Functional Genomics Approach to Exploiting Molecular Diversity

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(1) By utilizing genomic information, twenty four new triterpene synthase genes were cloned from higher plants, ferns and filamentous fungi. Their enzyme functions were identified by heterologous expression in yeast.(2) As the products of these triterpene synthases, three new triterpenes with novel skeletons were identified.(3) Lanosterol synthase genes were discovered for the first time from the plant kingdom and the presence of biosynthetic pathway of phytosterols via lanosterol was demonstrated.(4) By reacting triterpene synthases with non-physiological substrates and synthetic substrate analogues, five new polyprenoids were obtained(5) Each three novel oxidases and sugartransferases were cloned and their functions identified as the triterpene modifying enzymes.(6) Eleven new iterative polyketide synthase (i-PKS) genes were cloned from filamentous fungi. Their enzyme functions were identified by expression in a heterologous fungal strain.(7) Six new polyketide compounds with unique structures were identified as the products of these i-PKSs.(8) Four Type-III i-PKS genes were identified in the genome of Asp. oryzae and one of them was demonstrated to encode an orsellinic acid synthase.(9) As the higher structure for Type-I i-PKSs, a novel homo-tetramer model was proposed.(10) As the novel polyketide modifying enzymes, a chain-shortening enzyme, a dehydratase, an oxidase with cycloaddition activity and two prenyltrasferase were cloned and their enzyme functions identified.