Method of making polyhydroxyalkanoate copolymers from diverse substrates

A method of forming at least one of a polyhydroxyalkanoate (PHA) homopolymer and a PHA copolymer, comprising:

• cultivating a recombinant E. coli cell in a medium conducive for a production of the at least one of the PHA homopolymer and the PHA copolymer,
• wherein the recombinant E. coli cell comprises:
• a disrupted endogenous β-oxidation pathway having at least one of a genetically disrupted endogenous fadB gene and a genetically disrupted fadJ gene, and
• a nucleic acid construct comprising:
• (a) at least one phaJ4 gene, cDNA, or nucleic acid encoding active enoyl-CoA hydratase 2,
• (b) at least one phaA gene, cDNA, or nucleic acid encoding active β-ketothiolase,
• (c) at least one phaB gene, cDNA, or nucleic acid encoding active acetoacetyl-CoA reductase, and
• (d) at least one phaC1 gene, cDNA, or nucleic acid encoding active type II polyhydroxyalkanoate synthase,
• wherein said cultivating occurs under conditions suitable for bioconverting a substrate to that at least one of the PHA homopolymer and the PHA copolymer.

A genetically engineered E. coli cell comprising:

• a disrupted endogenous β-oxidation pathway comprising at least one of a genetically disrupted endogenous fadB gene and a genetically disrupted fadJ gene, and
• at least one nucleic acid comprising:
• at least one gene encoding active enoyl-CoA hydratase 2;
• at least one gene encoding active β-ketothiolase;
• at least one gene encoding active acetoacetyl-CoA reductase;
• and
• at least one gene encoding active type II polyhydroxyalkanoate synthase,
• wherein the at least one nucleic acid causes the genetically engineered E. coli cell to have increased activity of enoyl-CoA hydratase 2, β-ketothiolase, acetoacetyl-CoA reductase, and type II polyhydroxyalkanoate synthase, with respect to a wild type E. coli lacking the at least one nucleic acid construct.