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US 2011/0217742 A1Published application
Microorganisms and methods for the coproduction 1,4-butanediol and gamma-butyrolactone
AssigneeGenomatica, Inc.
InventorsJun Sun, Mark J. Burk, Anthony P. Burgard, Robin E. Osterhout, Wei Niu, John D. Trawick, Robert Haselbeck
Life of this patent
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Abstract
The invention provides non-naturally occurring microbial organisms comprising 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL) pathways comprising at least one exogenous nucleic acid encoding a 14-BDO and GBL pathway enzyme expressed in a sufficient amount to produce 14-BDO and GBL. The invention additionally provides methods of using such microbial organisms to produce 14-BDO and GBL.
Claims52
A method for producing 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL), comprising culturing a non-naturally occurring microbial organism under conditions and for a sufficient period of time to produce 14-BDO and 4-hydroxybutyryl-CoA followed by spontaneous lactonization of 4-hydroxybutyryl-CoA to form GBL, wherein said non-naturally occurring microbial organism comprise a microbial organism having a 14-BDO pathway and a 4-hydroxybutyryl-CoA pathway, said 14-BDO pathway comprising at least one exogenous nucleic acid encoding a 14-BDO pathway enzyme expressed in a sufficient amount to produce 14-BDO, said 14-BDO pathway comprising:
- a 4-hydroxybutyryl-CoA reductase (alcohol forming), a succinyl-CoA reductase (aldehyde forming), an alpha-ketoglutarate decarboxylase, a 4-hydroxybutyrate dehydrogenase, a 4-hydroxybutyrate reductase, a 1,4-butanediol dehydrogenase, a succinate reductase, a succinyl-CoA transferase, a succinyl-CoA hydrolase, a succinyl-CoA synthetase, a glutamate dehydrogenase, a glutamate transaminase, a glutamate decarboxylase, an 4-aminobutyrate dehydrogenase, an 4-aminobutyrate transaminase, a 4-hydroxybutyrate kinase, a phosphotrans-4-hydroxybutyrylase, a 4-hydroxybutyryl-CoA reductase (aldehyde forming), a 4-hydroxybutyryl-phosphate reductase, a succinyl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase, a 4-hydroxybutyryl-CoA synthetase, an alpha-ketoglutarate reductase, a 5-hydroxy-2-oxopentanoate dehydrogenase, a 5-hydroxy-2-oxopentanoate decarboxylase or a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation),
- said 4-hydroxybutyryl-CoA pathway comprising at least one exogenous nucleic acid encoding an 4-hydroxybutyryl-CoA pathway enzyme expressed in a sufficient amount to produce 4-hydroxybutyryl-CoA, said 4-hydroxybutyryl-CoA pathway comprising:
- a 4-hydroxybutyryl-CoA transferase, a succinyl-CoA reductase (aldehyde forming), an alpha-ketoglutarate decarboxylase, a 4-hydroxybutyrate dehydrogenase, a succinate reductase, a succinyl-CoA transferase, a succinyl-CoA hydrolase, a succinyl-CoA synthetase, a glutamate dehydrogenase, a glutamate transaminase, a glutamate decarboxylase, a 4-aminobutyrate dehydrogenase, a 4-aminobutyrate transaminase, a 4-hydroxybutyrate kinase, a phosphotrans-4-hydroxybutyrylase, a succinyl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA hydrolase, a 4-hydroxybutyryl-CoA synthetase, an alpha-ketoglutarate reductase, a 5-hydroxy-2-oxopentanoate dehydrogenase, or a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation).
8-19. (canceled)
26-34. (canceled)
37-40. (canceled)
43-45. (canceled)
A method for producing 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL), comprising culturing a non-naturally occurring microbial organism under conditions and for a sufficient period of time to produce 14-BDO and 4-hydroxybutyryl-CoA followed by spontaneous lactonization of 4-hydroxybutyryl-CoA to form GBL, wherein said non-naturally occurring microbial organism comprise a microbial organism having a 14-BDO pathway and a 4-hydroxybutyryl-CoA pathway, said 14-BDO pathway comprising a first set of exogenous nucleic acids encoding 14-BDO pathway enzymes expressed in a sufficient amount to produce 14-BDO, said first set is selected from the group consisting of:
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and a 1,4-butanediol dehydrogenase)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase, or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming), a 4-hydroxybutyrate kinase, a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase and 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation);
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation);
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- and
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate decarboxylase;
- and a 1,4-butanediol dehydrogenase],
- said 4-hydroxybutyryl-CoA pathway comprising a second set of exogenous nucleic acids encoding 4-hydroxybutyryl-CoA pathway enzymes expressed in a sufficient amount to produce 4-hydroxybutyryl-CoA, said second set selected from the group consisting of:
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- and a phosphotrans-4-hydroxybutyrylase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate kinase;
- and a phosphotrans-4-hydroxybutyrylase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- and a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase and a phosphotrans-4-hydroxybutyrylase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- and a phosphotrans-4-hydroxybutyrylase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- an 4-aminobutyrate dehydrogenase or an 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- and a phosphotrans-4-hydroxybutyrylase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- an 4-aminobutyrate dehydrogenase or an 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase];
- and
- an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase and a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation)].
215 more elements
51-58. (canceled)
A method for producing 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL), comprising culturing a non-naturally occurring microbial organism under conditions and for a sufficient period of time to produce 14-BDO and 4-hydroxybutyryl-phosphate followed by spontaneous lactonization of 4-hydroxybutyryl-phosphate to form GBL, wherein said non-naturally occurring microbial organism comprise a microbial organism having a 14-BDO pathway and a 4-hydroxybutyryl-phosphate pathway, said 14-BDO pathway comprising at least one exogenous nucleic acid encoding a 14-BDO pathway enzyme expressed in a sufficient amount to produce 14-BDO, said 14-BDO pathway comprising:
- a 4-hydroxybutyryl-CoA reductase (alcohol forming), a succinyl-CoA reductase (aldehyde forming), an alpha-ketoglutarate decarboxylase, a 4-hydroxybutyrate dehydrogenase, a 4-hydroxybutyrate reductase, a 1,4-butanediol dehydrogenase, a succinate reductase, a succinyl-CoA transferase, a succinyl-CoA hydrolase, a succinyl-CoA synthetase, a glutamate dehydrogenase, a glutamate transaminase, a glutamate decarboxylase, an 4-aminobutyrate dehydrogenase, an 4-aminobutyrate transaminase, a 4-hydroxybutyrate kinase, a phosphotrans-4-hydroxybutyrylase, a 4-hydroxybutyryl-CoA reductase (aldehyde forming), a 4-hydroxybutyryl-phosphate reductase, a succinyl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase, a 4-hydroxybutyryl-CoA synthetase, an alpha-ketoglutarate reductase, a 5-hydroxy-2-oxopentanoate dehydrogenase, a 5-hydroxy-2-oxopentanoate decarboxylase or a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation),
- said 4-hydroxybutyryl-phosphate pathway comprising at least one exogenous nucleic acid encoding an 4-hydroxybutyryl-phosphate pathway enzyme expressed in a sufficient amount to produce 4-hydroxybutyryl-phosphate, said 4-hydroxybutyryl-phosphate pathway comprising:
- a 4-hydroxybutyrate kinase, a succinyl-CoA reductase (aldehyde forming), an alpha-ketoglutarate decarboxylase, a 4-hydroxybutyrate dehydrogenase, a succinate reductase, a succinyl-CoA transferase, a succinyl-CoA hydrolase, a succinyl-CoA synthetase, a glutamate dehydrogenase, a glutamate transaminase, a glutamate decarboxylase, a 4-aminobutyrate dehydrogenase, a 4-aminobutyrate transaminase or a succinyl-CoA reductase (alcohol forming).
66-77. (canceled)
84-92. (canceled)
94-95. (canceled)
(canceled)
A method for producing 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL), comprising culturing a non-naturally occurring microbial organism under conditions and for a sufficient period of time to produce 14-BDO and 4-hydroxybutyryl-phosphate followed by spontaneous lactonization of 4-hydroxybutyryl-phosphate to form GBL, wherein said non-naturally occurring microbial organism comprise a microbial organism having a 14-BDO pathway and a 4-hydroxybutyryl-phosphate pathway, said 14-BDO pathway comprising a first set of exogenous nucleic acids encoding 14-BDO pathway enzymes expressed in a sufficient amount to produce 14-BDO, said first set is selected from the group consisting of:
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and a 1,4-butanediol dehydrogenase)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase, or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming), a 4-hydroxybutyrate kinase, a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and 1,4-butanediol dehydrogenase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase and 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a phosphotrans-4-hydroxybutyrylase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyrate kinase;
- a 4-hydroxybutyryl-phosphate reductase;
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- a 4-hydroxybutyryl-CoA transferase, a 4-hydroxybutyryl-CoA hydrolase or a 4-hydroxybutyryl-CoA synthetase;
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation);
- a 4-hydroxybutyryl-CoA reductase (aldehyde forming);
- and a 1,4-butanediol dehydrogenase];
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation);
- and a 4-hydroxybutyryl-CoA reductase (alcohol forming)];
- and
- [an alpha-ketoglutarate reductase;
- a 5-hydroxy-2-oxopentanoate dehydrogenase;
- a 5-hydroxy-2-oxopentanoate decarboxylase;
- and a 1,4-butanediol dehydrogenase],
- said 4-hydroxybutyryl-phosphate pathway comprising a second set of exogenous nucleic acids encoding 4-hydroxybutyryl-phosphate pathway enzymes expressed in a sufficient amount to produce 4-hydroxybutyryl-phosphate, said second set selected from the group consisting of:
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (aldehyde forming);
- a 4-hydroxybutyrate dehydrogenase;
- and 4-hydroxybutyrate kinase];
- [a succinyl-CoA transferase, a succinyl-CoA hydrolase or a succinyl-CoA synthetase;
- a succinyl-CoA reductase (alcohol forming);
- and a 4-hydroxybutyrate kinase];
- [a succinate reductase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyrate kinase];
- [an alpha-ketoglutarate decarboxylase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyrate kinase];
- and
- [a glutamate dehydrogenase or a glutamate transaminase;
- a glutamate decarboxylase;
- an 4-aminobutyrate dehydrogenase or an 4-aminobutyrate transaminase;
- a 4-hydroxybutyrate dehydrogenase;
- and a 4-hydroxybutyrate kinase].
191 more elements
103-157. (canceled)
Dependencies are inferred from the claim text.