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

A non-naturally occurring Escherichia coli organism, comprising a 1,4-butanediol (BDO) pathway comprising BDO pathway enzymes expressed in a sufficient amount to produce BDO, wherein said bacterial organism is genetically modified, and wherein said bacterial organism:

• (A) comprises a BDO pathway selected from:
• (i) a BDO pathway comprising:
• (a) alpha-ketoglutarate decarboxylase;
• or
• alpha-ketoglutarate dehydrogenase and CoA-dependent succinate semialdehyde dehydrogenase;
• or
• glutamate:
• succinate semialdehyde transaminase and glutamate decarboxylase;
• (b) 4-hydroxybutyrate dehydrogenase;
• (c) 4-hydroxybutyryl-CoA transferase;
• or
• 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase;
• (d) 4-hydroxybutyryl-CoA reductase;
• and
• (e) 4-hydroxybutyraldehyde reductase;
• or
• aldehyde/alcohol dehydrogenase, said aldehyde/alcohol dehydrogenase converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
• (ii) a BDO pathway comprising:
• (a) alpha-ketoglutarate decarboxylase;
• or
• succinyl-CoA synthetase and CoA-dependent succinate semialdehyde dehydrogenase;
• (b) 4-hydroxybutyrate dehydrogenase;
• (c) 4-hydroxybutyryl-CoA transferase;
• or
• 4-hydroxybutyrate kinase and phosphotrans-4-hydroxybutyrylase;
• and
• (d) aldehyde dehydrogenase, said aldehyde dehydrogenase converting 4-hydroxybutyryl-CoA to 4-hydroxybutyraldehyde;
• and alcohol dehydrogenase, said alcohol dehydrogenase converting 4-hydroxybutyraldehyde to 1,4-butanediol;
• or
• aldehyde/alcohol dehydrogenase, said aldehyde/alcohol dehydrogenase converting 4-hydroxybutyryl-CoA to 1,4-butanediol;
• (iii) a BDO pathway comprising:
• (a) 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;
• (b) 4-hydroxybutyrate dehydrogenase;
• and
• (c) 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;
• and
• (iv) a BDO pathway comprising:
• (a) 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;
• (b) deaminating 2-amino-5-hydroxypentanoic acid oxidoreductase or 2-amino-5-hydroxypentanoic acid transaminase;
• and
• (c) 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;
• and
• (B) comprises disruption of a gene encoding a protein in an aerobic respiratory control regulatory system;
• expresses an exogenous NADH insensitive citrate synthase;
• or expresses exogenous phosphoenolpyruvate carboxykinase.