSAASVPLALDVAVRDGRIKQGDLVM LEAMGGGFTWGASLIRI SEQIDNO:239 FabH >gi|407684813|ref|YP_ MSQQVVISGVGVWHPKDSITNEELVDSYNA 006799987.1| 3- YVDAFNEENKAQIESGDVAAMPYSSAEFIEK oxoacyl-ACPsynthase ASGIKSRYIYQKEGALDITRMKPKIAPRADDE [Alteromonas LSHQAEIAVEAAKLALASANVTADEIDAVIV macleodiistr.English SCAYTQRAYPAIAIEVQEALNIEGFGFDMLV Channel673] ACSAATFGMHRAYEMLSAKNATRVLVINPE LVSPQINYADRDSHFIFGDVATATVLELAET AKSEHVYDVLSTKALTKFSNNIRSNFGYMTR AEDVDPYGPDKLFHQAGRKVFKEVCPLAAA HIEAHLASHDITPEGVKRWWLHQANINMNT LICKRLLGRDADRTEAPIVLDEYANTASAGS VIAFGLNHEDLVAGDVGVLCSFGAGYSIGSL VIRKR SEQIDNO:240 FabH >gi|410479651|ref|YP_ MTPTMLNRSIILGTGSFAPANVLTNEDISRKV 006767288.1| 3- ETSDLWIRERTGIRERRIASSGESTSDLALEA oxoacyl-(acyl-carrier- GRNALRNAALSPADLDGIIVATATPDLTFPST protein)synthaseIII ACLVQARLGIPGTFAFDVNAVCSGFMYALKI [Leptospirillum ADSMIRSGQCETLLVIGAEVMSRFVDWSDRS ferriphilumML-04] TCILFGDGAGAVVLGKSGSPQTGGVGTVTL HADGRYWDLIHVPGGGSRSPVETEKPPGNA CTIRMKGSETFRMAVRSLEESVREVLKEEGI GVNELDWVVPHQANIRILEALSERLGIPLGH FVVNIDRYGNTSAASIPMALDEAVQDKRIQP GHRILLTAFGSGVTWGSGLVHWTQKAGGDR SEQIDNO:241 FabH >gi|410617776|ref|ZP_ MNSRIIGTGSYYPSEVRTNADLSLMVDTSDE 11328741.1| 3- WITDRTGIKERRIIGADETAASMGVEASKKA oxoacyl-[acyl-carrier- LEAAGIDAKSLDMIVCATTSGRYALPSTACEI protein]synthase3 QKALDIDGIPAFDVAAACAGYCYALSVADQ protein1[Glaciecola YIKSGMAKRILVVGTDCLSRMISPEDRTMVI polarisLMG21857] LFGDAAGATIIEASEEPGILSTHIHAAGSYGD LLAIGNPTRGDEASIHENWGSMKGNEVFRV AVTKLSEVVEETLAANNMQKSDLDWLVPH QANFRIIKATAKKLNMSLDQVVLTLERYGNT SAATVPTALDEAIRDGRIKRGQNLLLEAFGG GFAWASALVRY SEQIDNO:242 FabH >gi|417318270|ref|ZP_ MDTSDEWIRTRTGIEERRIARDDEYTHDLAY 12104859.1| 3- EAAKVAIKNAGLTPDDIDLFIVATVTQEATFP oxoacyl-(acylcarrier SVANIIQDRLGAKNAAGMDVEAACAGFTFG protein)synthaseIII VVTAAQFIKTGAYKNIVVVGADKLSKITNW [Listeria DDRTTAVLFGDGAGAVVMGPVSDDHGLLSF monocytogenesJ1- DLGSDGSGGKYLNLDENKKIYMNGREVFRF 220] AVRQMGEASLRVLERAGLEKEDLDLLIPHQ ANIRIMEASRERLNLPEEKLMKTVHKYGNTS SSSIALALVDAVEEGRIKDNDNVLLVGFGGG LTWGALIIRWGK SEQIDNO:243 FabH >gi|417747984|ref|ZP_ MKQIAATSGPTNIGLLSVGSYRPQRVVTNDE 12396438.1| 3- LCQNIDSSDEWIYSRTGIKTRRFAARDESTAS oxoacyl-(acyl-carrier- MATEAGREAIAKAGLEASDIDCVVVATSTHF protein)synthaseIII LQTPACGPAVAAALGATGVPAFDISAGCAGF [Mycobacterium GYALGVAADMVRGGTAGKVLVLGSEKLSP aviumsubsp. TVDMTDRSNCFIFADGAAGVVVGETPTQGIG paratuberculosisS397] PTVWGSDGTQATAIRQDIDWMDYLDRPTGP RPFLRLEGSAVFRWAAFEMGKVGQQAMDA AGVRPDEIDVFLPHQANSRINEILAKSLELRP DAVIANDIEHTGNTSAASIPLAMAEVLATGA AKAGDLALLIGYGAGLSYAAQVVRLPPG SEQIDNO:244 FabH >gi|422338672|ref|ZP_ MQSIGIKGIGYYVPENVFTNFDFEKIIDTSDE 16419632.1| 3- WIRTRTGIVERRFASKDQATSDLAREAALKA oxoacyl-(acyl-carrier- IENAKIKKEDVDMIILATTTPDYIAQGAACIV protein)synthaseIII QNKLGLTSIPCFDLNAACTGFIYGLEVAYSL [Fusobacterium VKSGLYKNVLVIGAETLSRIIDMQNRNTCVL nucleatumsubsp. FGDGAAAAIVGQVEEGYGFLGLSIGAEGEDD polymorphumF0401] MILKVPAGGSKKPNDEETIKNRENFVIMKGQ DVFKFAVSTLPKVTLDALEKAKLDVNDLSM VFPHQANLRIIESAAKRMKFPLEKFYMNLSR YGNTSSASVGIALGEAVEKGLVKKGDNIALT GFGGGLTYGSAIIKWAY SEQIDNO:245 FabH >gi|443491493|ref|YP_ MEHRPECCCGCALAQMPSPPEESVPLPPTVG 007369640.1| 3- ILGTAAFVPPRVVTNNQAGASAGIDDAWIFA oxoacyl-[acyl-carrier- RTGIRTRRWADPEQATSDLAVQAAEQALAN protein]synthaseIII, TAINAGQLGAIIVSTSTPDQPQPPTAAFVQNA FabH_1 LHANSAYAFDTNAVCSGFLFAINTAHALAQ [Mycobacterium RDSIHVLVIGADVYSRILDPTDRKTVCLFGD liflandii128FXT] GAGAVVVGPTTASSRHLRIVDTELHTFTQHI NLIGVPGGGSRQPLTTATLDAGQHYFHMDG RGVRDFVTTTVPEQVRKFLARHHLAVEDID HVVMHQANGRMLDEIYSLLDLRNATCHQTI DRFGNTGSASIPITLHHAYPELHGNILCIGFG GGMAAGITLLAAASGSAGDVGAHK SEQIDNO:246 FabH >gi|474659331|emb|C MHRVIISGLGVEIPEPSITNEELVASFNAWVD CV14840.1| Beta- TENVRRQASGEAPLAKSDSAFIVHASGVQTR ketoacyl-acyl-carrier- HVIEREGILDPTRMAPRIPARPDDALSLQAEF proteinsynthaseI GIASARKALDHAGLKPSDIDLVICSSSHQQRP [Mesorhizobiumsp. YPAIAIEMQEALGTKGAGFDMGLGCSSAAA STM4661] ALHMAVNLVRSGAHKRVLVTTPEIITGHLNF RDRQTHFIFGDASVSMIVEGLAKGDKRPGRF EVLDTRIWTQMSNNIRTNLGYHTRTAQDDP YMINLEGNLIKQVGNKVFKEVTVAGHKFIVE FLAEHGLTPEAIRRFWLHQANARMNAMILK LSFGHEVGHDRAPMVLERLGNTAGAGAIIAL SENHADMKPGDFGLLCAFGAGYSIGGALLR ML SEQIDNO:247 FabH >gi|21224866|ref|NP_ MHQGSRITAVGHYQPARILTNEDLAGMVDT 630645.1| 3-oxoacyl- SDEWIRSRVGIRTRRIAGPDEPVDELAGHAA ACPsynthase AKALASAGLTPADVDLVVVATSTAIDRSPNT [Streptomyces AARVAARLGIPGPAALDLNVVCAGFTHALA coelicolorA3(2)] TADHAVRAGSASRALVVGADKMSEVVDWT DRTTCVLVGDGAGAAVVEACAPGEEPGIGP VLWGSVPEMGNAVRIEGTPPRFAQEGQSVY RWATTRLPAIARQACERSGLEPADLAAVVL HQANLRIVEPLAAKIGAVNAVVARDVVESG NTSAASIPLALSKLAERGEITTGDPALLFGFG GNLSYAGQVVRCP SEQIDNO:248 FabH >gi|239623103|ref|ZP_ MTTRIIGTGSYVPEQIVTNNDLAQIVETNDE 04666134.1| 3- WIRSRTGIGERRIATTESTSYMAANAAMRAL oxoacyl-[acyl-carrier- EQSGVKPEEIDLILLGTSSPDYCFPNGACEVQ protein]synthaseIII GMIGAVNAACYDISAACTGFVYALNTAHAFI [Clostridiales SSGIYKTALVIGSDVLSKLIDWTDRGTCVLFG bacterium DGAGAVVVKADETGILGINMHSDGTKGNVL 1_7_47_FAA] TCGSRTNGNFLLGKKPELGYMTMDGQEVFK FAVRKVPECIKQVLDDAGVAAAEVRYFVIH QANYRIIESIAKRLKVSVDCFPVNMEHYGNT SGASVPLLLDEINRKGMLESGDKIVFSGFGA GLTWGATLLEW SEQIDNO:249 FabH >gi|254477647|ref|ZP_ MTRRAVIAGIGHYLPERIVENAEFEATLDTSD 05091033.1| 3- EWIRSRSGIERRHFAAEGETTSNMATKAAQN oxoacyl-(acyl-carrier- ALADAGMTADDIDAIVVATSTADLTFPSAAT protein)synthaseIII MVQAQLGMTKGFAFDVQAVCAGFVYALSN [Ruegeriasp.R11] ANALVASGQADKVLVIGAETFSKIMDWTDR STCVLFGDGAGALVLEAQEGAGTSDDRGIL ATDLNSDGRFKDLLYVDGGVSTQNTGHLRM QGNQVI-RHAVEKLASTAHTSLERAGLGADD VDWIVPHQANIRIIQGTAKKMGLPMDKVVV TVQDHGNTSAASIPLALSVGKARGQIKQGDL IVTEAIGGGLAWGSVVLRW SEQIDNO:250 FabH >gi|311113478|ref|YP_ MTTLKQYENNRYSRILGYGASRGEVIVHNN 003984700.1| 3- DIVEAINSSDEWIKQRTGISTRHRASENQTVN oxoacyl-(acyl-carrier- DLAIAAAHDALANSHVTGEQIDAVIISTISHP protein)synthaseIII YATPSLAVLVADAIGSRCPAYDISAACAGFC [Rothiadentocariosa YGIAQADAMVRSGMAQNVLVIGVEKLSDFI ATCC17931] DNTERSISFLLGDGAGAAVVSVSDEPGIAPTI WGSDGSRWGTVGMTHSLLDIRNRDFVVNPV QEDEKIWPTLRQDGPSVFRWAVWEMAKVA QQALESAGITPDELGALIPHQANARIIDQMA KTLKLPENVAIARDIADAGNTSAASVPLAAH RLLQEQPELSGKFALQIGFGAGLAYAAQVV VLP SEQIDNO:251 FabH >gi|312793335|ref|YP_ MKQNVKILSTGRFVPEKILSNYDLEKMVETS 004026258.1| 3- DEWITQRTGIKERRIVDGRTSTTDLAVQAAR oxoacyl-(acyl-carrier- NAMQKAGISPDEIDLVIVATVTPEMFFPSTAC protein)synthaseiii LVQKELKLKNAFAFDISAACSGFIYGMAVAT [Caldicellulosiruptor QFIQNGFCKTALVIGAEALSKITNWSDRSTC kristjanssonii VLFGDGAGAAILTASSEEGILGFELGSDGEN 177R1B] GLLLYCHAFGLSDLSYSQFKDMPNFRKIYM DGNEVYKFAVKIMPYAVEKVLEKVGLSSSDI DVFIPHQANIRIIESAAKRLKIPMEKVFVNLH KYGNTSAASIPIALDEAIEEGRIKKGDRIVLV GFGGGLTWASCAVKWI SEQIDNO:252 FabH >gi|320449672|ref|YP_ MSGILALGAYAPERVMKNEEFEAYLDTSDE 004201768.1| 3- WIVTRTGIRERRIAAEDEYTSDLAFKAVEDL oxoacyl-ACPsynthase LGRHPGALEGVDGVIVATNTPDALFPDTAAL [Thermusscotoductus VQARFGIQGFAYDLLAGCPGWLYALAQAHA SA-01] MVEAGLARKVLVVGAEALSKIVDWNDRAT AVLFGDAGGAAVVGKVSKGFGFRSFVLGAD GTGAKELYHACVAPRLPDGTSMRNRLYMN GREVFKFAVRVMNTATLEAIEKAGLTPEDIK VFVPHQANLRIIDAARERLGLPWERVVVNV DRYGNTSTASIPLALKEAVDEGRIREGDHVL LVSFGAGLTWAAAVITWGGA SEQIDNO:253 FabH >gi|322421910|ref|YP_ MIRAEILGTGGFVPARVVPNAHFNYLVDDA 004201133.1| 3- DQWIHSRTGIRERRFASAEEATSDLATNAAL oxoacyl-(acyl-carrier- LALENGDVDPLEIDCIIVSTSTPDMILPATAC protein)synthaseIII MVQKNIGAAKAFAFDMNAVCSSFIYGMEVA [Geobactersp.M18] DNLIRSGKYRKVLLIGADTYSKILDFDDKGS APLFGDGAGAVILGAGLSGKGILQSVMHSD GNGWELIQVPSSGSRKPVTAESIAAKENTFK MAGKSVFTFATDVIPRIISDLAERGGIRAEDI DHIIPHQANVRIIDFISRKTGIPKEKFLLNLDR YGNTAAASVGLALDENRRNGVIKSGELVLM MGFGGGLSWGGVLLKA SEQIDNO:254 FabH >gi|325677042|ref|ZP_ MPAPIATATPAAHAALLGLGVYRPRRVVPNS 08156713.1| 3- EIVDRIDSSDEWIRTRSGITARGWAEPDETIV oxoacyl-(acyl-carrier- SMSVAAARDALAAAGLVAEQIDAVVLATSS protein)synthaseIII QMVLGPSAGAVVATELGMQDTAAFDISAGC [Rhodococcusequi AGFCYALGNAASLVRAGQARHVLVIGVERL ATCC33707] SDLLDPTDRTCAFIFADGAGAVVVGPSDSEG IGPVAWGSDGSQTKAIKQDKDFMQYFAEVA AAEAAGGSTERPYIRMDGQAVFRWAITFLE KACRDALEKAGVTADDLDAFVPHQANSRIT DALIRTLGLPDSVAVARDIAESGNTSAASIPM AMEQLLRSGEARPGDTALLLGFGAGLAYAG QVVQLPAIS SEQIDNO:255 FabH >gi|326203621|ref|ZP_ MIKSTKSVGIIGTGSFVPEKVLTNNDLEKMV 08193485.1| 3- DTSDEWIIKRTGISERRILDHDTPNYTMGIEA oxoacyl-(acyl-carrier- ANRALEDAGLKAEDIDLLILSTEAPDYMSPS protein)synthaseIII MSCIIQGAIGAVNAIAFDLNAACTGFIYSLSV [Clostridium ARQFIANGVYRNALVIGCEGLSKIVDWKDR papyrosolvensDSM NTCILFGDASGAVVLGEVDEGYGILDSFLGS 2782] NGAEGMNITIPNLYLSEEEKAKRVNEKYNTL WMDGKEVFKFAVKAMSSATMHVLDNLNM DIKELDFIFPHQANTRIIDGAIKKLGITDDKIH YIINKYGNISSASIPVAMDEAKRDGKLKKGD NMVLVAFGGGLTWGSMAVKWSK SEQIDNO:256 FabH >gi|332670773|ref|YP_ MTRPTLTQATGPAHSRILGIGGVRGERVVPN 004453781.1| 3- DDLVGPIDSSDEWIRQRTGIVTRRRAGEGTD oxoacyl-(acyl-carrier- VLDLAEGAARAAIENAGLTGADIDAVILSTV protein)synthaseIII TYFHQTPAGAAIIADRIGATPAAAYDISAACA [Cellulomonasfimi GYCYGIGQADALVRAGAARHVLVIGAEKMS ATCC484] EFVDPTDRSISFLLGDGAGAVVIGPSDTPGIG PTVWGSDGAQAQAIRQTHSWLATRDEGAG WPTLRQEGQSVFKWAVWQMAPVAQKALD AAGVTADQIDAFVPHQANMRIIDQMIKQLKL PETVVVGRDIADTGNTSAASIPLATERLLREG QVSSGALALQIGFGAGLVYAAQVVVLP SEQIDNO:257 FabH >gi|340361349|ref|ZP_ MQYAKILGTGSYLPANRVSNDDLAKKVDTS 08683778.1| 3- DEWITTRTGIKFRHIADEGEKTSDLAAEASRR oxoacyl-[acyl-carrier- ALVAAGVTADEIDLIIVATATPDMQFPSTATI protein]synthaseIII VQQKLGIANGCPAFDVQAVCAGFMYALSTA [Neisseriamacacae NAYIKSGMAKKALVIGAETFSRIVDWNDRTT ATCC33926] CVLFGDGAGAVVLGASDEAGIIHSKLKADG NYLDLLNVPGQIANGQVCGSPYITMDGPGVF KFAVKMLAKIADEVISEAGYTPDQIDWLVPH QANKRIIDSTAKHLGLDMEKVILTVQEHGNT SAASIPLALDVGIQNGQIKRGQNLLLEGIGGG FAWGAVLVKY SEQIDNO:258 FabH >gi|345304635|ref|YP_ MPYAAITAVGHFLPEDRLTNADLEKMVDTS 004826537.1| 3- DEWIRTRTGIRERRILRDPNKATSYMATEAA oxoacyl-ACPsynthase RECLRKRGMDPEDVELIIVATVTPDMFFPAT III[Rhodothermus ACLVQANLGARNAWGFDLSAACSGFLFALS marinusSG0.5JP17- TAARFIESGKHKRVMVIGADKMSTITDYTDR 172] KNCILFGDAAAAVLLEPDPECGVIDSVEHCD GNNWELLCMLGGGSLNPPTHETVDRKMHY LHQEGRAVFKLAVEGMAQVAVEIMERNNLT ADDVRYLVPHQANLRIIDATARRMGLSPDK VMVNIDRYGNTTAATIPLCLYDWERQLRRG DNLILAAFGGGFTWGAIYLKWAYDGDKVA AAAEATAETSTENA SEQIDNO:259 FabH >gi|349685677|ref|ZP_ MTAKRSLLSGFGGYLPERIVTNDELASRLDT 08896819.1|3- SDEWIRGRTGIGQRHIAGENDTAVSMAAQA oxoacyl-[acyl-carrier- ARRALDYAGAAPDDVDAIIVATSTPDQAFPS protein]synthaseIII TAVRVQAELGMTSGFGFDLAAACSGFIYALS [Gluconacetobacter MADSLIRSGQARSALVIGSEVYSRILDWSDR oboediens174Bp2] GTCVLFGDGAGAAFLTAAGPDDGDAGILST HLHSDGQYGDLLYVDGATGQHDRPAHLRM QGRDVFRHAVGKLSASVDEALAANNLSHAD VNWLVPHQANLRIIDGVARKLALPAERVVV TVDRHANTSAASIPLALNEAVRDGRIRKGDL VLMEALGGGLTWGSALVRL SEQIDNO:260 FabH >gi|352106212|ref|ZP_ MTHVVITGTGLYTPEHAIDNAALVAAFNAW 08961263.1| 3- VDGENEQHAEAIERGEREPLANSSSEFIEKAS oxoacyl-(acylcarrier GIKSRYVLDASGILDPQRMRPKLPQRSNDEP protein)synthaseIII SLQCEMATEAAHQALAAAQVDAADIELVIV [Halomonassp. ACSNLERAYPAVAVEVQQTLGTSGYGFDMN HAL1] VACSSATFALETAANAIASGSVNRALVVNPE ICSAHLNFRDRDSHFIFGDACTAVVLENSAV AVADEQFEILGTRLVTKFSNAIRNNAGFLNR VTDSDPMALDKLFVQEGRRVFKEVCPMVAK LITDHLASLELNGSDLKRMWLHQANRHMN DLIARKVLGYDPSETQAPIILDRYANTSSAGS IIAFHLHREQFNQGDIGVICSFGAGYSAGSVV IRRV SEQIDNO:261 FabH >gi|375098553|ref|ZP_ MSTQDARGVAVLAGLGGWLPPRVVDNDEL 09744816.1| 3- SRRLDTSDEWIRTRTGIAKRHVVHTGLSTVD oxoacyl-(acyl-carrier- MAVEAGRRALESAGPYGENVDAVVLATSTP protein)synthaseIII DHVCPASAPQVAAELGLSGAAAFDVNAVCS [Saccharomonospora GFVYALATASGLISGGVAKRVLLVGADAFT cyaneaNA-134] TLLDPDDRTTVPIFGDGAGAVVLREGSADEL GAVGPFDLHSDGELAELLIVPAGGSRRKKSE NASDHFLKMQGPAVFRHATARMASSSRAVL EKAGWTTSDVDRFVGHQANVRILTATAKNL GLPADSLVVNIGHTGNTSAASIPLAMVDAAV DGMLQPGDRVLVTAFGAGLTWGSTVLRWP ELACAPLP SEQIDNO:262 FabH >gi|384154990|ref|YP_ MIYAAFRSIGAYIPPKIMSNADFEKIIDTSDE 005537805.1| 3- WITKRTGIKERRIANEGEASSDLGARAGELAI oxoacyl-ACPsynthase ERAGISKEEIDLVICATVTPDFLCMPSTACLIA [Arcobacterbutzleri AKLGLPNVMAFDVSAACTGFVYALNVAKA ED-1] FIESGMKKNVLIVGAEKYSAILDYTDRTTCFL FGDGAGAAIISATNDKNESIIDINCSSDGNYE DLIKTPGGGSKNPCSQEVLENKMACIKMKG NETFKLAVKTLTSDVKTMLEKHNLTNEDIN HFIPHQANYRIIKAVGEALDLSDEKTVVTVD KYGNTSAASIPMAMNYAFEQGKIKAGDTILF DAFGGGLTWGSALFKFAPIKR SEQIDNO:263 FabH >gi|385331603|ref|YP_ MIKAVISGTGLYTPPATISNDELVEAFNQYVE 005885554.1| 3- LFNAENADAIASGDVTPLQPSSSSFIEKASGIK oxoacyl-ACPsynthase RRHVIDKDGILDPNRMKPYIPDRSNEEPSVQ [Marinobacter CDMAVTACREALEQAGKSAEDVDAVIVACS adhaerensHP15] NLQRAYPAVSIEVQEALGIDGFAYDMNVAC SSATFGLQAAVNSVENGSARAVLVVSPEICS GHLNFRDRDSHFIFGDACTAILVEREEDTRE GQGFEILGTSLKTKFSNNIRNNFGFLNRADES GVGKPDKLFVQQGRKVFKEVSPLVAETIQK QLQSLSLAPDDLRRMWLHQANLNMNQLIAR KVLGRDATEEEAPVILDEYANTSSAGSIIAFH KNKDDLVSGDLGVICSFGAGYSIGSVVVRRR SEQIDNO:264 FabH >gi|400755130|ref|YP_ MFTPAITGTGVFTPSQTITNAELVAAFNAYA 006563498.1| 3- DKTNAENAKAIAAGEMEPLAHSSEEFILKAS oxoacyl-[acyl-carrier- GIEQRYVMDKSGVLDPEVMHPLLRQRGDDE protein]synthase3 PSIMAEMALDAAKKALAQAGKTAADVDTVI [Phaeobacter CAASNMERAYPALAIEIQDLLGIKGFAFDMN gallaeciensis2.10] VACSSATFGIQAAADMVRSGSIRSALVVNPEI CSGHLEWRDRDCHFIFGDVATATLIERSEDA TGAYFEILSTRCATSFSNNIRNNNGYLRRSRP DGVEDRRDMQFMQNGRKVFKEVLPMVSQH IAEHMEAEGVSNTDLKRLWLHQANKTMND FIGKKVLGRTPEAGEQPNILQDYANTSSAGSI IAFSKYSDDLSAGDLGLICSFGAGYSVGSVIL RRVA SEQIDNO:265 FabH >gi|423197564|ref|ZP_ MTSIVISGSGLYTPPFAVSNEALVAAFNQYV 17184147.1| DLYNEENASAIDAGQLPAKQHSSSEFIEKAS hypotheticalprotein GIKSRYLVSKEGVLDPDIMQPLLAERPDDKP HMPREF1171_02179 SIMVEMAVAAAEQALIAAGREPGEIDLVIVA [Aeromonas ASNMPRPYPALSIELQHYLGASGMAFDMNV hydrophilaSSU] ACSSATFGIKTAADMLAAGSARLALVVNPEI CSGHLNFRDRDSHFIFGDACTAVLLEREADC QVANPWQLVASKLVTQYSNNIRNNFGFLNR LSPRTRYGDDKLFRQQGRKVFKEVLPLVCD QIAGQLDEQGWAADSLSRLWLHQANLTMN QFIARKLLGHDASQQEAPVILDSYGNTSSAG SIIAFHLYNRDLPAGARGVLCSFGAGYSIGSL LLRRL SEQIDNO:266 FabH >gi|424853848|ref|ZP_ MGKQIATVAGGRQSALLGLGVYRPERVVTN 18278206.1| 3- DEICELIDSNDEWIQSRSGIRNRRFAAEDENV oxoacyl-[acyl-carrier- VTMSIAAGRKAIEASGIDPEQIGCVIVATSTY protein]synthase LLLTPPAAAVVADALGTNGPGAFDLGGGCA [Rhodococcusopacus GFCTALTVASDLVRGGSVDYALVVGVEKMS PD630] ITTDPTDRSTRFIFGDGAGAVVVGKSDVAGI GPVEWGSDGAQADAIVQDLDWYEYITTPGA TRPYIKMAGTAVFRWAAFEMGKVALRAVE KAGMSVDDLDAFVPHQANSRITEVIARSMK LPENVPVSDDIAESGNTSAASVPLAMEEMLQ SGATKPGDTALLLAFGAGLSYAAQVVTMPV LAKD SEQIDNO:267 FabH >gi|441509582|ref|ZP_ MSVIAANTGHQNVAMLGIGAYRPQRLVSND 20991498.1| 3- EVCEVLDSSDEWIFERSGVRNRRWISGDESA oxoacyl-[acyl-carrier- RSMAAAAAERAIENSGIAKEKIGALILATNS protein]synthaseIII WKTKIPHGGPIVAYDIGLNGIPAYDIAAGCG [Gordoniaaichiensis GFGYALGVAADTVRAGSAEYVLVVGVETM NBRC108223] SVVMEPTDRNTAFIFGDGAGAVVVGPSEAN GISPTVWGSDGENAEAIGQNYDIPEYMDRAQ EYQHKDPETDPVGRMVVTMQGPRVFRWAA ITLPKALTSVIERSGISADDIEVFVPHQANARI NELMKKNLGFPDDMPMANDIENTGNTSAAS IPLAMEEMLATGKAKGGQTALLLGFGAGLS YAGAVVTLPPAPKVSSFDDLG SEQIDNO:268 FabH >gi|47987537|gb|EN MGIRITGTGLFHPTESISNEELVESLNAYVEQ U26638.1| FNQENAEQIAAGEIEALRGSSPEFIEKASGIQR hypotheticalprotein RYVVEKSGILDPKRLRPRLQERSNDELSLQA F992_02187 EWGVIAAKQAMENAGVTAEDIDVVILACSN [Acinetobactersp. MQRAYPAVAIEIQSALGIQGYAYDMNVACS NIPH236] AATFGLKQAYDAVKCGARRVLLLNVEITSG HLDYRTRDAHFIFGDVATASIIEETETKSGYE ILDIHLFTQFSNNIRNNFGFLNRSEDAVVDDK LFRQDGRKVFKEVCPLVAKIITAQLEKLELTP EQVKRFWLHQANANMNELILKLVVGKEAD LERAPIILDEFANTSSAGVIIAMHRTGEQVNN GEYAVISSFGAGYSVGSIIVQKHIA SEQIDNO:269 PaFabG >gi|15598163|ref|NP_ MSLQGKVALVTGASRGIGQAIALELGRLGA 251657.1| 3-ketoacyl- VVIGTATSASGAEKIAETLKANGVEGAGLVL ACPreductase DVSSDESVAATLEHIQQHLGQPLIVVNNAGI [Pseudomonas TRDNLLVRMKDDEWFDVVNTNLNSLYRLS aeruginosaPAO1] KAVLRGMTKARWGRIINIGSVVGAMGNAG QTNYAAAKAGLEGFTRALAREVGSRAITVN AVAPGFIDTDMTRELPEAQREALLGQIPLGR LGQAEEIAKVVGFLASDGAAYVTGATVPVN GGMYMS SEQIDNO:270 fabG >gi|150963085|gb|AB MSLQGKVALVTGASRGIGQAIALELGRLGA R85110.1| 3-oxoacyl- VVIGTATSASGAEKIAETLKANGVEGAGLVL (acyl-carrier-protein) DVSSDESVAATLEHIQQHLGQPLIVVNNAGI reductase TRDNLLVRMKDDEWFDVVNTNLNSLYRLS [Pseudomonas KAVLRGMTKARWGRIINIGSVVGAMGNAG aeruginosaPA7] QTNYAAAKAGLEGFTRALAREVGSRAITVN AVAPGFIDTDMTRELPEAQREALLAQIPLGR LGQAEEIAKVVGFLASDGAAYVTGATVPVN GGMYMS SEQIDNO:271 hbd >gi|20162442|gb|AA MKKIFVLGAGTMGAGIVQAFAQKGCEVIVR M14586.1|AF494018_ DIKEEFVDRGIAGITKGLEKQVAKGKMSEED 53-hydroxybutyryl- KEAILSRISGTTDMKLAADCDLVVEAAIENM CoAdehydrogenase KIKKEIFAELDGICKPEAILASNTSSLSITEVAS [Clostridium ATKRPDKVIGMHFFNPAPVMKLVEIIKGIATS beijerinckii] QETFDAVKELSVAIGKEPVEVAEAPGFVVNG ILIPMINEASFILQEGIASVEDIDTAMKYGAN HPMGPLALGDLIGLDVCLAIMDVLFTETGDN KYRASSILRKYVRAGWLGRKSGKGFYDYSK SEQIDNO:272 crt >gi:1706153 MELNNVILEKEGKVAVVTINRPKALNALNS P52046|CRT_CLOAB DTLKEMDYVIGEIENDSEVLAVILTGAGEK 3-hydroxybutyryl- SFVAGADISEMKEMNTIEGRKFGILGNKVFR CoAdehydratase RLELLEKPVIAAVNGFALGGGCEIAMSCD Clostridium IRIASSNARFGQPEVGLGITPGFGGTQRLSRL acetobutylicum VGMGMAKQLIFTAQNIKADEALRIGLVN KVVEPSELMNTAKEIANKIVSNAPVAVKLSK QAINRGMQCDIDTALAFESEAFGECFSTE DQKDAMTAFIEKRKIEGFKNR SEQIDNO:273 ech >gi|74484320|gb|ABA MSDTEVPVLAEVRNRVGHLALNRPVGLNAL 10805.1| enoylCoA TLQMIRITWRQLHAWESDPEIVAVVLRANGE hydratase KAFCAGGDIRSLYDSYQAGDDLHHVFLEEK [Pseudomonasputida] YSLDQYIHGYPKPIVALMDGFVLGGGMGLV QGTALRVVTERVKMGMPETSIGYFPDVGGS YFLPRLPGELGLYLGITGIQIRAADALYARLA DWCLPSERISEFDRRLDQISWGYAPREILAGL LSSLASNRLLGAELKSLHPAIDEHFTQPDLSA IRASLQAERRPEYQDWAEQTVELLNNRSPLA MSATLKLLRLGRTLSLANCFELELHLERQWF AKGDLIEGVRALLIDKDKTPRWNPPTLEQLD TNRVNEFFDGFQPAT SEQIDNO:274 ech2 >gi|162287198|ref|NP_ MASPLRFDGRVVLVTGAGGGLGRAYALAFA 077368.21 ERGALVVVNDLGGDFKGVGKGSSAADKVV peroxisomal EEIRRRGGKAVANYDSVEAGEKLVKTALDT multifunctional FGRIDVVVNNAGILRDRSFSRISDEDWDIIQR enzymetype2[Rattus VHLRGSFQVTRAAWDHMKKQNYGRIIMTAS norvegicus] ASGIYGNFGQANYSAAKLGLLGLANTLVIEG RKNNIHCNTIAPNAGSRMTETVMPEDLVEAL KPEYVAPLVLWLCHESCEENGGLFEVGAGW IGKLRWERTLGAIVRKRNQPMTPEAVRDNW VKICDFSNASKPKSIQESTGGIIEVLHKIDSEGI SQNHTGQVASADASGFAGVVGHKLPSFSSS YTELQCIMYALGVGASVKNPKDLKFVYEGS ADFSCLPTFGVIVAQKSLMSGGLAEVPGLSIN FAKVLHGEQYLELYKPLPRSGELKCEAVIAD ILDKGSGIVIVMDVYSYSGKELICYNQFSVFV VGSGGFGGKRTSEKLKAAVAVPSRPPDAVL RDTTSLNQAALYRLSGDSNPLHIDPSFASIAG FEKPILHGLCTFGFSARHVLQQFADNDVSRF KAIKVRFAKPVYPGQTLQTEMWKEGNRIHF QTKVQETGDIVISNAYVDLVPTSGVSAQTPS EGGALQSALVFGEIGRRLKDVGREVVKKVN AVFEWHITKNGNVAAKWTIDLKNGSGEVYQ GPAKGSADTTITISDEDFMEVVLGKLNPQNA FFSGRLKARGNIMLSQKLQMILKDYAKL SEQIDNO:275 ter >gi|488758537|ref|WP_ MIVKPMVRNNICLNAHPQGCKKGVEDQIEY 002681770.1| trans- TKKRITAEVKAGAKAPKNVLVLGCSNGYGL 2-enoyl-CoA ASRITAAFGYGAATIGVSFEKAGSETKYGTP reductase[Treponema GWYNNLAFDEAAKREGLYSVTIDGDAFSDEI denticola] KAQVIEEAKKKGIKFDLIVYSLASPVRTDPDT GIMHKSVLKPFGKTFTGKTVDPFTGELKEISA EPANDEEAAATVKVMGGEDWERWIKQLSK EGLLEEGCITLAYSYIGPEATQALYRKGTIGK AKEHLEATAHRLNKENPSIRAFVSVNKGLVT RASAVIPVIPLYLASLFKVMKEKGNHEGCIE QITRLYAERLYRKDGTIPVDEENRIRIDDWEL EEDVQKAVSALMEKVTGENAESLTDLAGYR HDFLASNGFDVEGINYEAEVERFDRI SEQIDNO:276 ccr >gi|81309006|sp|Q538 MTVKDILDAIQSKDATSADFAALQLPESYRA 65.1|CCR_STRCU ITVHKDETEMFAGLETRDKDPRKSIHLDEVP RecName: VPELGPGEALVAVMASSVNYNSVWTSIFEPV Full= Crotonyl-CoA STFAFLERYGKLSPLTKRHDLPYHIIGSDLAG reductase VVLRTGPGVNAWQPGDEVVAHCLSVELESP DGHDDTMLDPEQRIWGFETNFGGLAEIALV KTNQLMPKPKHLTWEEAAAPGLVNSTAYRQ LVSRNGAAMKQGDNVLIWGASGGLGSYAT QFALAGGANPICVVSSPQKAEICRSMGAEAII DRNAEGYKFWKDEHTQDPKEWKRFGKRIRE LTGGEDIDIVFEHPGRETFGASVYVTRKGGTI TTCASTSGYMHEYDNRYLWMSLKRIIGSHF ANYREAYEANRLIAKGKIHPTLSKTYSLEET GQAAYDVHRNLHQGKVGVLCLAPEEGLGV RDAEMRAQHIDAINRFRNV Thioesterases SEQIDNO:277 tesA >gi|16128478|ref|NP_ MMNFNNVFRWHLPFLFLVLLTFRAAAADTL 415027.1| LILGDSLSAGYRMSASAAWPALLNDKWQSK multifunctionalacyl- TSVVNASISGDTSQQGLARLPALLKQHQPRW CoAthioesteraseIand VLVELGGNDGLRGFQPQQTEQTLRQILQDV proteaseIand KAANAEPLLMQIRLPANYGRRYNEAFSAIYP lysophospholipaseL1 KLAKEFDVPLLPFFMEEVYLKPQWMQDDGI [Escherichiacolistr. HPNRDAQPFIADWMAKQLQPLVNHDS K-12substr.MG1655] SEQIDNO:278 tesA acyl-CoAthioesterase AADTLLILGDSLSAGYRMSASAAWPALLND I,cytosolicform KWQSKTSVVNASISGDTSQQGLARLPALLK QHQPRWVLVELGGNDGLRGFQPQQTEQTLR QILQDVKAANAEPLLMQIRLPANYGRRYNE AFSAIYPKLAKEFDVPLLPFFMEEVYLKPQW MQDDGIHPNRDAQPFIADWMAKQLQPLVN HDS SEQIDNO:279 tesB >gi|16128437|ref|NP_ MSQALKNLLTLLNLEKIEEGLFRGQSEDLGL 414986.1| acyl-CoA RQVFGGQVVGQALYAAKETVPEERLVHSFH thioesteraseII SYFLRPGDSKKPIIYDVETLRDGNSFSARRVA [Escherichiacolistr. AIQNGKPIFYMTASFQAPEAGFEHQKTMPSA K-12substr.MG1655] PAPDGLPSETQIAQSLAHLLPPVLKDKFICDR PLEVRPVEFHNPLKGHVAEPHRQVWIRANG SVPDDLRVHQYLLGYASDLNFLPVALQPHGI GFLEPGIQIATIDHSMWFHRPFNLNEWLLYS VESTSASSARGFVRGEFYTQDGVLVASTVQE GVMRNHN SEQIDNO:280 yciA >gi|16129214|ref|NP_ MSTTHNVPQGDLVLRTLAMPADTNANGDIF 415769.1| acyl-CoA GGWLMSQMDIGGAILAKEIAHGRVVTVRVE esterase[Escherichia GMTFLRPVAVGDVVCCYARCVQKGTTSVSI colistr.K-12substr. NIEVWVKKVASEPIGQRYKATEALFKYVAV MG1655] DPEGKPRALPVE SEQIDNO:281 ybgC >gi|16128711|ref|NP_ MNTTLFRWPVRVYYEDTDAGGVVYHASYV 415264.1| acyl-CoA AFYERARTEMLRHHHFSQQALMAERVAFVV thioesterase,involved RKMTVEYYAPARLDDMLEIQTEITSMRGTSL inphospholipid VFTQRIVNAENTLLNEAEVLVVCVDPLKMK metabolism PRALPKSIVAEFKQ [Escherichiacolistr. K-12substr.MG1655] SEQIDNO:282 ybfF >gi|16128662|ref|NP_ MKLNIRAQTAQNQHNNSPIVLVHGLFGSLD 415212.1| acyl-CoA NLGVLARDLVNDHNIIQVDMRNHGLSPRDP esterase[Escherichia VMNYPAMAQDLVDTLDAQQIDKATFIGHSM colistr.K-12substr. GGKAVMALTALASDRIDKLVAIDIAPVDYH MG1655] VRRHDEIFAAINAVSESDAQTRQQAAAIMRQ HLNEEGVIQFLLKSFVDGEWRFNVPVLWDQ YPHIVGWEKIPAWDHPALFIPGGNSPYVSEQ YRDDLLAQFPQARAHVIAGAGHWVHAEKP DAVLRAIRRYLND SEQIDNO:283 fadM >gi|16128428|ref|NP_ MQTQIKVRGYHLDVYQHVNNARYLEFLEEA 414977.1| long-chain RWDGLENSDSFQWMTAHNIAFVVVNININY acyl-CoAthioesterase RRPAVLSDLLTITSQLQQLNGKSGILSQVITL III[Escherichiacoli EPEGQVVADALITFVCIDLKTQKALALEGEL str.K-12substr. REKLEQMVK MG1655] SEQIDNO:284 AtTE >gi|227217220|gb|EEI MKFKKKFKIGRMHVDPFNYISMRYLVALMN 82564.1| Acyl-ACP EVAFDQAEILEKDIDMKNLRWIIYSWDIQIEN thioesterase NIRLGEEIEITTIPTHMDKFYAYRDFIVESRGN [Anaerococcus ILARAKATFLLMDITRLRPIKIPQNLSLAYGK tetradiusATCC ENPIFDIYDMEIRNDLAFIRDIQLRRADLDNN 35098] FHINNAVYFDLIKETVDIYDKDISYIKLIYRNE IRDKKQIQAFARREDKSIDFALRGEDGRDYC LGKIKTNV SEQIDNO:285 CpTE >gi|1215718|gb|AAC4 MVAAAASSACFPVPSPGASPKPGKLGNWSSS 9179.1| thioesterase LSPSLKPKSIPNGGFQVKANASAHPKANGSA [Cupheapalustris] VTLKSGSLNTQEDTLSSSPPPRAFFNQLPDWS MLLTAITTVFVAPEKRWTMFDRKSKRPNML MDSFGLERVVQDGLVFRQSFSIRSYEICADR TASIETVMNHVQETSLNQCKSIGLLDDGFGR SPEMCKRDLIWVVTRMKIMVNRYPTWGDTI EVSTWLSQSGKIGMGRDWLISDCNTGEILVR ATSVYAMMNQKTRRFSKLPHEVRQEFAPHF LDSPPAIEDNDGKLQKFDVKTGDSIRKGLTP GWYDLDVNQHVSNVKYIGWILESMPTEVLE TQELCSLTLEYRRECGRDSVLESVTSMDPSK VGDRFQYRHLLRLEDGADIMKGRTEWRPKN AGTNGAISTGKT SEQIDNO:286 CperfTE >gi|110673483|gb|AB MGKAYEKVYEVTYGETDGRKDCRITSMMN G82470.1| acyl-ACP FFSDCCLSQEEKNSMNYADNSSETTWVFFD thioesterasefamily YEIIVNRYPRYREKIKVKTYVESIRKFYSNRV protein[Clostridium FEAYDMDGALVARADVLAFLINKKTRRPAR perfringensATCC ISDEEYEIHGLSKESSKLLRKKLNFEKFDKED 13124] LEMNFHIRYLDIDLNMHVSNIKYVEWILETV PVDIVLNYKMKKIKIKFEKEITYGHNVIIKSKI IKGEDEVKVLHKVENEEGESITLAETYWY SEQIDNO:287 LpTE >gi|28270407|emb|CA MATLGANASLYSEQHRITYYECDRTGRATL D63310.1| oleoyl- TTLIDIAVLASEDQSDALGLTTEMVQSHGVG [acyl-carrierprotein] WVVTQYAIDITRMPRQDEVVTIAVRGSAYN thioesterase(putative) PYFAYREFWIRDADGQQLAYITSIWVMMSQ [Lactobacillus TTRRIVKILPELVAPYQSEVVKRIPRLPRPISF plantarumWCFS1] EATDTTITKPYHVRFFDIDPNRHVNNAHYFD WLVDTLPATFLLQHDLVHVDVRYENEVKY GQTVTAHANILPSEVADQVTTSHLIEVDDEK CCEVTIQWRTLPEPIQ SEQIDNO:288 PA2801TE >gi|15597997|ref|NP_ MADRQLLHTAHIPVRWGDMDSYGHVNNTL 251491.1| hypothetical YFQYLEEARVAWFETLGIDLEGAAEGPVVL proteinPA2801 QSLHTYLKPVVHPATVVVELYAGRLGTSSL [Pseudomonas VLEHRLHTLEDPQGTYGEGHCKLVWVRHAE aeruginosaPAO1] NRSTPVPDSIRAAIA Waxestersynthases SEQIDNO:289 Maq1 >gi|120553111|ref|YP_ MTPLNPTDQLFLWLEKRQQPMHVGGLQLFS 957462.1| FPEGAPDDYVAQLADQLRQKTEVTAPFNQR hypotheticalprotein LSYRLGQPVWVEDEHLDLEHHFRFEALPTPG Maqu_0168 RIRELLSFVSAEHSHLMDRERPMWEVHLIEG [Marinobacter LKDRQFALYTKVHHSLVDGVSAMRMATRM aquaeoleiVT8] LSENPDEHGMPPIWDLPCLSRDRGESDGHSL WRSVTHLLGLSGRQLGTIPTVAKELLKTINQ ARKDPAYDSIFHAPRCMLNQKITGSRRFAAQ SWCLKRIRAVCEAYGTTVNDVVTAMCAAA LRTYLMNQDALPEKPLVAFVPVSLRRDDSSG GNQVGVILASLHTDVQEAGERLLKIHHGME EAKQRYRHMSPEEIVNYTALTLAPAAFHLLT GLAPKWQTFNVVISNVPGPSRPLYWNGAKL EGMYPVSIDMDRLALNMTLTSYNDQVEFGL IGCRRTLPSLQRMLDYLEQGLAELELNAGL SEQIDNO:290 Pcry1 >gi|93005078|ref|YP_ MRLLTAVDQLFLLLESRKQPMHVGGLFLFEL 579515.1| hypothetical PEDADISFVHQLVKQMQDSHVPPTFPFNQVL proteinPcryo_0247 EHMVFWKKDKNFDVEHHLHHVALPKPARV [Psychrobacter RELLMYVSREHGRLLDRAMPLWECHVIEGI cryohalolentisK5] QPESEGSPERFALYFKIHHSLVDGIAAMRLV KKSLSQSPNEPVTLPIWSLMARHRNQIDAILP KERSALRILKEQVSTIKPVFTELLDNFKNYND DSYVSTFDAPRSILNRRISASRRIAAQSYDIKR FNDIAERINISKNDVVLAVCAGAIRRYLISMD ALPSKPLIAFVPMSLRTDDSVAGNQLSFVLA NLGTHLDDPLSRIKLIHRSMNNGKRRFRRMN QAQVINYSVVSYAWEGINLATGLFPKKQAF NLIISNVPGSEKSLYWNGARLQSLYPASIVFN GQAMNITLASYLDKIEFGITACSKALPHVQD MLMLIEEELQLLEKVSKELEFNGITVEDKSG YKGNGKTKKLAP SEQIDNO:291 Rjos1 >gi|111018600|ref|YP_ MPVTDSIFLLGESREHPMHVGSLELFTPPEDA 701572.1| GPDYVKSMHETLLEHTDVDPAFRKKPAGPV hypotheticalprotein GSLGNLWWADESDVDLEYHVRHSALPAPY RHA1_ro01601 RVRELLTLTSRLHGTLLDRHRPLWEMYLIEG [Rhodococcusjostii LSDGRFAIYTKLHHSLMDGVSGLRLLMRTLS RHA1] TDPDVRDAPPPWNLPRRASANGAAPAPDLW SVMNGVRRTVGEVAGLAPASLRIARTAMGQ HDMRFPYEAPRTMLNVPIGGARRFAAQSWP LERVHAVRKVAGVSVNDVVMAMCAGALR GYLEEQNALPDEPLIAMVPVSLRDEQQADA GGNAVGVTLCNLATDVDDPAERLTAISASM SQGKELFGSLTSMQALAWSAVNMSPIALTPV PGFVRFTPPPFNVIISNVPGPRKTMYWNGSRL DGIYPTSVVLDGQALNITLTTNGGNLDFGVI GCRRSVPSLQRILFYLETALGELEAALL SEQIDNO:292 Abork1 >gi|110835603|ref|YP_ MKALSPVDQLFLWLEKRQQPMHVGGLQLFS 694462.1| FPEGAGPKYVSELAQQMRDYCHPVAPFNQR acyltransferase LTRRLGQYYWTRDKQFDIDHHFRHEALPKP [Alcanivorax GRIRELLSLVSAEHSNLLDRERPMWEAHLIE borkumensisSK2] GIRGRQFALYYKIHHSVMDGISAMRIASKTL STDPSEREMAPAWAFNTKKRSRSLPSNPVD MASSMARLTASISKQAATVPGLAREVYKVT QKAKKDENYVSIFQAPDTILNNTITGSRRFAA QSFPLPRLKVIAKAYNCTINTVVLSMCGHAL REYLISQHALPDEPLIAMVPMSLRQDDSTGG NQIGMILANLGTHICDPANRLRVIHDSVEEA KSRFSQMSPEEILNFTALTMAPTGLNLLTGL APKWRAFNVVISNIPGPKEPLYWNGAQLQG VYPVSIALDRIALNITLTSYVDQMEFGLIACR RTLPSMQRLLDYLEQSIRELEIGAGIK Miscellaneous SEQIDNO:293 prpE >gi|16759349|ref|NP_ MSFSEFYQRSINEPEAFWAEQARRIDWRQPF 454966.1| propionyl- TQTLDHSRPPFARWFCGGTTNLCHNAVDRW CoAsynthetase RDKQPEALALIAVSSETDEERTFTFSQLHDEV [Salmonellaenterica NAVAAMLLSLGVQRGDRVLVYMPMIAEAQI subsp.entericaserovar TLLACARIGAIHSVVFGGFASHSVAARIDDA Typhistr.CT18] RPALIVSADAGARGGKILPYKKLLDDAIAQA QHQPKHVLLVDRGLAKMSWVDGRDLDFST LRQQYLGASVPVAWLESNETSCILYTSGTTG KPKGVQRDVGGYAVALATSMDTIFGGKAG GVFFCASDIGWVVGHSYIVYAPLLAGMATIV YEGLPTYPDCGVWWKIVEKYQVNRMFSAPT AIRVLKKFPTAQIRNHDLSSLEALYLAGEPLD EPTASWVTETLGVPVIDNYWQTESGWPIMA LARALDDRPSRLGSPGVPMYGYNVQLLNEV TGEPCGINEKGMLVIEGPLPPGCIQTIWGDDA RFVKTYWSLFNRQVYATFDWGIRDAEGYYF ILGRTDDVINIAGHRLGTREIEESISGYPNVAE VAVVGIKDALKGQVAVAFVIPKQSDTLADR EAARDEEKAIMALVDNQIGHFGRPAHVWFV SQLPKTRSGKMLRRTIQAICEGRDPGDLTTID DPASLQQIRQAIEE SEQIDNO:294 phaA >gi|77464320|ref|YP_ MVIVSAARTAVGSFNGAFASTPAHDLGAAVI 353824.1| acetyl-CoA EAVVARAGIDKADVSETILGQVLTAGQGQN acetyltransferase PARQAHIKAGLPQESAAWSINQVCGSGLRA [Rhodobacter VALAAQHVQLGDASIVVAGGQENMSLSPHV sphaeroides2.4.1] AHLRAGQKMGDLSFIDSMIKDGLWDAFNGY HMGQTAENVAAKWQISRDMQDEFAVASQN KAEAAQKAGRFADEIVPFVIKTRKGDVTVD ADEYIRHGATLDAMAKLRPAFIKDGTVTAA NASGINDGAAAVLVMSAEEAEKRGLSPLARI ASYATAGLDPSIMGVGPIHASRKALEKAGW KVGDLDLVEANEAFAAQACAVNKDMGWD PSIVNVNGGAIAIGHPIGASGARVLNTLLFEM QRRNAKKGLATLCIGGGMGVAMCLERP SEQIDNO:295 phaB >gi|77464321|ref|YP_ MSKVALVTGGSRGIGAAISVALKNAGYTVA 353825.1| 3-oxoacyl- ANYAGNDEAARKFTEETGIKTYKWSVADYD ACPreductase ACAAGIAQVEAELGPVAVLVNNAGITRDSM [Rhodobacter FHKMTRDQWKEVIDTNLSGLFNMTHPVWS sphaeroides2.4.1] GMRDRKFGRIINISSINGQKGQAGQANYSAA KAGDLGFTKALAQEGARAGITVNAICPGYIA TEMVMAVPEKVRESIIAQIPTGRLGEPEEIAR CVVFLASDDAGFVTGSTITANGGQYFV SEQIDNO:296 phaC >gi|28916412|gb|AAO MSDKNNEDLKRQASENTLGLNPVIGIRGKDL 59383.1| PHA LTSARMVLAQALKQPFHSAKHVAHFGLELK synthase1 NVVFGQSELKPEDGDRRFADPAWSQNPLYR [Pseudomonas RYLQTYLAWRKELHDWIEHSSLSEQDASRG stutzeri] HFVINLMTEAMAPSNSMANPAAVKRFFETG GKSLLDGMSHLAKDMINNGGMPSQVNMAA FEVGKNLATTEGAVVFRNDVLELIQYKPITE SVHERPLLVVPPQINKFYVFDLSPDKSLARFL LRSQVQTFVVSWRNPTKAQREWGLSTYIAA LKEAIEVICAITGSKDVNMLGACSGGLTTAS LLGHYAALGEQKVHALTLLVSVLDTQLDTQ VALFADEKTLEAAKRRSYQAGVLEGSDMAK VFAWMRPNDLIWNYWVNNYLLGNEPPVFDI LYWNNDTTRLPAALHGEFIEMFQTNPLTRPG ALEVCGTPIDLKQVTCDFFCVAGTTDHITPW DSCYKSAHLFGGKCEFVLSNSGHIQSILNPPG NPKARYMTNSEMPADPKAWQESSTKHADS WWLHWQSWLAERSGKTKNAPTALGNKKFP AGEAAPGTYVHER SEQIDNO:297 phaC >gi|151442|gb|AAA25 MSNKNNDELQRQASENTLGLNPVIGIRRKDL 932.1| PHA- LSSARTVLRQAVRQPLHSAKHVAHFGLELK polymerase1 NVLLGKSSLAPESDDRRFNDPAWSNNPLYR [Pseudomonas RYLQTYLAWRKELQDWIGNSDLSPQDISRG oleovorans] QFVINLMTEAMAPTNTLSNPAAVKRFFETGG KSLLDGLSNLAKDLVNNGGMPSQVNMDAF EVGKNLGTSEGAVVYRNDVLELIQYKPITEQ VHARPLLVVPPQINKFYVFDLSPEKSLARYC LRSQQQTFIISWRNPTKAQREWGLSTYIDAL KEAVDAVLAITGSKDLNMLGACSGGITCTAL VGHYAALGENKVNALTLLVSVLDTTMDNQ VALFVDEQTLEAAKRHSYQAGVLEGSEMAK VFAWMRPNDLIWNYWVNNYLLGNEPPVFDI LFWNNDTTRLPAAFHGDLIEMFKSNPLTRPD ALEVCGTPIDLKQVKCDIYSLAGTNDHITPW QSCYRSAHLFGGKIEFVLSNSGHIQSILNPPG NPKARFMTGADRPGDPVAWQENATKHADS WWLHWQSWLGERAGELEKAPTRLGNRAYA AGEASPGTYVHER SEQIDNO:298 phaC >gi|9951348|gb|AG0 MSQKNNNELPKQAAENTLNLNPVIGIRGKDL 8441.1|AE004919_2 LTSARMVLLQAVRQPLHSARHVAHFSLELK poly(3- NVLLGQSELRPGDDDRRFSDPAWSQNPLYK hydroxyalkanoicacid) RYMQTYLAWRKELHSWISHSDLSPQDISRG synthase1 QFVINLLTEAMSPTNSLSNPAAVKRFFETGG [Pseudomonas KSLLDGLGHLAKDLVNNGGMPSQVDMDAF aeruginosaPAO1] EVGKNLATTEGAVVFRNDVLELIQYRPITES VHERPLLVVPPQINKFYVFDLSPDKSLARFCL RNGVQTFIVSWRNPTKSQREWGLTTYIEALK EAIEVVLSITGSKDLNLLGACSGGITTATLVG HYVASGEKKVNAFTQLVSVLDFELNTQVAL FADEKTLEAAKRRSYQSGVLEGKDMAKVFA WMRPNDLIWNYWVNNYLLGNQPPAFDILY WNNDTTRLPAALHGEFVELFKSNPLNRPGA LEVSGTPIDLKQVTCDFYCVAGLNDHITPWE SCYKSARLLGGKCEFILSNSGHIQSILNPPGNP KARFMTNPELPAEPKAWLEQAGKHADSWW LHWQQWLAERSGKTRKAPASLGNKTYPAG EAAPGTYVHER

IV. Methods and Modification Techniques Relating to Genes, Nucleotide Sequences, and Amino Acid Sequences

[0153] A. Amino Acid Sequence Variants

[0154] Some amino acids in amino acid sequences can be varied without significant effect on the structure or function of proteins. Variants included can constitute deletions, insertions, inversions, repeats, and type substitutions so long as the indicated enzyme activity is not significantly adversely affected. Guidance concerning which amino acid changes are likely to be phenotypically silent can be found, inter alia, in Bowie, J. U., et al., Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions, Science 247:1306-1310 (1990). In various embodiments polypeptides obtained by the expression of the polynucleotide molecules of the present invention may have at least approximately 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to one or more amino acid sequences encoded by the genes and/or nucleic acid sequences described herein for the fatty acid or fatty acid derived product tolerance-related and biosynthesis pathways.

[0155] It will be appreciated by those skilled in the art that amino acids homologous to those described herein are within the scope of the present invention. It will be appreciated that amino acid homology includes conservative substitutions, i.e. those that substitute a given amino acid in a polypeptide by another amino acid of similar characteristics. Typically seen as conservative substitutions are the following replacements: replacements of an aliphatic amino acid such as Ala, Val, Leu and Ile with another aliphatic amino acid; replacement of a Ser with a Thr or vice versa; replacement of an acidic residue such as Asp or Glu with another acidic residue; replacement of a residue bearing an amide group, such as Asn or Gln, with another residue bearing an amide group; exchange of a basic residue such as Lys or Arg with another basic residue; and replacement of an aromatic residue such as Phe or Tyr with another aromatic residue.

[0156] For all nucleic acid and amino acid sequences provided herein, it is appreciated that conservatively modified variants of these sequences are included, and are within the scope of the invention in its various embodiments. Functionally equivalent nucleic acid and amino acid sequences (functional variants), which may include conservatively modified variants as well as more extensively varied sequences, which are well within the skill of the person of ordinary skill in the art, and microorganisms comprising these, also are within the scope of various embodiments of the invention, as are methods and systems comprising such sequences and/or microorganisms. In various embodiments, nucleic acid sequences encoding sufficiently homologous proteins or portions thereof are within the scope of the invention. More generally, nucleic acids sequences that encode a particular amino acid sequence employed in the invention may vary due to the degeneracy of the genetic code, and nonetheless fall within the scope of the invention. Table 15 provides a summary of similarities among amino acids, upon which conservative and less conservative substitutions may be based, and also various codon redundancies that reflect this degeneracy.

TABLE-US-00016 TABLE15 AminoAcidConservativeSubstitutions Relation- AminoAcid ships DNAcodons Alanine N,Ali GCT,GCC,GCA,GCG Proline N CCT,CCC,CCA,CCG Valine N,Ali GTT,GTC,GTA,GTG Leucine N,Ali CTT,CTC,CTA,CTG,TTA, TTG Isoleucine N,Ali ATT,ATC,ATA Methionine N ATG Phenylalanine N,Aro TTT,TTC Tryptophan N TGG Glycine PU GGT,GGC,GGA,GGG Serine PU TCT,TCC,TCA,TCG,AGT, AGC Threonine PU ACT,ACC,ACA,ACG Asparagine PU,Ami AAT,AAC Glutamine PU,Ami CAA,CAG Cysteine PU TGT,TGC Asparticacid NEG,A GAT,GAC Glutamicacid NEG,A GAA,GAG Arginine POS,B CGT,CGC,CGA,CGG,AGA, AGG Lysine POS,B AAA,AAG Histidine POS CAT,CAC Tyrosine Aro TAT,TAC StopCodons TAA,TAG,TGA

[0157] Legend: side groups and other related properties: A=acidic; B=basic; Ali=aliphatic; Ami=amine; Aro=aromatic; N=nonpolar; PU=polar uncharged; NEG=negatively charged; POS=positively charged.

[0158] Also, variants and portions of particular nucleic acid sequences, and respective encoded amino acid sequences recited herein may be exhibit a desired functionality, e.g., enzymatic activity at a selected level, when such nucleic acid sequence variant and/or portion contains a 15 nucleotide sequence identical to any 15 nucleotide sequence set forth in the nucleic acid sequences recited herein including, without limitation, the sequence starting at nucleotide number 1 and ending at nucleotide number 15, the sequence starting at nucleotide number 2 and ending at nucleotide number 16, the sequence starting at nucleotide number 3 and ending at nucleotide number 17, and so forth. It will be appreciated that the invention also provides isolated nucleic acid that contains a nucleotide sequence that is greater than 15 nucleotides (e.g., 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more nucleotides) in length and identical to any portion of the sequence set forth in nucleic acid sequences recited herein. For example, the invention provides isolated nucleic acid that contains a 25 nucleotide sequence identical to any 25 nucleotide sequence set forth in any one or more (including any grouping of) nucleic acid sequences recited herein including, without limitation, the sequence starting at nucleotide number 1 and ending at nucleotide number 25, the sequence starting at nucleotide number 2 and ending at nucleotide number 26, the sequence starting at nucleotide number 3 and ending at nucleotide number 27, and so forth. Additional examples include, without limitation, isolated nucleic acids that contain a nucleotide sequence that is 50 or more nucleotides (e.g., 100, 150, 200, 250, 300, or more nucleotides) in length and identical to any portion of any of the sequences disclosed herein. Such isolated nucleic acids can include, without limitation, those isolated nucleic acids containing a nucleic acid sequence represented in any one section of discussion and/or examples, such as regarding a fatty acid or fatty acid derived product production pathways, nucleic acid sequences encoding enzymes of the fatty acid synthase system, or a fatty acid or fatty acid derived product tolerance. For example, the invention provides an isolated nucleic acid containing a nucleic acid sequence listed herein that contains a single insertion, a single deletion, a single substitution, multiple insertions, multiple deletions, multiple substitutions, or any combination thereof (e.g., single deletion together with multiple insertions). Such isolated nucleic acid molecules can share at least 60, 65, 70, 75, 80, 85, 90, 95, 97, 98, or 99 percent sequence identity with a nucleic acid sequence listed herein (i.e., in the sequence listing).

[0159] Additional examples include, without limitation, isolated nucleic acids that contain a nucleic acid sequence that encodes an amino acid sequence that is 50 or more amino acid residues (e.g., 100, 150, 200, 250, 300, or more amino acid residues) in length and identical to any portion of an amino acid sequence listed or otherwise disclosed herein.

[0160] In addition, the invention provides isolated nucleic acid that contains a nucleic acid sequence that encodes an amino acid sequence having a variation of an amino acid sequence listed or otherwise disclosed herein. For example, the invention provides isolated nucleic acid containing a nucleic acid sequence encoding an amino acid sequence listed or otherwise disclosed herein that contains a single insertion, a single deletion, a single substitution, multiple insertions, multiple deletions, multiple substitutions, or any combination thereof (e.g., single deletion together with multiple insertions). Such isolated nucleic acid molecules can contain a nucleic acid sequence encoding an amino acid sequence that shares at least 60, 65, 70, 75, 80, 85, 90, 95, 97, 98, or 99 percent sequence identity with an amino acid sequence listed or otherwise disclosed herein.

[0161] Examples of properties that provide the bases for conservative and other amino acid substitutions are exemplified in Table 15. Accordingly, one skilled in the art may make numerous substitutions to obtain an amino acid sequence variant that exhibits a desired functionality. BLASTP, CLUSTALP, and other alignment and comparison tools may be used to assess highly conserved regions, to which fewer substitutions may be made (unless directed to alter activity to a selected level, which may require multiple substitutions). More substitutions may be made in regions recognized or believed to not be involved with an active site or other binding or structural motif. In accordance with Table 15, for example, substitutions may be made of one polar uncharged (PU) amino acid for a polar uncharged amino acid of a listed sequence, optionally considering size/molecular weight (i.e., substituting a serine for a threonine). Guidance concerning which amino acid changes are likely to be phenotypically silent can be found, inter alia, in Bowie, J. U., et Al., Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions, Science 247:1306-1310 (1990). This reference is incorporated by reference for such teachings, which are, however, also generally known to those skilled in the art. Recognized conservative amino acid substitutions comprise (substitutable amino acids following each colon of a set): ala:ser; arg:lys; asn:gln or his; asp:glu; cys:ser; gln:asn; glu:asp; gly:pro; his:asn or gln; ile:leu or val; leu:ile or val; lys: arg or gln or glu; met:leu or ile; phe:met or leu or tyr; ser:thr; thr:ser; trp:tyr; tyr:trp or phe; val:ile or leu.

[0162] It is noted that codon preferences and codon usage tables for a particular species can be used to engineer isolated nucleic acid molecules that take advantage of the codon usage preferences of that particular species. For example, the isolated nucleic acid provided herein can be designed to have codons that are preferentially used by a particular microorganism of interest. Numerous software and sequencing services are available for such codon-optimizing of sequences.

[0163] The invention provides polypeptides that contain the entire amino acid sequence of an amino acid sequence listed or otherwise disclosed herein. In addition, the invention provides polypeptides that contain a portion of an amino acid sequence listed or otherwise disclosed herein. For example, the invention provides polypeptides that contain a 15 amino acid sequence identical to any 15 amino acid sequence of an amino acid sequence listed or otherwise disclosed herein including, without limitation, the sequence starting at amino acid residue number 1 and ending at amino acid residue number 15, the sequence starting at amino acid residue number 2 and ending at amino acid residue number 16, the sequence starting at amino acid residue number 3 and ending at amino acid residue number 17, and so forth. It will be appreciated that the invention also provides polypeptides that contain an amino acid sequence that is greater than 15 amino acid residues (e.g., 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more amino acid residues) in length and identical to any portion of an amino acid sequence listed or otherwise disclosed herein For example, the invention provides polypeptides that contain a 25 amino acid sequence identical to any 25 amino acid sequence of an amino acid sequence listed or otherwise disclosed herein including, without limitation, the sequence starting at amino acid residue number 1 and ending at amino acid residue number 25, the sequence starting at amino acid residue number 2 and ending at amino acid residue number 26, the sequence starting at amino acid residue number 3 and ending at amino acid residue number 27, and so forth. Additional examples include, without limitation, polypeptides that contain an amino acid sequence that is 50 or more amino acid residues (e.g., 100, 150, 200, 250, 300 or more amino acid residues) in length and identical to any portion of an amino acid sequence listed or otherwise disclosed herein. Further, it is appreciated that, per above, a 15 nucleotide sequence will provide a 5 amino acid sequence, so that the latter, and higher-length amino acid sequences, may be defined by the above-described nucleotide sequence lengths having identity with a sequence provided herein.

[0164] In addition, the invention provides polypeptides that an amino acid sequence having a variation of the amino acid sequence set forth in an amino acid sequence listed or otherwise disclosed herein. For example, the invention provides polypeptides containing an amino acid sequence listed or otherwise disclosed herein that contains a single insertion, a single deletion, a single substitution, multiple insertions, multiple deletions, multiple substitutions, or any combination thereof (e.g., single deletion together with multiple insertions). Such polypeptides can contain an amino acid sequence that shares at least 60, 65, 70, 75, 80, 85, 90, 95, 97, 98 or 99 percent sequence identity with an amino acid sequence listed or otherwise disclosed herein. A particular variant amino acid sequence may comprise any number of variations as well as any combination of types of variations.

[0165] As indicated herein, polypeptides having a variant amino acid sequence can retain enzymatic activity. Such polypeptides can be produced by manipulating the nucleotide sequence encoding a polypeptide using standard procedures such as site-directed mutagenesis or various PCR techniques. As noted herein, one type of modification includes the substitution of one or more amino acid residues for amino acid residues having a similar chemical and/or biochemical property. For example, a polypeptide can have an amino acid sequence set forth in an amino acid sequence listed or otherwise disclosed herein comprising one or more conservative substitutions.

[0166] More substantial changes can be obtained by selecting substitutions that are less conservative, and/or in areas of the sequence that may be more critical, for example selecting residues that differ more significantly in their effect on maintaining: (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation; (b) the charge or hydrophobicity of the polypeptide at the target site; or (c) the bulk of the side chain. The substitutions that in general are expected to produce the greatest changes in polypeptide function are those in which: (a) a hydrophilic residue, e.g., serine or threonine, is substituted for (or by) a hydrophobic residue, e.g., leucine, isoleucine, phenylalanine, valine or alanine; (b) a cysteine or proline is substituted for (or by) any other residue; (c) a residue having an electropositive side chain, e.g., lysine, arginine, or histidine, is substituted for (or by) an electronegative residue, e.g., glutamic acid or aspartic acid; or (d) a residue having a bulky side chain, e.g., phenylalanine, is substituted for (or by) one not having a side chain, e.g., glycine. The effects of these amino acid substitutions (or other deletions or additions) can be assessed for polypeptides having enzymatic activity by analyzing the ability of the polypeptide to catalyze the conversion of the same substrate as the related native polypeptide to the same product as the related native polypeptide. Accordingly, polypeptides having 5, 10, 20, 30, 40, 50 or less conservative substitutions are provided by the invention.

[0167] B. Determining Amino Acid Sequence Identity

[0168] As a practical matter, whether any particular polypeptide is at least 50%, 60%, 70%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identical to any reference amino acid sequence of any polypeptide described herein (which may correspond with a particular nucleic acid sequence described herein), such particular polypeptide sequence can be determined conventionally using known computer programs such the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711). When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set such that the percentage of identity is calculated over the full length of the reference amino acid sequence and that gaps in homology of up to 5% of the total number of amino acid residues in the reference sequence are allowed.

[0169] For example, in a specific embodiment the identity between a reference sequence (query sequence, i.e., a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, may be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). Preferred parameters for a particular embodiment in which identity is narrowly construed, used in a FASTDB amino acid alignment, are: Scoring Scheme=PAM (Percent Accepted Mutations) 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of the subject amino acid sequence, whichever is shorter. According to this embodiment, if the subject sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, a manual correction is made to the results to take into consideration the fact that the FASTDB program does not account for N- and C-terminal truncations of the subject sequence when calculating global percent identity. For subject sequences truncated at the N- and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are lateral to the N- and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence. A determination of whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of this embodiment. Only residues to the N- and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query residue positions outside the farthest N- and C-terminal residues of the subject sequence are considered for this manual correction. For example, a 90 amino acid residue subject sequence is aligned with a 100 residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence. This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected for.

[0170] C. Techniques for Making Genetic Modifications and Nucleic Acid Constructs

[0171] Various methods and techniques may be used in accordance with the present invention to modify microorganisms. Embodiments of the present invention may result from introduction of an expression vector into a host microorganism, wherein the expression vector contains a nucleic acid sequence coding for an enzyme that is, or is not, normally found in a host microorganism.

[0172] The ability to genetically modify a host cell is essential for the production of any genetically modified (recombinant) microorganism. The mode of gene transfer technology may be by electroporation, conjugation, transduction, or natural transformation. A broad range of host conjugative plasmids and drug resistance markers are available. The cloning vectors are tailored to the host microorganisms based on the nature of antibiotic resistance markers that can function in that host. Also, as disclosed herein, a genetically modified (recombinant) microorganism may comprise modifications other than via plasmid introduction, including modifications to its genomic DNA.

[0173] More generally, nucleic acid constructs can be prepared comprising an isolated polynucleotide encoding a polypeptide having enzyme activity operably linked to one or more (several) control sequences that direct the expression of the coding sequence in a microorganism, such as E. coli, under conditions compatible with the control sequences. The isolated polynucleotide may be manipulated to provide for expression of the polypeptide. Manipulation of the polynucleotide's sequence prior to its insertion into a vector may be desirable or necessary depending on the expression vector. The techniques for modifying polynucleotide sequences utilizing recombinant DNA methods are well established in the art.

[0174] The control sequence may be an appropriate promoter sequence, a nucleotide sequence that is recognized by a host cell for expression of a polynucleotide encoding a polypeptide of the present invention. The promoter sequence contains transcriptional control sequences that mediate the expression of the polypeptide. The promoter may be any nucleotide sequence that shows transcriptional activity in the host cell of choice including mutant, truncated, and hybrid promoters, and may be obtained from genes encoding extracellular or intracellular polypeptides either homologous or heterologous to the host cell. Examples of suitable promoters for directing transcription of the nucleic acid constructs, especially in an E. coli host cell, are the lac promoter (Gronenborn, 1976, Mol. Gen. Genet. 148: 243-250), tac promoter (DeBoer et a/., 1983, Proceedings of the National Academy of Sciences USA 80: 21-25), trc promoter (Brosius et al, 1985, J. Biol. Chem. 260: 3539-3541), T7 RNA polymerase promoter (Studier and Moffatt, 1986, J. MoI. Biol. 189: 113-130), phage promoter p.sub.L (Elvin et al., 1990, Gene 87: 123-126), tetA promoter (Skerra, 1994, Gene 151: 131-135), araBAD promoter (Guzman et al., 1995, J. Bacteriol. 177: 4121-4130), and rhaP.sub.BAD promoter (Haldimann et al., 1998, J. Bacteriol. 180: 1277-1286). Other promoters are described in Useful proteins from recombinant bacteria in Scientific American, 1980, 242: 74-94; and in Sambrook and Russell, Molecular Cloning: A Laboratory Manual, Third Edition 2001 (volumes 1-3), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

[0175] The control sequence may also be a suitable transcription terminator sequence, a sequence recognized by a host cell to terminate transcription. The terminator sequence is operably linked to the 3 terminus of the nucleotide sequence encoding the polypeptide. Any terminator that is functional in an E. coli cell may be used in the present invention. It may also be desirable to add regulatory sequences that allow regulation of the expression of the polypeptide relative to the growth of the host cell. Examples of regulatory systems are those that cause the expression of the gene to be turned on or off in response to a chemical or physical stimulus, including the presence of a regulatory compound. Regulatory systems in prokaryotic systems include the lac, tac, and trp operator systems.

[0176] For various embodiments of the invention the genetic manipulations and/or modifications may be described to include various genetic manipulations, including those directed to change regulation of, and therefore ultimate activity of, an enzyme or enzymatic activity of an enzyme identified in any of the respective pathways. Such genetic modifications, and any references herein to modulating a gene, may be directed to transcriptional, translational, and post-translational modifications that result in a change of enzyme activity and/or selectivity under selected and/or identified culture conditions and/or to provision of additional nucleic acid sequences such as to increase copy number and/or mutants of an enzyme related to fatty acid or fatty acid derived product production. Specific methodologies and approaches to achieve such genetic modification and/or modulation are well known to one skilled in the art, and include, but are not limited to: increasing expression of an endogenous genetic element; decreasing functionality of a repressor gene; introducing a heterologous genetic element; increasing copy number of a nucleic acid sequence encoding a polypeptide catalyzing an enzymatic conversion step to produce fatty acid or a fatty acid derived product; mutating a genetic element to provide a mutated protein to increase specific enzymatic activity; over-expressing; under-expressing; over-expressing a chaperone; knocking out a protease; altering or modifying feedback inhibition; providing an enzyme variant comprising one or more of an impaired binding site for a repressor and/or competitive inhibitor; knocking out a repressor gene; evolution, selection and/or other approaches to improve mRNA stability as well as use of plasmids having an effective copy number and promoters to achieve an effective level of improvement. Random mutagenesis may be practiced to provide genetic modifications that may fall into any of these or other stated approaches. The genetic modifications and/or modulation further broadly fall into additions (including insertions), deletions (such as by a mutation) and substitutions of one or more nucleic acids in a nucleic acid of interest. In various embodiments a genetic modification and/or modulation results in improved enzymatic specific activity and/or turnover number of an enzyme. Without being limited, changes may be measured by one or more of the following: K.sub.M; K.sub.cat; and K.sub.avidity.

[0177] In various embodiments, to function more efficiently, a microorganism may comprise one or more gene deletions. For example, in E. coli, the genes encoding the lactate dehydrogenase (ldhA), phosphate acetyltransferase (pta), pyruvate oxidase (poxB), and pyruvate-formate lyase (pflB) may be disrupted, including deleted. Such gene disruptions, including deletions, are not meant to be limiting, and may be implemented in various combinations in various embodiments. Gene deletions may be accomplished by mutational gene deletion approaches, and/or starting with a mutant strain having reduced or no expression of one or more of these enzymes, and/or other methods known to those skilled in the art. Gene deletions may be effectuated by any of a number of known specific methodologies, including but not limited to the RED/ET methods using kits and other reagents sold by Gene Bridges (Gene Bridges GmbH, Dresden, Germany, <<www.genebridges.com>>).

[0178] More particularly as to the latter method, use of Red/ET recombination, is known to those of ordinary skill in the art and described in U.S. Pat. Nos. 6,355,412 and 6,509,156, issued to Stewart et al. and incorporated by reference herein for its teachings of this method. Material and kits for such method are available from Gene Bridges (Gene Bridges GmbH, Dresden, Germany, <<www.genebridges.com>>), and the method may proceed by following the manufacturer's instructions. The method involves replacement of the target gene by a selectable marker via homologous recombination performed by the recombinase from -phage. The host microorganism expressing -red recombinase is transformed with a linear DNA product coding for a selectable marker flanked by the terminal regions (generally 50 bp, and alternatively up to about 300 bp) homologous with the target gene. The marker could then be removed by another recombination step performed by a plasmid vector carrying the FLP-recombinase, or another recombinase, such as Cre.

[0179] Targeted deletion of parts of microbial chromosomal DNA or the addition of foreign genetic material to microbial chromosomes may be practiced to alter a host cell's metabolism so as to reduce or eliminate production of undesired metabolic products. This may be used in combination with other genetic modifications such as described herein in this general example. In this detailed description, reference has been made to multiple embodiments and to the accompanying drawings in which is shown by way of illustration specific exemplary embodiments in which the invention may be practiced. These embodiments are described in sufficient detail to enable those skilled in the art to practice the invention, and it is to be understood that modifications to the various disclosed embodiments may be made by a skilled artisan.

[0180] Polypeptides and nucleic acids encoding polypeptides can be produced by standard DNA mutagenesis techniques, for example, M13 primer mutagenesis. Details of these techniques are provided in Sambrook and Russell, 2001. Nucleic acid molecules can contain changes of a coding region to fit the codon usage bias of the particular microorganism into which the molecule is to be introduced.

[0181] Alternatively, the coding region can be altered by taking advantage of the degeneracy of the genetic code to alter the coding sequence in such a way that, while the nucleic acid sequence is substantially altered, it nevertheless encodes a polypeptide having an amino acid sequence identical or substantially similar to the native amino acid sequence. For example, alanine is encoded in the open reading frame by the nucleotide codon triplet GCT. Because of the degeneracy of the genetic code, three other nucleotide codon tripletsGCA, GCC, and GCGalso code for alanine. Thus, the nucleic acid sequence of the open reading frame can be changed at this position to any of these three codons without affecting the amino acid sequence of the encoded polypeptide or the characteristics of the polypeptide. Based upon the degeneracy of the genetic code, nucleic acid variants can be derived from a nucleic acid sequence disclosed herein using standard DNA mutagenesis techniques as described herein, or by synthesis of nucleic acid sequences. Thus, for various embodiments the invention encompasses nucleic acid molecules that encode the same polypeptide but vary in nucleic acid sequence by virtue of the degeneracy of the genetic code.

[0182] The invention also provides an isolated nucleic acid that is at least about 12 bases in length (e.g., at least about 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 100, 250, 500, 750, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000 or 20000 bases in length) and hybridizes, under hybridization conditions, to the sense or antisense strand of a nucleic acid having a sequence listed or otherwise disclosed herein. The hybridization conditions can be moderately or highly stringent hybridization conditions.

V. Fermentation Process

[0183] In accordance with the present invention, the microorganisms described herein are used in a fermentation process to produce a desired chemical product, such as a fatty acid or fatty acid derivative, through the bioproduction pathways described herein. Without being limiting, such a process may be exemplified by providing into a vessel, such as a culture or bioreactor vessel, the following: (1) bio-production media, (2) nutrient media, such as a minimal media as known to those skilled in the art, and (3) an inoculum of a genetically modified microorganism so as to provide a population of such microorganism, such as a bacterium, and more particularly a member of the family Enterobacteriaceae, such as E. coli. In accordance with one aspect of the invention, the genetically modified microorganism comprises a metabolic pathway that converts malonyl-CoA to a selected chemical product. The inoculum is cultured in the vessel so that the cell density increases to a cell density suitable for reaching a production level of a fatty acid or fatty acid derived product that meets the desired overall productivity metrics. In various alternative embodiments, a population of these genetically modified microorganisms may be cultured to a first cell density in a first, preparatory vessel, and then transferred to the noted vessel so as to provide the selected cell density. Numerous multi-vessel culturing strategies are known to those skilled in the art.

[0184] A. Bio-Production Media (Carbon Sources)

[0185] Bio-production media, which is used in the present invention with recombinant microorganisms having a biosynthetic pathway for a fatty acid or fatty acid derived product, may contain suitable carbon sources or substrates for the intended metabolic pathways. Suitable substrates may include, but are not limited to, monosaccharides such as glucose and fructose, oligosaccharides such as lactose or sucrose, polysaccharides such as starch or cellulose or mixtures thereof and unpurified mixtures from renewable feedstocks such as cheese whey permeate, cornsteep liquor, sugar beet molasses, and barley malt. Additionally the carbon substrate may also be one-carbon substrates such as carbon dioxide, carbon monoxide, or methanol for which metabolic conversion into key biochemical intermediates has been demonstrated. In addition to one and two carbon substrates methylotrophicmicroorganisms are also known to utilize a number of other carbon containing compounds such as methylamine, glucosamine and a variety of amino acids for metabolic activity.

[0186] Although it is contemplated that all of the above mentioned carbon substrates and mixtures thereof are suitable in the present invention as a carbon source, common carbon substrates used as carbon sources are various monomeric and oligomeric sugars, such as for example glucose, fructose, and sucrose, as well as mixtures of any of these sugars. Other suitable substrates include xylose, arabinose, other cellulose-based C-5 sugars, high-fructose corn syrup, and various other sugars and sugar mixtures as are available commercially. Sucrose may be obtained from feedstocks such as sugar cane, sugar beets, cassava, bananas or other fruit, and sweet sorghum. Glucose and dextrose may be obtained through saccharification of starch based feedstocks including grains such as corn, wheat, rye, barley, and oats. Also, in some embodiments all or a portion of the carbon source may be glycerol. Alternatively, glycerol may be excluded as an added carbon source.

[0187] In one embodiment, the carbon source is selected from glucose, fructose, sucrose, dextrose, lactose, glycerol, and mixtures thereof. Variously and independently, the amount of these components in the carbon source may be greater than about 50%, greater than about 60%, greater than about 70%, greater than about 80%, greater than about 90%, or more, up to 100% or essentially 100% of the carbon source.

[0188] In addition, methylotrophicmicroorganisms are known to utilize a number of other carbon containing compounds such as methylamine, glucosamine and a variety of amino acids for metabolic activity. For example, methylotrophic yeast are known to utilize the carbon from methylamine to form trehalose or glycerol (Bellion et al., Microb. Growth C1 Compd. (Int. Symp.), 7th (1993), 415-32. Editor(s): Murrell, J. Collin; Kelly, Don P. Publisher: Intercept, Andover, UK). Similarly, various species of Candida will metabolize alanine or oleic acid (Sulter et al., Arch. Microbiol. 153:485-489 (1990)). Hence it is contemplated that the source of carbon utilized in embodiments of the present invention may encompass a wide variety of carbon-containing substrates.

[0189] In addition, fermentable sugars may be obtained from cellulosic and lignocellulosic biomass through processes of pretreatment and saccharification, as described, for example, in U.S. Patent Publication No. 2007/0031918A1, which is herein incorporated by reference. Biomass refers to any cellulosic or lignocellulosic material and includes materials comprising cellulose, and optionally further comprising hemicellulose, lignin, starch, oligosaccharides and/or monosaccharides. Biomass may also comprise additional components, such as protein and/or lipid. Biomass may be derived from a single source, or biomass can comprise a mixture derived from more than one source; for example, biomass could comprise a mixture of corn cobs and corn stover, or a mixture of grass and leaves. Biomass includes, but is not limited to, bioenergy crops, agricultural residues, municipal solid waste, industrial solid waste, sludge from paper manufacture, yard waste, wood and forestry waste. Examples of biomass include, but are not limited to, corn grain, corn cobs, crop residues such as corn husks, corn stover, grasses, wheat, wheat straw, barley, barley straw, hay, rice straw, switchgrass, waste paper, sugar cane bagasse, sorghum, soy, components obtained from milling of grains, trees, branches, roots, leaves, wood chips, sawdust, shrubs and bushes, vegetables, fruits, flowers and animal manure. Any such biomass may be used in a bio-production method or system to provide a carbon source. Various approaches to breaking down cellulosic biomass to mixtures of more available and utilizable carbon molecules, including sugars, include: heating in the presence of concentrated or dilute acid (e.g., <1% sulfuric acid); treating with ammonia; treatment with ionic salts; enzymatic degradation; and combinations of these. These methods normally follow mechanical separation and milling, and are followed by appropriate separation processes.

[0190] In various embodiments, any of a wide range of sugars, including, but not limited to sucrose, glucose, xylose, cellulose or hemicellulose, are provided to a microorganism, such as in an industrial system comprising a reactor vessel in which a defined media (such as a minimal salts media including but not limited to M9 minimal media, potassium sulfate minimal media, yeast synthetic minimal media and many others or variations of these), an inoculum of a microorganism providing one or more of the fatty acid or fatty acid derived biosynthetic pathway alternatives, and the a carbon source may be combined. The carbon source enters the cell and is catabolized by well-known and common metabolic pathways to yield common metabolic intermediates, including phosphoenolpyruvate (PEP). (See Molecular Biology of the Cell, 3rd Ed., B. Alberts et al. Garland Publishing, New York, 1994, pp. 42-45, 66-74, incorporated by reference for the teachings of basic metabolic catabolic pathways for sugars; Principles of Biochemistry, 3rd Ed., D. L. Nelson & M. M. Cox, Worth Publishers, New York, 2000, pp 527-658, incorporated by reference for the teachings of major metabolic pathways; and Biochemistry, 4th Ed., L. Stryer, W.H. Freeman and Co., New York, 1995, pp. 463-650, also incorporated by reference for the teachings of major metabolic pathways.)

[0191] Bio-based carbon can be distinguished from petroleum-based carbon according to a variety of methods, including without limitation ASTM D6866, or various other techniques. For example, carbon-14 and carbon-12 ratios differ in bio-based carbon sources versus petroleum-based sources, where higher carbon-14 ratios are found in bio-based carbon sources. In various embodiments, the carbon source is not petroleum-based, or is not predominantly petroleum based. In various embodiments, the carbon source is greater than about 50% non-petroleum based, greater than about 60% non-petroleum based, greater than about 70% non-petroleum based, greater than about 80% non-petroleum based, greater than about 90% non-petroleum based, or more. In various embodiments, the carbon source has a carbon-14 to carbon-12 ratio of about 1.010-14 or greater, for example, 2.010-14 or greater, 3.010-14 or greater, 4.010-14 or greater, 5.010-14 or greater, 6.010-14 or greater, 7.010-14 or greater, 8.010-14 or greater, 9.010-14 or greater, or 10.010-14 or greater.

[0192] The carbon source of any embodiment, comprising a C6 carbon source or C3 carbon source. The carbon source of any embodiment, comprising one or more cellulosic sugars, such as glucose, sucrose, fructose, dextrose, lactose, xylose, or any combination thereof. The carbon source of any embodiment, comprising less than about 50%, 40%, 30%, 20%, 10%, or 5% by mass of glycerol.

[0193] B. The Inoculum (Microorganisms)

[0194] The fermentation bioproduction process in accordance with the present invention may utilize an inoculum comprising any of the genetically modified microorganism described hereinabove. Features as described and claimed herein may be provided in a microorganism selected from the listing herein, or another suitable microorganism, that also comprises one or more natural, introduced, or enhanced fatty acid or fatty acid derived product bio-production pathways. Thus, in some embodiments the microorganism comprises an endogenous fatty acid or fatty acid derived product production pathway (which may, in some such embodiments, be enhanced), whereas in other embodiments the microorganism does not comprise an endogenous fatty acid or fatty acid derived product production pathway.

[0195] Varieties of these genetically modified microorganisms may comprise genetic modifications and/or other system alterations as may be described in other patent applications of one or more of the present inventor(s) and/or subject to assignment to the owner of the present patent application.

[0196] The examples describe specific modifications and evaluations to certain bacterial and yeast microorganisms. The scope of the invention is not meant to be limited to such species, but to be generally applicable to a wide range of suitable microorganisms. Generally, a microorganism used for the present invention may be selected from bacteria, cyanobacteria, filamentous fungi and yeasts.

[0197] For some embodiments, microbial hosts initially selected for bio-production of a selected chemical product should also utilize sugars including glucose at a high rate. Most microbes are capable of utilizing carbohydrates. However, certain environmental microbes cannot utilize carbohydrates to high efficiency, and therefore would not be suitable hosts for such embodiments that are intended for glucose or other carbohydrates as the principal added carbon source.

[0198] As the genomes of various species become known, the present invention easily may be applied to an ever-increasing range of suitable microorganisms. Further, given the relatively low cost of genetic sequencing, the genetic sequence of a species of interest may readily be determined to make application of aspects of the present invention more readily obtainable (based on the ease of application of genetic modifications to a microorganism having a known genomic sequence).

[0199] More particularly, based on the various criteria described herein, suitable microbial hosts for the bio-production of a chemical product generally may include, but are not limited to, any gram negative microorganisms, more particularly a member of the family Enterobacteriaceae, such as E. coli, or Oligotropha carboxidovorans, or Pseudomononas sp.; any gram positive microorganism, for example Bacillus subtilis, Lactobaccilus sp. or Lactococcus sp.; a yeast, for example Saccharomyces cerevisiae, Pichia pastoris or Pichia stipitis; and other groups or microbial species. More particularly, suitable microbial hosts for the bio-production of a fatty acid or fatty acid derived product generally include, but are not limited to, members of the genera Clostridium, Zymomonas, Escherichia, Salmonella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klebsiella, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Pichia, Candida, Hansenula and Saccharomyces. Hosts that may be particularly of interest include: Oligotropha carboxidovorans (such as strain OM5), Escherichia coli, Alcaligenes eutrophus (Cupriavidus necator), Bacillus licheniformis, Paenibacillus macerans, Rhodococcus erythropolis, Pseudomonas putida, Lactobacillus plantarum, Enterococcus faecium, Enterococcus gallinarium, Enterococcus faecalis, Bacillus subtilis and Saccharomyces cerevisiae.

[0200] More particularly, suitable microbial hosts for the bio-production of fatty acid or fatty acid derived product generally include, but are not limited to, members of the genera Clostridium, Zymomonas, Escherichia, Salmonella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klebsiella, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Pichia, Candida, Hansenula and Saccharomyces.

[0201] Hosts that may be particularly of interest include: Oligotropha carboxidovorans (such as strain OMS.sup.T), Escherichia coli, Alcaligenes eutrophus (Cupriavidus necator), Bacillus licheniformis, Paenibacillus macerans, Rhodococcus erythropolis, Pseudomonas putida, Lactobacillus plantarum, Enterococcus faecium, Enterococcus gallinarium, Enterococcus faecalis, Bacillus subtilis and Saccharomyces cerevisiae. Also, any of the known strains of these species may be utilized as a starting microorganism, as may any of the following species including respective strains thereofCupriavidus basilensis, Cupriavidus campinensis, Cupriavidus gilardi, Cupriavidus laharsis, Cupriavidus metallidurans, Cupriavidus oxalaticus, Cupriavidus pauculus, Cupriavidus pinatubonensis, Cupriavidus respiraculi, and Cupriavidus taiwanensis.

[0202] In some embodiments, the recombinant microorganism is a gram-negative bacterium. In some embodiments, the recombinant microorganism is selected from the genera Zymomonas, Escherichia, Pseudomonas, Alcaligenes, and Klebsiella. In some embodiments, the recombinant microorganism is selected from the species Escherichia coli, Cupriavidus necator, Oligotropha carboxidovorans, and Pseudomonas putida. In some embodiments, the recombinant microorganism is an E. coli strain.

[0203] In some embodiments, the recombinant microorganism is a gram-positive bacterium. In some embodiments, the recombinant microorganism is selected from the genera Clostridium, Salmonella, Rhodococcus, Bacillus, Lactobacillus, Enterococcus, Paenibacillus, Arthrobacter, Corynebacterium, and Brevibacterium. In some embodiments, the recombinant microorganism is selected from the species Bacillus licheniformis, Paenibacillus macerans, Rhodococcus erythropolis, Lactobacillus plantarum, Enterococcus faecium, Enterococcus gallinarium, Enterococcus faecalis, and Bacillus subtilis. In particular embodiments, the recombinant microorganism is a B. subtilis strain.

[0204] In some embodiments, the recombinant microorganism is yeast. In some embodiments, the recombinant microorganism is selected from the genera Pichia, Candida, Hansenula, Klebsiella, Issatchenkia, and Saccharomyces. In particular embodiments, the recombinant microorganism is Saccharomyces cerevisiae.

[0205] It is further appreciated, in view of the disclosure, that any of the above microorganisms may be used for production of chemical products other than fatty acid or fatty acid derived product.

[0206] The ability to genetically modify the host is essential for the production of any recombinant microorganism. The mode of gene transfer technology may be by electroporation, conjugation, transduction or natural transformation. A broad range of host conjugative plasmids and drug resistance markers are available. The cloning vectors are tailored to the host microorganisms based on the nature of antibiotic resistance markers that can function in that host.

[0207] C. Fermentation Nutrient Media and Culture Conditions

[0208] In addition to an appropriate carbon source, such as selected from one of the herein-disclosed types, bio-production media must contain suitable minerals, salts, cofactors, buffers and other components, known to those skilled in the art, suitable for the growth of the cultures and promotion of the enzymatic pathway necessary for chemical product bio-production under the present invention.

[0209] Another aspect of the invention regards media and culture conditions that comprise genetically modified microorganisms of the invention and optionally supplements.

[0210] Typically cells are grown at a temperature in the range of about 25 C. to about 40 C. in an appropriate medium, as well as up to 70 C. for thermophilic microorganisms. Suitable growth media in the present invention are common commercially prepared media such as Luria Bertani (LB) broth, Terrific Broth (TB), M9 minimal media, Sabouraud Dextrose (SD) broth, Yeast medium (YM) broth, (Ymin) yeast synthetic minimal media, and minimal media as described herein, such as M9 minimal media. Other defined or synthetic growth media may also be used, and the appropriate medium for growth of the particular microorganism will be known by one skilled in the art of microbiology or bio-production science. In various embodiments a minimal media may be developed and used that does not comprise, or that has a low level of addition of various components, for example less than 10, 5, 2 or 1 g/L of a complex nitrogen source including but not limited to yeast extract, peptone, tryptone, soy flour, corn steep liquor, or casein. These minimal medias may also have limited supplementation of vitamin mixtures including biotin, vitamin B12 and derivatives of vitamin B12, thiamin, pantothenate and other vitamins. Minimal media may also have limited simple inorganic nutrient sources containing less than 28, 17, or 2.5 mM phosphate, less than 25 or 4 mM sulfate, and less than 130 or 50 mM total nitrogen.

[0211] Suitable pH ranges for the bio-production are from pH 3.0 to pH 10.0, where pH 6.0 to pH 8.0 is a typical pH range for the initial condition. For example, the pH can be 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, or 10.0 However, the actual culture conditions for a particular embodiment are not meant to be limited by these pH ranges.

[0212] Bio-productions may be performed under aerobic, microaerobic, or anaerobic conditions, with or without agitation.

[0213] In various embodiments, specific supplements to a bioreactor vessel comprising such microorganism population may also be provided to further improve the methods and systems.

[0214] D. Bio-Production Reactors and Systems

[0215] Fermentation systems utilizing methods and/or compositions according to the invention are also within the scope of the invention.

[0216] Any of the recombinant microorganisms as described and/or referred to herein may be introduced into an industrial bio-production system where the microorganisms convert a carbon source into a fatty acid or fatty acid derived product in a commercially viable operation. The bio-production system includes the introduction of such a recombinant microorganism into a bioreactor vessel, with a carbon source substrate and bio-production media suitable for growing the recombinant microorganism, and maintaining the bio-production system within a suitable temperature range (and dissolved oxygen concentration range if the reaction is aerobic or microaerobic) for a suitable time to obtain a desired conversion of a portion of the substrate molecules to a selected chemical product. Industrial bio-production systems and their operation are well-known to those skilled in the arts of chemical engineering and bioprocess engineering.

[0217] Bio-productions may be performed under aerobic, microaerobic, or anaerobic conditions, with or without agitation. The operation of cultures and populations of microorganisms to achieve aerobic, microaerobic and anaerobic conditions are known in the art, and dissolved oxygen levels of a liquid culture comprising a nutrient media and such microorganism populations may be monitored to maintain or confirm a desired aerobic, microaerobic or anaerobic condition. When syngas is used as a feedstock, aerobic, microaerobic, or anaerobic conditions may be utilized. When sugars are used, anaerobic, aerobic or microaerobic conditions can be implemented in various embodiments.

[0218] Any of the recombinant microorganisms as described and/or referred to herein may be introduced into an industrial bio-production system where the microorganisms convert a carbon source into a selected chemical product in a commercially viable operation. The bio-production system includes the introduction of such a recombinant microorganism into a bioreactor vessel, with a carbon source substrate and bio-production media suitable for growing the recombinant microorganism, and maintaining the bio-production system within a suitable temperature range (and dissolved oxygen concentration range if the reaction is aerobic or microaerobic) for a suitable time to obtain a desired conversion of a portion of the substrate molecules to the selected chemical product.

[0219] In various embodiments, syngas components or sugars are provided to a microorganism, such as in an industrial system comprising a reactor vessel in which a defined media (such as a minimal salts media including but not limited to M9 minimal media, potassium sulfate minimal media, yeast synthetic minimal media and many others or variations of these), an inoculum of a microorganism providing an embodiment of the biosynthetic pathway(s) taught herein, and the carbon source may be combined. The carbon source enters the cell and is catabolized by well-known and common metabolic pathways to yield common metabolic intermediates, including phosphoenolpyruvate (PEP) or acetyl-CoA. (See Molecular Biology of the Cell, 3.sup.rd Ed., B. Alberts et al. Garland Publishing, New York, 1994, pp. 42-45, 66-74, incorporated by reference for the teachings of basic metabolic catabolic pathways for sugars; Principles of Biochemistry, 3.sup.rd Ed., D. L. Nelson & M. M. Cox, Worth Publishers, New York, 2000, pp. 527-658, incorporated by reference for the teachings of major metabolic pathways; and Biochemistry, 4.sup.th Ed., L. Stryer, W.H. Freeman and Co., New York, 1995, pp. 463-650, also incorporated by reference for the teachings of major metabolic pathways.).

[0220] Further to types of industrial bio-production, various embodiments of the present invention may employ a batch type of industrial bioreactor. A classical batch bioreactor system is considered closed meaning that the composition of the medium is established at the beginning of a respective bio-production event and not subject to artificial alterations and additions during the time period ending substantially with the end of the bio-production event. Thus, at the beginning of the bio-production event the medium is inoculated with the desired microorganism or microorganisms, and bio-production is permitted to occur without adding anything to the system. Typically, however, a batch type of bio-production event is batch with respect to the addition of carbon source and attempts are often made at controlling factors such as pH and oxygen concentration. In batch systems the metabolite and biomass compositions of the system change constantly up to the time the bio-production event is stopped. Within batch cultures cells moderate through a static lag phase to a high growth log phase and finally to a stationary phase where growth rate is diminished or halted. If untreated, cells in the stationary phase will eventually die. Cells in log phase generally are responsible for the bulk of production of a desired end product or intermediate.

[0221] A variation on the standard batch system is the fed-batch system. Fed-batch bio-production processes are also suitable in the present invention and comprise a typical batch system with the exception that the nutrients, including the substrate, are added in increments as the bio-production progresses. Fed-Batch systems are useful when catabolite repression is apt to inhibit the metabolism of the cells and where it is desirable to have limited amounts of substrate in the media. Measurement of the actual nutrient concentration in Fed-Batch systems may be measured directly, such as by sample analysis at different times, or estimated on the basis of the changes of measurable factors such as pH, dissolved oxygen and the partial pressure of waste gases such as CO.sub.2. Batch and fed-batch approaches are common and well known in the art and examples may be found in Thomas D. Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc., Sunderland, Mass., Deshpande, Mukund V., Appl. Biochem. Biotechnol., 36:227, (1992), and Biochemical Engineering Fundamentals, 2.sup.nd Ed. J. E. Bailey and D. F. Ollis, McGraw Hill, N.Y., 1986, herein incorporated by reference for general instruction on bio-production.

[0222] Although embodiments of the present invention may be performed in batch mode, or in fed-batch mode, it is contemplated that the invention would be adaptable to continuous bio-production methods. Continuous bio-production is considered an open system where a defined bio-production medium is added continuously to a bioreactor and an equal amount of conditioned media is removed simultaneously for processing. Continuous bio-production generally maintains the cultures within a controlled density range where cells are primarily in log phase growth. Two types of continuous bioreactor operation include a chemostat, wherein fresh media is fed to the vessel while simultaneously removing an equal rate of the vessel contents. The limitation of this approach is that cells are lost and high cell density generally is not achievable. In fact, typically one can obtain much higher cell density with a fed-batch process. Another continuous bioreactor utilizes perfusion culture, which is similar to the chemostat approach except that the stream that is removed from the vessel is subjected to a separation technique which recycles viable cells back to the vessel. This type of continuous bioreactor operation has been shown to yield significantly higher cell densities than fed-batch and can be operated continuously. Continuous bio-production is particularly advantageous for industrial operations because it has less down time associated with draining, cleaning and preparing the equipment for the next bio-production event. Furthermore, it is typically more economical to continuously operate downstream unit operations, such as distillation, than to run them in batch mode.

[0223] Continuous bio-production allows for the modulation of one factor or any number of factors that affect cell growth or end product concentration. For example, one method will maintain a limiting nutrient such as the carbon source or nitrogen level at a fixed rate and allow all other parameters to moderate. In other systems a number of factors affecting growth can be altered continuously while the cell concentration, measured by media turbidity, is kept constant. Methods of modulating nutrients and growth factors for continuous bio-production processes as well as techniques for maximizing the rate of product formation are well known in the art of industrial microbiology and a variety of methods are detailed by Brock, supra.

[0224] It is contemplated that embodiments of the present invention may be practiced using either batch, fed-batch or continuous processes and that any known mode of bio-production would be suitable. It is contemplated that cells may be immobilized on an inert scaffold as whole cell catalysts and subjected to suitable bio-production conditions for chemical product bio-production, or be cultured in liquid media in a vessel, such as a culture vessel. Thus, embodiments used in such processes, and in bio-production systems using these processes, include a population of genetically modified microorganisms of the present invention, a culture system comprising such population in a media comprising nutrients for the population, and methods of making a selected chemical product.

[0225] Embodiments of the invention include methods of making a selected chemical product in a bio-production system, some of which methods may include obtaining a fatty acid or fatty acid derived product after such bio-production event. For example, a method of making a fatty acid or fatty acid derived product may comprise: providing to a culture vessel a media comprising suitable nutrients; providing to the culture vessel an inoculum of a genetically modified microorganism comprising genetic modifications described herein such that the microorganism produces a selected chemical product from syngas and/or a sugar molecule; and maintaining the culture vessel under suitable conditions for the genetically modified microorganism to produce a selected chemical product.

[0226] It is within the scope of the present invention to produce, and to utilize in bio-production methods and systems, including industrial bio-production systems for production of a selected chemical product, a recombinant microorganism genetically engineered to modify one or more aspects effective to increase chemical product bio-production by at least 20 percent over control microorganism lacking the one or more modifications.

[0227] In various embodiments, the invention is directed to a system for bio-production of a chemical product as described herein, said system comprising: a fermentation tank suitable for microorganism cell culture; a line for discharging contents from the fermentation tank to an extraction and/or separation vessel; and an extraction and/or separation vessel suitable for removal of the chemical product from cell culture waste. In various embodiments, the system includes one or more pre-fermentation tanks, distillation columns, centrifuge vessels, back extraction columns, mixing vessels, or combinations thereof.

[0228] The following published resources are incorporated by reference herein for their respective teachings to indicate the level of skill in these relevant arts, and as needed to support a disclosure that teaches how to make and use methods of industrial bio-production of chemical product(s) produced under the invention, from sugar sources, and also industrial systems that may be used to achieve such conversion with any of the recombinant microorganisms of the present invention (Biochemical Engineering Fundamentals, 2.sup.nd Ed. J. E. Bailey and D. F. Ollis, McGraw Hill, N.Y., 1986, entire book for purposes indicated and Chapter 9, pages 533-657 in particular for biological reactor design; Unit Operations of Chemical Engineering, 5th Ed., W. L. McCabe et al., McGraw Hill, N.Y. 1993, entire book for purposes indicated, and particularly for process and separation technologies analyses; Equilibrium Staged Separations, P. C. Wankat, Prentice Hall, Englewood Cliffs, N.J. USA, 1988, entire book for separation technologies teachings).

[0229] F. Production Metrics

[0230] In some embodiments, the genetic modification increases microbial synthesis of a selected fatty acid or fatty acid derived chemical product above a rate or titer of a control microorganism lacking said at least one genetic modification to produce a selected chemical product. In some embodiments, the genetic modification is effective to increase enzymatic conversions to a selected chemical product by at least about 5 percent, at least about 10 percent, at least about 20 percent, at least about 30 percent, or at least about 50 percent above the enzymatic conversion of a control microorganism lacking the genetic modification. In various embodiments, bio-production of a selected chemical product may reach at least 1, at least 2, at least 5, at least 10, at least 20, at least 30, at least 40, and at least 50 g/liter titer, such as by using one of the methods disclosed herein.

[0231] As may be realized by appreciation of the advances disclosed herein as they relate to commercial fermentations of selected chemical products, embodiments of the present invention may be combined with other genetic modifications and/or method or system modulations so as to obtain a microorganism (and corresponding method) effective to produce at least 10, at least 20, at least 30, at least 40, at least 45, at least 50, at least 80, at least 100, or at least 120 grams of a chemical product (such as a fatty acid or fatty acid derivative) per liter of final (e.g., spent) fermentation broth while achieving this with specific and/or volumetric productivity rates as disclosed herein. The amount of a chemical product produced in a bio-production media generally can be determined using a number of methods known in the art, for example, high performance liquid chromatography (HPLC), gas chromatography (GC), or GC/Mass Spectroscopy (MS).

[0232] Unexpected increases in specific productivity by a population of a genetically modified microorganism may be achieved in methods and systems in which that microorganism has a microbial production pathway from malonyl-CoA to a selected chemical product as well as a reduction in the enzymatic activity of a selected enzyme of the microorganism's fatty acid synthase system (more particularly, its malonyl-ACP dependent fatty acid elongation enzymes), in addition to the increase activity of a microorganisms malonyl-CoA dependent fatty acyl-CoA production pathway.

[0233] In some embodiments a microbial chemical bio-production event (i.e., a fermentation event using a cultured population of a microorganism) proceeds using a genetically modified microorganism as described herein, wherein the specific productivity is between 0.01 and 0.60 grams of selected chemical product produced per gram of microorganism cell on a dry weight basis per hour (g chemical product/g DCW-hr). In various embodiments the specific productivity is greater than 0.01, greater than 0.05, greater than 0.10, greater than 0.15, greater than 0.20, greater than 0.25, greater than 0.30, greater than 0.35, greater than 0.40, greater than 0.45, or greater than 0.50 g chemical product/g DCW-hr. Specific productivity may be assessed over a 2, 4, 6, 8, 12 or 24 hour period in a particular microbial chemical production event. More particularly, the specific productivity for a chemical product is between 0.05 and 0.10, 0.10 and 0.15, 0.15 and 0.20, 0.20 and 0.25, 0.25 and 0.30, 0.30 and 0.35, 0.35 and 0.40, 0.40 and 0.45, or 0.45 and 0.50 g chemical product/g DCW-hr., 0.50 and 0.55, or 0.55 and 0.60 g chemical product/g DCW-hr. Various embodiments comprise culture systems demonstrating such productivity.

[0234] Also, in various embodiments of the present invention the volumetric productivity achieved may be about 0.25 g fatty acid (or other chemical product) per liter per hour (g (chemical product)/L-hr), may be greater than about 0.25 g fatty acid (or other chemical product)/L-hr, may be greater than about 0.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 1.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 1.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 2.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 2.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 3.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 3.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 4.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 4.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 5.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 5.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 6.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 6.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 7.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 7.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 8.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 8.50 g fatty acid (or other chemical product)/L-hr, may be greater than about 9.0 g fatty acid (or other chemical product)/L-hr, may be greater than about 9.50 g fatty acid (or other chemical product)/L-hr, or may be greater than about 10.0 g fatty acid (or other chemical product)/L-hr.

[0235] In some embodiments, specific productivity as measured over a 24-hour fermentation (culture) period may be greater than about 0.01, 0.05, 0.10, 0.20, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0 or 12.0 grams of chemical product per gram DCW of microorganisms (based on the final DCW at the end of the 24-hour period).

[0236] In various aspects and embodiments of the present invention, there is a resulting substantial increase in microorganism specific productivity that advances the fermentation art and commercial economic feasibility of microbial chemical production, such as of a fatty acid (but not limited thereto).

[0237] Stated in another manner, in various embodiments the specific productivity exceeds (is at least) 0.01 g chemical product/g DCW-hr, exceeds (is at least) 0.05 g chemical product/g DCW-hr, exceeds (is at least) 0.10 g chemical product/g DCW-hr, exceeds (is at least) 0.15 g chemical product/g DCW-hr, exceeds (is at least) 0.20 g chemical product/g DCW-hr, exceeds (is at least) 0.25 g chemical product/g DCW-hr, exceeds (is at least) 0.30 g chemical product/g DCW-hr, exceeds (is at least) 0.35 g chemical product/g DCW-hr, exceeds (is at least) 0.40 g chemical product/g DCW-hr, exceeds (is at least) 0.45 g chemical product/g DCW-hr, exceeds (is at least) 0.50 g chemical product/g DCW-hr, exceeds (is at least) 0.60 g chemical product/g DCW-hr. In accordance with certain embodiments of the present invention the chemical product is a fatty acid or a fatty acid derived product.

[0238] More generally, based on various combinations of the genetic modifications described herein, optionally in combination with supplementations described herein, specific productivity values for a fatty acid or fatty acid derived product, and for other chemical products described herein, may exceed 0.01 g chemical product/g DCW-hr, may exceed 0.05 g chemical product/g DCW-hr, may exceed 0.10 g chemical product/g DCW-hr, may exceed 0.15 g chemical product/g DCW-hr, may exceed 0.20 g chemical product/g DCW-hr, may exceed 0.25 g chemical product/g DCW-hr, may exceed 0.30 g chemical product/g DCW-hr, may exceed 0.35 g chemical product/g DCW-hr, may exceed 0.40 g chemical product/g DCW-hr, may exceed 0.45 g chemical product/g DCW-hr, and may exceed 0.50 g or 0.60 chemical product/g DCW-hr. Such specific productivity may be assessed over a 2, 4, 6, 8, 12 or 24 hour period in a particular microbial chemical production event.

[0239] The improvements achieved by embodiments of the present invention may be determined by percentage increase in specific productivity, or by percentage increase in volumetric productivity, compared with an appropriate control microorganism lacking the particular genetic modification combinations taught herein (with or without the supplements taught herein, added to a vessel comprising the microorganism population). For particular embodiments and groups thereof, such specific productivity and/or volumetric productivity improvements is/are at least 10, at least 20, at least 30, at least 40, at least 50, at least 100, at least 200, at least 300, at least 400, and at least 500 percent over the respective specific productivity and/or volumetric productivity of such appropriate control microorganism.

[0240] The specific methods and teachings of the specification, and/or cited references that are incorporated by reference, may be incorporated into the examples. Also, production of a chemical product may reach at least 1, at least 2, at least 5, at least 10, at least 20, at least 30, at least 40, and at least 50 g/liter titer in various embodiments.

[0241] The metrics may be applicable to any of the compositions, e.g., genetically modified microorganisms, methods, e.g., of producing chemical products, and systems, e.g., fermentation systems utilizing the genetically modified microorganisms and/or methods disclosed herein.

[0242] It is appreciated that iterative improvements using the strategies and methods provided herein, and based on the discoveries of the interrelationships of the pathways and pathway portions, may lead to even greater chemical product bio-production at the conclusion of a bio-production event.

VI. Products ProducedThe Chemical Product

[0243] The novel bioproduction pathways, fermentation processes and genetically modified microorganisms described herein are engineered to produce various chemical products of interest. One chemical product may be a fatty acid of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms. This group of chemical products includes: butyrate or butyric acid, valerate or valeric acid, caproate or caproic acid, enanthate or enanthic acid, caprylate or caprylic acid, pelargonate or pelargonic acid, caprate or capric acid, undecylate or undecylic acid, laurate or lauric acid, tridecylate or tridecylic acid, myristate or myristic acid, pentadecylate or pentadecylic acid, palmitate or palmitic acid, margarate or margaric acid, stearate or stearic acid, nonadecylate or nonadecylic acid, arachidate or arachidic acid. These fatty acid products may be produced from a fatty acyl-CoA intermediate via the activity of a fatty acyl-CoA thioesterase or wax ester synthase. Alternatively, these fatty acids may be produced from a fatty acyl-CoA intermediate via concerted activities of a fatty acyl-CoA phosphotransferase first producing a fatty acyl-phosphate and then the action of a fatty acid kinase operating to produce a fatty acid from the fatty acyl-phosphate.

[0244] Another chemical product may be a fatty aldehyde of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms. This group of chemical products includes: Butyraldehyde, Valeraldehyde, Caproaldehyde, Enanthaldehyde, Caprylaldehyde, Pelargonaldehyde, Capraldehyde, Undecylaldehyde, Lauraldehyde, Tridecylaldehyde, Myristaldehyde, Pentadecylaldehyde, Palmitaldehyde, Margaraldehyde, Stearaldehyde, Nonadecylaldehyde, and Arachidaldehyde. These aldehyde products may be produced from a fatty acyl-CoA intermediate via the activity of a fatty acyl-CoA reductase or acyl-CoA reductase. Production strains making fatty acids may also be used to produce fatty aldehydes.

[0245] Another chemical product may be a fatty alcohol of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms. This group of chemical products includes: butanol, amyl alcohol, hexanol, heptanol, octanol, nonanol, decanol, hendecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol, heptadecanol, octadecanol, nonadecanol, and eicosanol. These fatty acid products may be produced from a fatty aldehyde via the activity of an aldehyde reductase. Production strains making fatty acids may also be used to produce fatty alcohols by expressing genes encoding enzymes that convert fatty acyl-CoA or free fatty acids to fatty alcohols. Examples of these enzymes include an alcohol-forming acyl-CoA reductase (EC 1.2.1.-), or a long-chain-fatty-acyl-CoA reductase (EC 1.2.1.50) plus an alcohol dehydrogenase (EC 1.1.1.1), or a combination of an aldehyde dehydrogenase (EC 1.2.1.-) and an alcohol dehydrogenase. A polypeptide with fatty acyl-CoA reductase activity is provided by the fabG gene of Acinetobacter SP. ADP1, accession number YP_047869. A polypeptide with fatty-acyl reductase activity is provided by the FAR-N_SDR_e gene of Bombyx mori, accession number BAC79425. A polypeptide with aldehyde dehydrogenase is provided by the ALDH gene of Geobacillus thermodenitrificans NG80-2, accession number YP_001125970. A polypeptide with alcohol dehydrogenase activity is provided by the yqhD gene of E. coli, accession number AP_003562.1. Additional sources of these activities are known to the art and can be combined to generate a production strain that produces fatty alcohols.

[0246] Another chemical product may be an alpha olefin of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms.

[0247] Another chemical product may be an alkane of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms.

[0248] Another chemical product may be a diacid of any chain length from 4 to greater than 18 carbons, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, or more carbon atoms. These fatty acid derived products may be produced from a fatty acid via omega or terminal oxidation by enzymes known in the art.

[0249] Any of these may be described herein as a selected chemical product, or a chemical product of interest or as a fatty acid product or as a fatty acid derivative or fatty acid product derivative. Also, any grouping, including any sub-group, of the above listing may be considered what is referred to by selected chemical product, chemical product of interest, and the like. For any of these chemical products a microorganism may inherently comprise a biosynthesis pathway to such chemical product and/or may require addition of one or more heterologous nucleic acid sequences to provide or complete such a biosynthesis pathway, in order to achieve a desired production of such chemical product.

VII. Disclosed Embodiments are Non-Limiting

[0250] While various embodiments of the present invention have been shown and described herein, it is emphasized that such embodiments are provided by way of example only. Numerous variations, changes and substitutions may be made without departing from the invention herein in its various embodiments. Specifically, and for whatever reason, for any grouping of compounds, nucleic acid sequences, polypeptides including specific proteins including functional enzymes, metabolic pathway enzymes or intermediates, elements, or other compositions, or concentrations stated or otherwise presented herein in a list, table, or other grouping (such as metabolic pathway enzymes shown in a scheme), unless clearly stated otherwise, it is intended that each such grouping provides the basis for and serves to identify various subset embodiments, the subset embodiments in their broadest scope comprising every subset of such grouping by exclusion of one or more members (or subsets) of the respective stated grouping. Moreover, when any range is described herein, unless clearly stated otherwise, that range includes all values therein and all sub-ranges therein.

[0251] Also, and more generally, in accordance with disclosures, discussions, examples and embodiments herein, there may be employed conventional molecular biology, cellular biology, microbiology, and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. (See, e.g., Sambrook and Russell, Molecular Cloning: A Laboratory Manual, Third Edition 2001 (volumes 1-3), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Animal Cell Culture, R. I. Freshney, ed., 1986.) These published resources are incorporated by reference herein for their respective teachings of standard laboratory methods found therein. Such incorporation, at a minimum, is for the specific teaching and/or other purpose that may be noted when citing the reference herein. If a specific teaching and/or other purpose is not so noted, then the published resource is specifically incorporated for the teaching(s) indicated by one or more of the title, abstract, and/or summary of the reference. If no such specifically identified teaching and/or other purpose may be so relevant, then the published resource is incorporated in order to more fully describe the state of the art to which the present invention pertains, and/or to provide such teachings as are generally known to those skilled in the art, as may be applicable. However, it is specifically stated that a citation of a published resource herein shall not be construed as an admission that such is prior art to the present invention. Also, in the event that one or more of the incorporated published resources differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls. Subject matter in the Examples is incorporated into this section to the extent not already present.

EXAMPLES

Example 1NphT7 Mutants

[0252] The enzyme NphT7 is a 3-keto-acyl-CoA synthase that is active with acetyl-CoA as the primer and malonyl-CoA as the extender donor to generate a 3-keto-C4-CoA product; the native enzyme has no detectable activity on longer chain primers. Residue modifications were made to NphT7 to alter the acyl-CoA binding pocket to accept substrates with chain lengths greater than or equal to 4 carbons. These modifications are single amino acid changes, combinations of single amino acid changes, and targeted structural loop modifications that allow the condensation reaction of acyl-CoAs with chain lengths greater than or equal to 4 carbons, such as C4-CoA and C6-CoA, with malonyl-CoA. The modifications were made based on the following criteria:

[0253] (a) Examination of the crystal structure of a related enzyme, the fabH from Mycobacterium tuberculosis (structure 1U6S in the Protein DataBase) identified the residues in Table 16 that contact the acyl chain. The corresponding residues in NphT7 were identified based on homology.

TABLE-US-00017 TABLE 16 Residues of mtFabH that contact the acyl chain in the substrates. 1U6S (mtFabH) Asn B81 Thr B82 Leu B142 Thr B145 Phe B157 Ile B189 Ser B276 Val B205 Gln A86 Thr A87 Ile A196 Tyr 304

[0254] (b) Lid swap mutants. Comparison of the sequence and structural homologies between the mtFabH and NphT7 reveals a predicted L9 loop in NphT7 comprising residues 167-201. The amino acid sequence: GGLTDLIRVPAGGSRQPLDTDGLDAGLQYFAMD, makes up the L9 loops structure corresponding to the acyl-CoA lid. Saturated mutagenesis of the lid (Conversion of each amino acid in the lid to every other amino acid, and combinations of mutations within the lid) may change the lid structure to accept larger acyl-CoA chains.

[0255] Mutant nphT7 genes were constructed by oligonucleotide-directed mutagenesis and all mutants were verified to be correct by DNA sequencing. Parent and mutant nphT7 genes were cloned in pET28b vectors in frame with 6 His residues, transformed into E. coli BL21(DE3), and cultures in Terrific Broth containing 35 g/ml kanamycin were incubated at 37 C. until the OD.sub.600 was 0.4. Expression was induced by the addition of 0.1 mM IPTG. Cells were incubated at 18 C. and harvested after 18 hours by centrifugation at 4,000 rpm at 4 C. for 10 minutes. Pellets were stored at 80 C. prior to lysis. Lysates were prepared by resuspending cells in 50 mL Lysis Buffer (25 mM Tris, pH 8.0, 300 mM NaCl, 5 mM -mercaptoethanol, and benzonase nuclease) and lysing with a Microfluidizer (two passes). Soluble fractions were isolated by centrifugation at 12,000 RPM at 4 C. for 30 minutes. Expression was analyzed by SDS-PAGE (coomassie staining) and anti-His western blotting (4 g soluble/lane, maintained same volume for soluble/insoluble fraction). NphT7 enzymes were purified by Ni-NTA chromatography. Loop mutants mtloop1 and mtloop2 were additionally purified using DEAE-Sepharose chromatography.

[0256] 3-ketoacyl-CoA synthase activity was monitored by measuring the release of free CoA-SH using the 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB) reagent, malonyl-CoA as the donor substrate, and various primer substrates (acetyl-CoA, C4-CoA, C6-CoA, or C10-CoA). The increase in absorbance at 412 nm at 37 C. (TNB product formation; =14.14 mM.sup.1cm.sup.1 in phosphate buffer at pH 8.0) was used to determine the enzymatic activity. 3-ketoacyl-CoA synthase activity was also monitored by coupling the production of 3-ketoacyl-CoA to the subsequent formation of the 3-hydroxyacyl-CoA product by purified PaFabG and NADPH. Reactions were carried out at room temperature for 30 min, stopped by the addition of acetonitrile to 20% and incubating on ice for 15 minutes, and analyzed by UPLC-MS/MS.

[0257] Specific activities of various engineered NphT7 mutants are shown in Table 17. In addition, by measuring the products of the reactions using UPLC-mass spectrometry, it was demonstrated that the variant of NphT7 with the I147T, F217V mutations produces 3-keto-C6-CoA from C4-CoA, 3-keto-C8-CoA from C6-CoA, and 3-keto-C12-CoA from C10-CoA using malonyl-CoA as the extender donor (see FIG. 15; products of the NphT7 reaction were converted to the 3-OH-acyl-CoA by PaFabG to allow quantitation by UPLC-MS). As may be seen from these results, modification of selected residues of NphT7 alters the substrate preference from the almost exclusive preference for acetyl-CoA in the wildtype enzyme to variants that have significant activity on C4-CoA, for example variants I147F, F217V and I147S, F217V.

TABLE-US-00018 TABLE 17 Specific Activities for various NphT7 Mutants S.A. (mol/min./mg), purified Acetyl- Mutation(s) CoA C4-CoA C6-CoA C10-CoA N/A (wild-type) gi|299758082 5.52 0.003 0 ND F217V 0.544 0.722 0.03 0.003 I147T 2.05 0.015 0.010 ND I147T, F217V 0.251 0.47 0.025 0.003 I147F, F217V 0.461 1.39 0.054 0.02 I147M, F217V 0.41 1.103 0.049 0.013 I147S, F217V 0.855 1.87 0.075 0.009 Y144L, I147T, F216V 0.005 0.065 0.059 0.007 ND = Not Determined

Example 2Strategies for Identifying 3-Ketoacyl-CoA Synthase Candidates

[0258] NphT7 is an ideal place to begin forming a strategy for identifying other 3-ketoacyl-CoA synthase candidates because unlike type II FAS 3-ketoacyl-ACP synthases (KAS) that uses malonyl-ACP as an extender, it can perform the targeted reaction using malonyl-CoA and therefore, homologs of NphT7 would likely have maintained specificity for malonyl-CoA. In addition, KAS III from various organisms have been characterized by crystal structures and biochemical assays to define substrate binding sites and substrate specificities. Unlike NphT7, KAS III from various organisms have shown different specificity for longer or branched acyl-CoA . There is similar information available for KAS I and KAS II but unlike KAS III that utilizes acyl-CoA as a substrate for the condensation reaction, they require acyl-ACP as a substrate. Therefore, crystal structures of known KAS III along with biochemical data provide guidance in identifying conserved residues that recognize acyl-CoA and aid in identification of NphT7 homologs that utilize longer chain acyl-CoAs.

TABLE-US-00019 TABLE 18 Summary of substrate specificity of KAS III from different organisms KAS III (FabH homologs) Substrate Specificity.sup.a Reference E. coli FabH C2-C3 Choi et al., J Bact. 2000 B. subtilis FabH1 C2-C8* Choi et al., J Bact. 2000 B. subtilis FabH2 C2-C7* Choi et al., J Bact. 2000 S. aureus FabH C2-C16* Qui et al., Protein Science 2005 S. pneumoniae FabH C2-C4 Khandekar et. al., J. Bio. Chem. 2001 M. tuberculosis FabH C8-C16 Choi et al., J Bio. Chem. 2000 .sup.aSubstrate specificity determined by enzyme activity *Substrates include branched chain acyl-CoA

[0259] Okamura et al. (PNAS, 2010) defines the biochemical function of NphT7 and compares the amino acid sequence to other NphT7 homologs and E. coli KAS III, FabH (ecFabH). Mainly, the well characterized ecFabH is used to describe the similarities between all NphT7 (NphT7 and 6 NphT7 homologs) and the main differences to KAS III. The information provided by Okaramura et al. with addition of other reports describing other KAS III will be used to define rules for identifying potential 3-ketoacyl-CoA candidates.

[0260] The following five strategies for identifying 3-ketoacyl-CoA candidates were used:

1. BLASTp to identify NphT7 homologs

[0261] Rationale: [0262] a. Most likely to utilize malonyl-CoA as an extender for the condensation reaction
2. Identify homologs that contains (A/G)GGSR sequence motif

[0263] Rationale: [0264] a. The predicted L9 loops in the NphT7 homologs are inserted with additional sequence and share an (A/G)GGSR sequence motif [0265] b. Okamura et al. suggest (A/G)GGSR motif may serve as one of recognition sites for the CoA moiety of the extender substrate malonyl-CoA [0266] c. (A/G)GGSR motif and additional sequence are not found in KAS III homologs, thus indicating the sequence motif is specific to NphT7 homologs [0267] d. Reference [0268] i. Okamura et al., PNAS 2010
3. Select for NphT7 homologs that do not contain STPDXPQ sequence motif

[0269] Rationale: [0270] a. Phe87 residue that dictates primer substrate specificity in ecFabH (KAS III) is replaced by Gln in the NphT7 homologs. [0271] b. All NphT7 homologs share a STPDXPQ sequence motif with Gln being part of the sequence motif. [0272] c. KAS III homologs do not have conserved STPDXPQ motif. [0273] d. Reference [0274] i. Okamura et al., PNAS 2010
4. Identify homologs that contain only hydrophobic residues in the substrate binding site

[0275] Rationale: [0276] a. Phe87, Leu142, Phe157, Leu188, and Leu205 of ecFabH that form the hydrophobic pocket for recognition of the acetyl methyl group of acetyl-CoA are not conserved in NphT7 homologs [0277] i. NphT7 has 3 out 5 amino acids that are hydrophobic residues [0278] b. Most hydrophobic residues are conserved among KAS III homologs. [0279] c. Reference [0280] i. Okamura et al., PNAS 2010 [0281] ii. Qui et al., Protein Science 2005 [0282] iii. Scarsdale et al., JBC 2001 [0283] iv. Qui et al., JBC 1999
5. Identify different families of NphT7 homologs

[0284] Rationale: [0285] a. Phylogenetic tree created from multiple sequence alignment (MSA) of NphT7 homologs that have met the above requirements will be used to select candidates that would represent the most diverse group of NphT7 homologs that have evolved from different ancestors. [0286] i. The diversity would allow for the highest possibility of finding an NphT7 homolog with different specificity due to evolving from different ancestors

Result/Outcome

[0287] The following summarizes the results from the five strategies for identifying 3-ketoacyl-CoA candidates outlined above:

[0288] 1. Homology search of NphT7 [0289] a. BLAST search was performed with NphT7 as a reference sequence with maximum sequence target at 10,000 without a cutoff for e-value [0290] i. BLAST search resulted in 7141 homologs of NphT7

[0291] 2. Select for NphT7 homologs with (A/G)GGSR motif [0292] a. 309 NphT7 homologs had (A/G)GGSR motif.

[0293] 3. Elimination of homologs with STPDXPQ sequence motif [0294] a. 288 NphT7 homologs did not have STPDXPQ motif

[0295] 4. Selection based on conservation of hydrophobic residues in the substrate binding pocket [0296] a. Of the 288 homologs, 144 NphT7 homologs had hydrophobic residues at the 5 residues that are conserved between KAS III

[0297] 5. Selection based on evolutionary distance from NphT7 and known KAS III [0298] a. Phylogenetic tree constructed from MSA of NphT7 homologs, NphT7, ecFabH, mtFabH, bsFabH1, bsFabH2, saFabH, spFabH with (A/G)GGSR sequence motif indicate there are 6 different families of NphT7 homologs (Table 17). [0299] b. 22 3-ketoacyl-CoA synthase candidate were chosen to cover all 6 families [0300] i. 10 3-ketoacyl-CoA synthase candidates [0301] 1. With (A/G)GGSR sequence motif [0302] 2. Without STPDXPQ sequence motif [0303] 3. With conserved hydrophobic residues [0304] ii. 11 3-ketoacyl-CoA synthase candidates [0305] 1. With (A/G)GGSR sequence motif [0306] 2. Without STPDXPQ sequence motif [0307] iii. 1 3-ketoacyl-CoA synthase candidates [0308] 1. With (A/G)GGSR sequence motif

List of 3-Ketoacyl-CoA Synthase Candidates

[0309] [00236] lists the 3-ketoacyl-CoA synthase chosen based on the criteria described above. In addition, each synthase was aligned to NphT7, ecFabH, mtFabH, and saFabH to determine percent sequence identity, similarity and gap. Synthases 1-10 are chosen based on having all criteria met. Synthases 11-21 are chosen based on having all criteria met except for conserved hydrophobic residues. Synthase 22 is chosen based on having (A/G)GGSR sequence motif

TABLE-US-00020 TABLE 19 List of 3-ketoacyl-CoA synthases NphT7 ecFabH saFabH mtFabH % % % % % % % % % % % % Organism Protein ID Identity positive Gap Identity positive Gap Identity positive Gap Identity positive Gap 1 Rhodothermus YP_004823890.1 39 56 1 37 54 3 35 55 5 36 50 5 marinus (SEQ ID NO 198) SG0.5JP17-172 2 Streptomyces YP_007523119.1 38 51 0 42 55 3 38 56 5 38 52 2 davawensis JCM (SEQ ID NO 199) 4913 3 Chlamydophila YP_004377485.1 36 55 1 46 63 3 40 59 4 33 53 3 pecorum E58 (SEQ ID NO 200) 4 Clostridium ZP_23165498.1 36 54 1 43 60 2 44 64 4 34 53 7 ultunense Esp (SEQ ID NO 201) 5 Corallococcus YP_005368607.1 42 57 2 49 63 5 34 54 6 41 62 6 coralloides DSM (SEQ ID NO 202) 2259 6 Desmospora sp. ZP_08463153.1 43 60 1 50 67 2 38 57 1 52 72 4 8437 (SEQ ID NO 203) 7 Paenibacillus ZP_10239638.1 44 58 1 47 65 2 38 55 1 55 73 4 peoriae KCTC (SEQ ID NO 204) 3763 8 Pelosinus ZP_10324886.1 41 62 2 46 64 2 38 57 3 48 69 4 fermentans DSM (SEQ ID NO 205) 17108 9 Candidatus YP_828246.1 35 53 2 42 60 3 37 58 5 35 52 10 Solibacter (SEQ ID NO 206) usitatus Ellin6076 10 Desulfotomaculum ZP_08114352.1 40 59 1 46 66 2 47 69 4 37 55 6 nigrificans (SEQ ID NO 207) DSM 574 11 Saccharomonospora ZP_10013188.1 40 55 2 32 51 5 35 54 5 30 48 5 glauca K62 (SEQ ID NO 208) 12 Corallococcus ADI59524.1 29 47 2 33 48 7 27 47 6 25 41 8 coralloides (SEQ ID NO 209) 13 Legionella YP_001250982.1 32 45 6 32 50 5 31 53 6 27 45 3 pneumophila str. (SEQ ID NO 210) Corby 14 Streptomyces BAB69376.1 42 54 2 40 55 3 36 54 5 38 51 2 avermitilis (SEQ ID NO 211) 15 Verrucosispora YP_004406674.1 41 52 5 36 55 3 36 56 5 36 50 11 maris AB-18-032 (SEQ ID NO 212) 16 Rhodopirellula CAD74700.1 42 57 6 42 56 8 40 60 9 32 47 9 baltica SH 1 (SEQ ID NO 213) 17 Candidatus YP_003206328.1 43 58 2 48 66 3 44 65 4 39 54 3 Methylomirabilis (SEQ ID NO 214) oxyfera 18 Thermaerobacter YP_004101787.1 43 59 1 47 62 2 46 65 4 41 56 3 marianensis DSM (SEQ ID NO 215) 12885 19 Caldisericum exile YP_005472409.1 38 59 1 49 64 2 47 69 4 37 57 4 AZM16c01 (SEQ ID NO 216) 20 Indibacter ZP_11015628.1 30 51 2 40 59 3 39 59 6 33 50 4 alkaliphilus LW1 (SEQ ID NO 217) 21 Candidatus YP_001957829.1 34 52 3 36 57 4 37 57 5 32 50 3 Amoebophilus (SEQ ID NO 218) asiaticus 5a2 22 Flavobacterium ZP_10480443.1 34 52 2 38 56 3 38 57 4 31 53 5 sp. F52 (SEQ ID NO 219)

Example 3Combining NphT7 Variants and/or fabH Homologs and Thioesterases to Produce Fatty Acids with Specified Chain Lengths

[0310] While mutants of NphT7 were engineered that are capable of extending acyl-CoAs of chain length C4, C6, and C10, the specific activities of these enzymes are relatively low for the higher chain lengths. The extension by 2 carbon lengths of acyl-CoAs to form 3-keto-acyl-CoAs is a reaction also carried out by keto-acyl-CoA synthases known as KASIII enzymes, encoded by fabH gene homologs. A number of such gene homologs were synthesized using codons for optimal expression in E. coli by a commercial DNA synthesis provider (DNA2.0) and fused with 6 His residues at the N-terminus for purification of the proteins by affinity chromatography. The genes were expressed in E. coli and KAS activity was assayed using the DTNB assay for CoA-SH release from the condensation of malonyl-CoA with acyl-CoAs of varying chain lengths. Table 20 lists the enzyme homologs with sufficiently high level KAS activity to enable such enzymes to extend the acyl-CoAs of the various chain lengths noted in the table. As may be seen from the results in Table 20, FabH enzymes from different sources have different substrate chain-length preferences.

TABLE-US-00021 TABLE 20 High level KAS activity Enzymes Organisms Acetyl-CoA Streptomyces sp. (strain CL190) Pelosinus fermentans DSM 17108 Saccharomonospora glauca K62 Verrucosispora maris AB-18-032 Clostridiales bacterium 1_7_47_FAA C4-CoA Streptomyces sp. (strain CL190) Saccharomonospora glauca K62 Saccharomonospora azurea NA-128 Mesorhizobium sp. STM 4661 Clostridiales bacterium 1_7_47_FAA C6-CoA Gordonia aichiensis NBRC 108223 Arcobacter butzleri ED-1 Clostridiales bacterium 1_7_47_FAA Saccharomonospora glauca K62 Ralstonia solanacearum Po82 C8-CoA Gordonia aichiensis NBRC 108223 Gluconacetobacter oboediens 174Bp2 Arcobacter butzleri ED-1 Ralstonia solanacearum Po82 Phaeobacter gallaeciensis 2.10 C10-CoA Alishewanella aestuarii B11 Streptomyces sp. (strain CL190)

[0311] A further approach to chain length specificity can be achieved by targeting the release of fatty acids from the acyl-CoA precursor. The genes encoding a variety of thioesterases were synthesized using codons optimized for expression in E. coli by a commercial DNA synthesis provider (DNA2.0) and the genes expressed. Purification of the enzymes was enabled by affinity chromatography based on the N-terminal 6His affinity tag. The activity of this variety of thioesterases on acyl-CoAs of different chain lengths was assessed (FIG. 16). Thus while thioesterase PA2801TE has broad specificity from C6-CoA to C16-CoA, thioesterase tesA has no detectable activity on acyl-CoAs shorter than C10, and is minimally active on C10-CoA.

[0312] Thus the incorporation of an NphT7 variant, a FabH with the desired specificity as shown in Table 20, and the appropriate thioesterase as shown in FIG. 16 into a recombinant organism along with the enzymes that comprise an engineered fatty acid pathway enables the targeted production of fatty acids with specified chain lengths.

Example 4Shake Flask Free Fatty Acid (FFA) Productions

[0313] A number of genetically modified E. coli strains were evaluated for production of free fatty acids. These strains comprise an engineered host based on strain BW25113 and with the additional genetic modifications: ldhA::frt, pflB::frt, mgsA::frt, poxB::frt, pta-ack::frt; a temperature-sensitive allele of fabI (fabI S241F); and the additional modifications: tig::frt, atoDAEB::frt, AfadD::frt that minimize diversion of acyl-CoA substrates or fatty acid products. Genes encoding NphT7, one or more thioesterases, a keto-CoA reductase (KCR), a 3-hydroxy-acyl-CoA dehydratase (3HDh), and an enoyl-CoA reductase (EnCr) are also provided on plasmids. The genes present in samples 1-5 are depicted in Table 21.

TABLE-US-00022 TABLE 21 Genes Present in Samples 1-5 3- ketoacyl- Strain CoA Thio- Sample synthase esterases KCR 3HDh EnCr Sample 1 NphT7 none (control) (SEQ ID NO 1) Sample 2 NphT7 tesA Hbd Crt Ter (SEQ ID (SEQ ID (SEQ ID (SEQ ID (SEQ ID NO 1) NO 277) NO 271) NO 272) NO 275) Sample 3 NphT7 FadA FadB FadB Ter (SEQ ID (SEQ ID (SEQ ID (SEQ ID (SEQ ID NO 1) NO 182) NO 183) NO 183) NO 275) tesA (SEQ ID NO 278) Sample 4 NphT7 tesA FadB FadB Ter (SEQ ID (SEQ ID (SEQ ID (SEQ ID (SEQ ID NO 1) NO 278) NO 183) NO 183) NO 275) Sample 5 NphT7 tesA FadB FadB Ter (SEQ ID (SEQ ID (SEQ ID (SEQ ID (SEQ ID NO 1) NO 278) NO 183) NO 183) NO 275) fadE (SEQ ID NO 180)

[0314] The rate of producing C8-C18 FFA by these samples is shown in FIG. 17 and the titers for C6-C18 FFA production is shown in FIG. 18. The distribution of chain length specificity with strain sample 3 is shown in FIG. 19; 36% of the product is C14-C16 FFA. These results demonstrate that increased fatty acid production is achieved in these engineered strains, with a titer of 4.07 g/L by Sample 3.

[0315] Alternative KCRs, 3HDh, and EnCr enzymes may be used to provide the requisite activities to convert the keto-acyl-CoA product of NphT7, NphT7 mutants, or fabH homologs to the fully saturated product elongated by 2 carbons, viz. the reduction of the keto-acyl-CoA to 3-hydroxyacyl-CoA by KCR, the dehydration of the 3-hydroxyacyl-CoA to the enoyl-CoA by 3HDh, and the reduction of the enoyl-CoA to acyl-CoA by EnCr. For example, alternative KCRs including FadIJ, Hbd, and FadB. FadB has sufficient activity as a KCR up to C16 (See the Table 22 below).

TABLE-US-00023 TABLE 22 Activity of FadB on 3-hydroxyacyl-CoAs of different chain lengths Substrate Specific Activity (U/mg) 3-OHC4-CoA 0.4321 3-OHC6-CoA 0.585 3-OHC8-CoA 0.1255 3-OHC10-CoA 0.1777 3-OHC12-CoA 0.1935 3-OHC14-CoA 0.2564 3-OHC16-CoA 0.1158

[0316] Alternative 3HDhs including the bifunctional FadB, FabG, FadJ, and Hbd were tested for activity and product specificity. The results are shown in FIG. 20 expressed as percent of the activity achieved with the most preferred substrate.

[0317] To prevent consumption of the fatty acid product and to maintain chain length specificity, additional host genetic modifications to eliminate thioesterases may be required. These modifications include deletion or modulation of tesB (SEQ ID NO 279), yciA (SEQ ID NO 280), fadM (SEQ ID NO 283), ybgC (SEQ ID NO 281), and ybfF (SEQ ID NO 282).

Example 5. Production of 3-Keto-C.SUB.5.-CoA

[0318] It was demonstrated that odd chain length fatty acids can be produced using the genetically modified enzymes and methods of the present invention. In particular, it was demonstrated that the enzymes NphT7 and NphT7 mutants are active with propionyl-CoA as the primer and malonyl-CoA as the extender donor to generate C5 keto-acyl-CoA. The NphT7 variants and fabHs described herein would further extend the C5 keto-acyl-CoA to make longer chain odd-numbered fatty acid products.

[0319] Freshly purified His6-NphT7, His6-NphT7(I147T, F217V), His6-NphT7(I147S, F217V), and His.sub.6-Hbd were used in all the experiments in this example. NphT7 reactions (200 L) contained 100 mM Tris-HCl (pH 8), 5 mM MgCl.sub.2, 1 mM malonyl-CoA, 1 mM primer CoA (C.sub.2-, or C.sub.3-CoA), and various concentrations of wild-type NphT7 or mutant enzymes. Reactions without any primer CoA but with malonyl-CoA were also run. Formation of Mg.sup.2+-3-keto-acyl-CoA adduct was monitored at 303 nm, at 37 C. for 15 mM. NphT7-Hbd coupled reactions (200 L) contained 100 mM Tris-HCl (pH 8), 0.75 mM NADH, 1 mM malonyl-CoA, 1 mM primer CoA (C2 or C.sub.3-CoA), 10 g of partially purified Hbd, and various concentrations of wild-type NphT7 or mutant enzymes. Reactions without any primer CoA but with malonyl-CoA were also run. Oxidation of NADH was followed at 340 nm, at 37 C. for 15 min. At the end of the 15-minute enzyme reactions, 100 L of samples were removed from each reaction and immediately mixed with 25 L acetonitrile to terminate enzyme reactions. The mixtures were incubated on ice for at least 15 min, followed by centrifugation at 3,220g at 4 C. for 15 mM. Supernatants were saved for UPLC-MS/MS analyses for the detection of 3-keto- and 3-OH-C4 and C.sub.5-CoA. In certain runs, the Hbd enzyme was also used to determine if keto-CoA produced could be reduced to hydroxyacyl-CoA. The experimental results are shown in Table 23.

TABLE-US-00024 TABLE 23 Summary of Substrates and Enzymes use in Experiment Enzyme Amount of Products Produced (ppm) Synthase 3-keto-C4- 3-OHC4- 3-keto-C5- 3-OHC5- Amount CoA CoA CoA CoA Runs Substrate 1 Substrate 2 Synthase Hbd (mg) (851.6) (853.6) (865.6) (867.6) 1 C2-CoA M-CoA NphT7 No 0.00404 587.1 35.9 0.1 0.1 0.00202 393.3 25.8 0.1 0 0.00101 282.4 22.1 0 0.1 0.000505 123.3 17.2 0.1 0 0.000253 61.9 15.4 0.1 0 0 22.3 16.1 0 0 2 C3-CoA M-CoA NphT7 No 0.00404 20 9.4 79.8 5.2 0.00202 10.1 10.6 31.3 2.2 0.00101 6.2 9.6 13.2 1 0.000505 4.9 10.4 6.1 0.5 0.000253 3.2 10.8 3 0.2 0 3.6 11.5 1.1 0.1 3 M-CoA NphT7 No 0.00404 7.9 7.3 0.6 0.1 0.00202 5.9 6.6 0.5 0 0.00101 4.1 5.7 0.4 0 0.000505 2.3 7.8 0.4 0 0.000253 1.5 6.3 0.3 0 0 1 10.3 0.3 0 4 C2-CoA M-CoA NphT7 Yes 0.00404 25 421.3 0.1 0.1 0.00202 17.5 220.6 0.1 0 0.00101 16.8 87.8 0.1 0 0.000505 17.6 34.7 0.1 0.1 5 C3-CoA M-CoA NphT7 Yes 0.00404 4.1 15.4 1.6 46.8 0.00202 2.3 9.1 1.4 13.1 0.00101 1.8 8.2 1.4 3.8 0.000505 1.3 6.8 1.1 1.6 6 M-CoA NphT7 Yes 0.00404 0.9 23.9 0.1 0 0.00202 0.7 13 0.1 0 0.00101 0.7 7 0.1 0 0.000505 0.7 8.2 0.1 0 7 C2-CoA M-CoA NphT7(I147T, F217V) No 0.146 613.6 33.8 0.1 0.1 0.073 637.8 35 0.1 0.1 0.0365 695.1 38.7 0.1 0.1 0.01825 664.1 39.9 0.1 0.1 0 26.6 10.7 0.1 0 8 C3-CoA M-CoA NphT7(I147T, F217V) No 0.146 49.5 5.9 221.1 15.1 0.073 43.2 6.5 232.2 16.1 0.0365 42.5 6 250.7 17.9 0.01825 28.3 4.9 237.9 15.8 0 2.2 8.6 4.5 0.3 9 M-CoA NphT7(I147T, F217V) No 0.146 125.9 12.6 0.3 0.1 0.073 169.1 9.7 0.3 0.1 0.0365 176.7 10.1 0.3 0.1 0.01825 88 12.1 0.3 0.1 0 3.6 8.6 0.3 0 10 C2-CoA M-CoA NphT7(I147T, F217V) Yes 0.146 99.7 568.8 0.1 0.6 0.073 55.3 605.7 0.1 0.6 0.0365 43.9 553.3 0.1 0.4 0 16 15.9 0 0.4 11 C3-CoA M-CoA NphT7(I147T, F217V) Yes 0.146 9.5 55.1 16.5 348.7 0.073 6.3 45.5 14.9 330.3 0.0365 4.7 47.1 13.8 351 0 1.2 8.5 1.1 4.3 12 M-CoA NphT7(I147T, F217V) Yes 0.146 4.4 270.1 0.1 2.1 0.073 4.7 302.5 0.2 1.5 0.0365 1.6 148.6 0.1 1.1 0 0.8 8.6 0.1 0.7 13 C2-CoA M-CoA NphT7(I147S, F217V) No 0.01925 570.3 26.8 0.3 0.1 0.009625 487 24.9 0.2 0.1 0.004813 340.7 19.5 0.2 0.1 0.002406 232.7 15.5 0.2 0 0 20.9 10.2 0.2 0 14 C3-CoA M-CoA NphT7(I147S, F217V) No 0.01925 33.1 5.4 247.1 18.4 0.009625 14.6 6.2 173.5 11.8 0.004813 6.6 6.2 107.2 7.5 0.002406 3.9 6.6 67.1 4.7 0 1.5 7.5 2.5 0.2 15 M-CoA NphT7(I147S, F217V) No 0.01925 121.9 8.7 0.2 0.1 0.009625 88.3 8.6 0.2 0.1 0.004813 40.2 8.1 0.2 0.1 0.002406 13.6 6.3 0.1 0.1 0 1.5 6.6 0.2 0.1 16 C2-CoA M-CoA NphT7(I147S, F217V) Yes 0.01925 23.6 427.2 0.1 0.1 0.009625 22.3 452.9 0.1 0.1 0.004813 17.4 342.2 0.1 0.1 0 18.2 14 0.1 0.1 17 C3-CoA M-CoA NphT7(I147S, F217V) Yes 0.01925 3.4 36.8 10.9 333.6 0.009625 2.5 32.6 7.1 306.4 0.004813 2.1 18.5 2 204.4 0 1 7.3 0.9 3.3 18 M-CoA NphT7(I147S, F217V) Yes 0.01925 2.3 268 0.1 1.4 0.009625 0.7 92.8 0.1 0.8 0.004813 0.8 23.5 0.1 0.6 0 0.6 7.4 0.1 0.4

[0320] 3-keto-CS-CoA was produced by NphT7 only when C3- and malonyl-CoA were present simultaneously (Table 23Run 2). When NphT7 was coupled to Hbd, the majority of the 3-keto-CS-CoA was reduced to 3-OH-CS-CoA. These results indicated that wild-type NphT7 is capable of utilizing a C3-CoA as primer in synthesizing 3-keto-CS-CoA, and Hbd from Clostridium acetobutylicum is capable of reducing 3-keto-CS-CoA.

[0321] Reactions using either NphT7 (I147T, F217V) or NphT7 (I147S, F217V) mutants were similar to those obtained in wild-type NphT7 reactions. Both mutants could use C3-CoA as primer to produce 3-keto-CS-CoA, which was further reduced to 3-OH-C5-CoA in the presence of Hbd plus NADH (Table 23Runs 7-18). With acetyl-CoA plus malonyl-CoA or malonyl-CoA alone, only 3-keto-C4-CoA was produced by these enzymes. Higher concentrations of products were detected in either NphT7 (I147T, F217V) or NphT7 (I147S, F217V) because more enzymes were used in these 2 reactions than the reactions with wild-type NphT7.

[0322] When 3-keto-CS-CoA concentrations were plotted against the amount of enzymes in each reaction, specific activities (average over 15 min) of NphT7, NphT7 (I147T, F217V), and NphT7 (I147S, F217V) were 0.3, 0.2, and 0.27 U/mg, respectively.

[0323] Production of odd chain fatty acids, such as fatty acids of C5, C7, C9, C11, C13, C15, and C17 in length, is made possible by the construction of recombinant strains carrying genes expressing NphT7 and/or an NphT7 mutant, a fabH with the desired chain length specificity, a KCR, a 3HDh, and an EnCr, and a terminating enzyme such as a thioesterase or an ester synthase with the desired chain length specificity, and providing a source of propionyl-CoA as the primer and malonyl-CoA as the extender.

Example 7. Production of C4 and C6 Fatty Acid

[0324] It was demonstrated that C4 and C6 fatty acids can be produced using the genetically modified enzymes and methods of the present invention. In particular, it was demonstrated that C4 and C6 fatty acids can be produced by microorganisms genetically modified to encode certain NphT7 mutant enzymes in combination with PA2801TE thioesterase. These amino acid modifications enable the condensation reaction of acyl-CoA (C4-CoA and C6-CoA) with malonyl-CoA. In particular, the genetically modified microorganism comprises one or more heterologous 3-ketoacyl-CoA synthases selected from the group comprising wild-type NphT7, a variant of NphT7 with the I147T and F217V mutations, or a variant of NphT7 with I147S and F217V mutations, and any combination thereof, and at least one of: a) a heterologous KCR, such as fadB; b) a heterologous 3HDh, such as a fadB, c) a heterologous EnCr, such as ter; and d) a thioesterase PA2801TE.

[0325] The following genetically modified E. coli strains were evaluated for production of free fatty acids:

TABLE-US-00025 TABLE 24 Genetic Modifications of Test Strains Host Strain Synthase/thiolase KCR 3HDh EnCr thioesterase Genotype* Plasmid 1 Plasmid 2 A nphT7 (SEQ ID NO 1) fadB(SEQ ID Ter (SEQ ID tesA (SEQ ID 1 pACYC- pET- fadA (SEQ ID NO 182) NO 183) NO 275) NO 278) PpstsIH- PpstsIH- nphtT7-ter fadBA-TT TT-PpstsIH tesA B nphT7(SEQ ID NO 1) fadB(SEQ ID ter (SEQ ID tesA (SEQ ID 2 pACYC- pET- fadA(SEQ ID NO 182) NO 183) NO 275) NO 278) PpstsIH- PpstsIH- nphtT7-ter fadBA-TT- TT-PpstsIH tesA C nphT7(SEQ ID NO 1) fadB(SEQ ID ter (SEQ ID NONE 2 pACYC- pET- NO 183) NO 275) PpstsIH- PpstsIH- nphtT7-ter empty vector TT-PpstsIH- fadB D nphT7(SEQ ID NO 1) fadB(SEQ ID ter (SEQ ID PA2801TE (SEQ ID 2 pACYC- pET- NO 183) NO 275) NO 288) PpstsIH- PpstsIH- nphtT7-ter PA2801TE TT-PpstsIH fadB E nphT7(SEQ ID NO 1) fadB(SEQ ID ter (SEQ ID PA2801TE (SEQ ID 2 pACYC- pET- npht7(I147T, F217V) NO 183) NO 275) NO 288) PpstsIH- PpstsIH- nphtT7-ter NphT7(I147T, TT-PpstsIH F216V)- fadB PA2801TE F nphT7(SEQ ID NO 1) fadB(SEQ ID ter (SEQ ID PA28018TE (SEQ ID 2 pACYC- pET- npht7(I147T, F217V) NO 183) NO 275) NO 288) PpstsIH- PpstsIH- nphtT7-ter NphT7(I147T, TT-PpstsIH F216V)- fadB PA2801TE *Genotype 1: F-, (araD-araB)567, lacZ4787(::rrnB-3), LAM-, rph-1, (rhaD-rhaB)568, hsdR514, ldhA::frt, pflB::frt, mgsA::frt, poxB::frt, pta-ack::frt, fabI(ts)-(S241F)-zeoR, tig::frt, atoDAEB::frt, fadD::frt, tesB::frt, yciA::frt *Genotype 2: F-, (araD-araB)567, lacZ4787(::rrnB-3), LAM-, rph-1, (rhaD-rhaB)568, hsdR514, ldhA::frt, pflB::frt, mgsA::frt, poxB::frt, pta-ack::frt, fabI(ts)-(S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt, ybgC::frt, ybfF::frt

[0326] A single colony was incubated at 30 C. for 20 hours in 150 ml SM11 with 35 g/ml Kanamycin and 20 g/ml Chloramphenicol. The cultures were transferred to 50 mL conical tubes and centrifuged at 4,000 RPM for 15 minutes. The pellets were resuspended in fresh SM11 (with phosphate) media to an optical density of 20. The resuspensions of each strain were combined, and 2.5 ml (5%) of the combined resuspensions was used to inoculate 50 ml of SM11 without phosphate media. The culture was incubated for 4 hours at 30 C., and thereafter the temperature was shifted to 37 C. After an additional 20 hours, samples were taken and analyzed for the amount of free fatty acid present. The amounts of C4, C6, and total free fatty acid measured as C4-C18 produced are shown in FIG. 21. In addition, samples were taken at the 18 hour time point post the temperature shift and analyzed for the free fatty acid distribution. The results of this analysis are shown in FIG. 22.

Example 8. Production of C4, C6 and C8 Fatty Acid

[0327] C4, C6 and C.sub.8 fatty acids were produced using the genetically modified enzymes and methods of the present invention. In particular, C.sub.4, C.sub.6 and C.sub.8 fatty acids were produced by microorganisms genetically modified to encode certain NphT7 mutant enzymes in combination with thioesterases. These amino acid modifications enable the condensation reaction of acyl-CoA (C2-CoA, C4-CoA, and C6-CoA) with malonyl-CoA. The genetically modified microorganism comprises one or more heterologous 3-ketoacyl-CoA synthases selected from the group comprising wild-type NphT7, or a variant of NphT7 with I147S and F217V mutations, and any combination thereof, and at least one of: a) a heterologous KCR, such as fadB; b) a heterologous 3HDh, such as a fadB, c) a heterologous EnCr, such as ter; and d) a thioesterase tesA.

[0328] The following genetically modified E. coli strains were evaluated for production of free fatty acids:

TABLE-US-00026 TABLE 25 Genetic Modifications of Test Strains Strain Synthase KCR 3Hdh EnCr thioesterase Host Genotype Plasmid(s) G nphT7(SEQ fadB(SEQ ID ter(SEQ ID tesA (SEQ ID F-, (araD-araB)567, pACYC_PpstsIH- ID NO 1) NO 183) NO 275) NO 278) lacZ4787(::rrnB-3), npht7- LAM-, rph-1, (rhaD- ter_PpstsIH-tesA rhaB)568, hsdR514, pET-PpstsIH- ldhA::frt, pflB::frt, FadB mgsA::frt, poxB::frt, pta- ack::frt, fabI(ts)- (S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt H nphT7(SEQ fadB(SEQ ID ter(SEQ ID NONE F-, (araD-araB)567, pACYC_PpstsIH- ID NO 1) NO 183) NO 275) lacZ4787(::rrnB-3), nphT7-ter-TT- nphT7(I147S- LAM-, rph-1, (rhaD- PpstsIH-fadB F217V) rhaB)568, hsdR514, pET_PpstsIH- ldhA::frt, pflB::frt, His- mgsA::frt, nphT7(I147S- poxB::frt, pta- F217V) ack::frt, fabI(ts)- (S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt, ybgC::frt I nphT7(SEQ fadB(SEQ ID ter(SEQ ID NONE F-, (araD-araB)567, pACYC_PpstsIH- ID NO 1) NO 183) NO 275) lacZ4787(::rrnB-3), nphT7-ter-TT- nphT7(I147S- LAM-, rph-1, (rhaD- PpstsIH-fadB F217V) rhaB)568, hsdR514, pET_PpstsIH- ldhA::frt, pflB::frt, His- mgsA::frt, nphT7(I147S- poxB::frt, pta- F217V) ack::frt, fabI(ts)- (S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt, ybgC::frt, ybfF::frt J nphT7(SEQ fadB(SEQ ID ter(SEQ ID tesA (SEQ ID F-, (araD-araB)567, pACYC_PpstsIH- ID NO 1) NO 183) NO 275) NO 278) lacZ4787(::rrnB-3), nphT7-ter-TT- nphT7(I147S- LAM-, rph-1, (rhaD- PpstsIH-fadB F217V) rhaB)568, hsdR514, pET_PpstsIH- ldhA::frt, pflB::frt, His- mgsA::frt, nphT7(I147S- poxB::frt, pta- F217V) ack::frt, fabI(ts)- (S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt, ybgC::frt, aldB::PpstsIH-tesA- TT_loxP-BlsdR-loxP K nphT7(SEQ fadB(SEQ ID ter tesA (SEQ ID F-, (araD-araB)567, pACYC_PpstsIH- ID NO 1) NO 183) NO 278) lacZ4787(::rrnB-3), nphT7-ter-TT- nphT7(I147S- LAM-, rph-1, (rhaD- PpstsIH-fadB F217V) rhaB)568, hsdR514, pET_PpstsIH- ldhA::frt, pflB::frt, His- mgsA::frt, nphT7(I147S- poxB::frt, pta- F217V) ack::frt, fabI(ts)- (S241F)-zeoR, fabB(ts), fabF::frt, coaA*, fabD(ts), tig::frt, atoDAEB::frt, fadD::frt, yciA::frt, tesB::frt, fadBA::frt, fadIJ::frt, ybgC::frt, ybfF::frt, aldB::PpstsIH- tesA-TT_loxP-BlsdR- loxP coaA* denotes an allele of coaA (pantothenate kinase) which is resistant to feedback inhibition.
The same method outlined in Example 7 was used, however the culture was allowed to ferment for 68 hours post temperature shift and samples were taken and analyzed for the amount of free fatty acid present. The amounts of C4, C6, and C8 free fatty acid produced are shown in FIG. 23 and the amounts of total fatty acids (C4-C18) produced are shown in FIG. 24. The distribution of free fatty acids produced by the various strains is shown in FIG. 25. These results indicate that strains J and K produce C6-fatty acids with high specificity.

[0329] Production of fatty acids of chain length >C8, such as C10, C12, C14, C16, and C18 fatty acids in length with high specificity, is made possible by the construction of recombinant strains carrying genes expressing NphT7 and/or an NphT7 mutant, a fabH with the desired chain length specificity, a KCR, a 3HDh, and an EnCr, and a terminating enzyme such as a thioesterase or an ester synthase with the desired specificity, and providing a source of acetyl-CoA as the primer and malonyl-CoA as the extender.

[0330] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention.