Claims
- 1. A method for the biological production of polyhydroxyalkanoates containing 3-hydroxyhexanoate comprising synthesizing the polyhydroxyalkanoate in a transgenic organism having at least one transgene encoding an enzyme selected from the group consisting of PHB polymerase, PHA polymerase, β-ketothiolase, β-ketoacyl-CoA reductase, D-specific enoyl-CoA hydratase, crotonase, butyryl-CoA dehydrogenase, and 3-hydroxybutyryl-CoA dehydrogenase integrated into the chromosome.
- 2. The method of claim 1 wherein the organism is a bacteria or plant.
- 3. The method of claim 2 wherein the organism is a plant selected from the group consisting of oil crop plants and starch accumulating plants.
- 4. The method of claim 3 wherein the plant is selected from the group consisting of Brassica, sunflower, soybean, corn, safflower, flax, palm, coconut, potato, tapioca, cassava, alfalfa, grass, and tobacco.
- 5. The method of claim 2 wherein the organism is a bacteria selected from the group consisting of Escherichia, Klebsiella, Ralstonia, Alcaligenes, Pseudomonas, and Azotobacter.
- 6. The method of claim 1 wherein the organism is genetically engineered to express or overexpress a PHA polymerase incorporating C6 substrates.
- 7. The method of claim 6 wherein the enzyme is derived from Aeromonas caviae, Comamonas testosteroni, Thiocapsia pfenigii, Chromatium vinosum, Bacillus cereus, Nocardia carolina, Nocardia salmonicolor, Rhodococcus ruber, Rhodcoccus rhodocrous, and Rhodospirilum rubrum.
- 8. The method of claim 1 wherein the organisms are genetically engineered to redirect metabolites to production of 3-hydroxyhexanoyl-CoA.
- 9. The method of claim 8 wherein the organisms are genetically engineered using a D-specific enoyl-CoA hydratase gene.
- 10. The method of 9 wherein the hydratase gene is isolated from a bacteria selected from the group consisting of R. eutropha, Klebsiella aerogenes, P. putida, and Aeromonas caviae.
- 11. The method of claim 8 wherein the organisms are genetically engineered using a butyrate fermentation pathway.
- 12. The method of claim 11 wherein the butyrate fermentation pathway is from Clostridium acetobutylicium or Thermoanaerobacterium thermosaccharolyticum.
- 13. The method of claim 11 wherein the organisms are genetically engineered to convert butyrate to butyryl CoA or butyryl CoA to crotonyl CoA.
- 14. The method of claim 11 wherein the organisms are genetically engineered to express a broad range reductase that is active on C6 substrates.
- 15. The method of claim 11 wherein the organisms are genetically engineered to express a polymerase that accepts 3-hydroxyhexanoyl CoA.
- 16. The method of claim 11 wherein the organisms are genetically engineered to express a thiolase accepting acetoacetyl CoA.
- 17. The method of claim 11 wherein the organisms are genetically engineered to express an enzyme selected from the group consisting of thiolases specific for 3-ketohexanoyl CoA, reductase active on 3-ketohexanoyl CoA, PHA polymerase that accepts 3-hydroxybutyryl CoA and 3-hydroxyhexanoyl CoA.
- 18. The method of claim 8 wherein the organisms are genetically engineered using fatty acid biosynthetic enzymes.
- 19. The method of claim 18 wherein the fatty acid biosynthetic enzymes are enzymes converting acyl ACP to acyl CoA.
- 20. The method of claim 19 where the enzymes are selected from the group consisting of ACP-CoA transacylase, acyl ACP thioesterase, and acyl CoA synthase.
- 21. The method of claim 20 wherein the enzymes are acyl ACP thioesterase and acyl CoA synthase.
- 22. The method of claim 18 wherein the enzymes are derived from E. coli.
- 23. The method of claim 8 wherein the organisms are genetically engineered using a fatty acid oxidation complex.
- 24. The method of claim 23 wherein the fatty acid oxidation complex comprises enzymes selected from the group consisting of enzymes epimerizing S-3 hydroxyhexanoyl CoA and enzymes reducing 3-ketohexanoyl CoA.
- 25. The method of claim 24 wherein the enzymes are derived from Nocardia salmonicolor.
- 26. The method of claim 24 wherein the enzymes for epimerization are derived from Pseudomonas putida FaoAB complex.
- 27. The method of claim 23 wherein the organism that is genetically engineered accumulates 3-ketohexanoyl CoA due to a lack of a thiolase.
- 28. A method for producing polyhydroxybutyrate-co-3-hydroxyhexanoate comprising feeding an organism
butyrate or butanol, and another feedstock selected from the group consisting of glucose, sucrose, lactose, xylose, methanol, and combinations thereof.
- 29. A method for producing 3-hydroxyhexanoate copolymers comprising
identifying an organism capable of taking up butyrate and converting it to butyryl-CoA, fermenting the organism in the presence of butyrate such that PHBH is produced, and recovering the PHBH.
- 30. A method for producing 3-hydroxyhexanoate copolymers comprising
identifying bacteria capable of taking up butanol and converting it to butyryl-CoA, fermenting the organism in the presence of butanol such that PHA is produced, and recovering the PHA.
- 31. A genetically engineered organism for use in any of the methods of claims 1-30.
- 32. The organism of claim 31 wherein the organism is a bacteria.
- 33. The organism of claim 31 wherein the organism is a higher order plant.
- 34. A polyhydroxybutyrate-co-3-hydroxyhexanoate produced in a genetically engineered Escherichia coli K12.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] Priority is claimed to U.S. Provisional application Serial No. 60/072,198, filed Jan. 22, 1998.
Provisional Applications (1)
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Number |
Date |
Country |
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60072198 |
Jan 1998 |
US |