Effects of addition of elicitors on rimocidin biosynthesis in Streptomyces rimosus M527
Effects of addition of elicitors on rimocidin biosynthesis in Streptomyces rimosus M527
复制标题
添加诱导子对 Rimosus M527 链霉菌中利莫西丁生物合成的影响
DOI：
10.1007/s00253-020-10565-4
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发表时间：
2020-03-27
影响因子：
5
通讯作者：
Yu, Xiaoping
中科院分区：
文献类型：
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作者：
Song, Zhangqing;Ma, Zheng;Yu, Xiaoping
由 Streptomyces rimosus M527 产生的多烯大环内酯利莫西丁对多种植物真菌病原体非常有效，但产量较低。诱导是刺激生物活性次级代谢产物产量的有效方法。在这项研究中，大肠杆菌 JM109、枯草芽孢杆菌 WB600、酿酒酵母和尖孢镰刀菌培养液的生物量和滤液。 sp。黄瓜被用作诱导子以促进 S. rimosus M527 中利莫西丁的产生。添加酿酒酵母（A3）和尖孢镰刀菌的培养液和生物量。 sp。与5升发酵罐正常条件下的产量相比，黄瓜(B4)导致利莫西丁产量分别增加51.2%和68.3%。此外，定量RT-PCR分析显示，A3或B4诱导实验组中与雷莫西丁生物合成相关的基因簇中的10个基因（rimA至rimK）的转录均高于对照组。使用 β-葡萄糖醛酸酶 (GUS) 报告系统、GUS 酶活性测定和蛋白质印迹分析，我们发现 A3 或 B4 的诱导增加了 S. rimosus M527 中的蛋白质合成。这些结果表明，添加引发子是提高利莫西丁产量的有效方法。要点 • 证明了提高利莫西丁链霉菌 M527 中利莫西丁产量的有效策略。 • 利莫西丁的过量产生是结构基因表达升高以及蛋白质合成增加的结果。
The polyene macrolide rimocidin, produced by Streptomyces rimosus M527, is highly effective against a broad range of fungal plant pathogens, but at low yields. Elicitation is an effective method of stimulating the yield of bioactive secondary metabolites. In this study, the biomass and filtrate of a culture broth of Escherichia coli JM109, Bacillus subtilis WB600, Saccharomyces cerevisiae, and Fusarium oxysporum f. sp. cucumerinum were employed as elicitors to promote rimocidin production in S. rimosus M527. Adding culture broth and biomass of S. cerevisiae (A3) and F. oxysporum f. sp. cucumerinum (B4) resulted in an increase of rimocidin production by 51.2% and 68.3% respectively compared with the production under normal conditions in 5-l fermentor. In addition, quantitative RT-PCR analysis revealed that the transcriptions of ten genes (rimA to rimK) located in the gene cluster involved in rimocidin biosynthesis in A3 or B4 elicitation experimental group were all higher than those of a control group. Using a β-glucuronidase (GUS) reporter system, GUS enzyme activity assay, and Western blot analysis, we discovered that elicitation of A3 or B4 increased protein synthesis in S. rimosus M527. These results demonstrate that the addition of elicitors is a useful approach to improve rimocidin production.Key Points • An effective strategy for enhancing rimocidin production in S. rimosus M527 is demonstrated. • Overproduction of rimocidin is a result of higher expressed structural genes followed by an increase in protein synthesis.