Production of farnesol and geranylgeraniol

Abstract

The invention provides a biological method of producing farnesol or geranylgeraniol.

What is claimed is:  
     
         1 . A method for producing farnesol comprising: 
 (a) culturing a microorganism selected from the group consisting of  Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida utilis, Candida albicans, Ustilago maydis, Zymomonas mobilis, Staphylococcus aureus  and  Methylococcus capsulatus , in a fermentation medium to produce a product selected from the group consisting of farnesyl phosphate and farnesol, wherein the action of squalene synthase of said microorganism is reduced; and    (b) recovering said product.    
     
     
         2 . The method of  claim 1 , wherein said microorganism is genetically modified to decrease the action of squalene synthase.  
     
     
         3 . The method of  claim 2 , wherein said microorganism is further genetically modified to increase the action of HMG-CoA reductase.  
     
     
         4 . The method of  claim 3 , wherein the action of HMG-CoA reductase is increased by overexpression of HMG-CoA reductase or the catalytic domain thereof in the microorganism.  
     
     
         5 . The method of  claim 4 , wherein said microorganism is further genetically modified to overexpress a protein selected from the group consisting of acetoacetyl Co-A thiolose, HMG-CoA synthase, mevalonate kinase, phosphomevalonate kinase, phosphomevalonate decarboxylase, isopentenyl pyrophosphate isomerase, farnesyl pyrophosphate synthase, D-1-deoxyxylulose 5-phosphate synthase, and 1-deoxy-D-xylulose 5-phosphate reductoisomerase.  
     
     
         6 . The method of  claim 5 , wherein the microorganism has been genetically modified to overexpress farnesyl pyrophosphate synthase.  
     
     
         7 . The method of  claim 1 , wherein said microorganism is an erg9 mutant.  
     
     
         8 . The method of  claim 7 , wherein said microorganism comprises a erg9Δ::HIS3 deletion/insertion allele.  
     
     
         9 . The method of  claim 1 , wherein said recovering step comprises recovering said product from said microorganism.  
     
     
         10 . The method of  claim 1 , wherein said product is secreted into said fermentation medium by said microorganism and wherein said step of recovering comprises purification of said product from said fermentation medium.  
     
     
         11 . The method of  claim 1 , wherein said product is intracellular farnesyl phosphate and farnesol and said step of recovering comprises isolating,said farnesyl phosphate and farnesol from said microorganism.  
     
     
         12 . The method of  claim 1 , wherein said product is intracellular farnesyl phosphate and said step of recovering further comprises dephosphorylating said farnesyl phosphate to produce farnesol.  
     
     
         13 . The method of  claim 5 , wherein said genetic modification to increase the action of a protein comprises transformation of said microorganism with a recombinant nucleic acid molecule encoding said protein, wherein said recombinant nucleic acid molecule is operatively linked to a transcription control sequence.  
     
     
         14 . The method of  claim 4 , wherein said genetic modification to increase the action of HMG-CoA reductase comprises transformation of said microorganism with a recombinant nucleic acid molecule that is integrated into the genome of said microorganism.  
     
     
         15 . The method of  claim 1 , wherein said farnesyl phosphate is intracellular and said farnesol is extracellular and intracellular, wherein said step of recovering comprises a recovering step selected from the group consisting of recovering said farnesyl phosphate from said microorganism, recovering said farnesol from said fermentation medium and from said microorganism, and a combination thereof.  
     
     
         16 . A method for producing farnesol comprising: 
 (a) culturing a microorganism selected from the group consisting of  Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida utilis, Candida albicans, Ustilago maydis, Zymomonas mobilis, Staphylococcus aureus  and  Methylococcus capsulatus , in a fermentation medium comprising a squalene synthase inhibitor, to produce a product selected from the group consisting of farnesyl phosphate and farnesol; and    (b) recovering said product.    
     
     
         17 . The method of  claim 16 , wherein the microorganism has been further genetically modified to increase the activity of farnesylpyrophosphate phosphatase.