Microorganisms for the production of 1,4-butanediol
View Patent ↗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.
1. A host microbial organism transformed with nucleic acids encoding enzymes of a 1,4-butanediol (BDO) pathway, said microbial organism comprising exogenous nucleic acids encoding BDO pathway enzymes expressed in a sufficient amount to produce BDO, said BDO pathway enzymes comprising: a) 3-hydroxybutyryl-CoA dehydrogenase, said enzyme classified as EC 1.1.1.a and converting acetoacetyl-coenzyme A (CoA) to 3-hydroxybutyryl-CoA; b) 3-hydroxybutyryl-CoA dehydratase, said enzyme classified as EC 4.2.1.a and converting 3-hydroxybutyryl-CoA to crotonoyl-CoA; c) vinylacetyl-CoA A-isomerase, said enzyme classified as EC 5.3.3.3 and converting crotonoyl-CoA to vinylacetyl-CoA; d) 4-hydroxybutyryl-CoA dehydratase, said enzyme classified as EC 4.2.1.a and converting vinylacetyl-CoA to 4-hydroxybutyryl-CoA; and e) 4-hydroxybutyryl-CoA reductase (alcohol forming), said enzyme classified as EC 1.1.1.c and converting 4-hydroxybutyryl-CoA to 1,4-butanediol; or 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 and converting 4-hydroxybutanal to 1,4-butanediol.
2. The host microbial organism of claim 1 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase (alcohol forming).
3. The host microbial organism of claim 1 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase and 1,4-butanediol dehydrogenase.
4. The host microbial organism of claim 1 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA reductase, and 1,4-butanediol dehydrogenase.
5. The host microbial organism of claim 1 , wherein at least one exogenous nucleic acid is a heterologous nucleic acid.
6. The host microbial organism of claim 1 , wherein said host microbial organism is in a substantially anaerobic culture medium.
7. A method for producing BDO, comprising culturing the host microbial organism of claim 1 under conditions and for a sufficient period of time to produce BDO.
8. The method of claim 7 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase (alcohol forming).
9. The method of claim 7 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase and 1,4-butanediol dehydrogenase.
10. The method of claim 7 , wherein said BDO pathway comprises 4-hydroxybutyryl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA reductase, and 1,4-butanediol dehydrogenase.
11. The method of claim 7 , wherein said non naturally occurring host microbial organism is in a substantially anaerobic culture medium.
12. The method of claim 7 , wherein at least one exogenous nucleic acid is a heterologous nucleic acid.