Functional Analysis of Biosynthetic Enzymes for Natural Products

天然产物生物合成酶的功能分析

基本信息

批准号：
12045213
负责人：
EBIZUKA Yutaka
金额：
$ 11.26万
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 on Priority Areas
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Diverse structures of natural products are elaborated by biosynthetic enzymes, and those enzymes which work at the most upstream of biosynthetic pathways are thus regarded as the primary origin of structural diversty exhibited by natural products.Our attentions have been focused on these biosynthetic enzymes for future engineering of rationally designed biosynthetic machineries for production of new chemicals. In this study, gene cloning and functional analysis of some of these enzymes have been attempted.Continuous attempts for cloning of new polyketide synthases from filamentous fungi yielded several new clones.Enzyme functions of these clones were analyzed by heterologous expression in fungi and/or yeast. Construction of chimeric enzymes and site-directed mutants of these clones identified a new functional domain for aromatic pokyketide production.Over thirty triterpene synthase cDNAs have been cloned from various triterpene producing plants by homology based PCR methods.Their enzyme functions have been identified by heterologous expression in yeast.Functional analysis of the eight out of thirteen Arabidopsis thaliana triterpene synthase clones was attempted.Only two clones are product specific synthases for known triterpenes, while the others are multifunctional clones.In this four-year project, functional analysis of biosynthetic enzymes for polyketides and triterpenoids was extensively conducted.The results obtained in this study clearly show that functional genomics approach would provide a new methodology for the search of new natural products.Furthermore, by employing recombinant technology of these biosynthetic enzymes, development of practical production system for useful chemicals and construction of artificial chemical library comprising of "un-natural natural products" with diverse structures wouldbe expected.

天然产物的多种结构由生物合成酶进行了详细说明，因此，在生物合成途径上最上游的那些酶被视为自然产物所表现出的结构多样性的主要起源。我们的注意力集中在这些生物合成酶的生物合成酶的生物工具上，以实现生物合作设计的生物合作式机械工具。在这项研究中，已经尝试了其中一些酶的基因克隆和功能分析。从丝状真菌中克隆新的聚酮化合物合酶的连续尝试产生了几个新的克隆。这些克隆的酶函数在真菌和/或酵母中通过异型表达进行了分析。 Construction of chimeric enzymes and site-directed mutants of these clones identified a new functional domain for aromatic pokyketide production.Over thirty triterpene synthase cDNAs have been cloned from various triterpene producing plants by homology based PCR methods.Their enzyme functions have been identified by heterologous expression in yeast.Functional analysis of the eight out of thirteen Arabidopsis thaliana尝试尝试三萜合酶克隆。仅两个克隆是已知三萜的产品特定的合成酶，而其他克隆是多功能的克隆。在这个为期四年的项目中，对多酮化合物和三培养子的生物合成酶的功能分析进行了广泛的搜索。在此研究中，搜索的结果清楚地表明了一种新的方法，该方法是新的，这些方法是新的，这些方法是新的方法，即新的方法，该方法是新的方法，该方法是基因组学的，该方法是新的方法。采用这些生物合成酶的重组技术，开发实用生产系统，用于有用的化学品和人工化学图书馆的构建，其中包括具有多种结构的“非自然天然产物”的人工化学库。

项目成果

期刊论文数量（64）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Miho Izumikawa, Michio Murata, Kazuo Tachibana, Yutaka Ebizuka, Isao Fujii.: "Cloning of Modular Type I Polyketide Synthase Genes from Salinomycin Producing Strain of Streptomyces albus."Bioorg. Med. Chem.. 11. 3401-3405 (2003)

Miho Izumikawa、Michio Murata、Kazuo Tachibana、Yutaka Ebizuka、Isao Fujii.：“从产盐霉素的白色链霉菌菌株中克隆模块化 I 型聚酮合酶基因。”Bioorg。

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I.Fujii: "Enzymatic Synthesis of 1,3,6,8-Tetrahydroxynaphthalene solely from Maloxyl Coenzyme A by a Fungal Iterative Type I polykatide synthase PKSI."Biochemistry. 39. 8853-8858 (2000)

I.Fujii：“通过真菌迭代 I 型聚肽合酶 PKSI 仅从丙二酰辅酶 A 酶促合成 1,3,6,8-四羟基萘。”生物化学。

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Yutaka Ebizuka, Yuji Katsube, Takehiko Tsutsumi, Tetsuo Kushiro, Masaaki Shibuya: "Functional Genomics Approach to the Study of Triterpene Biosynthesis."Pure Appl. Chem.. 75. 369-374 (2003)

Yutaka Ebizuka、Yuji Katsube、Takehiko Tsutsumi、Tetsuro Kushiro、Masaaki Shibuya：“三萜生物合成研究的功能基因组学方法”。

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Akira Watanabe, Yutaka Ebizuka.: "A Novel Hexaketide Naphthalene Synthesized by a Chimeric Polyketide Synthase Composed of Fungal Pentaketide and Heptaketide Synthases."Tetrahedron Lett.. 43. 369-374 (2002)

Akira Watanabe、Yutaka Ebizuka.：“由真菌五肽和七肽合成酶组成的嵌合聚酮化合物合成酶合成的新型六酮化合物萘。”Tetrahedron Lett.. 43. 369-374 (2002)

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Hiroaki Hayashi, Pengu Huang, Keiichiro Inoue, Noboru Hiraoka, Yasumasa Ikeshiro, Kazufumi Yazaki, Shige Tanaka, Tetsuo Kushiro, Masaaki Shibuya, Yutaka Ebizuka.: "Molecular Cloning and Characterization of Isomultiflorenol Synthase, a New Triterpene Synth

Hiroaki Hayashi、Pengu Huang、Keiichiro Inoue、Noboru Hiraoka、Yasumasa Ikeshiro、Kazufumi Yazaki、Shige Tanaka、Tetsuro Kushiro、Masaaki Shibuya、Yutaka Ebizuka。：“异多花烯醇合成酶（一种新的三萜合成物）的分子克隆和表征