Sequential improvement of rimocidin production in Streptomyces rimosus M527 by introduction of cumulative drug-resistance mutations
Sequential improvement of rimocidin production in Streptomyces rimosus M527 by introduction of cumulative drug-resistance mutations
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通过引入累积耐药突变连续提高 Rimosus M527 中利莫西丁的产量
DOI：
10.1007/s10295-019-02146-w
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发表时间：
2019-01
影响因子：
3.4
通讯作者：
Yu X
中科院分区：
文献类型：
--
作者：
Zhao Y;Song Z;Ma Z;Bechthold A;Yu X
Rimocidin 是一种多烯大环内酯，对多种植物病原真菌具有很强的抑制活性。本研究采用发酵优化和核糖体工程技术来提高Streptomyces rimosus M527中利莫西丁的产量。发酵优化后，S. rimosus M527 中的利莫西丁产量在摇瓶实验中从 0.11 ± 0.01 增加到 0.23 ± 0.02 g/L，并使用 5 L 发酵罐达到 0.41 ± 0.05 g/L。发酵优化后，通过用不同浓度的庆大霉素 (Gen) 或利福霉素处理菌株，产生 S. rimosus M527 突变体。一种名为 S. rimosus M527-G37 的 Genr 突变体和一种名为 S. rimosus M527-R5 的 Rifr 突变体显示出利莫西丁产量增加。使用S. rimosus M527-G37和S. rimosus M527-R5选择双抗性（Genr和Rifr）突变体，并随后进行测试。源自 M527-G37 的一种突变体 S. rimosus M527-GR7 在利莫西丁产量方面取得了最大的累积改进。在5L发酵罐中，S. rimosus M527-GR7实现的最大瑞莫西丁产量分别比S. rimosus M527-G37和野生型菌株S. rimosus M527高出25.36%和62.89%。此外，突变株S. rimosus M527-G37和S. rimosus M527-GR7中与雷莫西丁生物合成相关的基因簇中的10个基因（rimA sr至rimK sr）的转录水平均不同程度高于亲本菌株M527。此外，在S. rimosus M527中表达单一雷莫西丁生物合成基因后，一些重组体显示雷莫西丁产量增加。 rimE 的表达导致最高产量。
Rimocidin is a polyene macrolide that exhibits a strong inhibitory activity against a broad range of plant-pathogenic fungi. In this study, fermentation optimization and ribosome engineering technology were employed to enhance rimocidin production in Streptomyces rimosus M527. After the optimization of fermentation, rimocidin production in S. rimosus M527 increased from 0.11 ± 0.01 to 0.23 ± 0.02 g/L during shake-flask experiments and reached 0.41 ± 0.05 g/L using 5-L fermentor. Fermentation optimization was followed by the generation of mutants of S. rimosus M527 through treatment of the strain with different concentrations of gentamycin (Gen) or rifamycin. One Genr mutant named S. rimosus M527-G37 and one Rifr mutant named S. rimosus M527-R5 showed increased rimocidin production. Double-resistant (Genr and Rifr) mutants were selected using S. rimosus M527-G37 and S. rimosus M527-R5, and subsequently tested. One mutant, S. rimosus M527-GR7, which was derived from M527-G37, achieved the greatest cumulative improvement in rimocidin production. In the 5-L fermentor, the maximum rimocidin production achieved by S. rimosus M527-GR7 was 25.36% and 62.89% greater than those achieved by S. rimosus M527-G37 and the wild-type strain S. rimosus M527, respectively. Moreover, in the mutants S. rimosus M527-G37 and S. rimosus M527-GR7 the transcriptional levels of ten genes (rimA sr to rimK sr) located in the gene cluster involved in rimocidin biosynthesis were all higher than those in the parental strain M527 to varying degrees. In addition, after expression of the single rimocidin biosynthetic genes in S. rimosus M527 a few recombinants showed an increase in rimocidin production. Expression of rimE led to the highest production.
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影响因子：
3.7
作者：
Escudero L;Al-Refai M;Nieto C;Laatsch H;Malpartida F;Seco EM
通讯作者：
Seco EM
影响因子：
6.7
作者：
Ren, Jun;Cui, Yuqiong;Xia, Huanzhang
通讯作者：
Xia, Huanzhang
影响因子：
5
作者：
Liu, Zhuo;Zhao, Xinqing;Bai, Fengwu
通讯作者：
Bai, Fengwu
影响因子：
3.1
作者：
Lu, Dandan;Ma, Zheng;Yu, Xiaoping
通讯作者：
Yu, Xiaoping
影响因子：
4.4
作者：
Tamehiro, N;Hosaka, T;Ochi, K
通讯作者：
Ochi, K