US2012136184A1PendingUtilityA1

Nucleic acid molecule

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Assignee: BALL ANDREWPriority: Sep 15, 2009Filed: Sep 29, 2011Published: May 31, 2012
Est. expirySep 15, 2029(~3.2 yrs left)· nominal 20-yr term from priority
Inventors:Andrew BallRobert MooreGregory P. KnowlesJian Qin
C07C 11/21C12P 5/02C12N 9/1085C12P 5/007C12N 15/52C12P 5/00C12N 15/63
45
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Claims

Abstract

The invention relates to an isolated nucleic acid molecule encoding a polypeptide capable of producing a triterpenoid hydrocarbon. The invention also relates to the encoded polypeptide, a vector comprising the nucleic acid molecule, a recombinant non-human organism comprising the nucleic acid molecule, and to methods of producing a triterpenoid hydrocarbon or an intermediate of biofuel using the nucleic acid molecule, polypeptide or recombinant organism.

Claims

exact text as granted — not AI-modified
1 . An isolated nucleic acid molecule encoding a polypeptide capable of producing a triterpenoid hydrocarbon, wherein the polypeptide comprises at least 29 contiguous amino acid residues of any one of SEQ ID NOs: 1 to 5 or 20 to 29. 
     
     
         2 . The nucleic acid molecule of  claim 1 , comprising any one of SEQ ID NOs: 6 to 9 or 30. 
     
     
         3 . An isolated polypeptide capable of producing a triterpenoid hydrocarbon, wherein the polypeptide comprises at least 29 contiguous amino acid residues of any one of SEQ ID NOs: 1 to 5 or 20 to 29. 
     
     
         4 . The polypeptide of  claim 3 , comprising any one of SEQ ID NOs: 1 to 5 or 20 to 29. 
     
     
         5 . A vector, comprising the nucleic acid molecule of  claim 1 . 
     
     
         6 . A recombinant non-human organism capable of producing a triterpenoid hydrocarbon, the organism comprising the nucleic acid molecule of  claim 1 . 
     
     
         7 . The recombinant organism of  claim 6 , wherein the organism is a microorganism. 
     
     
         8 . The recombinant organism of  claim 7 , wherein the microorganism is an alga or a bacterium. 
     
     
         9 . The recombinant organism of  claim 7 , wherein the microorganism is selected from the group consisting of:  Escherichia coli; Chlamydomonas reinhardtii; Saccharomyces cerevisiae ; and  Pichia  sp. 
     
     
         10 . A method for producing a triterpenoid hydrocarbon, comprising expressing the nucleic acid molecule of  claim 1 . 
     
     
         11 . A method for producing a triterpenoid hydrocarbon, comprising using the polypeptide of  claim 3 . 
     
     
         12 . A method for producing a triterpenoid hydrocarbon, comprising the step of growing the recombinant organism of  claim 6  under conditions sufficient for the organism to produce the triterpenoid hydrocarbon. 
     
     
         13 . The method of  claim 12 , wherein the triterpenoid hydrocarbon is squalene or botryococcene and the conditions comprise supplementing the recombinant organism with NADPH or NADH. 
     
     
         14 . The method of  claim 12 , wherein the triterpenoid hydrocarbon is dehydrosqualene or dehydrobotryococcene and the conditions comprise not supplementing the recombinant organism with, or minimising the concentration of, NADPH or NADH. 
     
     
         15 . A triterpenoid hydrocarbon produced by the method of  claim 12 . 
     
     
         16 . A method for producing an intermediate of a biofuel, a bioplastic, a pharmaceutical, a food additive, an industrial chemical or a specialty chemical production, comprising expressing the nucleic acid molecule of  claim 1 . 
     
     
         17 . A method for producing an intermediate of a biofuel, a bioplastic, a pharmaceutical, a food additive, an industrial chemical or a specialty chemical production, comprising using a polypeptide of  claim 3 . 
     
     
         18 . A method for producing an intermediate of a biofuel, a bioplastic, a pharmaceutical, a food additive, an industrial chemical or a specialty chemical production, comprising the steps of growing the recombinant organism of  claim 6  under conditions sufficient for the organism to produce a triterpenoid hydrocarbon, harvesting the triterpenoid hydrocarbon from the organism, and producing the intermediate from the triterpenoid hydrocarbon. 
     
     
         19 . An intermediate produced by the method of  claim 18 . 
     
     
         20 . The nucleic acid molecule of  claim 1 , wherein the triterpenoid hydrocarbon is produced from FPP and/or PSPP. 
     
     
         21 . The nucleic acid molecule of  claim 1 , wherein the triterpenoid hydrocarbon is botryococcene or squalene. 
     
     
         22 . A vector deposited as V10/022,706 or V10/022,707.

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