Research on the substrate specificity of plant P450 and its regulatory function in plant metabolism.

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The plant P450s are the group of oxidative enzymes in biosynthesis of the small molecule biologically active compounds in plant metabolism, and have important roles on the induction of those biological activities. Although the biologically active compounds have a diversity of chemical structures, the oxygen functionality in the structures is almost essential for the activity. Thus, the mechanisms to introduce those oxygens in the compounds are of quite interest to understand plant metabolism. For example, the phytoalexin, well-known antimicrobial compounds produced in the stressed plants, have a number of skeletons such as spirovetivanes and eremophilanes in Solanaceae species. The hydrocarbon skeleton itself does not show any remarkable activities, but once oxidized by P450 enzymes, the compounds become active. Deuterium labeled farnesol was synthesized and fed into green pepper seedlings, and the incorporation of the labeled precursor into the metabolites was confirmed. Furthermore, germacrene and spirovetivane hydrocarbon skeletons were synthesized in the way that deuterium or carbon-13 isotopes would be easily introduced. This research established the tools for metabolic experiments to explore the roles of these P450 enzymes.