Method of producing terpenes or terpenoids

Abstract

The present invention relates to a recombinant Deinococcus bacterium exhibiting enhanced 2-C-methyl-D-erythritol 4-phosphate/1-deoxy-D-xylulose 5-phosphate (MEP/DXP) pathway, and its use for producing terpene or terpenoid compounds.

1 . A method of producing a terpene or terpenoid comprising (i) culturing a recombinant  Deinococcus  bacterium that is genetically modified to overexpress a native, homologous or heterologous dxs gene encoding 1-deoxyxylulose 5-phosphate synthase (DXS) and to overexpress a native, homologous or heterologous idi gene encoding isopentenyl pyrophosphatase isomerase (IPP isomerase), under conditions suitable to produce the terpene or terpenoid and optionally (ii) recovering the terpene or terpenoid, wherein the 1-deoxyxylulose 5-phosphate synthase has at least 90% sequence identity to a polypeptide encoded by a dxs gene from a  Deinococcus  bacterium. 
     
     
         2 . The method of  claim 1 , wherein the 1-deoxyxylulose 5-phosphate synthase has at least 90% sequence identity to a polypeptide encoded by a dxs gene from a  Deinococcus  bacterium selected from the group consisting of  D. geothermalis, D. yunweiensis, D. apachensis, D. phoenicis, D. deserti, D. aquatilis, D. wulumuqiensis, D. radiodurans, D. gobiensis , D. sp. 2009 , D. maricopensis, D. peraridilitoris, D. proteolyticus, D. radiopugnans, D. murrayi, D. swuensis, D. solis  and  D. ficus.    
     
     
         3 . The method of  claim 1 , wherein the recombinant  Deinococcus  bacterium overexpresses a native, homologous or heterologous gene encoding farnesyl diphosphate synthase (FPP synthase). 
     
     
         4 . The method of  claim 3 , wherein the gene encoding FPP synthase encodes a polypeptide selected from the group consisting of:
 a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 47;   b) a polypeptide having an amino acid sequence having at least 60% sequence identity to SEQ ID NO: 47; and   c) a polypeptide encoded by a nucleotide sequence having at least 60% sequence identity to SEQ ID NO: 46.   
     
     
         5 . The method of  claim 3 , wherein the FPP synthase further exhibits dimethylallyltransferase activity and/or geranylgeranyl diphosphate synthase activity. 
     
     
         6 . The method of  claim 1 , wherein, in the recombinant  Deinococcus  bacterium, at least one gene selected from the group consisting of native, homologous or heterologous dxr, ispD, ispE, ispF, ispG and ispH genes, is overexpressed. 
     
     
         7 . The method of  claim 1 , wherein the recombinant  Deinococcus  bacterium further comprises a gene encoding a heterologous terpene synthase. 
     
     
         8 . The method of  claim 7 , wherein the heterologous terpene synthase is selected from the group consisting of monoterpene synthases, diterpene synthases, triterpene synthases and sesquiterpene synthases. 
     
     
         9 . The method of  claim 7 , wherein the heterologous terpene synthase is a monoterpene synthase, a cineole synthase, a limonene synthase or a linalool synthase. 
     
     
         10 . The method of  claim 7 , wherein the heterologous terpene synthase is a sesquiterpene synthase. 
     
     
         11 . The method of  claim 1 , wherein, in the recombinant  Deinococcus  bacterium, a gene encoding a lycopene beta-cyclase is inactivated. 
     
     
         12 . The method of  claim 1 , wherein the terpene or terpenoid is selected from monoterpenes, diterpenes, triterpenes, sesquiterpenes and carotenoids. 
     
     
         13 . The method of  claim 1 , wherein the terpene or terpenoid is a sesquiterpene. 
     
     
         14 . The method of  claim 1 , wherein the terpene or terpenoid is a carotenoid. 
     
     
         15 . The method of  claim 14 , wherein the carotenoid is lycopene or any other carotenoid compound derived from lycopene. 
     
     
         16 . The method of  claim 14 , wherein the carotenoid is deinoxanthine. 
     
     
         17 . A recombinant  Deinococcus  bacterium comprising genetic modifications to overexpress a native, homologous or heterologous dxs gene encoding 1-deoxyxylulose 5-phosphate synthase (DXS) and to overexpress a native, homologous or heterologous idi gene encoding isopentenyl pyrophosphatase isomerase (IPP isomerase), wherein the 1-deoxyxylulose 5-phosphate synthase has at least 90% sequence identity to a polypeptide encoded by a dxs gene from a  Deinococcus  bacterium. 
     
     
         18 . The recombinant  Deinococcus  bacterium of  claim 17 , wherein the 1-deoxyxylulose 5-phosphate synthase has at least 90% sequence identity to a polypeptide encoded by a dxs gene from a  Deinococcus  bacterium selected from the group consisting of  D. geothermalis, D. yunweiensis, D. apachensis, D. phoenicis, D. deserti, D. aquatilis, D. wulumuqiensis, D. radiodurans, D. gobiensis , D. sp. 2009 , D. maricopensis, D. peraridilitoris, D. proteolyticus, D. radiopugnans, D. murrayi, D. swuensis, D. solis  and  D. ficus.