Microorganisms for the production of 1,4-butanediol
AssigneeGenomatica, Inc.
InventorsMark J. Burk, Anthony P. Burgard, Robin E. Osterhout, Jun Sun
Life of this patent
Estimating the term…
Abstract
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.
Claims19
A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, said microbial organism comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise glutamate dehydrogenase, said enzyme classified as EC 1.4.1.a and converting alpha-ketoglutarate to glutamate;
- glutamate decarboxylase, said enzyme classified as EC 4.1.1.a and converting glutamate to 4-aminobutyrate;
- 4-aminobutyrate oxidoreductase (deaminating), said enzyme classified as EC 1.4.1.a and converting 4-aminobutyrate to succinate semialdehyde;
- and 4-hydroxybutyrate dehydrogenase, said enzyme classified as EC 1.1.1.a and converting succinate semialdehyde to 4-hydroxybutyrate;
- and wherein said BDO pathway enzymes further comprise:
- (a) 4-hydroxybutyrate kinase, said enzyme classified as EC 2.7.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-phosphate;
- phosphotrans-4-hydroxybutyrylase, said enzyme classified as EC 2.3.1.a and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutyryl-CoA;
- 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 converting 4-hydroxybutanal to 1,4-butanediol;
- (b) 4-hydroxybutyryl-CoA transferase, said enzyme classified as EC 2.8.3.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, 4-hydroxybutyryl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, or 4-hydroxybutyryl-CoA ligase, said enzyme classified as EC 6.2.1.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA;
- 4-hydroxybutyryl-CoA reductase;
- and 1,4-butanediol dehydrogenase;
- (c) 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, or 4-hydroxybutyryl-CoA ligase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
- (d) 4-hydroxybutyrate kinase;
- 4-hydroxybutanal dehydrogenase (phosphorylating), said enzyme classified as EC 1.2.1.d and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutanal;
- and 1,4-butanediol dehydrogenase;
- or
- (e) 4-hydroxybutyrate kinase;
- phosphotrans-4-hydroxybutyrylase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming).
A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, said microbial organism comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise alpha-ketoglutarate decarboxylase, said enzyme classified as EC 4.1.1.a and converting alpha-ketoglutarate to succinate semialdehyde, or glutamate dehydrogenase, said enzyme classified as EC 1.4.1.a and converting alpha-ketoglutarate to glutamate, glutamate decarboxylase, said enzyme classified as EC 4.1.1.a and converting glutamate to 4-aminobutyrate, and 4-aminobutyrate transaminase, said enzyme classified as EC 2.6.1.a and converting 4-aminobutyrate to succinate semialdehyde;
- and 4-hydroxybutyrate dehydrogenase, said enzyme classified as EC 1.1.1.a and converting succinate semialdehyde to 4-hydroxybutyrate;
- and wherein said BDO pathway enzymes further comprise:
- (a) 4-hydroxybutyryl-CoA ligase, said enzyme classified as EC 6.2.1.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA;
- 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 converting 4-hydroxybutanal to 1,4-butanediol;
- (b) 4-hydroxybutyryl-CoA transferase, said enzyme classified as EC 2.8.3.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, 4-hydroxybutyryl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, or 4-hydroxybutyryl-CoA ligase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
- (c) 4-hydroxybutyrate kinase, said enzyme classified as EC 2.7.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-phosphate;
- 4-hydroxybutanal dehydrogenase (phosphorylating), said enzyme classified as EC 1.2.1.d and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutanal;
- and 1,4-butanediol dehydrogenase;
- or
- (d) 4-hydroxybutyrate kinase;
- phosphotrans-4-hydroxybutyrylase, said enzyme classified as EC 2.3.1.a and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutyryl-CoA;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming).
A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, said microbial organism comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise alpha-ketoglutarate dehydrogenase, said enzyme classified as EC 1.2.1.c and converting alpha-ketoglutarate to succinyl-CoA;
- CoA-dependent succinic semialdehyde dehydrogenase, said enzyme classified as EC 1.2.1.b and converting succinyl-CoA to succinate semialdehyde;
- and 4-hydroxybutyrate dehydrogenase, said enzyme classified as EC 1.1.1.a and converting succinate semialdehyde to 4-hydroxybutyrate;
- and wherein said BDO pathway enzymes further comprise:
- (a) 4-hydroxybutyryl-CoA ligase, said enzyme classified as EC 6.2.1.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA;
- 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 converting 4-hydroxybutanal to 1,4-butanediol;
- (b) 4-hydroxybutyryl-CoA transferase, said enzyme classified as EC 2.8.3.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, 4-hydroxybutyryl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-CoA, or 4-hydroxybutyryl-CoA ligase;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
- (c) 4-hydroxybutyrate kinase, said enzyme classified as EC 2.7.2.a and converting 4-hydroxybutyrate to 4-hydroxybutyryl-phosphate;
- 4-hydroxybutanal dehydrogenase (phosphorylating), said enzyme classified as EC 1.2.1.d and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutanal;
- and 1,4-butanediol dehydrogenase;
- or
- (d) 4-hydroxybutyrate kinase;
- phosphotrans-4-hydroxybutyrylase, said enzyme classified as EC 2.3.1.a and converting 4-hydroxybutyryl-phosphate to 4-hydroxybutyryl-CoA;
- and 4-hydroxybutyryl-CoA reductase (alcohol forming).
Dependencies are inferred from the claim text.