The invention provides microorganisms and methods for the production of polyhydroxybutyrate (PHB) from gaseous substrates. In particular, the invention provides a non-naturally occurring Wood-Ljungdahl microorganism comprising (a) an enzyme that converts acetyl-CoA to acetoacetyl-CoA, (b) an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and (c) an enzyme that converts 3-hydroxybutyryl-CoA to polyhydroxybutyrate, and methods related thereto.

The present invention relates to a recombinant microorganism to which a gene coding for 2-hydroxyisocaproate-CoA transferase and a gene coding for polyhydroxyalkanoate synthase are introduced and which has a potential of producing polyhydroxyalkanoate bearing an aromatic monomer or a long-chain 2-HA monomer and a method for producing polyhydroxyalkanoate bearing an aromatic monomer or a long-chain 2-HA monomer, using the recombinant microorganism. According to the present invention, a biodegradable polymer bearing an aromatic monomer or a long-chain 2-HA monomer can be produced.

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 -keto acyl-CoA. Dehydrogenase converts alpha-functionalized -keto acyl-CoA to alpha-functionalized -hydroxy acyl-CoA. Dehydratase converts alpha-functionalized -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 -reduction degree.

The present disclosure provides thiolases and polypeptide variants of 3-hydroxybutyryl-CoA dehydrogenase, nucleic acids encoding the same, vectors comprising the nucleic acids, and cells comprising the polypeptide variants and/or thiolase, the nucleic acids, and/or the vectors. The present disclosure also provides methods of making and using the same, including methods for culturing cells, and for the production of various products, including 3-hydroxybutyryl-CoA (3-HB-CoA), 3-hydroxybutyraldehyde (3-HBal), 3-hydroxybutyrate (3-HB), 1,3-butanediol (1,3-BDO), and esters and amides thereof, and products made from any of these.

Provided is engineered microorganisms expressing acetoacetyl-CoA reductase variants and a method for improving the yield of PHA. Compared with the wild-type acetoacetyl-CoA reductase represented by SEQ ID NO. 31, the variant has one or more of the following mutations: (1) mutation of valine at position 141 to isoleucine or leucine; (2) mutation of methionine at position 12 to threonine, serine, alanine, leucine, lysine or isoleucine; (3) mutation of isoleucine at position 194 to valine, leucine or methionine; (4) mutation of glutamic acid at position 42 to lysine, glutamine, leucine, aspartic acid, proline, threonine, asparagine, or histidine; and (5) mutation of phenylalanine at position 55 to valine, alanine or isoleucine. The variants and their coding genes can promote the synthesis and accumulation of PHA by the strain and increase the yield of PHA.

The present disclosure provides thiolases and polypeptide variants of 3-hydroxybutyryl-CoA dehydrogenase, nucleic acids encoding the same, vectors comprising the nucleic acids, and cells comprising the polypeptide variants and/or thiolase, the nucleic acids, and/or the vectors. The present disclosure also provides methods of making and using the same, including methods for culturing cells, and for the production of various products, including 3-hydroxybutyryl-CoA (3-HB-CoA), 3-hydroxybutyraldehyde (3-HBal), 3-hydroxybutyrate (3-HB), 1,3-butanediol (1,3-BDO), and esters and amides thereof, and products made from any of these.

The present invention relates to a recombinant microorganism to which a gene coding for 2-hydroxyisocaproate-CoA transferase and a gene coding for polyhydroxyalkanoate synthase are introduced and which has a potential of producing polyhydroxyalkanoate bearing an aromatic monomer or a long-chain 2-HA monomer and a method for producing polyhydroxyalkanoate bearing an aromatic monomer or a long-chain 2-HA monomer, using the recombinant microorganism. According to the present invention, a biodegradable polymer bearing an aromatic monomer or a long-chain 2-HA monomer can be produced.

The invention provides microorganisms and methods for the production of polyhydroxybutyrate (PHB) from gaseous substrates. In particular, the invention provides a non-naturally occurring Wood-Ljungdahl microorganism comprising (a) an enzyme that converts acetyl-CoA to acetoacetyl-CoA, (b) an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and (c) an enzyme that converts 3-hydroxybutyryl-CoA to polyhydroxybutyrate, and methods related thereto.

The present invention provides a Clostridium species comprising a non-native gene capable of expressing (R)-3-hydroxybutyryl-Co A dehydrogenase. Also provided is a method of producing (R)-3-hydroxybutyric acid, or a salt thereof, and/or (R) 1,3-butanediol using such Clostridium.

The disclosure provides recombinant bacterial host cells that metabolize and convert glycerol or volatile fatty acids (VFAs) to poly(3-hydroxybutyrate-co-3-hydroxyvalerate) or PHBV. The disclosure further provides methods of producing PHBV using the recombinant bacteria disclosed herein.