title: Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production.
abstract: Limonene is a valuable monoterpene used in the production of several commodity chemicals and medicinal compounds. Among them, perillyl alcohol (POH) is a promising anti-cancer agent that can be produced by hydroxylation of limonene. We engineered E. coli with a heterologous mevalonate pathway and limonene synthase for production of limonene followed by coupling with a cytochrome P450, which specifically hydroxylates limonene to produce POH. A strain containing all mevalonate pathway genes in a single plasmid produced limonene at titers over 400mg/L from glucose, substantially higher than has been achieved in the past. Incorporation of a cytochrome P450 to hydroxylate limonene yielded approximately 100mg/L of POH. Further metabolic engineering of the pathway and in situ product recovery using anion exchange resins would make this engineered E. coli a potential production platform for any valuable limonene derivative.
authors.affiliation: Joint Bioenergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
authors.forename: Jorge
authors.initials: J
authors.lastname: Alonso-Gutierrez
authors.forename: Rossana
authors.initials: R
authors.lastname: Chan
authors.forename: Tanveer S
authors.initials: TS
authors.lastname: Batth
authors.forename: Paul D
authors.initials: PD
authors.lastname: Adams
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Petzold
authors.forename: Taek Soon
authors.initials: TS
authors.lastname: Lee
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism*; D053138:Cyclohexenes / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D006900:Hydroxylation / Q000235:genetics; D000077222:Limonene; D060847:Metabolic Engineering; D008798:Mevalonic Acid / Q000378:metabolism; D039821:Monoterpenes / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D053138:Cyclohexenes; D029968:Escherichia coli Proteins; D039821:Monoterpenes; D013729:Terpenes; C032208:perillyl alcohol; D003577:Cytochrome P-450 Enzyme System; D000077222:Limonene; D008798:Mevalonic Acid
keywords: Escherichia coli; Limonene; Metabolic engineering; Mevalonate pathway; Microbial production; Perillyl alcohol