Characterization of the Polyoxin Biosynthetic Gene Cluster from Streptomyces cacaoi and Engineered Production of Polyoxin H
Characterization of the Polyoxin Biosynthetic Gene Cluster from Streptomyces cacaoi and Engineered Production of Polyoxin H
复制标题
可可链霉菌多抗素生物合成基因簇的表征以及多抗素 H 的工程化生产。
DOI：
10.1074/jbc.m807534200
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发表时间：
2009-04-17
影响因子：
4.8
通讯作者：
Deng, Zixin
中科院分区：
文献类型：
--
作者：
Chen, Wenqing;Huang, Tingting;Deng, Zixin
从可可链霉菌（Streptomyces cacaoi）中克隆了一个多抗霉素生物合成所必需的基因簇（pol）。一个46，066-bp的区域被测序，39个推定的开放阅读框中的20个被定义为多抗霉素生物合成所必需的，如其在异源宿主变铅青链霉菌TK 24中的产生所证明的。波罗和PolA在多抗霉素合成中的作用通过体内实验证明，并且它们的功能分别通过体外实验明确地表征为O-氨基甲酰基转移酶和UMP-烯醇式乙酰转移酶，这使得能够产生与先前提出的略有不同的经修饰的化合物。这些研究为阐明多抗霉素生物合成的分子机制提供了坚实的基础，并为利用编码核苷类抗生素不同途径的基因进行组合生物合成奠定了基础。
A gene cluster (pol) essential for the biosynthesis of polyoxin, a nucleoside antibiotic widely used for the control of phytopathogenic fungi, was cloned from Streptomyces cacaoi. A 46,066-bp region was sequenced, and 20 of 39 of the putative open reading frames were defined as necessary for polyoxin biosynthesis as evidenced by its production in a heterologous host, Streptomyces lividans TK24. The role of PolO and PolA in polyoxin synthesis was demonstrated by in vivo experiments, and their functions were unambiguously characterized as O-carbamoyltransferase and UMP-enolpyruvyltransferase, respectively, by in vitro experiments, which enabled the production of a modified compound differing slightly from that proposed earlier. These studies should provide a solid foundation for the elucidation of the molecular mechanisms for polyoxin biosynthesis, and set the stage for combinatorial biosynthesis using genes encoding different pathways for nucleoside antibiotics.