title: Characterisation of a Recombinant Patchoulol Synthase Variant for Biocatalytic Production of Terpenes.
abstract: The patchoulol synthase (PTS) is a multi-product sesquiterpene synthases which is the central enzyme for biosynthesis of patchouli essential oil in the patchouli plant. Sesquiterpene synthases catalyse the formation of various complex carbon backbones difficult to approach by organic synthesis. Here, we report the characterisation of a recombinant patchoulol synthase complementary DNA (cDNA) variant (PTS var. 1), exhibiting significant amino acid exchanges compared to the native PTS. The product spectrum using the natural substrate E,E-farnesyl diphosphate (FDP) as well as terpenoid products resulting from conversions employing alternative substrates was analysed by GC-MS. In respect to a potential use as a biocatalyst, important enzymatic parameters such as the optimal reaction conditions, kinetic behaviour and the product selectivity were studied as well. Adjusting the reaction conditions, an increased patchoulol ratio in the recombinant essential oil was achieved. Nevertheless, the ratio remained lower than in plant-derived patchouli oil. As alternative substrates, several prenyl diposphates were accepted and converted in numerous compounds by the PTS var. 1, revealing its great biocatalytic potential.
authors.affiliation: , Hannover, Germany.
authors.forename: Thore
authors.initials: T
authors.lastname: Frister
authors.forename: Steffen
authors.initials: S
authors.lastname: Hartwig
authors.forename: Semra
authors.initials: S
authors.lastname: Alemdar
authors.forename: Katharina
authors.initials: K
authors.lastname: Schnatz
authors.forename: Laura
authors.initials: L
authors.lastname: Thöns
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheper
authors.forename: Sascha
authors.initials: S
authors.lastname: Beutel
mesh_terms: D055162:Biocatalysis; D008401:Gas Chromatography-Mass Spectrometry; D006863:Hydrogen-Ion Concentration; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D019686:Lamiaceae / Q000201:enzymology; D009154:Mutation / Q000235:genetics*; D009822:Oils, Volatile / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009822:Oils, Volatile; C483659:Patchouliol; D011994:Recombinant Proteins; D013729:Terpenes; D007535:Isomerases; C065995:patchoulol synthase