Microorganisms and methods for the biosynthesis of adipate, hexamethylenediamine and 6-aminocaproic acid
View Patent ↗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.
1. A non-naturally occurring microbial organism comprising a hexamethylenediamine pathway and at least two exogenous nucleic acids encoding a hexamethylenediamine pathway enzyme expressed in a sufficient amount to produce hexamethylenediamine, said hexamethylenediamine pathway comprising:
(A) 6-aminocaproate N-acetyltransferase;
(B) 6-acetamidohexanoate kinase;
(C) [(6-acetamidohexanoyl)oxy]phosphonate (6-AAHOP) oxidoreductase;
(D) 6-acetamidohexanal aminotransferase or 6-acetamidohexanal oxidoreductase (aminating); and
(E) 6-acetamidohexanamine N-acetyltransferase or 6-acetamidohexanamine hydrolase (amide).
2. The non-naturally occurring microbial organism of claim 1 , further comprising a 6-aminocaproic acid pathway.
3. The non-naturally occurring microbial organism of claim 2 , wherein said 6-aminocaproic acid pathway comprises:
(A) CoA-dependent aldehyde dehydrogenase; and
(B) transaminase or 6-aminocaproate dehydrogenase.
4. The non-naturally occurring microbial organism of claim 2 , wherein said 6-aminocaproic acid pathway comprises:
(A) 3-oxo-6-aminohexanoyl-CoA thiolase;
(B) 3-oxo-6-aminohexanoyl-CoA/acyl-CoA transferase, 3-oxo-6-aminohexanoyl-CoA synthase, or 3-oxo-6-aminohexanoyl-CoA hydrolase;
(C) 3-oxo-6-aminohexanoate reductase;
(D) 3-hydroxy-6-aminohexanoate dehydratase; and
(E) 6-aminohex-2-enoate reductase.
5. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an hexamethylenediamine pathway enzyme.
6. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an hexamethylenediamine pathway enzyme.
7. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises five exogenous nucleic acids each encoding an hexamethylenediamine pathway enzyme.
8. The non-naturally occurring microbial organism of claim 1 , wherein said hexamethylenediamine pathway comprises:
(A) 6-aminocaproate N-acetyltransferase;
(B) 6-acetamidohexanoate kinase;
(C) 6-AAHOP oxidoreductase;
(D) 6-acetamidohexanal aminotransferase; and
(E) 6-acetamidohexanamine N-acetyltransferase.
9. The non-naturally occurring microbial organism of claim 1 , wherein said hexamethylenediamine pathway comprises:
(A) 6-aminocaproate N-acetyltransferase;
(B) 6-acetamidohexanoate kinase;
(C) 6-AAHOP oxidoreductase;
(D) 6-acetamidohexanal aminotransferase; and
(E) 6-acetamidohexanamine hydrolase (amide).
10. The non-naturally occurring microbial organism of claim 1 , wherein said hexamethylenediamine pathway comprises:
(A) 6-aminocaproate N-acetyltransferase;
(B) 6-acetamidohexanoate kinase;
(C) 6-AAHOP oxidoreductase;
(D) 6-acetamidohexanal oxidoreductase (aminating); and
(E) 6-acetamidohexanamine N-acetyltransferase.
11. The non-naturally occurring microbial organism of claim 1 , wherein said hexamethylenediamine pathway comprises:
(A) 6-aminocaproate N-acetyltransferase;
(B) 6-acetamidohexanoate kinase;
(C) 6-AAHOP oxidoreductase;
(D) 6-acetamidohexanal oxidoreductase (aminating); and
(E) 6-acetamidohexanamine hydrolase (amide).
12. The non-naturally occurring microbial organism of claim 1 , wherein said at least two exogenous nucleic acid is a heterologous nucleic acid.
13. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in culture medium comprising dissolved oxygen in an amount that is less than 10% of saturation.
14. 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.
15. A method for producing hexamethylenediamine, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce hexamethylenediamine.
16. The method of claim 15 , wherein said non-naturally occurring microbial organism is in culture medium, wherein said culture medium is in an atmosphere of less than 1% oxygen.
17. The method of claim 16 , wherein said non-naturally occurring microbial organism is in culture medium comprising dissolved oxygen in an amount that is less than 10% of saturation.