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 .- 69 . (canceled)
70 . A non-naturally occurring eukaryotic organism, comprising one or more gene disruptions, said one or more gene disruptions occurring in genes encoding enzymes selected from the group consisting of a cytosolic pyruvate decarboxylase, a cytosolic ethanol-specific alcohol dehydrogenase, and a mitochondrial ethanol-specific alcohol dehydrogenase, wherein said one or more gene disruptions confers production of long chain alcohols in the mitochondrion of said organism.
71 . The organism of claim 70 , wherein production of long chain alcohols is growth-coupled.
72 . The organism of claim 70 , wherein production of long chain alcohols is not growth-coupled.
73 . The organism of claim 70 , wherein said one or more gene disruptions are in a gene selected from the group consisting of YLR044C, YLR134W, YGR087C, PDC3, YBR145W, YGL256W, YOL086C, YMR303, YMR083W, YPL088W, YAL061W, YMR318C, YCR105W, and YDL168W.
74 . The organism of claim 70 , further comprising one or more gene disruptions encoding an enzyme selected from the group consisting of a malate dehydrogenase, glycerol-3-phospate dehydrogenase shuttle, catalyzed by, the external NADH dehydrogenase, and internal NADH dehydrogenase.
75 . The organism of claim 74 , wherein said one or more gene disruptions are in a gene selected from the group consisting of YOL126C, YDL022W, YOL059W, YIL155C, YMR145C, YDL085W, and YML120C.
76 . The organism of claim 70 , 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.
77 . The organism of claim 76 , further comprising enhanced NADH transporting shuttle systems for transport of NADH from the cytosol into the mitochondrion.
78 . The organism of claim 76 , 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.
79 . The organism of claim 70 , wherein said strain is in a substantially anaerobic culture medium.
80 . The organism of claim 70 , wherein said strain is in a microaerobic culture medium.
81 . The organism of claim 70 , wherein said organism is a yeast or a fungus.
82 . The organism of claim 81 , 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.
83 . The organism of claim 82 , wherein said yeast is Saccharomyces cerevisiae.
84 . The organism of claim 81 , 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.
85 .- 95 . (canceled)
96 . A method for producing long chain alcohols, comprising culturing a non-naturally occurring eukaryotic organism according to claim 70 .
97 . The method of claim 96 , wherein production of long chain alcohols is growth-coupled.
98 . The method of claim 96 , wherein production of long chain alcohols is not growth-coupled.
99 . The method of claim 96 , wherein said one or more gene disruptions are in a gene selected from the group consisting of YLR044C, YLR134W, YGR087C, PDC3, YBR145W, YGL256W, YOL086C, YMR303, YMR083W, YPL088W, YAL061W, YMR318C, YCR105W, and YDL168W.
100 . The method of claim 96 , further comprising one or more gene disruptions encoding an enzyme selected from the group consisting of a cytosolic malate dehydrogenase, glycerol-3-phospate dehydrogenase shuttle, catalyzed by, the external NADH dehydrogenase, and internal NADH dehydrogenase.
101 . The method of claim 100 , wherein said one or more gene disruptions are in a gene selected from the group consisting of YOL126C, YDL022W, YOL059W, YIL155C, YMR145C, YDL085W, and YML120C.
102 . The method of claim 96 , 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.
103 . The method of claim 102 , further comprising enhanced NADH transporting shuttle systems for transport of NADH from the cytosol into the mitochondrion.
104 . The method of claim 102 , 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.
105 . The method of claim 96 , wherein said organism is a yeast or a fungus.
106 . The method of claim 105 , 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.
107 . The method of claim 106 , wherein said yeast is Saccharomyces cerevisiae.
108 . The method of claim 105 , 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.
109 . The method of claim 96 , wherein said strain is cultured in a substantially anaerobic medium.
110 . The method of claim 96 , wherein said strain is cultured in a microaerobic medium.
111 .- 113 . (canceled)