The present invention relates to methods of providing a biomass residuum and compositions thereof. In particular examples, the biomass residuum includes one or more high value amino acids, even after removal of mixed alcohol components. In particular, the methods include implementing pre-treatment conditions and employing fermentation conditions including modified organisms.

The present disclosure provides a recombinant microorganism for producing PHBV, a method for preparing the same, and a method for producing PHBV using the microorganism. The present disclosure may provide a recombinant microorganism capable of producing PHBV, which is a biodegradable plastic material with superior physical properties, directly from an inexpensive single carbon source with high efficiency without supplementation of organic acid. The present disclosure can enhance the utilization of PHA, which is expensive and has limited physical properties, and can also provide a technology more effective for industrialization using an inexpensive single carbon source. The PHBV produced according to an exemplary embodiment of the present disclosure can be used not only for general-purpose inexpensive products such as ecofriendly packing materials but also as a high-value-added medical biopolymer.

The present disclosure provides microbial organisms having decreased production of unwanted by-products (e.g, pyruvate-, CO.sub.2—, TCA-derived by-products; acetate; ethanol; and/or, alanine) to enhance carbon flux through acetyl-CoA, which can increase production of acetyl-CoA derived compounds (e.g, 1,3-BDO, MMA, and (3R)-hydroxybutyl (3R)-hydroxybutyrate, or any other acetyl-CoA derived compounds), and products made from any of these compounds. Also provided are one or more exogenous nucleic acids encoding enzymes that can decrease production of unwanted by-products (e.g, aldehyde dehydrogenase, acetyl-CoA synthase, amino acid dehydrogenase, alanine racemase, and/or citrate synthase), and/or one or more gene attenuations occurring in genes (e.g., acetolactate synthase) that result in decreased production of unwanted by-products. Various combinations of the exogenous nucleic acids and gene deletions are also provided in the present disclosure. Methods of making and using the same, including methods for culturing cells, and for the production of the various products are also provided.

A genetically engineered strain having high-yield of L-valine is disclosed. Starting from Escherichia coli W3110, an acetolactate synthase gene alsS of Bacillus subtilis is inserted into a genome thereof and overexpressed; a ppGpp 3′-pyrophosphate hydrolase mutant R290E/K292D gene spoTM of Escherichia coli is inserted into the genome and overexpressed; a lactate dehydrogenase gene ldhA, a pyruvate formate lyase I gene pflB, and genes frdA, frdB, frdC, frdD of four subunits of fumaric acid reductase are deleted from the genome; a leucine dehydrogenase gene bcd of Bacillus subtilis replaces a branched chain amino acid transaminase gene ilvE of Escherichia coli; and an acetohydroxy acid isomeroreductase mutant L67E/R68F/K75E gene ilvCM replaces the native acetohydroxy acid isomeroreductase gene ilvC of Escherichia coli. Furthermore, the L-valine fermentation method is improved by using a two-stage dissolved oxygen control. The L-valine titer and the sugar-acid conversion rate are increased.

Multi-carbon compounds such as ethanol, n-butanol, sec-butanol, isobutanol, tert-butanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters, 2,3-butanediol and the like, are important industrial commodity chemicals with a variety of applications. The present invention provides metabolically engineered host microorganisms which metabolize methane (CH.sub.4) as their sole carbon source to produce multi-carbon compounds for use in fuels (e.g., bio-fuel, bio-diesel) and bio-based chemicals. Furthermore, use of the metabolically engineered host microorganisms of the invention (which utilize methane as the sole carbon source) mitigate current industry practices and methods of producing multi-carbon compounds from petroleum or petroleum-derived feedstocks, and ameliorate much of the ongoing depletion of arable food source “farmland” currently being diverted to grow bio-fuel feedstocks, and as such, improve the environmental footprint of future bio-fuel, bio-diesel and bio-based chemical compositions.

The biological production of beta-hydroxyisovalerate (βHIV) using at least one non-natural enzyme. The non-natural enzyme for the biologically-derived βHIV provides more beta-hydroxyisovalerate synthase activity than the wild-type parent. The non-natural enzyme having one or more modifications of substrate-specificity positions. The non-natural enzyme can be expressed in a microorganism, such as a yeast or bacteria, wherein the microorganism comprises an active βHIV metabolic pathway for the production of βHIV. Alternatively, the non-natural enzyme can be a βHIV synthase used to produce βHIV in a cell-free environment. The biological derivation of βHIV eliminates toxic by-products and impurities that result from the chemical production of βHIV, such that βHIV produced by a non-natural enzyme prior to any isolation or purification process has not been in substantial contact with any halogen-containing component.

Genetically modified microorganisms that have the ability to convert carbon substrates into multicarbon products. Methods of making these genetically modified microorganisms and methods of using them. Vectors encoding enzymes for use in converting carbon substrates into multicarbon products.

The present invention relates to preparation of key drug intermediate, L-2-amino butyric acid (L-2-ABA) by a method of cell free system and biotransformation using genetically engineered strains from easily available economic substrates like citramalate or citraconate and enzymes like LeuCD, LeuB and ValDH or IlvE.

The present invention relates to methods of providing a biomass residuum and compositions thereof. In particular examples, the biomass residuum includes one or more high value amino acids, even after removal of mixed alcohol components. In particular, the methods include implementing pre-treatment conditions and employing fermentation conditions including modified organisms.

Genetically modified microorganisms that have the ability to convert carbon substrates into chemical products such as 2,3-BDO; 1,4-BDO; isobutyraldehyde; isobutanol; 1-butanol; n-butanol; ethanol; fatty alcohols; and fatty acid methyl ester are disclosed. For example, genetically modified methanotrophs that are capable of generating 2,3-BDO; 1,4-BDO; isobutyraldehyde; isobutanol; 1-butanol; n-butanol; ethanol; fatty alcohols; and fatty acid methyl ester at high titers from a methane source are disclosed. Methods of making these genetically modified microorganisms and methods of using them are also disclosed. These microorganisms and methods make use of molecular switches to regulate gene expression.