Genetic Engineering for Bio-active Natural Produtcs

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This study has been focussed on the biosynthesis of polyketide and terpenoids in the both countries. Plant plyketide synthases, chalcone and stilbene synthases were mainly investigated, in particular on their reaction mechanism in collaboration between Toyama Medical & Pharmaceutical University and the University of Tokyo. Heterologous expression system to produce a large amount of proteins were introduced from the University of Tokyo and the expressed proteins have been subjected in the trials of recrystallization. We have been investigated to obtain crystals with true substrates or reaction products. In the biosynthesis of isoprenoids, tetra- and bi-cyclic diterpenes of Scoparia dulcis have been shown to be biosynthesized via mevalonate independent pathway and geranylgeranyl diphophate synthase was cloned from this plant. It contained signal peptide targeting chloroplats supporting the diterpenes are biosynthesized in chloroplasts. cDNAs related to the biosynthesis of carotenoid were cloned from carrot and they also possess signal peptides. Characterization of the key reactions in tri-terpenoids biosynthesis was studied in the university of Tokyo. The group of USA mainly provide us technology transfer of gene cloning in particular polyketide biosynthesis enzymes. They contributeed one chapter in Comprehensive Natural Products Chemistry which was edited by the head investigator.