Methods for increasing product yields
View Patent ↗A non-naturally occurring microbial organism includes a microbial organism having a reductive TCA or Wood-Ljungdahl pathway in which at least one exogenous nucleic acid encoding these pathway enzymes is expressed in a sufficient amount to enhance carbon flux through acetyl-CoA. A method for enhancing carbon flux through acetyl-CoA includes culturing theses non-naturally occurring microbial organisms under conditions and for a sufficient period of time to produce a product having acetyl-CoA as a building block. Another non-naturally occurring microbial organism includes at least one exogenous nucleic acid encoding an enzyme expressed in a sufficient amount to enhance the availability of reducing equivalents in the presence of carbon monoxide or hydrogen, thereby increasing the yield of redox-limited products via carbohydrate-based carbon feedstock. A method for enhancing the availability of reducing equivalents in the presence of carbon monoxide or hydrogen includes culturing this organism for a sufficient period of time to produce a product.
1. A non-naturally occurring microbial organism comprising at least one exogenous nucleic acid encoding each of the following polypeptides:
(i) a carbon monoxide dehydrogenase,
(ii) a hydrogenase,
(iii) an NAD(P)H:ferredoxin oxidoreductase, and
(iv) a ferredoxin,
wherein the polypeptides are expressed in a sufficient amount to enhance the availability of reducing equivalents in the presence of carbon monoxide or hydrogen, thereby increasing the yield of redox-limited products via carbohydrate-based carbon feedstock.
2. The non-naturally occurring microbial organism of claim 1 further comprising one or more nucleic acids encoding an enzyme selected from the group consisting of a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a pyruvate carboxylase, and a malic enzyme.
3. The non-naturally occurring microbial organism of claim 2 further comprising one or more nucleic acids encoding an enzyme selected from the group consisting of a malate dehydrogenase, a fumarase, a fumarate reductase, a succinyl-CoA synthetase, and a succinyl-CoA transferase.
4. The non-naturally occurring microbial organism of claim 1 further comprising a 1,4-butanediol pathway, and wherein said microbial organism further comprises at least one exogenous nucleic acid encoding an enzyme selected from the group consisting of: 1) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase), 2) Succinyl-CoA reductase (aldehyde forming), 3) 4-Hydroxybutyrate dehydrogenase, 4) 4-Hydroxybutyrate kinase, 5) Phosphotrans-4-hydroxybutyrylase, 6) 4-Hydroxybutyryl-CoA reductase (aldehyde forming), 7) 1,4-butanediol dehydrogenase, 8) Succinate reductase, 9) Succinyl-CoA reductase (alcohol forming), 10) 4-Hydroxybutyryl-CoA transferase, 4-Hydroxybutyryl-CoA hydrolase, or 4-Hydroxybutyryl-CoA synthetase, 11) 4-Hydroxybutyrate reductase, 12) 4-Hydroxybutyryl-phosphate reductase, and 13) 4-Hydroxybutyryl-CoA reductase (alcohol forming).
5. The non-naturally occurring microbial organism of claim 1 further comprising a 1,3-butanediol pathway, and wherein said microbial organism further comprises at least one exogenous nucleic acid encoding an enzyme selected from the group consisting of: 1) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase), 2) Succinyl-CoA reductase (aldehyde forming), 3) 4-Hydroxybutyrate dehydrogenase, 4) 4-Hydroxybutyrate kinase, 5) Phosphotrans-4-hydroxybutyrylase, 6) 4-Hydroxybutyryl-CoA dehydratase, 7) Crotonase, 8) 3-Hydroxybutyryl-CoA reductase (aldehyde forming), 9) 3-Hydroxybutyraldehyde reductase, 10) Succinate reductase, 11) Succinyl-CoA reductase (alcohol forming), 12) 4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase, 13) 3-Hydroxybutyryl-CoA reductase (alcohol forming), 14) 3-Hydroxybutyryl-CoA hydrolase, or 3-Hydroxybutyryl-CoA synthetase, or 3-Hydroxybutyryl-CoA transferase, 15) 3-Hydroxybutyrate reductase.
6. The non-naturally occurring microbial organism of claim 1 further comprising a butanol pathway, and wherein said microbial organism further comprises at least one exogenous nucleic acid encoding an enzyme selected from the group consisting of: 1) Succinyl-CoA transferase, or Succinyl-CoA synthetase (or succinyl-CoA ligase), 2) Succinyl-CoA reductase (aldehyde forming), 3) 4-Hydroxybutyrate dehydrogenase, 4) 4-Hydroxybutyrate kinase, 5) Phosphotrans-4-hydroxybutyrylase, 6) 4-Hydroxybutyryl-CoA dehydratase, 7) Butyryl-CoA dehydrogenase, 8) Butyryl-CoA reductase (aldehyde forming), 9) Butyraldehyde reductase, 10) Succinate reductase, 11) Succinyl-CoA reductase (alcohol foaming), 12) 4-Hydroxybutyryl-CoA transferase, or 4-Hydroxybutyryl-CoA synthetase, 13) Butyryl-CoA reductase (alcohol forming), 14) Butyryl-CoA hydrolase, or Butyryl-CoA synthetase, or Butyryl-CoA transferase, 15) Butyrate reductase.
7. The non-naturally occurring microbial organism of claim 1 further comprising a 6-aminocaproic acid pathway.
8. The non-naturally occurring microbial organism of claim 1 further comprising a hexamethylenediamine pathway.
9. The non-naturally occurring microbial organism of claim 1 further comprising an adipic acid pathway.
10. The non-naturally occurring microbial organism of claim 1 further comprising a 1,3-propanediol pathway.
11. The non-naturally occurring microbial organism of claim 1 further comprising a glycerol pathway.
12. A method for enhancing the availability of reducing equivalents in the presence of carbon monoxide or hydrogen thereby increasing the yield of a redox-limited product via carbohydrate-based carbon feedstock, the method comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce the product.