Microorganisms and methods for the biosynthesis of adipate, hexamethylenediamine and 6-aminocaproic acid

Patent metadata

Field	Value
Publication number	US-11208673-B2
Application number	US-201816213671-A
Country	US
Kind code	B2
Filing date	Dec 7, 2018
Priority date	May 7, 2009
Publication date	Dec 28, 2021
Grant date	Dec 28, 2021

Abstract

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

First claim

What is claimed is:

1. A non-naturally occurring microbial organism, comprising a microbial organism having a 6-aminocaproic acid (6-ACA) pathway comprising at least two exogenous nucleic acids encoding 6-ACA pathway enzymes expressed in a sufficient amount to produce 6-ACA from glutamate, said 6-ACA pathway enzymes comprising: a glutamyl-CoA transferase or a glutamyl-CoA ligase; a beta-ketothiolase; an 3-oxo-6-aminopimeloyl-CoA oxidoreductase; a 3-hydroxy-6-aminopimeloyl-CoA dehydratase; a 6-amino-7-carboxyhept-2-enoyl-CoA reductase; 6-aminopimeloyl-CoA hydrolase, transferase and/or ligase; and a 2-aminopimelate decarboxylase.

2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises a set of exogenous nucleic acids encoding 6-ACA pathway enzymes expressed in a sufficient amount to produce 6-ACA, said set of exogenous nucleic acids comprising: a glutamyl-CoA transferase or a glutamyl-CoA ligase; a beta-ketothiolase; a 3-oxo-6-aminopimeloyl-CoA oxidoreductase; a 3-hydroxy-6-aminopimeloyl-CoA dehydratase; a 6-amino-7-carboxyhept-2-enoyl-CoA reductase; 6-aminopimeloyl-CoA hydrolase, transferase and/or ligase; and a 2-aminopimelate decarboxylase.

3. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least three exogenous nucleic acids.

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

5. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in culture medium, wherein said culture medium is in an atmosphere of less than 1% oxygen or comprises dissolved oxygen in an amount that is less than 10% of saturation.

6. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least four exogenous nucleic acids.

7. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least five exogenous nucleic acids.

8. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least six exogenous nucleic acids.

9. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least seven exogenous nucleic acids.

10. The non-naturally occurring microbial organism of claim 1 , wherein said 6-ACA pathway comprises at least eight exogenous nucleic acids.

11. A method for producing 6-aminocaproic acid (6-ACA), comprising culturing a non-naturally occurring microbial organism of claim 5 under conditions and for a sufficient period of time to produce 6-ACA.

12. The non-naturally occurring microbial organism of claim 1 , part (iv), further comprising a 2-amino-7-oxosubarate pathway comprising at least one exogenous nucleic acid encoding a 2-amino-7-oxosubarate pathway enzyme expressed in a sufficient amount to produce 2-amino-7-oxosubarate, said 2-amino-7-oxosubarate pathway comprising a 2-amino-5-hydroxy-7-oxosubarate aldolase, a 2-amino-5-hydroxy-7-oxosubarate dehydratase, or a 2-amino-5-ene-7-oxosubarate reductase.

13. The non-naturally occurring microbial organism of claim 2 , part (v), further comprising a 2-amino-7-oxosubarate pathway comprising a second set of exogenous nucleic acids encoding 2-amino-7-oxosubarate pathway enzymes expressed in a sufficient amount to produce 2-amino-7-oxosubarate, said second set of exogenous nucleic acids encoding a 2-amino-5-hydroxy-7-oxosubarate aldolase; a 2-amino-5-hydroxy-7-oxosubarate dehydratase; and a 2-amino-5-ene-7-oxosubarate reductase.

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

Assignees

• Genomatica Inc

Inventors

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

Classifications

• C12P13/04

  Alpha- or beta- amino acids {(other amino acids C12P13/005)} · CPC title

• C12P13/001

  Amines; Imines · CPC title

• C12P17/10 Primary

  Nitrogen as only ring hetero atom · CPC title

• C12P13/02

  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 title

• C12P7/40

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title