Molecular genetic analysis of enediyne antibiotic biosynthesis genes

烯二炔类抗生素生物合成基因的分子遗传学分析

• 批准号: 06807163
• 负责人: FUJII Isao
• 金额: $ 0.38万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06807163/
• 关键词: Enediyne antibiti; Dynemicin; Micromonospora chersina; biosynthesis gene; cloning; Micromonospora chersina

Dynemicin is an enediyne antitumor antibiotic with unique hybrid structure of anthraquinone and enediyne structures. To elucidate its biosynthesis mechanism, cloning of type-II polyketide biosynthesis gene was carried out from Micromonospora chersina, a dynemicin producing actinomycete. By screening with actI probe, ca.20kb fragment was cloned from M.chersina genomic DNA library. The nucleotide sequence of 14.3 kb region was determined and open reading frame (ORF) analysis was carried out. In this region, 12 ORFs were found. Data base homology search indicated that ORF2,4,5,6 and 8 encode cyclase, beta-ketoacyl synthase, chain length factor, CoA-ligase and acyltransferase, respectively. Thus, the cloned gene consisted of typical type-II polyketide synthase gene cluster with unique ORFs such as oxygenases and addtitional beta-ketoacyl synthase. Further analysis is on the way to identify whether this cloned gene cluster is involved in dynemicin biosynthesis in M.chersina.

动力霉素是一种烯二炔类抗肿瘤抗生素，具有蒽醌和烯二炔结构的独特杂化结构。为了阐明其生物合成机制，从产动力素放线菌樱桃小单孢菌中克隆了II型聚酮生物合成基因。通过ActI探针筛选，从切氏木霉基因组DNA文库中克隆出约20kb的片段。测定了14.3kb的核苷酸序列，并进行了开放阅读框架(ORF)分析。在该地区共发现12个ORF。数据库同源性搜索表明，ORF2、4、5、6和8分别编码环化酶、β-酮酰基合成酶、链长因子、辅酶A连接酶和酰基转移酶。因此，克隆的基因由典型的II型聚酮合成酶基因簇组成，具有独特的ORF，如加氧酶和附加的β-酮酰基合成酶。进一步的分析正在进行中，以确定这个克隆的基因簇是否参与了切氏木霉动力米星的生物合成。

项目成果

C. R. Hutchinson: "Polyketide syuthase gene manupilation: A Strueture-function approach in engineeving novel antibiotics" Annu. Rev. Microbiol.49. 201-238 (1995)

C. R. Hutchinson：“聚酮化合物合成酶基因操作：设计新型抗生素的结构功能方法”Annu。

C.R.Hutchinson: "Polyketidesyuthase gene manupilation:A Structure-function approach in engineering novel antibiotics" Annu.Rev.Microbiol. 49. 201-238 (1995)

C.R.Hutchinson：“聚酮化合物合成酶基因操作：新型抗生素工程中的结构功能方法”Annu.Rev.Microbiol。

C.R.Hutchinson and Isao Fujii: "Polyketide synthase gene manipulation : A Structure-Function Approach in Engineering Novel Antibiotics" Annu. Rev. Microbiol.49. 201-238 (1995)

C.R.Hutchinson 和 Isao Fujii：“聚酮合酶基因操作：工程新型抗生素的结构功能方法”Annu。