TAILIEUCHUNG - Báo cáo khoa học: A fatty-acid-metabolizing enzyme fromArabidopsis

A fatty-acid-metabolizing enzyme fromArabidopsis thaliana, CYP94C1, belonging to the cytochrome P450 family was cloned and characterized. CYP94C1 was heterologously expressed in aSaccharomyces cerevisiae strain (WAT11) engineered for P450 expression. When recombinant yeast microsomes were incubated with lauric acid (C12:0) for 15 min, one major metabolite was formed. | ễFEBS Journal Characterization of a methyl jasmonate and wounding-responsive cytochrome P450 of Arabidopsis thaliana catalyzing dicarboxylic fatty acid formation in vitro Sylvie Kandel1 Vincent Sauveplane1 Vincent Compagnon1 Rochus Franke2 Yves Millet1 Lukas Schreiber2 Daniele Werck-Reichhart1 and Franck Pinot1 1 Institut de Biologie Moleculaire des Plantes CNRS-Universite Louis Pasteur Strasbourg France 2 Institut fur Zellulare und Molekulare Botanik Universitat Bonn Germany Correspondence F. Pinot IBMP-CNRS UPR 2357 Institut de Botanique 28 Rue Goethe F-67083 Strasbourg Cedex France Fax 33 3 90 24 19 21 Tel 33 3 90 24 18 37 E-mail Received 4 July 2007 revised 31 July 2007 accepted 6 August 2007 doi A fatty-acid-metabolizing enzyme from Arabidopsis thaliana CYP94C1 belonging to the cytochrome P450 family was cloned and characterized. CYP94C1 was heterologously expressed in a Saccharomyces cerevisiae strain WAT11 engineered for P450 expression. When recombinant yeast microsomes were incubated with lauric acid C12 0 for 15 min one major metabolite was formed. The product was purified and identified by GC MS as 12-hydroxylauric acid. Longer incubation 40 min led to the formation of an additional metabolite identified by GC MS as dodecadioic acid. This diacid was also produced by incubation with 12-hydroxylauric acid. These compounds were not produced by incubating microsomes from yeast transformed with a void plasmid demonstrating the involvement of CYP94C1. This new enzyme also metabolized fatty acids of varying aliphatic chain lengths C12 to C18 and in-chain modifications for example degree of unsaturation or the presence of an epoxide as an additional polar functional group. Transcription of the gene encoding CYP94C1 is enhanced by stress treatment with the hormone methyl jasmonate and wounding. Treatment with methyl jasmonate also induced lauric acid metabolism in microsomes prepared from .

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