Microorganisms and methods for production of 4-hydroxybutyrate, 1,4-butanediol and related compounds

1. A non-naturally occurring microbial organism, said microbial organism having a 4-hydroxybutyrate pathway and being capable of producing 4-hydroxybutyrate, wherein said microbial organism comprises genetic modifications, said genetic modifications comprising:

(A) a genetic modification that increases expression of phosphoenolpyruvate carboxylase;

(B) a genetic modification that increases expression of alpha-ketoglutarate dehydrogenase; and

(C) a genetic modification that decreases expression of an acyl coenzyme A thioesterase.

2. The microbial organism of claim 1 , wherein:

(K) the microbial organism has decreased production of ethanol, acetate, pyruvate or alanine, or a combination thereof, relative to a parent microbial organism in the absence of said genetic modification;

(L) the microbial organism has decreased production of glutamate relative to a parent microbial organism in the absence of said genetic modification; and

(M) the microbial organism has decreased production of gamma-butyrolactone relative to a parent microbial organism in the absence of said genetic modification.

3. The microbial organism of claim 1 , wherein said microbial organism further comprises a 4-hydroxybutyryl-CoA pathway.

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

5. The method of claim 4 , wherein:

(K) the microbial organism has decreased production of ethanol, acetate, pyruvate or alanine, or a combination thereof, relative to a parent microbial organism in the absence of said genetic modification;

(L) the microbial organism has decreased production of glutamate relative to a parent microbial organism in the absence of said genetic modification; and

(M) the microbial organism has decreased production of gamma-butyrolactone relative to a parent microbial organism in the absence of said genetic modification.

6. The method of claim 4 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

7. The microbial organism of claim 1 , wherein said microbial organism further comprises a genetic modification selected from:

(D) a genetic modification that increases expression of a non-phosphotransferase (PTS) glucose uptake system;

(E) a genetic modification that increases expression of a gamma-butyrolactone esterase;

(F) a genetic modification that decreases expression of succinyl-CoA synthetase;

(G) a genetic modification that decreases expression of an alcohol dehydrogenase

(H) a genetic modification that decreases expression of a non-energy-producing NADH dehydrogenase;

(I) a genetic modification that decreases expression of a cytochrome oxidase; and

(J) a combination of two or more of the genetic modifications of parts (C)-(I).

8. The microbial organism of claim 7 , wherein:

(N) the microbial organism of part (D) has a genetic modification comprising increased expression of a permease, glucokinase, or a glucose facilitator, or a combination thereof;

(O) the microbial organism of part (E) has decreased production of gamma-butyrolactone relative to a parent microbial organism in the absence of said genetic modification;

(P) the microbial organism of part (F) has increased production of 4-hydroxybutyrate relative to a parent microbial organism in the absence of said genetic modification;

(Q) the microbial organism of part (G) has decreased backflux from a downstream product of the 4-hydroxybutyrate pathway relative to a parent microbial organism in the absence of said genetic modification;

(R) the microbial organism of part (H) has suppressed depletion of the NADH pool or increased energy efficiency in the microbial organism, or a combination thereof, relative to a parent microbial organism in the absence of said genetic modification;

(S) the microbial organism of part (I) has increased energy efficiency relative to a parent microbial organism in the absence of said genetic modification; or

(T) the microbial organism of part (I) has increased tolerance to a range of oxygen concentrations relative to a parent microbial organism in the absence of said genetic modification.

9. The method of claim 4 , wherein said microbial organism further comprises a genetic modification selected from:

(C) a genetic modification that increases expression of a non-phosphotransferase (PTS) glucose uptake system;

(D) a genetic modification that increases expression of a gamma-butyrolactone esterase;

(E) a genetic modification that decreases expression of succinyl-CoA synthetase;

(F) a genetic modification that decreases expression of an alcohol dehydrogenase

(G) a genetic modification that decreases expression of a non-energy-producing NADH dehydrogenase;

(H) a genetic modification that decreases expression of a cytochrome oxidase; and

(I) a combination of two or more of the genetic modifications of parts (C)-(I).

10. The method of claim 9 , wherein:

(N) the microbial organism of part (C) has a genetic modification comprising increased expression of a permease, glucokinase, or a glucose facilitator, or a combination thereof;

(O) the microbial organism of part (D) has decreased production of gamma-butyrolactone relative to a parent microbial organism in the absence of said genetic modification;

(P) the microbial organism of part (E) has increased production of 4-hydroxybutyrate relative to a parent microbial organism in the absence of said genetic modification;

(Q) the microbial organism of part (F) has decreased backflux from a downstream product of the 4-hydroxybutyrate pathway relative to a parent microbial organism in the absence of said genetic modification;

(R) the microbial organism of part (G) has suppressed depletion of the NADH pool or increased energy efficiency in the microbial organism, or a combination thereof, relative to a parent microbial organism in the absence of said genetic modification;

(S) the microbial organism of part (H) has increased energy efficiency relative to a parent microbial organism in the absence of said genetic modification; or

(T) the microbial organism of part (H) has increased tolerance to a range of oxygen concentrations relative to a parent microbial organism in the absence of said genetic modification.

11. The microbial organism of claim 1 , wherein the microbial organism has a genetic modification comprising at least two genetic modifications that decrease expression of at least two acyl coenzyme A thioesterases.

12. The method of claim 4 , wherein the microbial organism has a genetic modification comprising at least two genetic modifications that decrease expression of at least two acyl coenzyme A thioesterases.