MICROORGANISMS FOR PRODUCING 1,4-BUTANEDIOL AND METHODS RELATED THERETO

• US 20130034884A1
• Filed: 06/21/2012
• Published: 02/07/2013
• Est. Priority Date: 06/22/2011
• Status: Abandoned Application

• Current Assignee: Genomatica Incorporated
• Original Assignee: Genomatica Incorporated
• Inventors: BURGARD, Anthony P., Osterhout, Robin E., Sun, Jun, Pharkya, Priti
• Application Number:
  US13/530,053
• Publication Number:
  US 20130034884A1
• Time in Patent Office: Days
• US Class Current: 435/126
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12P 7/16 Butanols
• CPC Class Codes: C12P 7/18 polyhydric
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids
• CPC Class Codes: C12Y 102/99006 Carboxylate reductase (1.2.99.6)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

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.

Claims

1. A non-naturally occurring microbial organism, comprising a microbial organism having a 1,4-butanediol pathway comprising at least one exogenous nucleic acid encoding a 1,4-butanediol pathway enzyme expressed in a sufficient amount to produce 1,4-butanediol;

said non-naturally occurring microbial organism further comprising;

(i) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from an ATP-citrate lyase, a citrate lyase, a citryl-CoA synthetase, a citryl-CoA lyase, a fumarate reductase, isocitrate dehydrogenase, and an alpha-ketoglutarate;

ferredoxin oxidoreductase;

(ii) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from a pyruvate;

ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H₂ hydrogenase;

or

(iii) at least one exogenous nucleic acid encodes an enzyme selected from a CO dehydrogenase, an H₂ hydrogenase, and combinations thereof;

wherein said 1,4-butanediol pathway comprises a pathway selected from;

(a) 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, and α

-ketoglutarate decarboxylase;

(b) 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate;

CoA transferase, 4-butyrate kinase, phosphotransbutyrylase, α

-ketoglutarate decarboxylase, aldehyde dehydrogenase, alcohol dehydrogenase and an aldehyde/alcohol dehydrogenase;

(c) (i) an α

-ketoglutarate decarboxylase, or an α

-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase, or a glutamate;

succinate semialdehyde transaminase and a glutamate decarboxylase;

(ii) a 4-hydroxybutanoate dehydrogenase;

(iii) a 4-hydroxybutyryl-CoA;

acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase;

(iv) an aldehyde dehydrogenase; and

(v) an alcohol dehydrogenase;

(d) 4-aminobutyrate CoA transferase, 4-aminobutyryl-CoA hydrolase, 4-aminobutyrate-CoA ligase, 4-aminobutyryl-CoA oxidoreductase (deaminating), 4-aminobutyryl-CoA transaminase, and 4-hydroxybutyryl-CoA dehydrogenase;

(e) 4-aminobutyrate CoA transferase, 4-aminobutyryl-CoA hydrolase, 4-aminobutyrate-CoA ligase, 4-aminobutyryl-CoA reductase (alcohol forming), 4-aminobutyryl-CoA reductase, 4-aminobutan-1-ol dehydrogenase, 4-aminobutan-1-ol oxidoreductase (deaminating) and 4-aminobutan-1-ol transaminase;

(f) 4-aminobutyrate kinase, 4-aminobutyraldehyde dehydrogenase (phosphorylating), 4-aminobutan-1-ol dehydrogenase, 4-aminobutan-1-ol oxidoreductase (deaminating), 4-aminobutan-1-ol transaminase, [(4-aminobutanolyl)oxy]phosphonic acid oxidoreductase (deaminating), [(4-aminobutanolyl)oxy]phosphonic acid transaminase, 4-hydroxybutyryl-phosphate dehydrogenase, and 4-hydroxybutyraldehyde dehydrogenase (phosphorylating);

(g) alpha-ketoglutarate 5-kinase, 2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating), 2,5-dioxopentanoic acid reductase, alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, alpha-ketoglutaryl-CoA ligase, alpha-ketoglutaryl-CoA reductase, 5-hydroxy-2-oxopentanoic acid dehydrogenase, alpha-ketoglutaryl-CoA reductase (alcohol forming), 5-hydroxy-2-oxopentanoic acid decarboxylase, and 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(h) glutamate CoA transferase, glutamyl-CoA hydrolase, glutamyl-CoA ligase, glutamate 5-kinase, glutamate-5-semialdehyde dehydrogenase (phosphorylating), glutamyl-CoA reductase, glutamate-5-semialdehyde reductase, glutamyl-CoA reductase (alcohol forming), 2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating), 2-amino-5-hydroxypentanoic acid transaminase, 5-hydroxy-2-oxopentanoic acid decarboxylase, 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(i) 3-hydroxybutyryl-CoA dehydrogenase, 3-hydroxybutyryl-CoA dehydratase, vinylacetyl-CoA Δ

-isomerase, or 4-hydroxybutyryl-CoA dehydratase;

(j) homoserine deaminase, homoserine CoA transferase, homoserine-CoA hydrolase, homoserine-CoA ligase, homoserine-CoA deaminase, 4-hydroxybut-2-enoyl-CoA transferase, 4-hydroxybut-2-enoyl-CoA hydrolase, 4-hydroxybut-2-enoyl-CoA ligase, 4-hydroxybut-2-enoate reductase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybut-2-enoyl-CoA reductase;

(k) succinyl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(l) glutamate dehydrogenase, 4-aminobutyrate oxidoreductase (deaminating), 4-aminobutyrate transaminase, glutamate decarboxylase, 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(m) 4-aminobutyrate kinase;

4-aminobutyraldehyde dehydrogenase (phosphorylating);

4-aminobutan-1-ol dehydrogenase; and

4-aminobutan-1-ol oxidoreductase (deaminating) or 4-aminobutan-1-ol transaminase;

(n) 4-aminobutyrate kinase;

[(4-aminobutanolyl)oxy]phosphonic acid oxidoreductase (deaminating) or [(4-aminobutanolyl)oxy]phosphonic acid transaminase;

4-hydroxybutyryl-phosphate dehydrogenase; and

4-hydroxybutyraldehyde dehydrogenase (phosphorylating);

(o) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase (alcohol forming); and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(p) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase (alcohol forming); and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(q) alpha-ketoglutarate 5-kinase;

2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating);

2,5-dioxopentanoic acid reductase; and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(r) alpha-ketoglutarate 5-kinase;

2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating);

2,5-dioxopentanoic acid reductase; and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(s) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase;

5-hydroxy-2-oxopentanoic acid dehydrogenase; and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(t) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase;

5-hydroxy-2-oxopentanoic acid dehydrogenase; and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(u) glutamate CoA transferase, glutamyl-CoA hydrolase, or glutamyl-CoA ligase;

glutamyl-CoA reductase (alcohol forming);

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(v) glutamate 5-kinase;

glutamate-5-semialdehyde dehydrogenase (phosphorylating);

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(w) glutamate CoA transferase, glutamyl-CoA hydrolase, or glutamyl-CoA ligase;

glutamyl-CoA reductase;

glutamate-5-semialdehyde reductase;

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(x) glutamate 5-kinase;

glutamate-5-semialdehyde dehydrogenase (phosphorylating);

glutamate-5-semialdehyde reductase;

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(y) homoserine deaminase;

4-hydroxybut-2-enoyl-CoA transferase, 4-hydroxybut-2-enoyl-CoA hydrolase, 4-hydroxybut-2-enoyl-CoA ligase;

4-hydroxybut-2-enoyl-CoA reductase;

(z) homoserine CoA transferase, homoserine-CoA hydrolase, or homoserine-CoA ligase;

homoserine-CoA deaminase; and

4-hydroxybut-2-enoyl-CoA reductase;

(aa) homoserine deaminase;

4-hydroxybut-2-enoate reductase; and

4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, or 4-hydroxybutyryl-CoA ligase;

(bb) (i) alpha-ketoglutarate decarboxylase;

or alpha-ketoglutarate dehydrogenase and CoA-dependent succinate semialdehyde dehydrogenase;

or glutamate;

succinate semialdehyde transaminase and glutamate decarboxylase;

(ii) 4-hydroxybutyrate dehydrogenase;

(iii) 4-hydroxybutyryl-CoA transferase;

or 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase;

(iv) 4-hydroxybutyryl-CoA reductase; and

(v) 4-hydroxybutyraldehyde reductase;

or aldehyde/alcohol dehydrogenase;

(cc) (i) alpha-ketoglutarate decarboxylase;

or succinyl-CoA synthetase and CoA-dependent succinate semialdehyde dehydrogenase;

(ii) 4-hydroxybutyrate dehydrogenase;

(iii) 4-hydroxybutyryl-CoA transferase;

or 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase; and

(iv) aldehyde dehydrogenase; and

alcohol dehydrogenase;

or aldehyde/alcohol dehydrogenase;

(dd) (i) alpha-ketoglutarate decarboxylase;

or glutamate dehydrogenase;

glutamate decarboxylase; and

deaminating 4-aminobutyrate oxidoreductase or 4-aminobutyrate transaminase;

or alpha-ketoglutarate dehydrogenase and CoA-dependent succinate semialdehyde dehydrogenase;

(ii) 4-hydroxybutyrate dehydrogenase; and

(iii) 4-hydroxybutyrate kinase;

phosphotrans-4-hydroxybutyrylase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyrate kinase;

phosphorylating 4-hydroxybutanal dehydrogenase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyrate kinase;

phosphotrans-4-hydroxybutyrylase; and

alcohol forming 4-hydroxybutyryl-CoA reductase;

or 4-hydroxybutyryl-CoA transferase or 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA ligase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyryl-CoA transferase or 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA ligase; and

alcohol forming 4-hydroxybutyryl-CoA reductase;

(ee) (i) glutamate CoA transferase or glutamyl-CoA hydrolase or glutamyl-CoA ligase;

glutamyl-CoA reductase; and

glutamate-5-semialdehyde reductase;

or glutamate CoA transferase or glutamyl-CoA hydrolase or glutamyl-CoA ligase; and

alcohol forming glutamyl-CoA reductase;

or glutamate 5-kinase;

phosphorylating glutamate-5-semialdehyde dehydrogenase; and

glutamate-5-semialdehyde reductase;

(ii) deaminating 2-amino-5-hydroxypentanoic acid oxidoreductase or 2-amino-5-hydroxypentanoic acid transaminase; and

(iii) 5-hydroxy-2-oxopentanoic acid decarboxylase; and

4-hydroxybutyraldehyde reductase;

or decarboxylating 5-hydroxy-2-oxopentanoic acid dehydrogenase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or decarboxylating 5-hydroxy-2-oxopentanoic acid dehydrogenase and alcohol forming 4-hydroxybutyryl-CoA reductase;

(ff) succinyl-CoA reductase (aldehyde forming);

4-hydroxybutyrate dehydrogenase; and

4-hydroxybutyrate reductase; and

optionally 1,4-butandiol dehydrogenase;

(gg) alpha-ketoglutarate decarboxylase;

4-hydroxybutyrate dehydrogenase; and

4-hydroxybutyrate reductase; and

optionally 1,4-butandiol dehydrogenase;

(hh) succinate reductase;

4-hydroxybutyrate dehydrogenase, and 4-hydroxybutyrate reductase; and

optionally 1,4-butandiol dehydrogenase;

(ii) alpha-ketoglutarate decarboxylase, or glutamate dehydrogenase or glutamate transaminase and glutamate decarboxylase and 4-aminobutyrate dehydrogenase or 4-aminobutyrate transaminase;

4-hydroxybutyrate dehydrogenase; and

4-hydroxybutyrate reductase; and

optionally 1,4-butandiol dehydrogenase;

(jj) alpha-ketoglutarate reductase;

5-hydroxy-2-oxopentanoate dehydrogenase; and

5-hydroxy-2-oxopentanoate decarboxylase; and

optionally 1,4-butandiol dehydrogenase;

(kk) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase; and

4-hydroxybutyryl-CoA reductase (alcohol forming);

(ll) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase;

4-hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(mm) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase;

4-Hydroxybutyryl-CoA transferase, 4-Hydroxybutyryl-CoA synthetase, 4-Hydroxybutyryl-CoA hydrolase, or Phosphotrans-4-hydroxybutyrylase/4-Hydroxybutyrate kinase;

4-Hydroxybutyrate reductase; and

1,4-butanediol dehydrogenase;

(nn) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylasel;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(oo) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

4-Hydroxybutyryl-phosphate reductase; and

1,4-butanediol dehydrogenase;

(pp) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate reductase; and

1,4-butanediol dehydrogenase;

(qq) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(rr) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(ss) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(tt) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(uu) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

4-Hydroxybutyryl-phosphate reductase; and

1,4-butanediol dehydrogenase;

(vv) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate reductase; and

1,4-butanediol dehydrogenase;

(ww) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(xx) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(yy) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(zz) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(aaa) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

4-Hydroxybutyryl-phosphate reductase; and

1,4-butanediol dehydrogenase;

(bbb) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate reductase; and

1,4-butanediol dehydrogenase;

(ccc) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(ddd) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(eee) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(fff) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(ggg) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyrate kinase;

4-Hydroxybutyryl-phosphate reductase; and

1,4-butanediol dehydrogenase;

(hhh) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyrate reductase; and

1,4-butanediol dehydrogenase;

(iii) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming);

(jjj) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase;

4-Hydroxybutyryl-CoA reductase (aldehyde forming); and

1,4-butanediol dehydrogenase;

(kkk) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA reductase (alcohol forming); and

(lll) any of the pathways that produce 1,4-butanediol as shown in any of FIG. 1, 8-13, 58, 62, 63 or 72-74.

2. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism comprising (i) further comprises an exogenous nucleic acid encoding an enzyme selected from a pyruvate:

ferredoxin oxidoreductase, an aconitase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, an acetate kinase, a phosphotransacetylase, an acetyl-CoA synthetase, an NAD(P)H;

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

3. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism comprising (ii) further comprises an exogenous nucleic acid encoding an enzyme selected from an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, NAD(P):

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

4. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism comprising (iii) further comprises an exogenous nucleic acid encoding an enzyme selected from NAD(P):

ferredoxin oxidoreductase and ferredoxin.

7. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism:

(i) comprising pathways (a), (b) or (c) further comprises an enzyme selected from succinyl-CoA synthetase, exogenous CoA-dependent succinic semialdehyde dehydrogenase or exogenous succinyl-CoA synthetase and exogenous CoA-dependent succinic semialdehyde dehydrogenase;

(ii) comprising pathway (d), (g), (h), (i), (j) further comprises an enzyme selected from 4-hydroxybutyryl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA reductase, or 1,4-butanediol dehydrogenase;

(iii) comprising pathway (e) or (f) further comprises 1,4-butanediol dehydrogenase;

(iv) comprising pathway (k) further comprises succinyl-CoA reductase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyrate kinase, phosphotrans-4-hydroxybutyrylase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutyryl-CoA reductase (alcohol forming), or 1,4-butanediol dehydrogenase;

(v) comprising pathway (1) further comprises alpha-ketoglutarate decarboxylase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyrate kinase, phosphotrans-4-hydroxybutyrylase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutyryl-CoA reductase (alcohol forming), or 1,4-butanediol dehydrogenase.

8. The non-naturally occurring microbial organism of claim 1, wherein said microbial organism comprises two, three, four or five exogenous nucleic acids each encoding enzymes of (i), (ii) or (iii).

9. The non-naturally occurring microbial organism of claim 8, wherein said microbial organism comprising (i) comprises three exogenous nucleic acids encoding ATP-citrate lyase or citrate lyase, a fumarate reductase, and an alpha-ketoglutarate:

ferredoxin oxidoreductase;

wherein said microbial organism comprising (ii) comprises five exogenous nucleic acids encoding pyruvate;

ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H₂ hydrogenase;

or

wherein said microbial organism comprising (iii) comprises two exogenous nucleic acids encoding CO dehydrogenase and H₂ hydrogenase.

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

5-6. -6. (canceled)

10. (canceled)

12. A non-naturally occurring microbial organism, comprising a microbial organism having a 4-hydroxybutyrate pathway comprising at least one exogenous nucleic acid encoding a 4-hydroxybutyrate pathway enzyme 4-hydroxybutyrate expressed in a sufficient amount to produce 4-hydroxybutyrate;

said non-naturally occurring microbial organism further comprising;

(i) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from an ATP-citrate lyase, citrate lyase, a citryl-CoA synthetase, a citryl-CoA lyase, a fumarate reductase, isocitrate dehydrogenase and an alpha-ketoglutarate;

ferredoxin oxidoreductase;

(ii) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from a pyruvate;

ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H₂ hydrogenase;

or

(iii) at least one exogenous nucleic acid encodes an enzyme selected from a CO dehydrogenase, an H₂ hydrogenase, and combinations thereof;

(a) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase; and

4-Hydroxybutyryl-CoA transferase, 4-Hydroxybutyryl-CoA synthetase, 4-Hydroxybutyryl-CoA hydrolase, or Phosphotrans-4-hydroxybutyrylase/4-Hydroxybutyrate kinase;

(b) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase; and

4-Hydroxybutyryl-CoA transferase, hydrolase or synthetase;

(c) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyrate kinase;

(d) Succinate reductase; and

4-Hydroxybutyrate dehydrogenase;

(e) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming); and

4-Hydroxybutyrate dehydrogenase;

(f) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase); and

4-Hydroxybutyrate dehydrogenase;

(g) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase); and

Succinyl-CoA reductase (alcohol forming);

(h) acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase, a 3-hydroxybutyryl-CoA dehydrogenase, a crotonase, a crotonyl-CoA hydratase, a 4-hydroxybutyryl-CoA transferase, a phosphotrans-4-hydroxybutyrylase, and a 4-hydroxybutyrate kinase;

(i) 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, and α

-ketoglutarate decarboxylase;

(j) (i) an α

-ketoglutarate decarboxylase, or an α

-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase, or a glutamate;

succinate semialdehyde transaminase and a glutamate decarboxylase;

(ii) a 4-hydroxybutanoate dehydrogenase;

(k) succinyl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(l) glutamate dehydrogenase, 4-aminobutyrate oxidoreductase (deaminating), 4-aminobutyrate transaminase, glutamate decarboxylase, 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(m) homoserine deaminase;

4-hydroxybut-2-enoyl-CoA transferase, 4-hydroxybut-2-enoyl-CoA hydrolase, 4-hydroxybut-2-enoyl-CoA ligase;

4-hydroxybut-2-enoyl-CoA reductase;

(n) homoserine CoA transferase, homoserine-CoA hydrolase, or homoserine-CoA ligase;

homoserine-CoA deaminase; and

4-hydroxybut-2-enoyl-CoA reductase;

(o) homoserine deaminase;

4-hydroxybut-2-enoate reductase; and

4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, or 4-hydroxybutyryl-CoA ligase;

(p) succinyl-CoA reductase (aldehyde forming); and

4-hydroxybutyrate dehydrogenase;

(q) alpha-ketoglutarate decarboxylase; and

4-hydroxybutyrate dehydrogenase;

(r) succinate reductase; and

4-hydroxybutyrate dehydrogenase;

(s) alpha-ketoglutarate decarboxylase, or glutamate dehydrogenase or glutamate transaminase and glutamate decarboxylase and 4-aminobutyrate dehydrogenase or 4-aminobutyrate transaminase; and

4-hydroxybutyrate dehydrogenase; and

(t) a 4-hydroxybutyrate pathway selected from any of the pathways that produce 4-hydroxybutyrate as shown in any of FIG. 1, 8-13, 58, 62, 63 or 72-74.

13. The non-naturally occurring microbial organism of claim 12, wherein said microbial organism comprising (i) further comprises an exogenous nucleic acid encoding an enzyme selected from a pyruvate:

ferredoxin oxidoreductase, an aconitase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, an acetate kinase, a phosphotransacetylase, an acetyl-CoA synthetase, an NAD(P)H;

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

14. The non-naturally occurring microbial organism of claim 12, wherein said microbial organism comprising (ii) further comprises an exogenous nucleic acid encoding an enzyme selected from an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, NAD(P):

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

15. The non-naturally occurring microbial organism of claim 12, wherein said microbial organism comprising (iii) further comprises an exogenous nucleic acid encoding an enzyme selected from NAD(P):

ferredoxin oxidoreductase and ferredoxin.

21. A method for producing 4-hydroxybutyrate, comprising culturing the non-naturally occurring microbial organism of claim 12 under conditions and for a sufficient period of time to produce 4-hydroxybutyrate.

16-18. -18. (canceled)

20. (canceled)

22. A non-naturally occurring microbial organism, comprising a microbial organism having a gamma-butyrolactone pathway comprising at least one exogenous nucleic acid encoding a gamma-butyrolactone pathway enzyme expressed in a sufficient amount to produce gamma-butyrolactone;

said non-naturally occurring microbial organism further comprising;

(i) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from an ATP-citrate lyase, citrate lyase, a citryl-CoA synthetase, a citryl-CoA lyase, a fumarate reductase, isocitrate dehydrogenase and an alpha-ketoglutarate;

ferredoxin oxidoreductase;

(ii) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from a pyruvate;

ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H₂ hydrogenase;

or

(iii) at least one exogenous nucleic acid encodes an enzyme selected from a CO dehydrogenase, an H₂ hydrogenase, and combinations thereof;

(a) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase; and

spontaneous or enzyme catalyzed;

(b) Acetoacetyl-CoA thiolase or acetoacetyl-CoA synthase;

3-Hydroxybutyryl-CoA dehydrogenase;

Crotonase;

Crotonyl-CoA hydratase;

Phosphotrans-4-hydroxybutyrylase;

amd spontaneous or enzyme catalyzed;

(c) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(d) Succinate reductase;

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(e) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(f) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (aldehyde forming);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase;

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(g) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(h) Alpha-ketoglutarate decarboxylase or (Glutamate dehydrogenase and/or Glutamate transaminase;

Glutamate decarboxylase;

4-aminobutyrate dehydrogenase and/or 4-aminobutyrate transaminase);

4-Hydroxybutyrate dehydrogenase;

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(i) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyrate kinase;

Phosphotrans-4-hydroxybutyrylase;

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(j) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase);

Succinyl-CoA reductase (alcohol forming);

4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase; and

4-Hydroxybutyryl-CoA hydrolase or spontaneous;

(k) alpha-ketoglutarate reductase;

5-hydroxy-2-oxopentanoate dehydrogenase; and

5-hydroxy-2-oxopentanoate dehydrogenase (decarboxylation)

(l) 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, and α

-ketoglutarate decarboxylase;

(m) 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate;

CoA transferase, 4-butyrate kinase, phosphotransbutyrylase, α

-ketoglutarate decarboxylase;

(n) (i) an α

-ketoglutarate decarboxylase, or an α

-ketoglutarate dehydrogenase and a CoA-dependent succinic semialdehyde dehydrogenase, or a glutamate;

succinate semialdehyde transaminase and a glutamate decarboxylase;

(ii) a 4-hydroxybutanoate dehydrogenase;

(iii) a 4-hydroxybutyryl-CoA;

acetyl-CoA transferase, or a butyrate kinase and a phosphotransbutyrylase;

(o) 4-aminobutyrate CoA transferase, 4-aminobutyryl-CoA hydrolase, 4-aminobutyrate-CoA ligase, 4-aminobutyryl-CoA oxidoreductase (deaminating), 4-aminobutyryl-CoA transaminase, and 4-hydroxybutyryl-CoA dehydrogenase;

(p) 4-aminobutyrate CoA transferase, 4-aminobutyryl-CoA hydrolase, 4-aminobutyrate-CoA ligase, 4-aminobutyryl-CoA reductase (alcohol forming), 4-aminobutyryl-CoA reductase, 4-aminobutan-1-ol dehydrogenase, 4-aminobutan-1-ol oxidoreductase (deaminating) and 4-aminobutan-1-ol transaminase;

(q) 4-aminobutyrate kinase, 4-aminobutyraldehyde dehydrogenase (phosphorylating), 4-aminobutan-1-ol dehydrogenase, 4-aminobutan-1-ol oxidoreductase (deaminating), 4-aminobutan-1-ol transaminase, [(4-aminobutanolyl)oxy]phosphonic acid oxidoreductase (deaminating), [(4-aminobutanolyl)oxy]phosphonic acid transaminase, 4-hydroxybutyryl-phosphate dehydrogenase, and 4-hydroxybutyraldehyde dehydrogenase (phosphorylating);

(r) alpha-ketoglutarate 5-kinase, 2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating), 2,5-dioxopentanoic acid reductase, alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, alpha-ketoglutaryl-CoA ligase, alpha-ketoglutaryl-CoA reductase, 5-hydroxy-2-oxopentanoic acid dehydrogenase, alpha-ketoglutaryl-CoA reductase (alcohol forming), 5-hydroxy-2-oxopentanoic acid decarboxylase, and 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(s) glutamate CoA transferase, glutamyl-CoA hydrolase, glutamyl-CoA ligase, glutamate 5-kinase, glutamate-5-semialdehyde dehydrogenase (phosphorylating), glutamyl-CoA reductase, glutamate-5-semialdehyde reductase, glutamyl-CoA reductase (alcohol forming), 2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating), 2-amino-5-hydroxypentanoic acid transaminase, 5-hydroxy-2-oxopentanoic acid decarboxylase, 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(t) 3-hydroxybutyryl-CoA dehydrogenase, 3-hydroxybutyryl-CoA dehydratase, vinylacetyl-CoA Δ

-isomerase, or 4-hydroxybutyryl-CoA dehydratase;

(u) homoserine deaminase, homoserine CoA transferase, homoserine-CoA hydrolase, homoserine-CoA ligase, homoserine-CoA deaminase, 4-hydroxybut-2-enoyl-CoA transferase, 4-hydroxybut-2-enoyl-CoA hydrolase, 4-hydroxybut-2-enoyl-CoA ligase, 4-hydroxybut-2-enoate reductase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybut-2-enoyl-CoA reductase;

(v) succinyl-CoA reductase (alcohol forming), 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(w) glutamate dehydrogenase, 4-aminobutyrate oxidoreductase (deaminating), 4-aminobutyrate transaminase, glutamate decarboxylase, 4-hydroxybutyryl-CoA hydrolase, 4-hydroxybutyryl-CoA ligase, or 4-hydroxybutanal dehydrogenase (phosphorylating);

(x) 4-aminobutyrate kinase;

4-aminobutyraldehyde dehydrogenase (phosphorylating);

4-aminobutan-1-ol dehydrogenase; and

4-aminobutan-1-ol oxidoreductase (deaminating) or 4-aminobutan-1-ol transaminase;

(y) 4-aminobutyrate kinase;

[(4-aminobutanolyl)oxy]phosphonic acid oxidoreductase (deaminating) or [(4-aminobutanolyl)oxy]phosphonic acid transaminase;

4-hydroxybutyryl-phosphate dehydrogenase; and

4-hydroxybutyraldehyde dehydrogenase (phosphorylating);

(z) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase (alcohol forming); and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(aa) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase (alcohol forming); and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(bb) alpha-ketoglutarate 5-kinase;

2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating);

2,5-dioxopentanoic acid reductase; and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(cc) alpha-ketoglutarate 5-kinase;

2,5-dioxopentanoic semialdehyde dehydrogenase (phosphorylating);

2,5-dioxopentanoic acid reductase; and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(dd) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase;

5-hydroxy-2-oxopentanoic acid dehydrogenase; and

5-hydroxy-2-oxopentanoic acid decarboxylase;

(ee) alpha-ketoglutarate CoA transferase, alpha-ketoglutaryl-CoA hydrolase, or alpha-ketoglutaryl-CoA ligase;

alpha-ketoglutaryl-CoA reductase;

5-hydroxy-2-oxopentanoic acid dehydrogenase; and

5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(ff) glutamate CoA transferase, glutamyl-CoA hydrolase, or glutamyl-CoA ligase;

glutamyl-CoA reductase (alcohol forming);

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(gg) glutamate 5-kinase;

glutamate-5-semialdehyde dehydrogenase (phosphorylating);

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(hh) glutamate CoA transferase, glutamyl-CoA hydrolase, or glutamyl-CoA ligase;

glutamyl-CoA reductase;

glutamate-5-semialdehyde reductase;

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(ii) glutamate 5-kinase;

glutamate-5-semialdehyde dehydrogenase (phosphorylating);

glutamate-5-semialdehyde reductase;

2-amino-5-hydroxypentanoic acid oxidoreductase (deaminating) or 2-amino-5-hydroxypentanoic acid transaminase; and

5-hydroxy-2-oxopentanoic acid decarboxylase or 5-hydroxy-2-oxopentanoic acid dehydrogenase (decarboxylation);

(jj) homoserine deaminase;

4-hydroxybut-2-enoyl-CoA transferase, 4-hydroxybut-2-enoyl-CoA hydrolase, 4-hydroxybut-2-enoyl-CoA ligase;

4-hydroxybut-2-enoyl-CoA reductase;

(kk) homoserine CoA transferase, homoserine-CoA hydrolase, or homoserine-CoA ligase;

homoserine-CoA deaminase; and

4-hydroxybut-2-enoyl-CoA reductase;

(ll) homoserine deaminase;

4-hydroxybut-2-enoate reductase; and

4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA hydrolase, or 4-hydroxybutyryl-CoA ligase;

(mm) (i) alpha-ketoglutarate decarboxylase;

or alpha-ketoglutarate dehydrogenase and CoA-dependent succinate semialdehyde dehydrogenase;

or glutamate;

succinate semialdehyde transaminase and glutamate decarboxylase;

(ii) 4-hydroxybutyrate dehydrogenase;

(iii) 4-hydroxybutyryl-CoA transferase;

or 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase; and

(iv) 4-hydroxybutyryl-CoA reductase;

(nn) (i) alpha-ketoglutarate decarboxylase;

or succinyl-CoA synthetase and CoA-dependent succinate semialdehyde dehydrogenase;

(ii) 4-hydroxybutyrate dehydrogenase;

(iii) 4-hydroxybutyryl-CoA transferase;

or 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase;

(oo) (i) alpha-ketoglutarate decarboxylase;

or glutamate dehydrogenase;

glutamate decarboxylase; and

deaminating 4-aminobutyrate oxidoreductase or 4-aminobutyrate transaminase;

or alpha-ketoglutarate dehydrogenase and CoA-dependent succinate semialdehyde dehydrogenase;

(ii) 4-hydroxybutyrate dehydrogenase; and

(iii) 4-hydroxybutyrate kinase;

phosphotrans-4-hydroxybutyrylase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyrate kinase;

phosphorylating 4-hydroxybutanal dehydrogenase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyrate kinase;

phosphotrans-4-hydroxybutyrylase; and

alcohol forming 4-hydroxybutyryl-CoA reductase;

or 4-hydroxybutyryl-CoA transferase or 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA ligase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or 4-hydroxybutyryl-CoA transferase or 4-hydroxybutyryl-CoA hydrolase or 4-hydroxybutyryl-CoA ligase; and

alcohol forming 4-hydroxybutyryl-CoA reductase;

(pp) (i) glutamate CoA transferase or glutamyl-CoA hydrolase or glutamyl-CoA ligase;

glutamyl-CoA reductase; and

glutamate-5-semialdehyde reductase;

or glutamate CoA transferase or glutamyl-CoA hydrolase or glutamyl-CoA ligase; and

alcohol forming glutamyl-CoA reductase;

or glutamate 5-kinase;

phosphorylating glutamate-5-semialdehyde dehydrogenase; and

glutamate-5-semialdehyde reductase;

(ii) deaminating 2-amino-5-hydroxypentanoic acid oxidoreductase or 2-amino-5-hydroxypentanoic acid transaminase; and

(iii) 5-hydroxy-2-oxopentanoic acid decarboxylase; and

4-hydroxybutyraldehyde reductase;

or decarboxylating 5-hydroxy-2-oxopentanoic acid dehydrogenase;

4-hydroxybutyryl-CoA reductase; and

4-hydroxybutyraldehyde reductase;

or decarboxylating 5-hydroxy-2-oxopentanoic acid dehydrogenase and alcohol forming 4-hydroxybutyryl-CoA reductase;

(qq) a gamma-butyrolactone pathway comprising a pathway selected from any of the pathways that produce 4-hydroxybutyryl-CoA or 4-hydroxybutyryl phosphate as shown in FIG. 1, 8-13, 58, 62-63 or 72-74, wherein gamma-butyrolactone is produced enzymatically or by spontaneous chemical conversion.

23. The non-naturally occurring microbial organism of claim 22, wherein said microbial organism comprising (i) further comprises an exogenous nucleic acid encoding an enzyme selected from a pyruvate:

ferredoxin oxidoreductase, an aconitase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, an acetate kinase, a phosphotransacetylase, an acetyl-CoA synthetase, an NAD(P)H;

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

24. The non-naturally occurring microbial organism of claim 22, wherein said microbial organism comprising (ii) further comprises an exogenous nucleic acid encoding an enzyme selected from an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, NAD(P):

ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

25. The non-naturally occurring microbial organism of claim 22, wherein said microbial organism comprising (iii) further comprises an exogenous nucleic acid encoding an enzyme selected from NAD(P):

ferredoxin oxidoreductase and ferredoxin.

32. A method for producing gamma-butyrolactone, comprising culturing the non-naturally occurring microbial organism of claim 22 under conditions and for a sufficient period of time to produce gamma-butyrolactone.

26-31. -31. (canceled)

33. A carboxylic acid reductase, comprising an amino acid sequence having an amino acid substitution selected from E16K;

Q95L;

L100M;

A1011T;

K823E;

T941S;

H15Q;

D198E;

G446C;

S392N;

F699L;

V8831;

F467S;

T987S;

R12H;

V295G;

V295A;

V295S;

V295T;

V295G;

V295V;

V295L;

V2951;

V295M;

V295P;

V295F;

V295Y;

V295W;

V295D;

V295E;

V295N;

V295Q;

V295H;

V295K;

V295R;

M296G;

M296A;

M296S;

M296T;

M296C;

M296V;

M296L;

M2961;

M296M;

M296P;

M296F;

M296Y;

M296W;

M296D;

M296E;

M296N;

M296Q;

M296H;

M296K;

M296R;

G297G;

G297A;

G297S;

G297T;

G297C;

G297V;

G297L;

G2971;

G297M;

G297P;

G297F;

G297Y;

G297W;

G297D;

G297E;

G297N;

G297Q;

G297H;

G297K;

G297R;

G391G;

G391A;

G391S;

G391T;

G391C;

G391V;

G391L;

G391I;

G391M;

G391P;

G391F;

G391Y;

G391W;

G391D;

G391E;

G391N;

G391Q;

G391H;

G391K;

G391R;

G421G;

G421A;

G421S;

G421T;

G421C;

G421V;

G421L;

G421I;

G421M;

G421P;

G421F;

G421Y;

G421W;

G421D;

G421E;

G421N;

G421Q;

G421H;

G421K;

G421R;

D413G;

D413A;

D413S;

D413T;

D413C;

D413V;

D413L;

D413I;

D413M;

D413P;

D413F;

D413Y;

D413W;

D413D;

D413E;

D413N;

D413Q;

D413H;

D413K;

D413R;

G414G;

G414A;

G414S;

G414T;

G414C;

G414V;

G414L;

G414I;

G414M;

G414P;

G414F;

G414Y;

G414W;

G414D;

G414E;

G414N;

G414Q;

G414H;

G414K;

G414R;

Y415G;

Y415A;

Y415S;

Y415T;

Y415C;

Y415V;

Y415L;

Y415I;

Y415M;

Y415P;

Y415F;

Y415Y;

Y415W;

Y415D;

Y415E;

Y415N;

Y415Q;

Y415H;

Y415K;

Y415R;

G416G;

G416A;

G416S;

G416T;

G416C;

G416V;

G416L;

G416I;

G416M;

G416P;

G416F;

G416Y;

G416W;

G416D;

G416E;

G416N;

G416Q;

G416H;

G416K;

G416R;

S417G;

5417A;

S417S;

S417T;

S417C;

S417V S417L;

S417I;

S417M;

S417P;

S417F;

S417Y;

S417W;

S417D;

S417E;

S417N;

S417Q;

S417H;

S417K; and

S417R, or combinations thereof.