Recombinant microorganisms and uses therefor
View Patent ↗The invention provides, inter alia, methods for the production of acetone, isopropanol and/or precursors of acetone and/or isopropanol by microbial fermentation of substrates comprising CO, genetically modified microorganisms of use in such methods, nucleic acids suitable for preparation of genetically modified microorganisms, a novel alcohol dehydrogenase and nucleic acids encoding same.
1. A recombinant microorganism comprising an exogenous thiolase (EC 2.3.1.9), an exogenous CoA transferase (EC 2.8.3.9), and an exogenous decarboxylase (EC 4.1.1.4 or EC 4.1.1.74), wherein the recombinant microorganism is derived from a parental microorganism selected from the group consisting of Clostridium autoethanogenum, Clostridium ljungdahlii , and Clostridium ragsdalei , wherein the recombinant microorganism produces isopropanol.
2. The recombinant microorganism of claim 1 , wherein the thiolase is Clostridium acetobutylicum ThlA or Clostridium beijerinckii ThlA.
3. The recombinant microorganism of claim 2 , wherein the Clostridium acetobutylicum ThlA comprises SEQ ID NO: 42.
4. The recombinant microorganism of claim 1 , wherein the CoA transferase is Clostridium acetobutylicum CtfA and CtfB or Clostridium beijerinckii CtfA and CtfB.
5. The recombinant microorganism of claim 4 , wherein the Clostridium beijerinckii CtfA comprises SEQ ID NO: 43 and CtfB comprises SEQ ID NO: 44.
6. The recombinant microorganism of claim 1 , wherein the decarboxylase is acetoacetate decarboxylase (EC 4.1.1.4).
7. The recombinant microorganism of claim 6 , wherein the acetoacetate decarboxylase is Clostridium acetobutylicum Adc or Clostridium beijerinckii Adc.
8. The recombinant microorganism of claim 7 , wherein the Clostridium beijerinckii Adc comprises SEQ ID NO: 45.
9. The recombinant microorganism of claim 1 , wherein the decarboxylase is ketoisovalerate decarboxylase (EC 4.1.1.74).
10. The recombinant microorganism of claim 9 , wherein the ketoisovalerate decarboxylase is Lactococcus lactis KivD.
11. The recombinant microorganism of claim 10 , wherein the Lactococcus lactis KivD comprises SEQ ID NO: 73.
12. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises an endogenous or exogenous alcohol dehydrogenase.
13. The recombinant microorganism of claim 12 , wherein the endogenous or exogenous alcohol dehydrogenase is alcohol dehydrogenase (Adh) (EC 1.1.1.2) or alcohol dehydrogenase (Adh2) (EC 1.1.1.1).
14. The recombinant microorganism of claim 12 , wherein the exogenous alcohol dehydrogenase is Saccharomyces cerevisiae Adh2 (EC 1.1.1.1).
15. The recombinant microorganism of claim 1 , wherein the recombinant microorganism is a carboxydotrophic and acetogenic bacterium.
16. The recombinant microorganism of claim 1 , wherein the parental microorganism does not produce acetone or isopropanol.
17. The recombinant microorganism of claim 1 , wherein the parental microorganism is Clostridium autoethanogenum or Clostridium ljungdahlii.
18. The recombinant microorganism of claim 1 , wherein the parental microorganism is Clostridium autoethanogenum DSM23693 or Clostridium ljungdahlii DSM13528.
19. The recombinant microorganism of claim 1 , wherein the recombinant microorganism consumes a gaseous substrate comprising one or more of CO, CO 2 , and H 2 .
20. The recombinant microorganism of claim 19 , wherein the gaseous substrate comprises 20-70% CO.
21. The recombinant microorganism of claim 19 , wherein the gaseous substrate is derived from an industrial process selected from the group consisting of ferrous metal products manufacturing, non-ferrous products manufacturing, petroleum refining, coal gasification, biomass gasification, electric power production, carbon black production, and coke manufacturing.