Microorganisms and methods for the production of caprolactone

Patent metadata

Field	Value
Publication number	US-10351887-B2
Application number	US-201715634726-A
Country	US
Kind code	B2
Filing date	Jun 27, 2017
Priority date	Nov 2, 2011
Publication date	Jul 16, 2019
Grant date	Jul 16, 2019

Abstract

The invention provides non-naturally occurring microbial organisms containing caprolactone pathways having at least one exogenous nucleic acid encoding a butadiene pathway enzyme expressed in a sufficient amount to produce caprolactone. The invention additionally provides methods of using such microbial organisms to produce caprolactone by culturing a non-naturally occurring microbial organism containing caprolactone pathways as described herein under conditions and for a sufficient period of time to produce caprolactone.

First claim

What is claimed is:

1. A non-naturally occurring microbial organism having a caprolactone pathway and a Wood-Ljungdahl pathway, wherein said non-naturally occurring microbial organism comprises at least one exogenous nucleic acid encoding a caprolactone pathway enzyme expressed in a sufficient amount to produce caprolactone.

2. The non-naturally occurring microbial organism of claim 1 , wherein said caprolactone pathway comprises a pathway selected from:
(1) 1D, 1A, 1B, and 1C;
(2) 1E, 1B, 1C and 1D;
(3) 1F, 1A, 1B, 1C and 1D;
(4) 1F, 1E, 1B, 1C and 1D;
(5) 1A, 1B and 1G;
(6) 1E, 1B and 1G;
(7) 1F, 1A, 1B and 1G;
(8) 1F, 1E, 1B and 1G;
(9) 1A, 1B, 1C, 1J and 1I;
(10) 1E, 1B, 1C, 1J and 1I;
(11) 1F, 1A, 1B, 1C, 1J and 1I;
(12) 1F, 1E, 1B, 1C, 1J and 1I;
(13) 1A, 1B, 1H and 1I;
(14) 1E, 1B, 1H and 1I;
(15) 1F, 1A, 1B, 1H and 1I;
(16) 1F, 1E, 1B, 1H and 1I;
(17) 2A, 2B, 2C, 2D, 2E and 2F;
(18) 2A, 2B, 2C, 2D and 2G;
(19) 2A, 2B, 2C, 2D, 2E, 2J and 2I;
(20) 2A, 2B, 2C, 2D, 2H and 2I;
(21) 4A, 4B, 4C, 4D and 3A; and
(22) 5A, 5B, 5C an 3A,
wherein 1A is an adipyl-CoA reductase, wherein 1B is an adipate semialdehyde reductase, wherein 1C is a 6-hydroxyhexanoyl-CoA transferase or a 6-hydroxyhexanoyl-CoA synthetase, wherein 1D is a 6-hydroxyhexanoyl-CoA cyclase or spontaneous cyclization, wherein 1E is an adipate reductase, wherein 1F is an adipyl-CoA transferase, an adipyl-CoA synthetase or an adipyl-CoA hydrolase, wherein 1G is a 6-hydroxyhexanoate cyclase, wherein 1H is a 6-hydroxyhexanoate kinase, wherein 1I is a 6-hydroxyhexanoyl phosphate cyclase or spontaneous cyclization, wherein 1J is a phosphotrans-6-hydroxyhexanoylase, wherein 2A is a 4-hydroxybutyryl-CoA:acetyl-CoA acyltransferase, wherein 2B is a 3-oxo-6-hydroxyhexanoyl-CoA reductase, wherein 2C is a 3,6-dihydroxyhexanoyl-CoA dehydratase, wherein 2D is a 6-hydroxyhex-2-enoyl-CoA reductase, wherein 2E is a 6-hydroxyhexanoyl-CoA transferase, a 6- hydroxyhexanoyl-CoA synthetase or a 6-hydroxyhexanoyl-CoA hydrolase, wherein 2F is a 6-hydroxyhexanoate cyclase, wherein 2G is a 6-hydroxyhexanoyl-CoA cyclase or spontaneous cyclization, wherein 2H is a phosphotrans-6-hydroxyhexanoylase, wherein 2I is a 6-hydroxyhexanoyl phosphate cyclase or spontaneous cyclization, wherein 2J is a 6-hydroxyhexanoate kinase, wherein 3A is a cyclohexanone monooxygenase, wherein 4A is an adipate semialdehyde dehydratase, wherein 4B is a cyclohexane-1,2-dione reductase, wherein 4C is a 2-hydroxycyclohexanone reductase, wherein 4D is a cyclohexane-1,2-diol dehydratase, wherein 5A is a 2-ketocyclohexane-1-carboxoyl-CoA hydrolase (acting on C-C), wherein 5B is a 2-ketocyclohexane-1-carboxoyl-CoA transferase, a 2-ketocyclohexane-1-carboxoyl-CoA synthetase or a 2-ketocyclohexane-1-carboxoyl-CoA hydrolase, and wherein 5C is a 2-ketocyclohexane-1-carboxylate decarboxylase.

3. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two, three, four, five, six or seven exogenous nucleic acids each encoding a caprolactone pathway enzyme.

4. The non-naturally occurring microbial organism of claim 2 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(22).

5. The non-naturally occurring microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

6. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

7. The non-naturally occurring microbial organism of claim 1 , wherein said Wood-Ljungdahl pathway has a set of Wood-Ljungdahl pathway enzymes that convert syngas to acetyl-CoA.

8. The non-naturally occurring microbial organism of claim 7 , wherein said set of Wood-Ljungdahl pathway enzymes comprises ferredoxin oxidoreductase, formate dehydrogenase, formyltetrahydrofolate synthetase, methenyltetrahydrofolate cyclodehydratase, methylenetetrahydrofolate dehydrogenase and methylenetetrahydrofolate reductase, methyltetrahydrofolate:corrinoid protein methyltransferase, corrinoid iron-sulfur protein, nickel-protein assembly protein, ferredoxin, acetyl-CoA synthase, carbon monoxide dehydrogenase and nickel-protein assembly protein.

9. The non-naturally occurring microbial organism of claim 7 , wherein said non-naturally occurring microbial organism further comprises at least one exogenous nucleic acid encoding said Wood-Ljungdahl pathway enzymes.

10. A method for producing caprolactone, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce caprolactone.

11. Culture medium comprising bioderived caprolactone produced by a non-naturally occurring microbial organism of claim 1 .

12. The culture medium of claim 11 , wherein said culture medium is separated from a non-naturally occurring microbial organism having a caprolactone pathway.

13. Bioderived caprolactone produced by a non-naturally occurring microbial organism of claim 1 .

14. A composition comprising said bioderived caprolactone of claim 13 and a compound other than said bioderived caprolactone.

15. A biobased polymer comprising said bioderived caprolactone of claim 13 .

16. A biobased resin comprising said bioderived caprolactone of claim 13 .

17. A molded product obtained by molding a biobased polymer of claim 15 .

18. A molded product obtained by molding a biobased resin of claim 16 .

19. A process for producing a biobased polymer of claim 15 comprising chemically reacting said bioderived caprolactone with itself or another compound in a polymer producing reaction.

20. A process for producing a biobased resin of claim 16 comprising chemically reacting said bioderived caprolactone with itself or another compound in a resin producing reaction.

Assignees

• Genomatica Inc

Inventors

• Burgard Anthony P
• Osterhout Robin E
• Pharkya Priti
• Burk Mark J

Classifications

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C07D313/04

  not condensed with other rings · CPC title

• C08G63/08

  Lactones or lactides · CPC title

• C12P17/08 Primary

  containing a hetero ring of at least seven ring members, e.g. zearalenone, macrolide aglycons · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title