Microorganisms for the production of 1,4-Butanediol
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-007/18
C12N-009/04
C12N-009/88
C12N-009/90
C12N-015/52
C12N-015/70
C12N-015/81
출원번호
US-0449187
(2012-04-17)
등록번호
US-9175297
(2015-11-03)
발명자
/ 주소
Burk, Mark J.
Burgard, Anthony P.
Osterhout, Robin E.
Sun, Jun
출원인 / 주소
Genomatica, Inc.
대리인 / 주소
Jones Day
인용정보
피인용 횟수 :
1인용 특허 :
96
초록▼
The invention provides non-naturally occurring microbial organisms comprising a 1,4-butanediol (BDO) pathway comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO. The invention additionally provides methods of using such microbi
The invention provides non-naturally occurring microbial organisms comprising a 1,4-butanediol (BDO) pathway comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO. The invention additionally provides methods of using such microbial organisms to produce BDO.
대표청구항▼
1. A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, wherein said microbial organism is transformed with at least one heterologous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein s
1. A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, wherein said microbial organism is transformed with at least one heterologous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise: (a) 4-aminobutyrate CoA transferase, said enzyme classified as EC 2.8.3.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA; 4-aminobutyryl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA; or 4-aminobutyrate-CoA ligase, said enzyme classified as EC 6.2.1.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA;(b) 4-aminobutyryl-CoA oxidoreductase (deaminating), said enzyme classified as EC1.4.1.a and converting 4-aminobutyryl-CoA to 4-oxobutyryl-CoA; or 4-aminobutyryl-CoA transaminase, said enzyme classified as EC 2.6.1.a and converting 4-aminobutyryl-CoA to 4-oxobutyryl-CoA;(c) 4-hydroxybutyryl-CoA dehydrogenase, said enzyme classified as EC 1.1.1.a and converting 4-oxobutyryl-CoA to 4-hydroxybutyryl-CoA, with the proviso that said non-naturally occurring microorganism is not a wild type microorganism inherently having any of the above genes; and(d) 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol; or 4-hydroxybutyryl-CoA reductase, said enzyme classified as EC 1.2.1.b and converting 4-hydroxybutyryl-CoA to 4-hydroxybutanal, and 1,4-butanediol dehydrogenase, said enzyme classified as EC 1.1.1.a and converting 4-hydroxybutanal to 1,4-butanediol. 2. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism is transformed with heterologous nucleic acids encoding one enzyme of each step of: (a) 4-aminobutyrate CoA transferase; 4-aminobutyryl-CoA hydrolase; or 4-aminobutyrate-CoA ligase;(b) 4-aminobutyryl-CoA oxidoreductase (deaminating); or 4-aminobutyryl-CoA transaminase;(c) 4-hydroxybutyryl-CoA dehydrogenase; and(d) 4-hydroxybutyryl-CoA reductase (alcohol forming); or 4-hydroxybutyryl-CoA reductase and 1,4-butanediol dehydrogenase. 3. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyrate CoA transferase. 4. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA hydrolase. 5. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyrate-CoA ligase. 6. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA oxidoreductase (deaminating). 7. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA transaminase. 8. The non-naturally occurring microbial organism of claim 2, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-hydroxybutyryl-CoA reductase (alcohol forming). 9. The non-naturally occurring microbial organism of claim 2, wherein said microbial is transformed with a heterologous nucleic acid encoding 4-hydroxybutyryl-CoA reductase and 1,4-butanediol dehydrogenase. 10. A method for producing BDO, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce BDO. 11. The method of claim 10, wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 12. A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, wherein said microbial organism is transformed with at least one heterologous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise: (a) 4-aminobutyrate CoA transferase, said enzyme classified as EC 2.8.3.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA; 4-aminobutyryl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA; or 4-aminobutyrate-CoA ligase, said enzyme classified as EC 6.2.1.a and converting 4-aminobutyrate to 4-aminobutyryl-CoA;(b) 4-aminobutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-aminobutyryl-CoA to 4-aminobutan-1-ol; or 4-aminobutyryl-CoA reductase, said enzyme classified as EC 1.2.1.b and converting 4-aminobutyryl-CoA to 4-aminobutanal, and 4-aminobutan-1-ol dehydrogenase, said enzyme classified as EC 1.1.1.a and converting 4-aminobutanal to 4-aminobutan-1-ol;(c) 4-aminobutan-1-ol oxidoreductase (deaminating), said enzyme classified as EC 1.4.1.a and converting 4-aminobutan-1-ol to 4-hydroxybutanal; or 4-aminobutan-1-ol transaminase, said enzyme classified as EC 2.6.1.a and converting 4-aminobutan-1-ol to 4-hydroxybutanal; and(d) 1,4-butanediol dehydrogenase, said enzyme classified as EC 1.1.1.a and converting 4-hydroxybutanal to 1,4-butanediol. 13. The non-naturally occurring microbial organism of claim 12, wherein said microbial organism is transformed with heterologous nucleic acids encoding at least one enzyme of each step of: (a) 4-aminobutyrate CoA transferase; 4-aminobutyryl-CoA hydrolase; or 4-aminobutyrate-CoA ligase;(b) 4-aminobutyryl-CoA reductase (alcohol forming); or 4-aminobutyryl-CoA reductase and 4-aminobutan-1-ol dehydrogenase;(c) 4-aminobutan-1-ol oxidoreductase (deaminating); or 4-aminobutan-1-ol transaminase; and(d) 1,4-butanediol dehydrogenase. 14. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyrate CoA transferase. 15. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA hydrolase. 16. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyrate-CoA ligase. 17. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA reductase (alcohol forming). 18. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutyryl-CoA reductase and 4-aminobutan-1-ol dehydrogenase. 19. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutan-1-ol oxidoreductase (deaminating). 20. The non-naturally occurring microbial organism of claim 13, wherein said microbial organism is transformed with a heterologous nucleic acid encoding 4-aminobutan-1-ol transaminase. 21. A method for producing BDO, comprising culturing the non-naturally occurring microbial organism of claim 12 under conditions and for a sufficient period of time to produce BDO. 22. The method of claim 21, wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.
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