MICROORGANISMS FOR THE PRODUCTION OF ADIPIC ACID AND OTHER COMPOUNDS

• US 20130095540A1
• Filed: 06/15/2012
• Published: 04/18/2013
• Est. Priority Date: 03/27/2008
• Status: Active Grant

• Current Assignee: Genomatica Incorporated
• Original Assignee: Genomatica Incorporated
• Inventors: BURGARD, Anthony P., Pharkya, Priti, Osterhout, Robin E.
• Granted Patent: US 8,592,189 B2
• Time in Patent Office: Days
• US Class Current: 435/142
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/001 acting on the CH-CH group of donors (1.3)
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/1096 transferring nitrogenous groups (2.6)
• CPC Class Codes: C12N 9/80 acting on amide bonds in linear amides (3.5.1)
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 13/001 Amines; Imines
• CPC Class Codes: C12P 13/005 Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
• CPC Class Codes: C12P 13/02 Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes peptides C12P21/00 or C07K
• CPC Class Codes: C12P 17/10 Nitrogen as only ring hetero atom
• CPC Class Codes: C12P 7/44 Polycarboxylic acids
• CPC Class Codes: C12P 7/46 Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
• CPC Class Codes: C12P 7/62 Carboxylic acid esters
• CPC Class Codes: C12Y 101/01035 3-Hydroxyacyl-CoA dehydrogenase (1.1.1.35)
• CPC Class Codes: C12Y 102/0101 Acetaldehyde dehydrogenase (acetylating) (1.2.1.10)
• CPC Class Codes: C12Y 103/01031 2-Enoate reductase (1.3.1.31)
• CPC Class Codes: C12Y 206/01 Transaminases (2.6.1)
• CPC Class Codes: C12Y 402/01 Hydro-lyases (4.2.1)

Abstract

The invention provides a non-naturally occurring microbial organism having an adipate, 6-aminocaproic acid or caprolactam pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective adipate, 6-aminocaproic acid or caprolactam pathway. The invention additionally provides a method for producing adipate, 6-aminocaproic acid or caprolactam. The method can include culturing an adipate, 6-aminocaproic acid or caprolactam producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an adipate, 6-aminocaproic acid or caprolactam pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce adipate, 6-aminocaproic acid or caprolactam.

Claims

1. A non-naturally occurring microbial organism, comprising a microbial organism having an adipate pathway comprising at least one exogenous nucleic acid encoding an adipate pathway enzyme expressed in a sufficient amount to produce adipate, said adipate pathway comprising succinyl-CoA:

acetyl-CoA acyl transferase;

3-hydroxyacyl-CoA dehydrogenase;

3-hydroxyadipyl-CoA dehydratase;

5-carboxy-2-pentenoyl-CoA reductase; and

adipyl-CoA synthetase, phosphotransadipylase/adipate kinase, adipyl-CoA;

acetyl-CoA transferase or adipyl-CoA hydrolase.

2. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism comprises two, three, four or five exogenous nucleic acids each encoding an adipate pathway enzyme.

6. The non-naturally occurring microbial organism of claim 2, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.

7. The non-naturally occurring microbial organism of claim 2, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.

8. The non-naturally occurring microbial organism of claim 2, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA;

acetyl-CoA transferase.

9. The non-naturally occurring microbial organism of claim 2, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.

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

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

3-5. -5. (canceled)

12. A method for producing adipate, comprising culturing a non-naturally occurring microbial organism having an adipate pathway, said pathway comprising at least one exogenous nucleic acid encoding an adipate pathway enzyme expressed in a sufficient amount to produce adipate, under conditions and for a sufficient period of time to produce adipate, said adipate pathway comprising succinyl-CoA:

acetyl-CoA acyl transferase;

3-hydroxyacyl-CoA dehydrogenase;

3-hydroxyadipyl-CoA dehydratase;

5-carboxy-2-pentenoyl-CoA reductase; and

adipyl-CoA synthetase, phosphotransadipylase/adipate kinase, adipyl-CoA;

acetyl-CoA transferase or adipyl-CoA hydrolase.

13. The method of claim 12, wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

14. The method of claim 12, wherein said microbial organism comprises two, three, four or five exogenous nucleic acids each encoding an adipate pathway enzyme.

18. The method of claim 14, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.

19. The method of claim 14, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.

20. The method of claim 14, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA;

acetyl-CoA transferase.

21. The method of claim 14, wherein said five exogenous nucleic acids encode succinyl-CoA:

acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.

22. The method of claim 12, wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

15-17. -17. (canceled)

23-112. -112. (canceled)