Hydrocarbon-forming oxidative decarbonylase enzyme, hydrocarbons produced thereby, and method of use

Abstract

The present disclosure relates to oxidative decarbonylase enzymes, methods of making hydrocarbons with such enzymes, hydrocarbons produced therefrom and uses thereof. More particularly, the present disclosure relates to isolated polypeptide sequences that are cytochrome P450 enzymes with oxidative decarbonylase activity and methods of their use to generate hydrocarbon products, such as biofuels.

We claim: 
     
         1 . An isolated polypeptide sequence having an amino acid sequence with at least 95% identity to the amino acid sequence of cytochrome P450 CYP4G1 (SEQ ID NO: 2) and having cytochrome P450 monooxygenase activity. 
     
     
         2 . The isolated polypeptide sequence of  claim 1 , wherein the amino acid sequence has at least 99% identity to the amino acid sequence of SEQ ID NO: 2. 
     
     
         3 . The isolated polypeptide sequence of  claim 1 , wherein the polypeptide sequence is as set forth by SEQ ID NO: 2. 
     
     
         4 . An isolated polynucleotide sequence that encodes the polypeptide sequence of  claim 1  or a polypeptide sequence having an amino acid sequence with at least 95% identity to the amino acid sequence of cytochrome P450 CYP4G2 (SEQ ID NO: 1) and having cytochrome P450 monooxygenase activity SEQ ID NO: 1. 
     
     
         5 . A method of producing a polypeptide having oxidative decarbonylase activity comprising:
 introducing the polynucleotide of  claim 4  into an isolated host cell;   culturing the host cell; and   expressing from the host cell a polypeptide having oxidative decarbonylase activity.   
     
     
         6 . The method of  claim 5 , further comprising isolating the polypeptide with oxidative decarbonylase activity following expressing the polypeptide with oxidative decarbonylase activity. 
     
     
         7 . The method of  claim 5 , further comprising measuring oxidative decarbonylase activity of the expressed polypeptide. 
     
     
         8 . A method of catalyzing hydrocarbon formation comprising contacting a sample comprising a fatty aldehyde with the polypeptide of  claim 1 . 
     
     
         9 . A method of catalyzing hydrocarbon formation comprising contacting a sample comprising a fatty aldehyde with the polypeptide of  claim 3 . 
     
     
         10 . A transgenic host cell produced by inserting the polynucleotide of  claim 4  into the host cell. 
     
     
         11 . The transgenic host cell of  claim 10 , wherein the transgenic host cell is a yeast cell, an algae cell, a plant cell, an animal cell or a bacterial cell. 
     
     
         12 . A method for producing a hydrocarbon comprising culturing the cell of  claim 10  under conditions sufficient to produce hydrocarbons. 
     
     
         13 . The method of  claim 12 , further comprising isolating a hydrocarbon from the cell or from medium in which the cell is cultured. 
     
     
         14 . The method  claim 12 , wherein the cell is cultured in the presence of at least one substrate of oxidative decarbonylase activity. 
     
     
         15 . The method of  claim 14 , wherein the substrate is a fatty aldehyde. 
     
     
         16 . A biofuel comprising hydrocarbons produced by the method of  claim 12 . 
     
     
         17 . A biofuel comprising hydrocarbons produced by the method of  claim 8 . 
     
     
         18 . A recombinant nucleic acid sequence comprising a promoter operably linked to a nucleic acid sequence encoding a peptide selected from:
 a) a sequence as set forth by SEQ ID NO: 1 or 2; or   b) a sequence sharing 95% sequence identity with SEQ ID NO: 1 or 2.   
     
     
         19 . A cell comprising the recombinant nucleic acid sequence of  claim 18 . 
     
     
         20 . A method of making a hydrocarbon comprising culturing the cell of  claim 19  under conditions sufficient to produce a hydrocarbon.