Primary alcohol producing organisms

Abstract

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)