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.
Claims20
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 (a) glutamate CoA transferase, said enzyme classified as EC 2.8.3.a and converting glutamate to glutamyl-CoA;
- (b) glutamyl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting glutamate to glutamyl-CoA;
- (c) glutamyl-CoA ligase, said enzyme classified as EC 6.2.1.a and converting glutamate to glutamyl-CoA;
- (d) glutamate 5-kinase, said enzyme classified as EC 2.7.2.a and converting glutamate to glutamate-5-phosphate;
- (e) glutamate-5-semialdehyde dehydrogenase (phosphorylating), said enzyme classified as EC 1.2.1.d and converting glutamate-5-phosphate to glutamate-5-semialdehyde;
- (f) glutamyl-CoA reductase, said enzyme classified as EC 1.2.1.b and converting glutamyl-CoA to glutamate-5-semialdehyde;
- (g) glutamate-5-semialdehyde reductase, said enzyme classified as EC 1.1.1.a and converting glutamate-5-semialdehyde to 2-amino-5-hydroxypentanoic acid;
- (h) glutamyl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting glutamyl-CoA to 2-amino-5-hydroxypentanoic acid;
- (i) 2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating), said enzyme classified as EC 1.4.1.a and converting 2-amino-5-hydroxypentanoic acid to 5-hydroxy-2-oxopentanoic acid;
- (i) 2-amino-5-hydroxypentanoic acid transaminase, said enzyme classified as EC 2.6.1.a and converting 2-amino-5-hydroxypentanoic acid to 5-hydroxy-2-oxopentanoic acid;
- (k) 5-hydroxy-2-oxopentanoic acid decarboxylase, said enzyme classified as EC 4.1.1.a and converting 5-hydroxy-2-oxopentanoic acid to 4-hydroxybutanal;
- and (1) 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation), said enzyme classified as EC 1.2.1.c and converting 5-hydroxy-2-oxopentanoic acid to 4-hydroxybutyryl-CoA, and wherein said microbial organism further comprises at least one exogenous nucleic acid encoding an enzyme selected from (m) 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
- (n) 4-hydroxybutyryl-CoA reductase, said enzyme classified as EC 1.2.1.b and converting 4-hydroxybutyryl-CoA to 4-hydroxybutanal;
- and (o) 1,4-butanediol dehydrogenase, said enzyme classified as EC 1.1.1.a and converting 4-hydroxybutanal to 1,4-butanediol.
A non-naturally occurring microbial organism having nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, said microbial organism comprising at least four exogenous nucleic acids, each encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO, wherein said BDO pathway enzymes comprise (a) glutamate CoA transferase, said enzyme classified as EC 2.8.3.a and converting glutamate to glutamyl-CoA;
- (b) glutamyl-CoA hydrolase, said enzyme classified as EC 3.1.2.a and converting glutamate to glutamyl-CoA;
- (c) glutamyl-CoA ligase, said enzyme classified as EC 6.2.1.a and converting glutamate to glutamyl-CoA;
- (d) glutamate 5-kinase, said enzyme classified as EC 2.7.2.a and converting glutamate to glutamate-5-phosphate;
- (e) glutamate-5-semialdehyde dehydrogenase (phosphorylating), said enzyme classified as EC 1.2.1.d and converting glutamate-5-phosphate to glutamate-5-semialdehyde;
- (f) glutamyl-CoA reductase, said enzyme classified as EC 1.2.1.b and converting glutamyl-CoA to glutamate-5-semialdehyde;
- (g) glutamate-5-semialdehyde reductase, said enzyme classified as EC 1.1.1.a and converting glutamate-5-semialdehyde to 2-amino-5-hydroxypentanoic acid;
- (h) glutamyl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting glutamyl-CoA to 2-amino-5-hydroxypentanoic acid;
- (i) 2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating), said enzyme classified as EC 1.4.1.a and converting 2-amino-5-hydroxypentanoic acid to 5-hydroxy-2-oxopentanoic acid;
- (i) 2-amino-5-hydroxypentanoic acid transaminase, said enzyme classified as EC 2.6.1.a and converting 2-amino-5-hydroxypentanoic acid to 5-hydroxy-2-oxopentanoic acid;
- (k) 5-hydroxy-2-oxopentanoic acid decarboxylase, said enzyme classified as EC 4.1.1.a and converting 5-hydroxy-2-oxopentanoic acid to 4-hydroxybutanal;
- and (1) 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation), said enzyme classified as EC 1.2.1.c and converting 5-hydroxy-2-oxopentanoic acid to 4-hydroxybutyryl-CoA.
Dependencies are inferred from the claim text.