Heterologous production of 10-methylstearic acid

Abstract

Nucleic acids and cells comprising a methyltransferase gene and/or a reductase gene are disclosed. These nucleic acids and cells may be used to produce branched (methyl)lipids, such as 10-methylstearate.

What is claimed is: 
     
         1 .- 29 . (canceled) 
     
     
         30 . A nucleic acid comprising a recombinant methyltransferase gene and a constitutive promoter operably-linked to the recombinant methyltransferase gene, wherein the constitutive promoter is a eukaryotic promoter, and wherein the recombinant methyltransferase gene encodes an amino acid sequence with at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:64, SEQ ID NO:70, or SEQ ID NO: 76. 
     
     
         31 . The nucleic acid of  claim 30 , wherein
 the recombinant methyltransferase gene is codon-optimized for expression in yeast, algae, or plants.   
     
     
         32 . The nucleic acid of  claim 30 , further comprising a recombinant reductase gene, wherein the recombinant reductase gene encodes an amino acid sequence with at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO: 10, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO:24, SEQ ID NO:28, SEQ ID NO:34, SEQ ID NO:40, SEQ ID NO:44, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:62, SEQ ID NO:68, or SEQ ID NO: 74. 
     
     
         33 . (canceled) 
     
     
         34 . The nucleic acid of  claim 32 , wherein
 the recombinant reductase gene is codon-optimized for expression in yeast, algae, or plants.   
     
     
         35 .- 36 . (canceled) 
     
     
         37 . The nucleic acid of  claim 32 , wherein the recombinant methyltransferase gene and the recombinant reductase gene are part of a single open reading frame that encodes a fusion protein. 
     
     
         38 - 49 . (canceled) 
     
     
         50 . The nucleic acid of  claim 30 , wherein the constitutive promoter is a bacterial or yeast promoter. 
     
     
         51 . The nucleic acid of  claim 50 , wherein the constitutive promoter is a  Yarrowia, Arxula , or  Saccharomyces  promoter. 
     
     
         52 . The nucleic acid of  claim 32 , further comprising a second constitutive promoter operably-linked to the recombinant reductase gene, wherein the second constitutive promoter is a eukaryotic promoter. 
     
     
         53 . The nucleic acid of  claim 52 , wherein the second constitutive promoter is a bacterial or yeast promoter. 
     
     
         54 . The nucleic acid of  claim 53 , wherein the second constitutive promoter is a  Yarrowia, Arxula , or  Saccharomyces  promoter. 
     
     
         55 . The nucleic acid of  claim 30 , wherein the amino acid sequence has at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:76. 
     
     
         56 . The nucleic acid of  claim 55 , wherein the amino acid sequence comprises the amino acid sequence set forth in SEQ ID NO:76. 
     
     
         57 . The nucleic acid of  claim 55 , wherein the recombinant methyltransferase gene comprises a nucleic acid sequence having at least 95% sequence identity with the nucleotide sequence set forth in SEQ ID NO:75. 
     
     
         58 . The nucleic acid of  claim 57 , wherein the recombinant methyltransferase gene comprises the nucleotide sequence set forth in SEQ ID NO:75. 
     
     
         59 . The nucleic acid of  claim 32 , wherein the amino acid sequence encoded by the recombinant reductase gene has at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:74. 
     
     
         60 . The nucleic acid of  claim 59 , wherein the amino acid sequence encoded by the recombinant reductase gene comprises the amino acid sequence set forth in SEQ ID NO:74. 
     
     
         61 . The nucleic acid of  claim 59 , wherein the recombinant reductase gene comprises a nucleic acid sequence having at least 95% sequence identity with the nucleotide sequence set forth in SEQ ID NO:73. 
     
     
         62 . The nucleic acid of  claim 61 , wherein the recombinant reductase gene comprises the nucleotide sequence set forth in SEQ ID NO:73. 
     
     
         63 . The nucleic acid of  claim 37 , wherein the nucleic acid comprises a nucleotide sequence having at least 95% sequence identity with the nucleotide sequence set forth in SEQ ID NO:97 or wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:97. 
     
     
         64 . The nucleic acid of  claim 37 , wherein the nucleic acid comprises a nucleotide sequence having at least 95% sequence identity with the nucleotide sequence set forth in SEQ ID NO:98 or wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID.