title: Cloning and functional characterization of two monoterpene acetyltransferases from glandular trichomes of L. x intermedia.
abstract: Two alcohol acetyltransferases, LiAAT-3 and LiAAT-4, from L. x intermedia were cloned, expressed in bacteria, and functionally characterized. Two monoterpene acetyltransferase cDNA clones (LiAAT-3 and LiAAT-4) were isolated from L. x intermedia glandular trichomes, expressed in bacteria to produce, and functionally characterize the encoded proteins in vitro. The recombinant LiAAT-3 and LiAAT-4 proteins had molecular weights of ca. 47 and 49 kDa, respectively, as evidenced by SDS-PAGE. The K m (mM) values for the recombinant LiAAT-3 and LiAAT-4 were 1.046 and 0.354 for lavandulol, 1.31 and 0.279 for geraniol, and 0.87 and 0.113 for nerol, respectively. The V max (pkat/mg) values for LiAAT-3 and LiAAT-4 were 92.13 and 105.1 for lavandulol, 81.07 and 52.17 for geraniol, and 15.02 and 15.8 for nerol, correspondingly. Catalytic efficiencies (mM(-1) min(-1)) for LiAAT-3 and LiAAT-4 were 0.27 and 0.85 for lavandulol, 0.19 and 0.54 for geraniol, and 0.052 and 0.4 for nerol, respectively. These kinetic properties are in the range of those reported for other plant acetyltransferases, and indicate that LiAAT-4 has a better catalytic efficiency than LiAAT-3, with lavandulol serving as the preferred substrate for both enzymes. Transcripts for both genes were abundant in L. angustifolia and L. x intermedia flowers, where monoterpene acetates are produced, and were undetectable (or present in trace quantities) in L. latifolia flowers, which do not accumulate significant amounts of these metabolites.
authors.affiliation: Department of Biology, University of British Columbia, Okanagan Campus, 1177 Research Road, Kelowna, BC, V1V 1V7, Canada.
authors.forename: Lukman S
authors.initials: LS
authors.lastname: Sarker
authors.forename: Soheil S
authors.initials: SS
authors.lastname: Mahmoud
mesh_terms: D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D027523:Lavandula / Q000201:enzymology* / Q000235:genetics; D039821:Monoterpenes / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D064201:Trichomes / Q000201:enzymology* / Q000235:genetics
chemical_list: D039821:Monoterpenes; D010940:Plant Proteins; D000123:Acetyltransferases