Targeted activation of silent natural product biosynthesis pathways by reporter-guided mutant selection
Targeted activation of silent natural product biosynthesis pathways by reporter-guided mutant selection
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DOI：
10.1016/j.ymben.2014.12.006
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发表时间：
2015-03-01
影响因子：
8.4
通讯作者：
Yang, Kegian
中科院分区：
文献类型：
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作者：
Guo, Fang;Xiang, Sihai;Yang, Kegian
不断增加的基因组测序数据揭示了许多潜在的途径，这些途径可能编码具有有趣生物活性的新的次级代谢产物。然而，这种隐藏的潜力的利用受到了许多这些基因簇在实验室条件下保持沉默（或表达差）的观察的阻碍。在这里，我们提出了一个有效的和广泛适用的方法，有针对性地激活沉默基因簇在本地生产者的突变体选择（RGMS）。该策略具有基因组规模随机突变产生遗传多样性的优势，以及报告基因指导的选择系统用于识别所需的靶点激活突变体的优势。它首先在委内瑞拉链霉菌ISP 5230中重新激活杰多霉素生物合成中得到验证，实现了高效率的激活。同样的策略应用于链霉菌PGA64中迄今未激活的pga基因簇，导致两个新的蒽醌氨基糖苷类抗生素，gaudimycin D和E的鉴定。(C)2015年国际代谢工程学会。爱思唯尔公司出版All rights reserved.
The continuously increasing genome sequencing data has revealed numerous cryptic pathways, which might encode novel secondary metabolites with interesting biological activities. However, utilization of this hidden potential has been hindered by the observation that many of these gene clusters remain silent (or poorly expressed) under laboratory conditions. Here we present reporter-guided mutant selection (RGMS) as an effective and widely applicable method for targeted activation of silent gene clusters in the native producers. The strategy Lakes advantage of genome-scale random mutagenesis for generation of genetic diversity and a reporter-guided selection system for the identification of the desired target-activated mutants. It was first validated in the re-activation of jadomycin biosynthesis in Streptomyces venezuelae ISP5230, where high efficiency of activation was achieved. The same strategy was then applied to a hitherto unactivable pga gene cluster in Streptomyces sp. PGA64 leading to the identification of two new anthraquinone aminoglycosides, gaudimycin D and E. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.