C12Y103/01044

Iterative platform for the synthesis of alpha functionalized products

The use of microorganisms to make alpha-functionalized chemicals and fuels, (e.g. alpha-functionalized carboxylic acids, alcohols, hydrocarbons, amines, and their beta-, and omega-functionalized derivatives), by utilizing an iterative carbon chain elongation pathway that uses functionalized extender units. The core enzymes in the pathway include thiolase, dehydrogenase, dehydratase and reductase. Native or engineered thiolases catalyze the condensation of either unsubstituted or functionalized acyl-CoA primers with an alpha-functionalized acetyl-CoA as the extender unit to generate alpha-functionalized β-keto acyl-CoA. Dehydrogenase converts alpha-functionalized β-keto acyl-CoA to alpha-functionalized β-hydroxy acyl-CoA. Dehydratase converts alpha-functionalized β-hydroxy acyl-CoA to alpha-functionalized enoyl-CoA. Reductase converts alpha-functionalized enoyl-CoA to alpha-functionalized acyl-CoA. The platform can be operated in an iterative manner (i.e. multiple turns) by using the resulting alpha-functionalized acyl-CoA as primer and the aforementioned alpha-functionalized extender unit in subsequent turns of the cycle. Termination pathways acting on any of the four alpha-functionalized CoA thioester intermediates terminate the platform and generate various alpha-functionalized carboxylic acids, alcohols and amines with different β-reduction degree.

Enzymatic methods for butanol production

A process for producing butanol is provided, involving: A) mixing water, lactate, an enzyme mixture comprising at least one enzyme, at least one cofactor and at least one coenzyme, to prepare a reaction mixture; B) catalytically reacting the reaction mixture for an amount of time sufficient to cause conversion of lactate into butanol; and wherein the conversion of lactate into butanol in B) is associated with a regeneration system of NAD (P).sup.+/NAD (P) H and/or acetyl-CoA/CoA.

HIGH YIELD ROUTE FOR THE PRODUCTION OF COMPOUNDS FROM RENEWABLE SOURCES
20220389433 · 2022-12-08 ·

Provided herein are methods, compositions, and non-naturally occurring microbial organism for preparing compounds such as α-butanol, butyric acid, succinic acid, 1,4-butanediol, 1-pentanol, pentanoic acid, glutaric acid, 1,5-pentanediol, 1-hexanol, hexanoic acid, adipic acid, 1,6-hexanediol, 6-hydroxy hexanoic acid, ε-Caprolactone, 6-amino-hexanoic acid, ε-Caprolactam, hexamethylenediamine, linear fatty acids and linear fatty alcohols that are between 7-25 carbons long, linear alkanes and linear α-alkenes that are between 6-24 carbons long, sebacic acid and dodecanedioic acid comprising: a) converting a C.sub.N aldehyde and pyruvate to a C.sub.N+3 β-hydroxyketone intermediate through an aldol addition; and b) converting the C.sub.N+3 β-hydroxyketone intermediate to the compounds through enzymatic steps, or a combination of enzymatic and chemical steps.

MICROORGANISMS AND METHODS FOR PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS AND RELATED COMPOUNDS
20220333143 · 2022-10-20 ·

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

METHOD FOR PRODUCING HETEROGENOUS CANNABICHROMENE FROM SACCHAROMYCES CEREVISIAE
20230167473 · 2023-06-01 ·

A recombinant host cell capable of biosynthesizing cannabichromenic acid and a construction method thereof, and a method for biosynthesizing cannabichromenic acid through the recombinant host cell. Saccharomyces cerevisiae is taken as a host. First, cannabigerolic acid synthase and cannabichromenic acid synthase are over-expressed in the host; then, a metabolic pathway of a precursor compound, olivetolic acid, synthesizing cannabichromenic acid from saccharides is constructed in the host, a metabolic pathway for hexanoic acid to olivetolic acid is further constructed in the host, an endogenous mevalonate pathway of the host and a metabolic pathway of acetyl-CoA are optimized, cannabichromenic acid synthase is rationally designed, highly active cannabichromenic acid synthase is screened out, and finally, a cannabichromene pathway is located to peroxisomes and lipid droplets by using the cell compartmentalization principle to obtain recombinant Saccharomyces cerevisiae capable of biosynthesizing cannabichromenic acid.

ENGINEERED TRANSAMINASE AND METHODS OF MAKING AND USING

Disclosed are transaminase (TA) enzymes and nucleic acids encoding them. In some cases, the transaminase enzymes are non-natural, engineered transaminases. Also disclosed are biosynthetic methods and engineered microorganisms that enhance or improve the biosynthesis of 6-aminocaproate, hexamethylenediamine, caproic acid, caprolactone, or caprolactam. The engineered microorganisms include exogenous TA and in some cases engineered TA.

ENGINEERED MICROORGANISMS AND METHODS FOR IMPROVED ALDEHYDE DEHYDROGENASE ACTIVITY
20220235385 · 2022-07-28 ·

Disclosed are biosynthetic methods and engineered microorganism that enhance or improve the biosynthesis of hexamethylenediamine, caproic acid or caprolactam. The engineered microorganisms include selected aldehyde dehydrogenase activity.

Microorganisms and methods for production of specific length fatty alcohols and related compounds

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

<i>Saccharomyces cerevisiae </i>strain with high yield of ethyl butyrate and construction method and application of <i>Saccharomyces cerevisiae </i>strain

A Saccharomyces cerevisiae strain with high yield of ethyl butyrate and a construction method and an application thereof are provided. The strain is obtained by over-expressing in the starting strain acetyl coenzyme A acyl transferase gene Erg10, 3-hydroxybutyryl coenzyme A dehydrogenase gene Hbd, 3-hydroxybutyryl coenzyme A dehydratase gene Crt, trans-2-enoyl coenzyme A reductase gene Ter, and alcohol acyl transferase gene AAT. Compared to the starting bacteria not producing ethyl butyrate, the yield of ethyl butyrate of the constructed strain reaches 77.33±3.79 mg/L, the yield of the ethyl butyrate of the strain with double copy expression of gene Ter and gene AAT reaches 99.65±7.32 mg/L, increased by 28.9% compared with the EST strain, and 40.93±3.18 mg/L of ethyl crotonate is unexpectedly produced.

ITERATIVE PLATFORM FOR THE SYNTHESIS OF ALPHA FUNCTIONALIZED PRODUCTS

The use of microorganisms to make alpha-functionalized chemicals and fuels, (e.g. alpha-functionalized carboxylic acids, alcohols, hydrocarbons, amines, and their beta-, and omega-functionalized derivatives), by utilizing an iterative carbon chain elongation pathway that uses functionalized extender units. The core enzymes in the pathway include thiolase, dehydrogenase, dehydratase and reductase. Native or engineered thiolases catalyze the condensation of either unsubstituted or functionalized acyl-CoA primers with an alpha-functionalized acetyl-CoA as the extender unit to generate alpha-functionalized β-keto acyl-CoA. Dehydrogenase converts alpha-functionalized β-keto acyl-CoA to alpha-functionalized β-hydroxy acyl-CoA. Dehydratase converts alpha-functionalized β-hydroxy acyl-CoA to alpha-functionalized enoyl-CoA. Reductase converts alpha-functionalized enoyl-CoA to alpha-functionalized acyl-CoA. The platform can be operated in an iterative manner (i.e. multiple turns) by using the resulting alpha-functionalized acyl-CoA as primer and the aforementioned alpha-functionalized extender unit in subsequent turns of the cycle. Termination pathways acting on any of the four alpha-functionalized CoA thioester intermediates terminate the platform and generate various alpha-functionalized carboxylic acids, alcohols and amines with different β-reduction degree.