Microorganisms for the production of adipic acid and other compounds

Patent metadata

Field	Value
Publication number	US-10415042-B2
Application number	US-201615184940-A
Country	US
Kind code	B2
Filing date	Jun 16, 2016
Priority date	Mar 27, 2008
Publication date	Sep 17, 2019
Grant date	Sep 17, 2019

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.

First claim

What is claimed is:

1. A non-naturally occurring microbial organism having a 6-aminocaproic acid pathway and an adipyl-CoA pathway, wherein said 6-aminocaproic acid pathway comprises at least one exogenous nucleic acid encoding a 6-aminocaproic acid pathway enzyme expressed in a sufficient amount to produce 6-aminocaproic acid, wherein said 6-aminocaproic acid pathway comprises a CoA-dependent aldehyde dehydrogenase; and a transaminase or a 6-aminocaproate dehydrogenase, wherein said transaminase converts adipate semialdehyde to 6-aminocaproic acid, wherein said adipyl-CoA pathway comprises at least one exogenous nucleic acid encoding an adipyl-CoA pathway enzyme expressed in a sufficient amount to produce adipyl-CoA, and wherein said adipyl-CoA pathway comprises alpha-ketoadipyl-CoA synthetase, phosphotransketoadipylase/alpha-ketoadipate kinase or alpha-ketoadipyl-CoA:acetyl-CoA transferase; 2-hydroxyadipyl-CoA dehydrogenase; 2-hydroxyadipyl-CoA dehydratase; and a 5-carboxy-2-pentenoyl-CoA reductase.

2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises one or two exogenous nucleic acids each encoding a 6-aminocaproic acid pathway enzyme.

3. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism further has a caprolactam pathway comprising at least one exogenous nucleic acid encoding a caprolactam pathway enzyme expressed in a sufficient amount to produce caprolactam, said caprolactam pathway comprising an amidohydrolase.

4. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises one, two, three, or four exogenous nucleic acids each encoding an adipyl-CoA pathway enzyme.

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 microbial organism is a species of bacteria, yeast, or fungus.

7. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism is a species of bacteria selected from the group consisting of Escherichia coli, Klebsiella oxytoca, Anaerobiospirillum succiniciproducens, Actinobacillus succinogenes, Mannheimia succiniciproducens, Rhizobium etli, Bacillus subtilis, Corynebacterium glutamicum, Gluconobacter oxydans, Zymomonas mobilis, Lactococcus lactis, Lactobacillus plantarum, Streptomyces coelicolor, Clostridium acetobutylicum, Pseudomonas fluorescens , and Pseudomonas putida.

8. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism is a species of yeast or fungus selected from the group consisting of Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces marxianus, Aspergillus terreus, Aspergillus niger and Pichia pastoris.

9. The non-naturally occurring microbial organism of claim 8 , wherein the microbial organism is a species of Saccharomyces cerevisiae.

10. A method for producing 6-aminocaproic acid, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce 6-aminocaproic acid.

11. The method of claim 10 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

12. The method of claim 10 , wherein said method further comprises separating the 6-aminocaproic acid from other components in the culture.

13. The method of claim 12 , wherein the separating comprises extraction, continuous liquid-liquid extraction, pervaporation, membrane filtration, membrane separation, reverse osmosis, electrodialysis, distillation, crystallization, centrifugation, extractive filtration, ion exchange chromatography, size exclusion chromatography, absorption chromatography, or ultrafiltration.

14. A method for producing caprolactam, comprising culturing the non-naturally occurring microbial organism of claim 3 under conditions and for a sufficient period of time to produce caprolactam.

15. The method of claim 14 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

16. The method of claim 14 , wherein said method further comprises separating the caprolactam from other components in the culture.

17. The method of claim 14 , wherein the separating comprises extraction, continuous liquid-liquid extraction, pervaporation, membrane filtration, membrane separation, reverse osmosis, electrodialysis, distillation, crystallization, centrifugation, extractive filtration, ion exchange chromatography, size exclusion chromatography, absorption chromatography, or ultrafiltration.

Assignees

• Genomatica Inc

Inventors

• Burgard Anthony P
• Pharkya Priti
• Osterhout Robin E

Classifications

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12P7/44

  Polycarboxylic acids · CPC title

• C12Y102/0101

  Acetaldehyde dehydrogenase (acetylating) (1.2.1.10) · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12N9/80

  acting on amide bonds in linear amides {(3.5.1)} · CPC title