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首页> 外文期刊>The Journal of Antibiotics: An International Journal >Identification and heterologous expression of the actinoallolide biosynthetic gene cluster
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Identification and heterologous expression of the actinoallolide biosynthetic gene cluster

机译：Actinoallolide生物合成基因簇的鉴定和异源表达

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摘要

Actinoallolides are anti-trypanosomal macrolides isolated from the secondary metabolites of two endophytic actinomycete strains, Actinoallomurus fulvus MK10-036 and K09-0307. A putative actinoallolide biosynthetic gene cluster was predicted from the genome sequence of the strain K09-0307. The gene cluster spans a contiguous 53 kb DNA region that comprises seven genes encoding three PKSs (aalA1, aalA2, and aalA3), cytochrome P450 (aalB), acyl-CoA dehydrogenase (aalC), crotonyl-CoA reductase (aalD), and TetR family regulator (aalR). The entire gene cluster was cloned into a plasmid pYIK1 by assembling DNA fragments, which were obtained from two cosmids containing left and right parts of the gene cluster. Following the introduction of an ermE* promoter at 100bp upstream from the start codon of aalAl , the gene cluster was introduced into Streptomyces coelicolor M1152. Subsequent LC-MS analysis revealed production of actinoallolide A in the culture broth. Thus, the actinoallolide biosynthetic gene cluster was identified by heterologous expression in Streptomyces.
机译：ActinoPlolides是抗锥体蛋白酶大环内酯分离出的两种内生菌菌株的二次代谢物，ACTINALOMURUS FULVUS MK10-036和K09-0307。从菌株K09-0307的基因组序列预测了备向的Actinoallide生物合成基因簇。该基因簇跨越连续的53kb DNA区域，其包含编码三个PKS的七种基因（AALA1，AALA2和AALA3），细胞色素P450（AALB），酰基-COA脱氢酶（AALC），克拉替哥伦比 - COA还原酶（AALD）和四缩略家庭调节器（AALR）。通过组装DNA片段，将整个基因簇克隆到质粒pIIK1中，该DNA片段从含有基因簇的左右部分的含有左右部分的两个粘膜获得。在引入ERME *启动子在Aalal起始密码子上游的100bp上游后，将基因簇引入链霉菌菌溶胶M1152中。随后的LC-MS分析显示培养液中的Actinoallolide A的产生。因此，通过链霉菌中的异源表达鉴定ActinoAllolide生物合成基因簇。

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    Inahashi Yuki; Shiraishi Taro; Take Akira; Matsumoto Atsuko; Takahashi Yoko; Omura Satoshi; Kuzuyama Tomohisa; Nakashima Takuji;

  • 作者单位

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

    Univ Tokyo Biotechnol Res Ctr Lab Cell Biotechnol Bunkyo Ku 1-1-1 Yayoi Tokyo 1138657 Japan;

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

    Univ Tokyo Biotechnol Res Ctr Lab Cell Biotechnol Bunkyo Ku 1-1-1 Yayoi Tokyo 1138657 Japan;

    Kitasato Univ Kitasato Inst Life Sci Minato Ku 5-9-1 Shirokane Tokyo 1088641 Japan;

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  • 正文语种 eng
  • 中图分类 药学;
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