title: Engineering of a Nepetalactol-Producing Platform Strain of Saccharomyces cerevisiae for the Production of Plant Seco-Iridoids.
abstract: The monoterpene indole alkaloids (MIAs) are a valuable family of chemicals that include the anticancer drugs vinblastine and vincristine. These compounds are of global significance-appearing on the World Health Organization's list of model essential medicines-but remain exorbitantly priced due to low in planta levels. Chemical synthesis and genetic manipulation of MIA producing plants such as Catharanthus roseus have so far failed to find a solution to this problem. Synthetic biology holds a potential answer, by building the pathway into more tractable organisms such as Saccharomyces cerevisiae. Recent work has taken the first steps in this direction by producing small amounts of the intermediate strictosidine in yeast. In order to help improve on these titers, we aimed to optimize the early biosynthetic steps of the MIA pathway to the metabolite nepetalactol. We combined a number of strategies to create a base strain producing 11.4 mg/L of the precursor geraniol. We also show production of the critical intermediate 10-hydroxygeraniol and demonstrate nepetalactol production in vitro. Lastly we demonstrate that activity of the iridoid synthase toward the intermediates geraniol and 10-hydroxygeraniol results in the synthesis of the nonproductive intermediates citronellol and 10-hydroxycitronellol. This discovery has serious implications for the reconstruction of the MIA in heterologous organisms.
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.
authors.forename: Alex
authors.initials: A
authors.lastname: Campbell
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bauchart
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Gold
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Biological Sciences, Brock University , 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Luca
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.
authors.forename: Vincent J J
authors.initials: VJ
authors.lastname: Martin
mesh_terms: D000080462:Acyclic Monoterpenes; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism*; D030008:Catharanthus / Q000378:metabolism*; D026121:Indole Alkaloids / Q000378:metabolism*; D039823:Iridoids / Q000378:metabolism*; D039821:Monoterpenes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism; D014748:Vinca Alkaloids / Q000378:metabolism
chemical_list: D000080462:Acyclic Monoterpenes; D019086:Bridged Bicyclo Compounds, Heterocyclic; D026121:Indole Alkaloids; D039823:Iridoids; D039821:Monoterpenes; D013729:Terpenes; D014748:Vinca Alkaloids; C497341:nepetalactol; C021093:strictosidine; C007836:geraniol; C007078:citronellol
keywords: 10-hydroxygeraniol; Saccharomyces cerevisiae; iridodial; monoterpene indole alkaloids; nepetalactol; seco-iridoid