title: Production of FAME biodiesel in E. coli by direct methylation with an insect enzyme.
abstract: Most biodiesel currently in use consists of fatty acid methyl esters (FAMEs) produced by transesterification of plant oils with methanol. To reduce competition with food supplies, it would be desirable to directly produce biodiesel in microorganisms. To date, the most effective pathway for the production of biodiesel in bacteria yields fatty acid ethyl esters (FAEEs) at up to ~1.5 g/L. A much simpler route to biodiesel produces FAMEs by direct S-adenosyl-L-methionine (SAM) dependent methylation of free fatty acids, but FAME production by this route has been limited to only ~16 mg/L. Here we employ an alternative, broad spectrum methyltransferase, Drosophila melanogaster Juvenile Hormone Acid O-Methyltransferase (DmJHAMT). By introducing DmJHAMT in E. coli engineered to produce medium chain fatty acids and overproduce SAM, we obtain medium chain FAMEs at titers of 0.56 g/L, a 35-fold increase over titers previously achieved. Although considerable improvements will be needed for viable bacterial production of FAMEs and FAEEs for biofuels, it may be easier to optimize and transport the FAME production pathway to other microorganisms because it involves fewer enzymes.
authors.affiliation: Department of Chemistry and Biochemistry UCLA-DOE Institute Molecular Biology Institute, University of California, Los Angeles, USA.
authors.forename: Saken
authors.initials: S
authors.lastname: Sherkhanov
authors.affiliation: Department of Chemistry and Biochemistry UCLA-DOE Institute Molecular Biology Institute, University of California, Los Angeles, USA.
authors.forename: Tyler P
authors.initials: TP
authors.lastname: Korman
authors.affiliation: Department of Chemistry and Biochemistry UCLA-DOE Institute Molecular Biology Institute, University of California, Los Angeles, USA.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Clarke
authors.affiliation: Department of Chemistry and Biochemistry UCLA-DOE Institute Molecular Biology Institute, University of California, Los Angeles, USA.
authors.forename: James U
authors.initials: JU
authors.lastname: Bowie
mesh_terms: D000818:Animals; D056804:Biofuels; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D057075:Enzyme Assays / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D007218:Industrial Microbiology / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D012436:S-Adenosylmethionine / Q000378:metabolism
chemical_list: D056804:Biofuels; D029721:Drosophila Proteins; D005227:Fatty Acids; D012436:S-Adenosylmethionine; D008780:Methyltransferases; C530958:juvenile hormone acid O-methyltransferase, Drosophila