Reconstitution of Kinamycin Biosynthesis within the Heterologous Host Streptomyces albus J1074
Reconstitution of Kinamycin Biosynthesis within the Heterologous Host Streptomyces albus J1074
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DOI：
10.1021/acs.jnatprod.7b00652
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发表时间：
2018-01-01
影响因子：
5.1
通讯作者：
Jiang, Ming
中科院分区：
文献类型：
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作者：
Liu, Xiangyang;Liu, Dongxu;Jiang, Ming
重氮芴类化合物如卡那霉素和洛麦维他星具有独特的分子结构和引人注目的药物生物活性。然而，对这个天然产物家族的生物合成细节的完整理解尚未完全阐明。此外，缺乏遗传和技术上可行的生产宿主限制了这些化合物的全部药用潜力。在这里，我们报告的捕获完整的卡那霉素基因簇从链霉菌galtieri Sgt26细菌人工染色体克隆，证实了成功生产的卡那霉素在异源宿主链霉菌白J1074。序列分析和一系列的基因缺失实验揭示了该簇的边界，它跨越75 kb的DNA。为了探索生物合成的最后一步，将kinamcyin F乙酰化为kinamycin D，基因敲除和互补实验确定了参与最终乙酰化转化的单个基因产物。本研究为分离自沙门氏菌的卡那霉素基因簇提供了完整的遗传信息。galtieri Sgt26的生物合成，并建立了异源生物合成作为生产平台，用于化合物形成和利用的持续机制评估。
Diazofluorene compounds such as kinamycin and lomaiviticin feature unique molecular structures and compelling medicinal bioactivities. However, a complete understanding of the biosynthetic details for this family of natural products has yet to be fully elucidated. In addition, a lack of genetically and technically amenable production hosts has limited access to the full medicinal potential of these compounds. Here, we report the capture of the complete kinamycin gene cluster from Streptomyces galtieri Sgt26 by bacterial artificial chromosome cloning, confirmed by successful production of kinamycin in the heterologous host Streptomyces albus J1074. Sequence analysis and a series of gene deletion experiments revealed the boundary of the cluster, which spans 75 kb DNA. To probe the last step in biosynthesis, acetylation of kinamcyin F to kinamycin D, gene knockout, and complementation experiments identified a single gene product involved with final acetylation conversions. This study provides full genetic information for the kinamycin gene cluster from S. galtieri Sgt26 and establishes heterologous biosynthesis as a production platform for continued mechanistic assessment of compound formation and utilization.