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Functional Genomics Approach to Exploiting Molecular Diversity

利用分子多样性的功能基因组学方法

基本信息

批准号：
15101007
负责人：
EBIZUKA Yutaka
金额：
$ 60.9万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (S)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2007
项目状态：
已结题

项目摘要

(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.

(1)利用基因组信息，从高等植物、蕨类植物和丝状真菌中克隆了24个新的三萜合酶基因。通过在酵母中异源表达鉴定了它们的酶功能。(2)作为这些三萜合酶的产物，鉴定了三种具有新颖骨架的新三萜。(3)首次从植物界发现了羊毛甾醇合酶基因，证明了通过羊毛甾醇生物合成植物甾醇途径的存在。(4)通过三萜合酶与 (5)从丝状真菌中克隆了11个新的迭代聚酮合酶(i-PKS)基因，并鉴定了它们的功能为三萜修饰酶。(5)克隆了11个新的迭代聚酮合酶(i-PKS)基因。通过在异源真菌菌株中的表达鉴定了它们的酶功能。(7)鉴定了六种结构独特的新聚酮化合物是这些i-PKS的产物。(8)在Asp基因组中鉴定了四个III型i-PKS基因。 (9)作为I型i-PKS的高级结构，提出了一种新型同源四聚体模型。(10)作为新型聚酮修饰酶，克隆了一种短链酶、一种脱水酶、一种具有环加成活性的氧化酶和两种异戊烯基转移酶，并克隆了它们的酶确定的功能。