Diterpene-Producing Unicellular Organism

Abstract

The present invention is directed to a unicellular organism system, such as a yeast, for producing geranylgeranyl pyrophosphate and a diterpene in vivo. The yeast cell preferably comprises an inducible nucleic acid sequence encoding geranylgeranyl pyrophosphate synthase, an inducible nucleic acid sequence encoding a soluble form of HMG-CoA reductase, a nucleic acid sequence of an allele that confers an increase in sterol metabolic flux and, in the diterpene-producing cell, a diterpene synthase.

1 - 18 . (canceled)  
     
     
         19 . A unicellular organism for producing a diterpene precursor, comprising: 
 an exogenous nucleic acid sequence encoding a geranylgeranyl pyrophosphate synthase under the control of an inducible promoter operable in said organism;    an exogenous nucleic acid sequence encoding a soluble form of HMG-CoA reductase under control of an inducible promoter operable in said organism and    an exogenous nucleic acid sequence that confers to said cell an increase in sterol metabolic flux as compared to native sterol metabolic flux levels.    
     
     
         20 . The unicellular organism of  claim 19 , wherein said nucleic acid sequence encoding said geranylgeranyl pyrophosphate synthase is SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21.  
     
     
         21 . The unicellular organism of  claim 19 , wherein said organism is a yeast.  
     
     
         22 . A unicellular organism for producing a diterpene precursor, comprising: 
 an exogenous nucleic acid sequence encoding a geranylgeranyl pyrophosphate synthase under the control of an inducible promoter operable in said organism;    an exogenous nucleic acid sequence encoding a soluble form of HMG-CoA reductase under control of an inducible promoter operable in said organism and    a upc2-1 nucleic acid sequence.    
     
     
         23 . The unicellular organism of  claim 22 , wherein said nucleic acid sequence encoding said geranylgeranyl pyrophosphate synthase is SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4; SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21.  
     
     
         24 . The unicellular organism of  claim 22 , wherein said organism is a yeast.  
     
     
         25 - 79 . (canceled)