Transgenic systems for the manufacture of poly(2-hydroxy-butyrate-co-3-hydroxyhexanoate)
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-015/82
C12N-015/87
A01H-001/00
출원번호
US-0703906
(2003-11-07)
등록번호
US-7504556
(2009-03-17)
발명자
/ 주소
Madison,Lara
Huisman,Gjalt W.
Peoples,Oliver P.
출원인 / 주소
Metabolix, Inc.
대리인 / 주소
Pabst Patent Group LLP
인용정보
피인용 횟수 :
13인용 특허 :
25
초록▼
Methods for engineering transgenic organisms that synthesize polyhydroxyalkanoates (PHAs) containing 3-hydroxyhexanoate as comonomer have been developed. These processes are based on genetically engineered bacteria such as Escherichia coli or in plant crops as production systems which include PHA bi
Methods for engineering transgenic organisms that synthesize polyhydroxyalkanoates (PHAs) containing 3-hydroxyhexanoate as comonomer have been developed. These processes are based on genetically engineered bacteria such as Escherichia coli or in plant crops as production systems which include PHA biosynthetic genes from PHA producers. In a preferred embodiment of the method, additional genes are introduced in wild type or transgenic polyhydroxybutyrate (PHB) producers, thereby creating new strains that synthesize 3HH monomers which are incorporated into PHAs. The 3HH monomer preferably is derived in microbial systems using butanol or butyrate as feedstocks, which are precursors of 3-hydroxyhexanoyl-CoA. Pathways for in vivo production of butyrol-CoA specifically encompassing butyryl-CoA dehydrogenase activity are provided.
대표청구항▼
We claim: 1. A method for the biological production of polyhydroxyalkanoates containing 3-hydroxyhexanoate in plants, comprising synthesizing the polyhydroxyalkanoate in plants expressing an enzyme selected from the group consisting of PHB polymerase, PHA polymerase, and β-ketoacyl-CoA reducta
We claim: 1. A method for the biological production of polyhydroxyalkanoates containing 3-hydroxyhexanoate in plants, comprising synthesizing the polyhydroxyalkanoate in plants expressing an enzyme selected from the group consisting of PHB polymerase, PHA polymerase, and β-ketoacyl-CoA reductase, wherein the plants are genetically engineered by integration into the chromosome of one or more genes encoding one or more enzymes forming a fatty acid oxidation complex, 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. 2. The method of claim 1 wherein the plant is selected from the group consisting of oil crop plants and starch accumulating plants. 3. The method of claim 2 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. 4. The method of claim 1 wherein the plant is genetically engineered to express or overexpress a PHA polymerase incorporating C6 substrates. 5. The method of claim 4 wherein the enzyme is derived from Aeromonas caviae, Comamonas testosteroni, Thiocapsia pfenigii, Chromatium vinosum, Bacillus cereus, Nocardia carolina, Nocardia salmonicolor, Rhodococcus ruber, Rhodococcus rhodocrous, and Rhodospirilum rubrum. 6. The method of claim 1 wherein the plant is genetically engineered to redirect metabolites to production of 3-hydroxyhexanoyl-CoA. 7. The method of claim 6 wherein the plant is genetically engineered using a D-specific enoyl-CoA hydratase gene. 8. The method of claim 7 wherein the hydratase gene is isolated from a bacteria selected from the group consisting of R. eutropha, Klebsiella aerogenes, P. putida, and Aeromonas caviae. 9. The method of claim 6 wherein the plant is genetically engineered using a butyrate fermentation pathway. 10. The method of claim 9 wherein the butyrate fermentation pathway is from Clostridium acetobutylicium or Thermoanaerobacterium thermosaccharolyticum. 11. The method of claim 9 wherein the plant is genetically engineered to convert butyrate to butyryl CoA or butyryl CoA to crotonyl CoA. 12. The method of claim 9 wherein the plant is genetically engineered to express a broad range reductase that is active on C6 substrates. 13. The method of claim 9 wherein the plant is genetically engineered to express a polymerase that accepts 3-hydroxyhexanoyl CoA. 14. The method of claim 9 wherein the plant is genetically engineered to express a thiolase accepting acetoacetyl CoA. 15. The method of claim 9 wherein the plant is 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. 16. The method of claim 6 wherein the plant is genetically engineered to integrate one or more nucleic acids encoding fatty acid biosynthetic enzymes derived from E. coli. 17. The method of claim 16 wherein the fatty acid biosynthetic enzymes are enzymes converting acyl ACP to acyl CoA. 18. The method of claim 17 where the enzymes are selected from the group consisting of ACP-CoA transacylase, acyl ACP thioesterase, and acyl CoA synthase. 19. The method of claim 18 wherein the enzymes are acyl ACP thioesterase and acyl CoA synthase. 20. The method of claim 1 wherein the fatty acid oxidation complex comprises enzymes epimerizing S-3 hydroxyhexanoyl CoA and enzymes reducing 3-ketohexanoyl CoA. 21. The method of claim 20 wherein the enzymes are derived from Nocardia salmonicolor. 22. The method of claim 20 wherein the enzymes for epimerization are derived from Pseudomonas putida FaoAB complex.
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이 특허에 인용된 특허 (25)
Hori Yoji (Kanagawa JPX) Yamaguchi Akio (Kanagawa JPX), Biodegradable optically active copolymer and process for producing the same.
Huisman Gjalt W. ; Skraly Frank ; Martin David P. ; Peoples Oliver P., Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids.
Reusch Rosetta N. (Okemos MI) Reusch William H. (Okemos MI), Branched polyhydroxyalkanoate polymer salt electrolytic compositions and method of preparation.
Sanders Johan P. M. (BV Delft NLX) Van Den Berg Johannes A. (AD Reeuwijk NLX) Andreoli Peter M. (EJ Rotterdam NLX) Vos Yvonne J. (MB Capelle a/d IJssel NLX) Van EE Jan H. (EZ Nieuwerkerk a/d IJssel N, Molecular cloning and expression in industrial Bacillus species.
Peoples Oliver P. (Arlington MA) Gerngross Tillman U. (Cambridge MA) Sinskey Anthony J. (Boston MA), Overproduction and purification of soluble PHA synthase.
Blauhut Wilfried (Linz ATX) Gierlinger Wolfgang (Linz ATX) Strempfl Friedrich (Linz ATX), Process for obtaining a polyhydroxyalkanoate from the cell material of a microogranism.
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