Chimeric isoprenoid synthases and uses thereof

Abstract

Disclosed is a chimeric isoprenoid synthase polypeptide including a first domain from a first isoprenoid synthase joined to a second domain from a second, heterologous, isoprenoid synthase, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced in the absence of the second domain of the second, heterologous, isoprenoid synthase. Also disclosed is a chimeric isoprenoid synthase polypeptide including an asymmetrically positioned heterologous domain, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced when the domain is positioned at its naturally-occurring site in the isoprenoid synthase polypeptide.

1 . A chimeric isoprenoid synthase polypeptide comprising a first isoprenoid synthase polypeptide joined to a second, different, isoprenoid synthase polypeptide such that the reactions catalyzed by the chimeric isoprenoid synthase polypeptide are catalyzed with altered specificity, selectivity, or catalytic efficiency as compared to an isoprenoid synthase polypeptide lacking the second isoprenoid synthase polypeptide. 
     
     
         2 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the reactions catalyzed by the chimeric isoprenoid synthase polypeptide are catalyzed with altered specificity as compared to an isoprenoid synthase polypeptide lacking the second isoprenoid synthase polypeptide. 
     
     
         3 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the reactions catalyzed by the chimeric isoprenoid synthase polypeptide are catalyzed with altered selectivity as compared to an isoprenoid synthase polypeptide lacking the second isoprenoid synthase polypeptide. 
     
     
         4 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the reactions catalyzed by the chimeric isoprenoid synthase polypeptide are catalyzed with altered catalytic efficiency as compared to an isoprenoid synthase polypeptide lacking the second isoprenoid synthase polypeptide. 
     
     
         5 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the chimeric isoprenoid synthase polypeptide catalyzes at least two different isoprenoid reaction products. 
     
     
         6 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the first isoprenoid synthase polypeptide is from a plant isoprenoid synthase and the second, different, isoprenoid synthase polypeptide is from a second isoprenoid synthase. 
     
     
         7 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the first isoprenoid synthase polypeptide is from tobacco and the second isoprenoid synthase polypeptide is from  Hyoscyamus.    
     
     
         8 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the isoprenoid synthase polypeptide catalyzes the production of an antifungal agent. 
     
     
         9 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the isoprenoid synthase polypeptide catalyzes the production of an antibacterial agent. 
     
     
         10 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the isoprenoid synthase polypeptide catalyzes the production of an antitumor agent. 
     
     
         11 . The chimeric isoprenoid synthase polypeptide of claim l wherein the isoprenoid synthase polypeptide catalyzes the production of an antimalarial agent. 
     
     
         12 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the isoprenoid synthase polypeptide catalyzes the production of an insecticide. 
     
     
         13 . The chimeric isoprenoid synthase polypeptide of  claim 1  wherein the isoprenoid synthase polypeptide catalyzes the production of a compound suitable for the production of a flavoring or fragrance. 
     
     
         14 . DNA encoding a chimeric isoprenoid synthase polypeptide comprising a first isoprenoid synthase polypeptide joined to a second, different, isoprenoid synthase polypeptide such that the reactions catalyzed by the chimeric isoprenoid synthase polypeptide are catalyzed with altered specificity, selectivity, or catalytic efficiency as compared to an isoprenoid synthase polypeptide lacking the second isoprenoid synthase polypeptide. 
     
     
         15 . A vector comprising the DNA of  claim 14 . 
     
     
         16 . A cell comprising the DNA of  claim 14 . 
     
     
         17 . The cell of  claim 16 , wherein the cell is a bacterial cell. 
     
     
         18 . The cell of  claim 17 , wherein the cell is an  Escherichia coli  cell. 
     
     
         19 . The cell of  claim 16 , wherein the cell is a eukaryotic cell. 
     
     
         20 . The cell of  claim 19 , wherein the cell is a yeast or fungal cell. 
     
     
         21 . The cell of  claim 20 , wherein the cell is a  Saccharomyces cerevisiae  cell. 
     
     
         22 . The cell of  claim 19 , wherein the cell is a plant cell. 
     
     
         23 . The cell of  claim 19 , wherein the cell is an algal cell. 
     
     
         24 . The cell of  claim 19 , wherein the cell is a mammalian cell. 
     
     
         25  A transgenic plant comprising the DNA of  claim 14 . 
     
     
         26 . A vector comprising the DNA of  claim 14  and a dominant selectable marker. 
     
     
         27 . The vector of  claim 26  further comprising additional elements selected from the group consisting of a promoter regulatory region, a transcription initiation start site, a ribosome binding site, an RNA processing signal, a transcription termination site, and a polyadenylation agent. 
     
     
         28 . The vector of  claim 15  comprising a transcription initiation regulatory region. 
     
     
         29 . The vector of  claim 15  comprising a transcription termination regulatory region.