Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors

1. A non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways, said pathways comprising exogenous nucleic acids encoding a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase, wherein said exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO).

2. The non-naturally occurring microbial organism of claim 1 , wherein said pathways comprise a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

3. The non-naturally occurring microbial organism of claim 2 , further comprising a CoA-dependent succinic semialdehyde dehydrogenase.

4. The non-naturally occurring microbial organism of claim 2 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

5. The non-naturally occurring microbial organism of claim 2 , wherein the microbial organism is in a substantially anaerobic culture medium.

6. The non-naturally occurring microbial organism of claim 1 , wherein said pathways comprise a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

7. The non-naturally occurring microbial organism of claim 6 , further comprising a CoA-dependent succinic semialdehyde dehydrogenase.

8. The non-naturally occurring microbial organism of claim 6 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

9. The non-naturally occurring microbial organism of claim 6 , wherein the microbial organism is in a substantially anaerobic culture medium.

10. A non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways, said pathways comprising exogenous nucleic acids encoding a) an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase, wherein said exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO).

11. The non-naturally occurring microbial organism of claim 10 , wherein said pathways comprise a) an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

12. The non-naturally occurring microbial organism of claim 11 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

13. The non-naturally occurring microbial organism of claim 11 , wherein the microbial organism is in a substantially anaerobic culture medium.

14. The non-naturally occurring microbial organism of claim 10 , wherein said pathways comprise a) an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

15. The non-naturally occurring microbial organism of claim 14 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

16. The non-naturally occurring microbial organism of claim 14 , wherein the microbial organism is in a substantially anaerobic culture medium.

17. A non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways, said pathways comprising exogenous nucleic acids encoding a) a glutamate:succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase, wherein said exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO).

18. The microbial organism of claim 17 , wherein said pathways comprise a) a glutamate: succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

19. The non-naturally occurring microbial organism of claim 18 , further comprising a CoA-dependent succinic semialdehyde dehydrogenase.

20. The non-naturally occurring microbial organism of claim 18 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

21. The non-naturally occurring microbial organism of claim 18 , wherein the microbial organism is in a substantially anaerobic culture medium.

22. The non-naturally occurring microbial organism of claim 17 , wherein said pathways comprise a) a glutamate:succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

23. The non-naturally occurring microbial organism of claim 22 , further comprising a CoA-dependent succinic semialdehyde dehydrogenase.

24. The non-naturally occurring microbial organism of claim 22 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

25. The non-naturally occurring microbial organism of claim 22 , wherein the microbial organism is in a substantially anaerobic culture medium.

26. A method for the production of 1,4-BDO comprising, culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce 1,4-BDO.

27. The method of claim 26 , wherein said pathways comprise a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

28. The method of claim 27 , wherein the microbial organism further comprises a CoA-dependent succinic semialdehyde dehydrogenase.

29. The method of claim 27 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

30. The method of claim 27 , wherein the microbial organism is in a substantially anaerobic culture medium.

31. The method of claim 26 , wherein said pathways comprise a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

32. The method of claim 31 , wherein the microbial organism further comprises a CoA-dependent succinic semialdehyde dehydrogenase.

33. The method of claim 31 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

34. The method of claim 31 , wherein the microbial organism is in a substantially anaerobic culture medium.

35. A method for the production of 1,4-BDO comprising, culturing the non-naturally occurring microbial organism of claim 10 under conditions and for a sufficient period of time to produce 1,4-BDO.

36. The method of claim 35 , wherein said pathways comprise a) an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

37. The method of claim 36 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

38. The method of claim 36 , wherein the microbial organism is in a substantially anaerobic culture medium.

39. The method of claim 35 , wherein said pathways comprise a) an α-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

40. The method of claim 39 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

41. The method of claim 39 , wherein the microbial organism is in a substantially anaerobic culture medium.

42. A method for the production of 1,4-BDO comprising, culturing the non-naturally occurring microbial organism of claim 17 under conditions and for a sufficient period of time to produce 1,4-BDO.

43. The method of claim 42 , wherein said pathways comprise a) a glutamate:succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

44. The method of claim 43 , wherein the microbial organism further comprises a CoA-dependent succinic semialdehyde dehydrogenase.

45. The method of claim 43 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

46. The method of claim 43 , wherein the microbial organism is in a substantially anaerobic culture medium.

47. The method of claim 42 , wherein said pathways comprise a) a glutamate:succinate semialdehyde transaminase and a glutamate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase; and e) an alcohol dehydrogenase.

48. The method of claim 47 , wherein the microbial organisms further comprises a CoA-dependent succinic semialdehyde dehydrogenase.

49. The method of claim 47 , wherein said exogenous nucleic acids comprise at least one heterologous nucleic acid.

50. The method of claim 47 , wherein the microbial organism is in a substantially anaerobic culture medium.