Microorganisms for the production of adipic acid and other compounds
View Patent ↗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.
1. An isolated host microbial organism having an adipate pathway and comprising at least one exogenous nucleic acid introduced into said host, wherein said exogenous nucleic acid encodes 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, or adipyl-CoA hydrolase.
2. The isolated host microbial organism of claim 1 , wherein said host microbial organism comprises two, three, four or five exogenous nucleic acids each encoding an adipate pathway enzyme.
3. The isolated host 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.
4. The isolated host 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.
5. The isolated host 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.
6. The isolated host microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
7. The isolated host microbial organism of claim 1 , wherein said host microbial organism is in a substantially anaerobic culture medium.
8. The isolated host microbial organism of claim 1 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.
9. The isolated host microbial organism of claim 1 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.
10. The isolated host microbial organism of claim 1 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.
11. The isolated host microbial organism of claim 2 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.
12. The isolated host microbial organism of claim 2 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.
13. The isolated host microbial organism of claim 2 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.
14. A method for producing adipate comprising culturing the isolated host microbial organism of claim 1 under conditions and for a sufficient period of time to produce adipate.
15. The method of claim 14 , wherein said host microbial organism is in a substantially anaerobic culture medium.
16. The method of claim 14 , wherein said host microbial organism comprises two, three, four or five exogenous nucleic acids each encoding an adipate pathway enzyme.
17. The method of claim 16 , 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.
18. The method of claim 16 , 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.
19. The method of claim 16 , 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.
20. The method of claim 14 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
21. The method of claim 14 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.
22. The method of claim 14 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.
23. The method of claim 14 , wherein said adipate pathway comprises succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.
24. The method of claim 16 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA synthetase.
25. The method of claim 16 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and phosphotransadipylase/adipate kinase.
26. The method of claim 16 , wherein said two, three or four exogenous nucleic acids each encode an adipate pathway enzyme selected from succinyl-CoA:acetyl-CoA acyl transferase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxyadipyl-CoA dehydratase, 5-carboxy-2-pentenoyl-CoA reductase, and adipyl-CoA hydrolase.