Microorganisms for the production of 1,4-butanediol, 4-hydroxybutanal, 4-hydroxybutyryl-coa, putrescine and related compounds, and methods related thereto
View Patent ↗The invention provides non-naturally occurring microbial organisms comprising a 1,4-butanediol (BDO), 4-hydroxybutyryl-CoA, 4-hydroxybutanal or putrescine pathway comprising at least one exogenous nucleic acid encoding a BDO, 4-hydroxybutyryl-CoA, 4-hydroxybutanal or putrescine pathway enzyme expressed in a sufficient amount to produce BDO, 4-hydroxybutyryl-CoA, 4-hydroxybutanal or putrescine and further optimized for expression of BDO. The invention additionally provides methods of using such microbial organisms to produce BDO, 4-hydroxybutyryl-CoA, 4-hydroxybutanal or putrescine.
1. A non-naturally occurring microbial organism, comprising a 4-hydroxybutanal pathway, said microbial organism comprising at least one exogenous nucleic acid encoding a 4-hydroxybutanal pathway enzyme expressed in a sufficient amount to produce 4-hydroxybutanal, said 4-hydroxybutanal pathway comprising succinyl-CoA reductase (aldehyde forming), which catalyzes the conversion of succinyl-CoA to succinate semialdehyde; 4-hydroxybutyrate dehydrogenase, which catalyzes the conversion of succinate semialdehyde to 4-hydroxybutyrate; and carboxylic acid reductase, which catalyzes the ATP-dependent reduction of 4-hydroxybutyrate to 4-hydroxybutanal, wherein said at least one exogenous nucleic acid comprises a nucleic acid encoding said carboxylic acid reductase.
2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises three exogenous nucleic acids encoding succinyl-CoA reductase (aldehyde forming); 4-hydroxybutyrate dehydrogenase; and carboxylic acid reductase.
3. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two exogenous nucleic acids encoding succinyl-CoA reductase (aldehyde forming) and carboxylic acid reductase.
4. The non-naturally occurring microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
5. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two exogenous nucleic acids encoding 4-hydroxybutyrate dehydrogenase and carboxylic acid reductase.
6. A method for producing 4-hydroxybutanal, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce 4-hydroxybutanal.
7. The method of claim 6 , wherein said microbial organism comprises two exogenous nucleic acids encoding succinyl-CoA reductase (aldehyde forming) and carboxylic acid reductase.
8. The method of claim 6 , wherein said microbial organism comprises three exogenous nucleic acids encoding succinyl-CoA reductase (aldehyde forming); 4-hydroxybutyrate dehydrogenase; and carboxylic acid reductase.
9. The method of claim 6 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.
10. The method of claim 6 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
11. The method of claim 6 , wherein said microbial organism comprises two exogenous nucleic acids encoding 4-hydroxybutyrate dehydrogenase and carboxylic acid reductase.