Inactivation of genes of the mep pathway

Abstract

The invention relates to cells and organisms as well as to methods for producing said cells and organisms, according to which intermediates of the mevalonate-independent pathway for isoprenoid biosynthesis (MEP pathway) are enriched by deleting or inactivating genes. The derivatives can also be enriched by using enzyme inhibitors. The enriched intermediates may be used as substrates in enzyme activity tests. The inventive cells and organisms and the enriched intermediates can further be used in the production of medicaments.

1 . A process for preparing intermediates of the MEP pathway or derivatives of these intermediates, comprising:
 modifying a gene involved in the MEP pathway in selected cells or organisms to a degree sufficient to delete, inactivate or change the gene, thereby reducing or modifying the enzymatic activity of a gene product derived from the modified gene with respect to the MEP pathway and producing an accumulation of an intermediate of the MEP pathway or a derivative of the intermediate within the selected cell or organism.   
     
     
         2 . A process according to  claim 1 , wherein the gene is the lytE gene. 
     
     
         3 . A process for preparing intermediates of the MEP pathway or derivatives of these intermediates, comprising exposing selected cells and organisms to an enzyme inhibitor. 
     
     
         4 . A process according to  claim 3 , wherein the enzyme inhibitor is capable of inhibiting the LytB enzyme. 
     
     
         5 . A process according to  claim 1 , wherein the organisms are selected from a group consisting of bacteria, algae, plants and protozoa. 
     
     
         6 . A process according to  claim 1  further comprising concentrating an intermediate of the MEP pathway. 
     
     
         7 . A process according to  claim 1  wherein the intermediate or derivative of the intermediate is suitable for preparing substrates for a GcpE enzyme or LytB enzyme. 
     
     
         8 . A process according to  claim 1  wherein the intermediate or derivative of the intermediate is capable of activating gamma/delta T cells. 
     
     
         9 . A process according to  claim 1  wherein the intermediate or derivative of the intermediate obtained is 3-formyl-1-butypyrophosphate. 
     
     
         10 . A method of activating gamma/delta T-cells comprising contacting gamma/delta T-cells with bacterial cells in which the lytB gene has been deleted or inactivated.