Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-007/18
C12N-001/21
출원번호
US-0348564
(2012-01-11)
등록번호
US-8357520
(2013-01-22)
발명자
/ 주소
Burk, Mark J.
Van Dien, Stephen J.
Burgard, Anthony P.
Niu, Wei
출원인 / 주소
Genomatica, Inc.
인용정보
피인용 횟수 :
4인용 특허 :
95
초록▼
The invention provides a non-naturally occurring microbial biocatalyst including a microbial organism having a 4-hydroxybutanoic acid (4-HB) biosynthetic pathway having at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semi
The invention provides a non-naturally occurring microbial biocatalyst including a microbial organism having a 4-hydroxybutanoic acid (4-HB) biosynthetic pathway having at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, or α-ketoglutarate decarboxylase, wherein the exogenous nucleic acid is expressed in sufficient amounts to produce monomeric 4-hydroxybutanoic acid (4-HB). Also provided is a non-naturally occurring microbial biocatalyst including a microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (BDO) biosynthetic pathways, the pathways include at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate:CoA transferase, 4-butyrate kinase, phosphotransbutyrylase, α-ketoglutarate decarboxylase, aldehyde dehydrogenase, alcohol dehydrogenase or an aldehyde/alcohol dehydrogenase, wherein the exogenous nucleic acid is expressed in sufficient amounts to produce 1,4-butanediol (BDO). Additionally provided are methods for the production of 4-HB and BDO.
대표청구항▼
1. A non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways, said pathways comprising exogenous nucleic acids encoding a) an α-ketoglutarate decarboxylase, or an α-ketoglutarate dehydrogenase and a CoA-dependent succinic sem
1. A non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways, said pathways comprising exogenous nucleic acids encoding a) an α-ketoglutarate decarboxylase, or an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase, or a glutamate:succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase, wherein said exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO). 2. The non-naturally occurring microbial organism of claim 1, wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid. 3. The non-naturally occurring microbial organism of claim 1, wherein the microbial organism is in a substantially anaerobic culture medium. 4. A method for the production of 1,4-BDO comprising, culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce 1,4-BDO. 5. The method of claim 4, wherein at least one exogenous nucleic acid comprises a heterologous nucleic acid. 6. The method of claim 4, wherein the microbial organism is in a substantially anaerobic culture medium. 7. The non-naturally occurring microbial organism of claim 1, comprising the α-ketoglutarate decarboxylase. 8. The non-naturally occurring microbial organism of claim 1, comprising the α-ketoglutarate dehydrogenase and the CoA-dependent succinic semialdehyde dehydrogenase. 9. The non-naturally occurring microbial organism of claim 1, comprising the glutamate:succinate semialdehyde transaminase and the glutamate decarboxylase. 10. The non-naturally occurring microbial organism of claim 1, comprising the 4-hydroxybutyryl-CoA:acetyl-CoA transferase. 11. The non-naturally occurring microbial organism of claim 1, comprising the butyrate kinase and the phosphotransbutyrylase. 12. The method of claim 4, wherein the microbial organism comprises the α-ketoglutarate decarboxylase. 13. The method of claim 4, wherein the microbial organism comprises the α-ketoglutarate dehydrogenase and the CoA-dependent succinic semialdehyde dehydrogenase. 14. The method of claim 4, wherein the microbial organism comprises the glutamate:succinate semialdehyde transaminase and the glutamate decarboxylase. 15. The method of claim 4, wherein the microbial organism comprises the 4-hydroxybutyryl-CoA:acetyl-CoA transferase. 16. The method of claim 4, wherein the microbial organism comprises the butyrate kinase and the phosphotransbutyrylase. 17. The non-naturally occurring microbial organism of claim 7, further comprising a CoA-dependent succinic semialdehyde dehydrogenase. 18. The non-naturally occurring microbial organism of claim 9, further comprising a CoA-dependent succinic semialdehyde dehydrogenase. 19. The method of claim 12, wherein the microbial organism further comprises a CoA-dependent succinic semialdehyde dehydrogenase. 20. The method of claim 14, wherein the microbial organism further comprises a CoA-dependent succinic semialdehyde dehydrogenase.
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