Methods And Compositions For Producing Hydrocarbons

Abstract

Compositions and methods for producing aldehydes, alkanes, and alkenes are described herein. The aldehydes, alkanes, and alkenes can be used in biofuels.

1 .- 43 . (canceled) 
     
     
         44 . A recombinant microorganism for the production of a hydrocarbon, wherein the recombinant microorganism expresses:
 a polynucleotide sequence encoding a decarbonylase;   a polynucleotide sequence encoding a ferredoxin; and   a polynucleotide sequence encoding a ferredoxin reductase.   
     
     
         45 . The recombinant microorganism of  claim 44 , wherein the decarbonylase comprises:
 an amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; or   an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36.   
     
     
         46 . The recombinant microorganism of  claim 44 , wherein the ferredoxin comprises:
 an amino acid sequence set forth in SEQ ID NO: 90, 92, or 94; or   an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 90, 92, or 94.   
     
     
         47 . The recombinant microorganism of  claim 44 , wherein the ferredoxin reductase comprises:
 an amino acid sequence set forth in SEQ ID NO:88; or   an amino acid sequence having at least 85% sequence identity to SEQ ID NO:88.   
     
     
         48 . The recombinant microorganism of  claim 44 , comprising a polynucleotide sequence encoding an acyl-ACP reductase (AAR). 
     
     
         49 . The recombinant microorganism of  claim 48 , wherein the AAR comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 66, 68, 70, 72, 74, 76, 78, 80, or 82. 
     
     
         50 . The recombinant microorganism of  claim 44 , comprising a polynucleotide sequence encoding a carboxylic acid reductase (CAR). 
     
     
         51 . The recombinant microorganism of  claim 50 , wherein the CAR comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:86. 
     
     
         52 . The recombinant microorganism of  claim 50 , further comprising a polynucleotide sequence encoding a thioesterase. 
     
     
         53 . The recombinant microorganism of  claim 52 , wherein the thioesterase is TesA without the leader sequence (TesA), TesB, FatA, FatA1, FatB, FatB2, or FatB3. 
     
     
         54 . The recombinant microorganism of  claim 44 , wherein the recombinant microorganism is selected from the group consisting of a bacterium, yeast, fungus, and algae. 
     
     
         55 . The recombinant microorganism of  claim 54 , wherein the recombinant microorganism is a bacterium. 
     
     
         56 . The recombinant microorganism of  claim 55 , wherein the bacterium is  E. coli.    
     
     
         57 . The recombinant microorganism of  claim 55 , wherein the bacterium is a cyanobacterium. 
     
     
         58 . The recombinant microorganism of  claim 44 , wherein the recombinant microorganism produces a hydrocarbon that is a C 3  to C 21  alkane or alkene. 
     
     
         59 . The recombinant microorganism of  claim 58 , wherein the alkane or alkene is tridecane, methyltridecane, nonadecane, methylnonadecane, heptadecane, methylheptadecane, pentadecane, methylpentadecane, pentadecene, heptadecene, methylpentadecene, or methylheptadecene. 
     
     
         60 . A cell culture comprising a recombinant microorganism for the production of a hydrocarbon, wherein the recombinant microorganism expresses:
 a polynucleotide sequence encoding a decarbonylase;   a polynucleotide sequence encoding a ferredoxin; and   a polynucleotide sequence encoding a ferredoxin reductase.   
     
     
         61 . The cell culture of  claim 60 , wherein the hydrocarbon is a C 3  to C 21  alkane or alkene. 
     
     
         62 . A method of producing a hydrocarbon, the method comprising:
 culturing a recombinant microorganism in a culture medium comprising a carbon source under conditions effective to produce the hydrocarbon, wherein the recombinant microorganism expresses:
 a polynucleotide sequence encoding a decarbonylase; 
 a polynucleotide sequence encoding a ferredoxin; and 
 a polynucleotide sequence encoding a ferredoxin reductase. 
   
     
     
         63 . The method of  claim 62 , further comprising purifying the hydrocarbon from the culture medium. 
     
     
         64 . The method of  claim 62 , wherein the recombinant microorganism further expresses:
 an aldehyde biosynthetic polypeptide that is an acyl-ACP reductase (AAR) that catalyzes the conversion of an acyl-ACP to a fatty aldehyde, or is a carboxylic acid reductase (CAR) that catalyzes the conversion of a fatty acid to a fatty aldehyde; and   optionally, a thioesterase that catalyzes the conversion of an acyl-ACP to a fatty acid.   
     
     
         65 . The method of  claim 62 , wherein the hydrocarbon is a C 3 -C 21  alkane or alkene.