Production of polyhydroxyalkanoates from polyols
View Patent ↗Recombinant processes are provided whereby additional genes are introduced into E. coli which have been genetically engineered to produce PHA so that the improved strains produce PHA homopolymers and copolymers directly from diols. In preferred embodiments, PHAs containing 4-hydroxybutyrate monomers are produced directly from 1,4-butanediol; PHAs containing 5-hydroxyvalerate are produced from 1,5-pentanediol; PHAs containing 6-hydroxyhexanoate (6HH) are produced from 1,6-hexanediol; PHAs containing 3-hydroxypropionate are produced from 1,3-propanediol; PHAs containing 2-hydroxypropionate (lactate) are produced from 1,2-propanediol (propylene glycol); PHAs containing 2-hydroxyethanoate (glycolate) are produced from 1,2-ethanediol (ethylene glycol). Genes encoding these same enzyme activities can be introduced or their expression amplified in wild type PHA producers to improve the production of PHA homopolymers and copolymers directly from diol and other alcohol feedstocks. The PHA polymers are readily recovered and industrially useful as polymers or as starting materials for a range of chemical intermediates.
1. Genetically engineered Escherichia coli ( E. coli ) for use in producing polyhydroxyalkanoates comprising 4-hydroxybutyrate monomers,
wherein the E. coli expresses enzymes selected from the group consisting of β-ketothiolase, acetoacetyl-CoA reductase and PHA synthase, and at least one enzyme selected from the group consisting of acyl-CoA transferase and acyl-CoA synthetase, and
wherein the E. coli is genetically engineered to express both aldehyde dehydrogenase and diol oxidoreductase, so that when the E. coli is provided with 1,4-butanediol and cultured under conditions where the 1,4-butanediol can be converted to 4-hydroxybutyryl-CoA monomer which can then be polymerized to form polyhydroxyalkanoates comprising 4-hydroxybutyrate, the E. coli produces polyhydroxyalkanoates comprising 4-hydroxybutyrate monomers.
2. A system for making polyhydroxyalkanoates comprising 4-hydroxybutyrate monomers, the system comprising
an E. coli expressing enzymes selected from the group consisting of β-ketothiolase, acetoacetyl-CoA reductase, and PHA synthase and at least one enzyme selected from the group consisting of acyl-CoA transferase and acyl-CoA synthetase,
wherein the E. coli is genetically engineered to express both aldehyde dehydrogenase and diol oxidoreductase, and
wherein the E. coli can convert 1,4-butanediol into 4-hydroxybutyryl-CoA monomer which can then be polymerized to form polyhydroxyalkanoates comprising 4-hydroxybutyrate.
3. The genetically engineered E. coli of claim 1 , wherein the 4-hydroxybutyrate monomer is polymerized to form poly-(4-hydroxybutyrate).
4. The genetically engineered E. coli of claim 1 , wherein the 4-hydroxybutyrate monomer is polymerized to form poly-(3-hydroxybutyrate-co-4-hydroxybutyrate).
5. The system of claim 2 , wherein the 4-hydroxybutyrate monomer is polymerized to form poly-(4-hydroxybutyrate).
6. The system of claim 2 , wherein the 4-hydroxybutyrate monomer is polymerized to form poly-(3-hydroxybutyrate-co-4-hydroxybutyrate).
7. The genetically engineered E. coli of claim 4 , wherein 4-hydroxybutyrate incorporation into the poly-(3-hydroxybutyrate-co-4-hydroxybutyrate) polymer by the E. coli expressing both aldehyde dehydrogenase and diol oxidoreductase is enhanced when compared to incorporation of 4-hydroxybutyrate into a poly-(3-hydroxybutyrate-co-4-hydroxybutyrate) by E. coli expressing only aldehyde dehydrogenase.
8. A method for producing polyhydroxyalkanoates comprising 4-hydroxybutyrate monomers comprising
providing genetically engineered Escherichia coli ( E. coli ) of claim 1 ,
providing 1,4-butanediol which can be converted into hydroxyalkanoate monomers by enzymes expressed by the E. coli , and
culturing the E. coli under conditions wherein the hydroxyalkanoate monomers are polymerized to form polyhydroxyalkanoates comprising 4-hydroxybutyrate monomers.
9. The method of claim 8 wherein the hydroxyalkanoate monomer is 4-hydroxybutyrate.