Chimeric isoprenoid synthases and uses thereof

Abstract

Provided 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 provided 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.

What is claimed is: 
     
         1 . A method for producing a chimeric isoprenoid synthase polypeptide, comprising:
 (a) providing a cell comprising DNA encoding the chimeric isoprenoid synthase polypeptide, wherein the encoded synthase comprises a first isoprenoid synthase polypeptide domain joined to a second, different, isoprenoid synthase polypeptide domain such that the chimeric isoprenoid synthase polypeptide produces one or more isoprenoid products that are not produced in the absence of the second domain of the second synthase; and   (b) culturing the cell under conditions in which the chimeric isoprenoid synthase encoded by the DNA is expressed.   
     
     
         2 . The method of  claim 1 , wherein the encoded chimeric isoprenoid synthase comprises a first isoprenoid synthase polypeptide domain joined to a second, different, isoprenoid synthase polypeptide domain, interrupted by or including a ratio determinant domain, whereby the chimeric isoprenoid synthase polypeptide encoded by the DNA catalyzes the production of at least one isoprenoid synthase reaction product that is not produced in the absence of the second isoprenoid synthase polypeptide and another isoprenoid synthase reaction product that is produced in the absence of the second synthase polypeptide domain. 
     
     
         3 . The method of  claim 1 , wherein the encoded chimeric isoprenoid synthase catalyzes the production of at least one isoprenoid synthase reaction product that is not produced in the absence of the second synthase domain. 
     
     
         4 . The method of  claim 1 , wherein the isoprenoid reaction product(s) is/are cyclic. 
     
     
         5 . The method of  claim 1 , wherein the second domain also determines the ratio of isoprenoid reaction products. 
     
     
         6 . The method of  claim 1 , wherein the chimeric isoprenoid synthase polypeptide catalyzes the production of at least two different isoprenoid reaction products. 
     
     
         7 . The method 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.   
     
     
         8 . The method of  claim 1 , wherein one or both of the isoprenoid synthases is a diterpene synthase. 
     
     
         9 . A method for production of an isoprenoid product, comprising:
 (a) providing a cell comprising DNA encoding a chimeric isoprenoid synthase polypeptide, wherein the encoded synthase comprises a first isoprenoid synthase polypeptide domain joined to a second, different, isoprenoid synthase polypeptide domain, such that the chimeric isoprenoid synthase polypeptide produces one or more isoprenoid products that are not produced in the absence of the second domain of the second synthase; and   (b) culturing the cell under conditions in which the encoded chimeric isoprenoid synthase catalyzes the production of an isoprenoid product.   
     
     
         10 . The method of  claim 9 , wherein the encoded chimeric synthase comprises a first isoprenoid synthase polypeptide domain joined to a second, different, isoprenoid synthase polypeptide domain, interrupted by or including a ratio determinant domain, whereby the chimeric isoprenoid synthase polypeptide encoded by the DNA catalyzes the production of at least one isoprenoid synthase reaction product that is not produced in the absence of the second isoprenoid synthase polypeptide, and another isoprenoid synthase reaction product that is produced in the absence of the second synthase polypeptide domain. 
     
     
         11 . The method of  claim 9 , wherein the encoded chimeric synthase catalyzes the production of at least one isoprenoid synthase reaction product that is not produced in the absence of the second isoprenoid synthase domain. 
     
     
         12 . The method of  claim 9 , wherein one or both of the isoprenoid synthases is a diterpene synthase. 
     
     
         13 . The method of  claim 9 , wherein the second domain also determines the ratio of isoprenoid reaction products. 
     
     
         14 . The method of  claim 9 , wherein the chimeric isoprenoid synthase polypeptide catalyzes the production of at least two different isoprenoid reaction products. 
     
     
         15 . The method of  claim 9 , 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.   
     
     
         16 . The method of  claim 1 , wherein the chimeric isoprenoid synthase is recovered. 
     
     
         17 . The method of  claim 9 , wherein the isoprenoid product is recovered. 
     
     
         18 . A chimeric isoprenoid synthase polypeptide, comprising a first isoprenoid synthase polypeptide domain joined to a second, different, isoprenoid synthase polypeptide domain such that the chimeric isoprenoid synthase polypeptide produces one or more isoprenoid products that are not produced in the absence of the second domain of the second synthase. 
     
     
         19 . The chimeric isoprenoid synthase polypeptide of  claim 18 , wherein the chimeric synthase catalyzes the production of at least one isoprenoid synthase reaction product that is not produced in the absence of the second isoprenoid synthase domain. 
     
     
         20 . The chimeric isoprenoid synthase polypeptide of  claim 18 , wherein one or both of the isoprenoid synthases is a diterpene synthase. 
     
     
         21 . The chimeric isoprenoid synthase polypeptide of  claim 18 , wherein the second domain also determines the ratio of isoprenoid reaction products. 
     
     
         22 . The chimeric isoprenoid synthase polypeptide of  claim 18 , wherein the chimeric isoprenoid synthase polypeptide catalyzes production of at least two different isoprenoid reaction products. 
     
     
         23 . A host cell, comprising the chimeric isoprenoid synthase polypeptide of  claim 18 . 
     
     
         24 . A host cell, comprising nucleic acid encoding the chimeric isoprenoid synthase polypeptide of  claim 18 . 
     
     
         25 . The host cell of  claim 23  that is a bacterial, yeast cell or a plant cell. 
     
     
         26 . The host cell of  claim 24  that is a bacterial, yeast cell or a plant cell. 
     
     
         27 . The method of  claim 1 , further comprising isolating the isoprenoid synthase. 
     
     
         28 . The method of  claim 9 , further comprising isolating an isoprenoid product.