Organisms for the production of isopropanol,
A non-naturally occurring microbial organism having an isopropanol pathway includes at least one exogenous nucleic acid encoding an isopropanol pathway enzyme. The pathway includes an enzyme selected from a 4-hydroxybutyryl-CoA dehydratase, a crotonase, a 3-hydroxybutyryl-CoA dehydrogenase, an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, an acetoacetyl-CoA hydrolase, an acetoacetate decarboxylase, and an acetone reductase. A non-naturally occurring microbial organism having an n-butanol pathway includes at least one exogenous nucleic acid encoding an n-butanol pathway enzyme. Other non-naturally occurring microbial organism have n-butanol or isobutanol pathways. The organisms are cultured to produce isopropanol, n-butanol, or isobutanol.
1. A non-naturally occurring microbial organism having an isopropanol pathway, wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes:
(i) a 4-hydroxybutyryl-CoA dehydratase, a crotonase, or a 3-hydroxybutyryl-CoA dehydrogenase,
(ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase,
(iii) an acetoacetate decarboxylase, and
(iv) an acetone reductase.
2. The non-naturally occurring microbial organism of claim 1 , wherein said exogenous nucleic acids are heterologous nucleic acid.
3. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.
4. A method for producing isopropanol comprising culturing a non-naturally occurring microbial organism according to claim 1 , under conditions and for a sufficient period of time to produce isopropanol.
5. The method of claim 4 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.
6. The method of claim 4 , wherein said exogenous nucleic acids are heterologous nucleic acids.
7. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a 4-hydroxybutyryl-CoA dehydratase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
8. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a crotonase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
9. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
10. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase and (b) a crotonase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
11. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase and (b) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
12. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a crotonase, and (b) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
13. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase, (b) a crotonase, and (c) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
14. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a 4-hydroxybutyryl-CoA dehydratase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
15. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a crotonase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
16. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
17. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase and (b) a crotonase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
18. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase and (b) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
19. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a crotonase, and (b) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA:acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.
20. The method of claim 4 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the following isopropanol pathway enzymes (i) (a) a 4-hydroxybutyryl-CoA dehydratase, (b) a crotonase, and (c) a 3-hydroxybutyryl-CoA dehydrogenase, (ii) an acetoacetyl-CoA synthetase, an acetyl-CoA: acetoacetate-CoA transferase, or an acetoacetyl-CoA hydrolase, (iii) an acetoacetate decarboxylase, and (iv) an acetone reductase.