Methods and compositions for producing olefins

Abstract

Compositions and methods for producing olefins are described herein. The olefins can be used to produced biofuels.

1 - 209 . (canceled) 
     
     
         210 . A microbial host cell engineered to produce an olefin, comprising a heterologous polynucleotide sequence encoding a polypeptide having fatty acid decarboxylase activity. 
     
     
         211 . The microbial host cell of  claim 210 , wherein the polypeptide has an amino acid motif that is selected from the group consisting of (1) F-X-X-E-[VI]-[RK]-R-X-Y-P-F-{F}-P-X-[LIV]; (2) P-X(6)-[LI]-X(4)-{G}-[VI]-[HQM]-X-[MLV]-D-G-X(2)-H-X(2)-R-K; (3) [AC]-[AG]-[IV]-[DE]-[IL]-X-N-X(2)-R-P-X-[VI]-A-X(3)-[FY]-X(2)-F-X(3)-A-[LMV]-X-[DE]; and (4) {RK}-X(6)-[LMV]-X(4)-[AC]-[AG]-[IV]-[DE]-[IL]-X-N-X(2)-R-P-X-[VI]-A-X(3)-[FY]-X(2)-F-X(3)-A-[LMV]-X-[DE]. 
     
     
         212 . The microbial host cell of  claim 211 , wherein the microbial host cell is a recombinant microbial host cell. 
     
     
         213 . The recombinant microbial host cell of  claim 212 , wherein the recombinant microbial host cell expresses an increased level of the polypeptide having fatty acid decarboxylase activity relative to a corresponding wild-type microbial host cell. 
     
     
         214 . The recombinant microbial host cell of  claim 212 , wherein a gene encoding an acyl-CoA synthase is attenuated or deleted in the recombinant microbial host cell that expresses a reduced level of an acyl-CoA synthase relative to a corresponding wild type microbial host cell. 
     
     
         215 . The recombinant microbial host cell of  claim 214 , wherein the gene encoding the acyl-CoA synthase is fadD. 
     
     
         216 . The recombinant microbial host cell of  claim 212 , wherein the recombinant microbial host cell is engineered to express, overexpress, or attenuate a gene encoding a thioesterase relative to a corresponding wild type microbial host cell. 
     
     
         217 . The microbial host cell of  claim 210 , wherein the microbial host cell is selected from the group consisting of  Escherichia Coli, Bacillus subtilis  and  Saccharomyces cerevisiae.    
     
     
         218 . The microbial host cell of  claim 217 , wherein the microbial host cell is  Escherichia Coli.    
     
     
         219 . A cell culture comprising the microbial host cell of  claim 210 , wherein an olefin is produced in the cell culture when the microbial host cell is cultured in a medium containing a substrate under conditions effective to express the polypeptide having fatty acid decarboxylase activity. 
     
     
         220 . The cell culture of  claim 219 , wherein the olefin is a terminal olefin. 
     
     
         221 . The cell culture of  claim 220 , wherein the terminal olefin is selected from the group consisting of a C 5 -C 25  terminal olefin, a C 13 -C 21  terminal olefin, nonadecene, methylnonadecene, heptadecene, methylheptadecene, and pentadecene, an unsaturated terminal olefin, a monounsaturated terminal olefin, a straight chain terminal olefin, a branched chain terminal olefin, and a terminal olefin having a cyclic moiety. 
     
     
         222 . The cell culture of  claim 219 , wherein the olefin is secreted by the microbial host cell. 
     
     
         223 . The cell culture of  claim 219 , wherein the substrate is a carbon source. 
     
     
         224 . The cell culture of  claim 223 , wherein the carbon source is a fatty acid. 
     
     
         225 . The cell culture of  claim 224 , wherein the fatty acid is selected from the group consisting of a C 6 -C 26  fatty acid, a C 14 -C 22  fatty acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, behenic acid, methyl eicosanoic acid, eicosanoic acid, methyl octadecanoic acid, stearic acid, and palmitic acid, an unsaturated fatty acid, a monounsaturated fatty acid, a saturated fatty acid, a straight chain fatty acid, and a cyclic moiety. 
     
     
         226 . The cell culture of  claim 223 , wherein the carbon source is a carbohydrate. 
     
     
         227 . The cell culture of  claim 226 , wherein the carbohydrate is glucose. 
     
     
         228 . A method of producing an olefin comprising culturing the bacterial host cell of  claim 210 , under conditions sufficient to allow expression of the polypeptide having fatty acid decarboxylase activity. 
     
     
         229 . The method of  claim 228 , further comprising culturing the microbial host cell in a culture medium in the presence of at least one biological substrate for the polypeptide having fatty acid decarboxylase activity. 
     
     
         230 . The method of  claim 229 , further comprising isolating the olefin from the culture medium. 
     
     
         231 . An olefin produced by the method of  claim 230 , wherein the olefin has a δ 13 C of about −15.4 or greater, about −15.4 to about −10.9, or about −13.92 to about −13.84. 
     
     
         232 . An olefin produced by the method of  claim 230 , wherein the olefin has an f M   14 C of at least about 1.003, at least about 1.01, at least about 1.5, or about 1.111 to about 1.124.