Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-001/20
C12P-007/18
C12P-007/42
C12N-009/04
C12N-009/88
C12N-009/00
C12N-009/02
C12P-007/52
C12P-017/04
출원번호
US-0286135
(2011-10-31)
등록번호
US-8889399
(2014-11-18)
발명자
/ 주소
Burk, Mark J.
Van Dien, Stephen J.
Burgard, Anthony P.
Niu, Wei
출원인 / 주소
Genomatica, Inc.
대리인 / 주소
Jones Day
인용정보
피인용 횟수 :
0인용 특허 :
95
초록▼
The invention provides a non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways. The pathways include exogenous nucleic acids encoding a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydrox
The invention provides a non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways. The pathways include exogenous nucleic acids encoding a) an α-ketoglutarate 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 the exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO). Also provide is a method for the production of 1,4-BDO. The method includes culturing the non-naturally occurring microbial organism having 4-HB and 1,4-BDO biosynthetic pathways substantially anaerobic conditions for a sufficient period of time to produce 1,4-BDO.
대표청구항▼
1. A non-naturally occurring microbial organism having increased production of 1,4-butanediol, which is prepared by introducing exogenous genes or increasing expression of endogenous genes encoding enzymes converting succinate into 4-hydroxybutyrate, 4-hydroxybutyrate into 4-hydroxybutyryl-CoA direc
1. A non-naturally occurring microbial organism having increased production of 1,4-butanediol, which is prepared by introducing exogenous genes or increasing expression of endogenous genes encoding enzymes converting succinate into 4-hydroxybutyrate, 4-hydroxybutyrate into 4-hydroxybutyryl-CoA directly or through 4-hydroxybutyryl phosphate, and 4-hydroxybutyryl-CoA into 1,4-butanediol directly or through 4-hydroxybutyraldehyde, in a microorganism capable of producing succinate, wherein said enzyme that converts 4-hydroxybutyrate into 4-hydroxybutyryl-CoA directly is a 4-hydroxybutyrate:CoA transferase, wherein said enzymes that converts 4-hydroxybutyrate into 4-hydroxybutyryl-CoA through 4-hydroxybutyryl phosphate is a butyryl kinase and a phosphotransbutyrylase, wherein said enzyme that converts 4-hydroxybutyryl-CoA into 1,4-butanediol directly is an aldehyde/alcohol dehydrogenase, wherein said enzymes that converts 4-hydroxybutyryl-CoA into 1,4-butanediol through 4-hydroxybutyraldehyde is an aldehyde dehydrogenase and an alcohol dehydrogenase. 2. The non-naturally occurring microbial organism of claim 1, wherein said microorganism capable of producing succinate is selected from the group consisting of bacteria, yeast and fungus that exhibit higher production of succinate compared to a wild-type microbial organism of the same species. 3. The non-naturally occurring microbial organism of claim 2, wherein the bacteria is a bacteria applicable for fermentation. 4. The non-naturally occurring microbial organism of claim 3, wherein the bacteria have genetic alterations in genes encoding lactate dehydrogenase and pyruvate formate-lyase, and produce succinate in a higher concentration compared to a wild-type microbial organism of the same species without substantial production of other organic acids in an anaerobic condition. 5. The non-naturally occurring microbial organism of claim 3, wherein the bacteria have genetic alterations in genes encoding lactate dehydrogenase, pyruvate formate-lyase, phosphotransacetylase and acetate kinase, and produce succinate in a higher concentration compared to a wild-type microbial organism of the same species without substantial production of other organic acids in an anaerobic condition. 6. The non-naturally occurring microbial organism of claim 3, wherein the bacteria have genetic alterations in genes encoding lactate dehydrogenase, pyruvate formate-lyase, and phosphopyruvate carboxylase, and produce succinate in a higher concentration compared to a wild-type microbial organism of the same species without substantial production of other organic acids in an anaerobic condition. 7. The non-naturally occurring microbial organism of claim 3, wherein the bacteria are selected from the group consisting of Anaerobiospirillum succiniciproducens, Actinobacillus succinogenes, Mannheimia succiniciproducens, Corynebacterium glutamicum, and E. coli. 8. The non-naturally occurring microbial organism of claim 7, wherein the bacteria is Mannheimia succiniciproducens. 9. The non-naturally occurring microbial organism of claim 7, wherein the bacteria is Mannheimia succiniciproducens having higher succinate production compared to a wild-type microbial organism of the same species. 10. The non-naturally occurring microbial organism of claim 7, wherein the bacteria is E. coli comprising genetic alterations in genes that result in production of succinate in a higher concentration compared to a wild-type microbial organism of the same species without substantial production of other organic acids in an anaerobic condition. 11. The non-naturally occurring microbial organism of claim 10, wherein the genetic alterations are nucleic acid deletions. 12. The non-naturally occurring microbial organism of claim 1, where the gene encoding the enzyme converting succinate into 4-hydroxybutrate is isolated from Clostridium kluyveri. 13. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding the enzyme converting succinate into 4-hydroxybutyrate is selected from the group consisting of genes encoding succinyl-CoA transferase (Cat1), succinate semialdehyde dehydrogenase (SucD), 4-hydroxybutyrate dehydrogenase (4hbD) and the 4-hydroxybutyrate dehydrogenase of Ralstonia eutropha. 14. The non-naturally occurring microbial organism of claim 13, wherein the gene encoding succinyl-CoA transferase has a nucleotide coding sequence of the Cat1 gene of Clostridium kluyveri DSM555, the gene encoding succinate semialdehyde dehydrogenase has a nucleotide coding sequence of the SucD gene of Clostridium kluyveriDSM555, the gene encoding 4-hydroxybutyrate dehydrogenase has a nucleotide coding sequence of the 4hbD gene of Clostridium kluyveri DSM555, and the gene encoding 4-hydroxybutyrate dehydrogenase of Ralstonia eutropha has a nucleotide coding sequence of the 4-hbD gene of Ralstonia eutropha H16. 15. The non-naturally occurring microbial organism of claim 13, wherein the microbial organism comprises a gene encoding succinyl-CoA transferase (Cat1); a gene encoding succinate semialdehyde dehydrogenase (SucD); and a gene encoding 4-hydroxybutyrate dehydrogenase (4hbD) or a gene encoding 4-hydroxybutyrate dehydrogenase of Ralstonia eutropha. 16. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding the enzyme converting 4-hydroxybutyrate into 1,4-butanediol is isolated from Clostridium acetobutylicum. 17. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding 4-hydroxybutyrate:CoA transferase has a nucleotide coding sequence of the Cat2 gene of Clostridium kluyveri DSM555. 18. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding phosphotransbutyrylase has a nucleotide coding sequence of the ptb gene of Clostridium acetobutylicum ATCC 824 and the gene encoding butyryl kinase have a nucleotide coding sequence of the bukl gene of Clostridium acetobutylicum ATCC 824. 19. The non-naturally occurring microbial organism of claim 1, wherein the alcohol dehydrogenase is butyl-CoA dehydrogenase isolated from Clostridium acetobutylicum. 20. The non-naturally occurring microbial organism of claim 19, wherein the gene encoding butyl-CoA dehydrogenase has a nucleotide coding sequence of the adhE gene of Clostridium acetobutylicum ATCC 824 or the adhE2 gene of Clostridium acetobutylicum ATCC 824. 21. The non-naturally occurring microbial organism of claim 1, wherein the microbial organism has a genetic alteration in a gene that encodes an enzyme that converts succinate semialdehyde into succinate. 22. The non-naturally occurring microbial organism of claim 21, wherein the gene that encodes an enzyme that converts succinate semialdehyde into succinate is a gene encoding succinic semialdehyde dehydrogenase (GabD). 23. The non-naturally occurring microbial organism of claim 22, wherein the gene encoding succinic semialdehyde dehydrogenase (GabD) has a nucleotide coding sequence of the GabD gene of E. coli. 24. The non-naturally occurring microbial organism of claim 1, wherein a gene encoding an enzyme that augments the synthesis of succinate is further introduced or endogenous expression is increased in the microbial organism. 25. A non-naturally occurring microbial organism having increased production of 1,4-butanediol, which is prepared by introducing exogenenous genes or increasing expression of endogenous genes encoding succinyl-CoA transferase (Cat1); semialdehyde dehydrogenase (SucD);4-hydroxybutyrate dehydrogenase (4hbD) or 4-hydroxybutyrate dehydrogenase of Ralstonia eutropha; 4-hydroxybutyrate:CoA transferase, or phosphotransbutyrylase and butyrate kinase I; and butyl-CoA dehydrogenase, in a microorganism capable of producing succinate. 26. The non-naturally occurring microbial organism of claim 25, wherein a gene encoding succinic semialdehyde dehydrogenase (GabD) is disrupted in the microbial organism. 27. The non-naturally occurring microbial organism of claim 25, wherein a gene encoding an enzyme that augments the synthesis of succinate is further introduced or endogenous expression is increased in the microbial organism. 28. A method for preparing 1,4-butanediol, comprising: culturing the non-naturally occurring microbial organism of claim 1, in a medium containing a carbon source; and obtaining 1,4-butanediol from the medium. 29. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding aldehyde dehydrogenase has a nucleotide coding sequence of the bld gene of Clostridium saccharoperbutylacetonicum. 30. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding alcohol dehydrogenase has a nucleotide coding sequence of a gene selected from the group consisting of the bdhB gene of Clostridium acetobutylicum ATCC 824, the adhEm gene isolated from metalibrary of anaerobic sewage digester microbial consortia, the adhE gene of Clostridium thermocellum, the ald gene of Clostridium beijerinckii, the bdhA gene of Clostridium acetobutylicum ATCC 824, and the bdh gene of Clostridium saccharoperbutylacetonicum. 31. The non-naturally occurring microbial organism of claim 1, wherein the gene encoding aldehyde/alcohol dehydrogenase has a nucleotide coding sequence of a gene selected from the group consisting of the adhE gene of Clostridium acetobutylicum ATCC 824, the adhE2 gene of Clostridium acetobutylicum ATCC 824, the adhE gene of E. coli, the yqhD gene of E. coli, the adhE gene of Clostridium tetani, the adhE gene of Clostridium perfringens, and the adhE gene of Clostridium difficile. 32. The non-naturally occurring microbial organism of claim 1, wherein the enzymes convert 4-hydroxybutyrate into 4-hydroxybutyryl-CoA directly and 4-hydroxybutyryl-CoA into 1,4-butanediol directly. 33. The non-naturally occurring microbial organism of claim 1, wherein the enzymes convert 4-hydroxybutyrate into 4-hydroxybutyryl-CoA through 4-hydroxybutyryl phosphate and 4-hydroxybutyryl-CoA into 1,4-butanediol directly. 34. The non-naturally occurring microbial organism of claim 1, wherein the enzymes convert 4-hydroxybutyrate into 4-hydroxybutyryl-CoA directly and 4-hydroxybutyryl-CoA into 1,4-butanediol through 4-hydroxybutyraldehyde. 35. The non-naturally occurring microbial organism of claim 1, wherein the enzymes convert 4-hydroxybutyrate into 4-hydroxybutyryl-CoA through 4-hydroxybutyryl phosphate and 4-hydroxybutyryl-CoA into 1,4-butanediol through 4-hydroxybutyraldehyde.
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Huisman Gjalt W. ; Skraly Frank ; Martin David P. ; Peoples Oliver P., Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids.
Huisman, Gjalt W.; Skraly, Frank; Martin, David P.; Peoples, Oliver P., Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids.
Tobin Allan J. (Los Angeles CA) Erlander Mark G. (Tarzana CA) Kaufman Daniel L. (Santa Monica CA) Clare-Salzler Michael J. (Los Angeles CA), Cloned glutamic acid decarboxylase peptides.
Burdette Douglas S. ; Zeikus Joseph G., Cloning and expression of the gene encoding thermoanaerobacter ethanolicus 39E secondary-alcohol dehydrogenase and enzy.
Burk, Mark J.; Van Dien, Stephen J.; Burgard, Anthony P.; Niu, Wei, Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors.
Frommer Wolf-Bernd (Berlin DEX) Riesmeier Jorg (Berlin DEX), DNA sequences with oligosaccharide transporter, plasmids, bacteria and plants containing a transporter as well as a proc.
Davis,S. Christopher; Grate,John H.; Gray,David R.; Gruber,John M.; Huisman,Gjalt W.; Ma,Steven K.; Newman,Lisa M.; Sheldon,Roger; Wang,Li A, Enzymatic processes for the production of 4-substituted 3-hydroxybutyric acid derivatives.
Davis,S. Christopher; Grate,John H.; Gray,David R.; Gruber,John M.; Huisman,Gjalt W.; Ma,Steven K.; Newman,Lisa M.; Sheldon,Roger; Wang,Li A, Enzymatic processes for the production of 4-substituted 3-hydroxybutyric acid derivatives and vicinal cyano, hydroxy substituted carboxylic acid esters.
Cao,Yongwei; Hinkle,Gregory J.; Slater,Steven C.; Chen,Xianfeng; Goldman,Barry S., Expression of microbial proteins in plants for production of plants with improved properties.
Rieping, Mechthild; Bastuck, Christine; Hermann, Thomas; Thierbach, Georg, Fermentation process for the preparation of L-amino acids using strains of the family Enterobacteriaceae.
Tobin Allan J. ; Erlander Mark G. ; Kaufman Daniel L., Hybridomas and monoclonal antibodies that specifically bind to glutamic acid decarboxylase peptides.
Broecker Franz Josef (Ludwigshafen DT) Schwarzmann Matthias (Limburgerhof DT), Manufacture of butanediol and/or tetrahydrofuran from maleic and/or succinic anhydride via g
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Donnelly Mark I. ; Sanville-Millard Cynthia ; Chatterjee Ranjini, Method for construction of bacterial strains with increased succinic acid production.
Allan J. Tobin ; Mark G. Erlander ; Daniel L. Kaufman, Methods and kits useful for determining the status of and detecting pancreatic B-cell associated autoimmune diseases.
Gruys Kenneth James ; Mitsky Timothy Albert ; Kishore Ganesh Murthy ; Slater Steven Charles ; Padgette Stephen Rogers ; Stark David Martin, Methods of optimizing substrate pools and biosynthesis of poly-.beta.-hydroxybutyrate-co-poly-.beta.-hydroxyvalerate in.
Gruys Kenneth James ; Mitsky Timothy Albert ; Kishore Ganesh Murthy ; Slater Steven Charles ; Padgette Stephen Rogers ; Stark David Martin, Methods of optimizing substrate pools and biosynthesis of poly-.beta.-hydroxybutyrate-co-poly-.beta.-hydroxyvalerate in.
Timothy A. Mitsky ; Steven C. Slater ; Steven E. Reiser ; Ming Hao ; Kathryn L. Houmiel, Multigene expression vectors for the biosynthesis of products via multienzyme biological pathways.
Steinbuchel Alexander,DEX ; Liebergesell Mathias,DEX ; Valentin Henry ; Pries Andreas,DEX, PHA E and PHA C components of poly(hydroxy fatty acid) synthase from thiocapsa pfennigii.
Tobin Allan J. (Los Angeles CA) Erlander Mark G. (Tarzana CA) Kaufman Daniel L. (Santa Monica CA) Clare-Salzler Michael J. (Gainesville FL), Peptides derived from glutamic acid decarboxylase.
Cannon, Maura; Cannon, Francis C.; McCool, Gabriel J.; Valentin, Henry E.; Gruys, Kenneth J., Polyhydroxyalkanoate biosynthesis associated proteins and coding region in bacillus megaterium.
Matsuyama Akinobu (Niigata JPX) Nikaido Teruyuki (Niigata JPX) Kobayashi Yoshinori (Niigata JPX), Process for producing optically active 1,3-butanediol by reduction of 4-hydroxy-2-butanone.
Emptage,Mark; Haynie,Sharon L.; Laffend,Lisa A.; Pucci,Jeff P.; Whited,Gregory, Process for the biological production of 1,3-propanediol with high titer.
Emptage,Mark; Haynie,Sharon L.; Laffend,Lisa A.; Pucci,Jeff P.; Whited,Gregory Marshall, Process for the biological production of 1,3-propanediol with high titer.
Gottschalk G. (Nrtenhardenberg DEX) Averhoff Beate (Gottingen DEX), Process for the microbiological preparation of 1,3-propane-diol from glycerol by citrobacter.
Rieping,Mechthild; Hermann,Thomas, Process for the production of L-amino acids using strains of the family Enterobacteriaceae that contain an attenuated fruR gene.
Somerville Christopher R. (Portola Valley CA) Nawrath Christiane (Palo Alto CA) Poirier Yves (Palo Alto CA), Processes for producing polyhydroxybutyrate and related polyhydroxyalkanoates in the plastids of higher plants.
Jao Go Pan KR; Soo An Shin KR; Chan Kyu Park KR; Pil Kim KR; Dong Eun Chang KR; Jae Eun Kim KR, Pta LDHA double mutant Escherichia coli SS373 and the method of producing succinic acid therefrom.
Hespell Robert B. ; Wyckoff Herbert A. ; Dien Bruce S. ; Bothast Rodney J., Stabilization of pet operon plasmids and ethanol production in bacterial strains lacking lactate dehydrogenase and pyruvate formate lyase activities.
Somerville Christopher R. (Okemos MI) Poirier Yves (East Lansing MI) Dennis Douglas E. (Weyers Cave VA), Transgenic plants producing polyhydroxyalkanoates.
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