Primary alcohol producing organisms
The invention provides a non-naturally occurring microbial organism having a microbial organism having at least one exogenous gene insertion and/or one or more gene disruptions that confer production of primary alcohols. A method for producing long chain alcohols includes culturing these non-naturally occurring microbial organisms.
1. A non-naturally occurring eukaryotic organism, comprising at least two gene disruptions, said at least two gene disruptions comprise a first gene disruption occurring in a gene encoding an enzyme selected from the group consisting of a cytosolic pyruvate decarboxylase, a cytosolic ethanol-specific alcohol dehydrogenase, and a mitochondrial ethanol-specific alcohol dehydrogenase, and a second gene disruption occurring in a gene encoding an enzyme selected from the group consisting of a cytosolic malate dehydrogenase, a glycerol-3-phosphate dehydrogenase shuttle, an external NADH dehydrogenase, and an internal mitochondrial NADH dehydrogenase, wherein said at least two gene disruptions confers production of long chain alcohols in the mitochondrion of said organism.
2. The organism of claim 1 , wherein production of long chain alcohols is growth-coupled.
3. The organism of claim 1 , wherein production of long chain alcohols is not growth-coupled.
4. The organism of claim 1 , wherein said first gene disruption is in a gene selected from the group consisting of YLR044C, YLR134W, YGR087C, PDC3, YBR145W, YGL256W, YOL086C, YMR303, YMR083W, YPL088W, YAL061W, YMR318C, YCR105W, and YDL168W.
5. The organism of claim 1 , wherein said second gene disruption is in a gene selected from the group consisting of YOL126C, YDL022W, YOL059W, YIL155C, YMR145C, YDL085W, and YML120C.
6. The organism of claim 1 , further comprising an exogenous nucleic acid encoding an enzyme in the mitochondrion selected from the group consisting of a pyruvate dehydrogenase, a pyruvate: NADP oxidoreductase, a pyruvate formate lyase, an acylating acetaldehyde dehydrogenase, an acetate CoA ligase, and an AMP-forming acetyl CoA synthetase;
or a gene regulatory region thereof.
7. The organism of claim 6 , further comprising enhanced NADH transporting shuttle systems for transport of NADH from the cytosol into the mitochondrion.
8. The organism of claim 6 , further comprising an exogenous nucleic acid encoding an enzyme in the mitochondrion selected from the group consisting of a transhydrogenase, formate dehydrogenase, a pyruvate decarboxylase, and a pyruvate oxidase; or a gene regulatory region thereof.
9. The organism of claim 1 , wherein said strain is in a substantially anaerobic culture medium.
10. The organism of claim 1 , wherein said strain is in a microaerobic culture medium.
11. The organism of claim 1 , wherein said organism is a yeast or a fungus.
12. The organism of claim 11 , wherein said yeast is selected from the group consisting of Saccharomyces spp. including Saccharomyces cerevisiae and Schizosaccharomyces pombe, Kluyveromyces spp. including Kluyveromyces lactis and Kluyveromyces marxianus , and Pichia spp. including Pichia pastoris.
13. The organism of claim 12 , wherein said yeast is Saccharomyces cerevisiae.
14. The organism of claim 11 , wherein said fungus is selected from the group consisting of Aspergillus spp., including Aspergillus terreus and Aspergillus niger , and Rhizopus spp., including Rhizopus arrhizus and Rhizopus oryzae.
15. A method for producing long chain alcohols, comprising culturing a non-naturally occurring eukaryotic organism according to claim 1 .
16. The method of claim 15 , wherein production of long chain alcohols is growth-coupled.
17. The method of claim 15 , wherein production of long chain alcohols is not growth-coupled.
18. The method of claim 15 , wherein said first gene disruption is in a gene selected from the group consisting of YLR044C, YLR134W, YGR087C, PDC3, YBR145W, YGL256W, YOL086C, YMR303, YMR083W, YPL088W, YAL061W, YMR318C, YCR105W, and YDL168W.
19. The method of claim 15 , wherein said second gene disruption is in a gene selected from the group consisting of YOL126C, YDL022W, YOL059W, YIL155C, YMR145C, YDL085W, and YML120C.
20. The method of claim 15 , further comprising an exogenous nucleic acid encoding an enzyme in the mitochondrion selected from the group consisting of a pyruvate dehydrogenase, a pyruvate: NADP oxidoreductase, a pyruvate formate lyase, an acylating acetaldehyde dehydrogenase, an acetate CoA ligase, and an AMP-forming acetyl CoA synthetase;
or a gene regulatory region thereof.
21. The method of claim 20 , further comprising enhanced NADH transporting shuttle systems for transport of NADH from the cytosol into the mitochondrion.
22. The method of claim 20 , further comprising an exogenous nucleic acid encoding an enzyme in the mitochondrion selected from the group consisting of a transhydrogenase, formate dehydrogenase, a pyruvate decarboxylase, and a pyruvate oxidase;
or a gene regulatory region thereof.
23. The method of claim 15 , wherein said organism is a yeast or a fungus.
24. The method of claim 23 , wherein said yeast is selected from the group consisting of Saccharomyces spp. including Saccharomyces cerevisiae and Schizosaccharomyces pombe, Kluyveromyces spp. including Kluyveromyces lactis and Kluyveromyces marxianus , and Pichia spp. including Pichia pastoris.
25. The method of claim 24 , wherein said yeast is Saccharomyces cerevisiae.
26. The method of claim 23 , wherein said fungus is selected from the group consisting of Aspergillus spp., including Aspergillus terreus and Aspergillus niger , and Rhizopus spp., including Rhizopus arrhizus and Rhizopus oryzae.
27. The method of claim 15 , wherein said strain is cultured in a substantially anaerobic medium.
28. The method of claim 15 , wherein said strain is cultured in a microaerobic medium.