Improved milbemycin production by engineering two Cytochromes P450 in Streptomyces bingchenggensis

文献类型： 外文期刊

第一作者： Wang, Haiyan

作者： Wang, Haiyan;Cheng, Xu;Li, Shanshan;Zhang, Yanyan;Xiang, Wensheng;Wang, Haiyan;Liu, Yuqing;Wang, Xiangjing

作者机构：

关键词： Milbemycins; Post-PKS step; CYPs; Cytochromes P450; Streptomyces bingchenggenis

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期：

页码：

收录情况： SCI

摘要： Milbemycins and their semisynthetic derivatives are recognized as effective and eco-friendly pesticides, whereas the high price limits their widespread applications in agriculture. One of the pivotal questions is the accumulation of milbemycin-like by-products, which not only reduces the yield of the target products milbemycin A3/A4, but also brings difficulty to the purification. With other analogous by-products abolished, alpha 9/alpha 10 and beta-family milbemycins remain to be eliminated. Herein, we solved these issues by engineering of post-modification steps. First, Cyp41, a CYP268 family cytochrome P450, was identified to participate in alpha 9/alpha 10 biosynthesis. By deleting cyp41, milbemycin alpha 9/alpha 10 was eliminated with an increase of milbemycin A3/A4 titer from 2382.5 +/- 55.7 mg/L to 2625.6 +/- 64.5 mg/L. Then, MilE, a CYP171 family cytochrome P450, was determined to be responsible for the generation of the furan ring between C6 and C8a of milbemycins. By further overexpression of milE, the production of beta-family milbemycins was reduced by 77.2%. Finally, the titer of milbemycin A3/A4 was increased by 53.1% to 3646.9 +/- 69.9 mg/L. Interestingly, overexpression of milE resulted in increased transcriptional levels of milbemycin biosynthetic genes and production of total milbemycins, which implied that the insufficient function of MilE was a limiting factor to milbemycin biosynthesis. Our research not only provides an efficient engineering strategy to improve the production of a commercially important product milbemycins, but also offers the clues for future study about transcriptional regulation of milbemycin biosynthesis.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序