title: Development of an orthogonal fatty acid biosynthesis system in E. coli for oleochemical production.
abstract: Here we report recombinant expression and activity of several type I fatty acid synthases that can function in parallel with the native Escherichia coli fatty acid synthase. Corynebacterium glutamicum FAS1A was the most active in E. coli and this fatty acid synthase was leveraged to produce oleochemicals including fatty alcohols and methyl ketones. Coexpression of FAS1A with the ACP/CoA-reductase Maqu2220 from Marinobacter aquaeolei shifted the chain length distribution of fatty alcohols produced. Coexpression of FAS1A with FadM, FadB, and an acyl-CoA-oxidase from Micrococcus luteus resulted in the production of methyl ketones, although at a lower level than cells using the native FAS. This work, to our knowledge, is the first example of in vivo function of a heterologous fatty acid synthase in E. coli. Using FAS1 enzymes for oleochemical production have several potential advantages, and further optimization of this system could lead to strains with more efficient conversion to desired products. Finally, functional expression of these large enzyme complexes in E. coli will enable their study without culturing the native organisms.
authors.affiliation: Joint BioEnergy Institute, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Haushalter
authors.affiliation: Joint BioEnergy Institute, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Groff
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States; Joint Genome Institute, Walnut Creek, CA 94598, United States.
authors.forename: Samuel
authors.initials: S
authors.lastname: Deutsch
authors.affiliation: QB3 Institute, University of California-Berkeley, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States.
authors.forename: Lionadi
authors.initials: L
authors.lastname: The
authors.affiliation: QB3 Institute, University of California-Berkeley, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States.
authors.forename: Ted A
authors.initials: TA
authors.lastname: Chavkin
authors.affiliation: QB3 Institute, University of California-Berkeley, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States.
authors.forename: Simon F
authors.initials: SF
authors.lastname: Brunner
authors.affiliation: QB3 Institute, University of California-Berkeley, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States; Synthetic Biology Engineering Research Center, University of California, Berkeley, CA 94720, United States.
authors.forename: Leonard
authors.initials: L
authors.lastname: Katz
authors.affiliation: Joint BioEnergy Institute, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States; QB3 Institute, University of California-Berkeley, 5885 Hollis Street, 4th Floor, Emeryville, CA 94608, United States; Synthetic Biology Engineering Research Center, University of California, Berkeley, CA 94720, United States; Department of Chemical & Biomolecular Engineering, Department of Bioengineering, University of California, Berkeley, CA 94720, United States. Electronic address: jdkeasling@lbl.gov.
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000096:biosynthesis* / Q000235:genetics; D005227:Fatty Acids / Q000096:biosynthesis* / Q000235:genetics; D053520:Marinobacter / Q000201:enzymology / Q000235:genetics*; D016982:Micrococcus luteus / Q000201:enzymology / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D064429:Fatty Acid Synthases
keywords: Fatty acid synthase; Fatty alcohols; Methyl ketones; Synthetic biology