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Combined application of plasma mutagenesis and gene engineering leads to 5-oxomilbemycins A3/A4 as main components from Streptomyces bingchenggensis

  • Applied genetics and molecular biotechnology
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Abstract

Milbemycin oxime has been commercialized as effective anthelmintics in the fields of animal health, agriculture, and human infections. Currently, milbemycin oxime is synthesized by a two-step chemical reaction, which involves the ketonization of milbemycins A3/A4 to yield the intermediates 5-oxomilbemycins A3/A4 using CrO3 as catalyst. Due to the low efficiency and environmental unfriendliness of the ketonization of milbemycins A3/A4, it is imperative to develop alternative strategies to produce 5-oxomilbemycins A3/A4. In this study, the atmospheric and room temperature plasma (ARTP) mutation system was first employed to treat milbemycin-producing strain Streptomyces bingchenggensis, and a mutant strain BC-120-4 producing milbemycins A3, A4, B2, and B3 as main components was obtained, which favors the construction of genetically engineered strains producing 5-oxomilbemycins. Importantly, the milbemycins A3/A4 yield of BC-120-4 reached 3,890 ± 52 g/l, which was approximately two times higher than that of the initial strain BC-109-6 (1,326 ± 37 g/l). The subsequent interruption of the gene milF encoding a C5-ketoreductase responsible for the ketonization of milbemycins led to strain BCJ60 (∆milF) with the production of 5-oxomilbemycins A3/A4 and the elimination of milbemycins A3, A4, B2, and B3. The high 5-oxomilbemycins A3/A4 yield (3,470 ± 147 g/l) and genetic stability of BCJ60 implied the potential use in industry to prepare 5-oxomilbemycins A3/A4 for the semisynthesis of milbemycins oxime.

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Acknowledgments

This research was supported by grants from the National Outstanding Youth Foundation (No. 31225024), the National Key Project for Basic Research (No. 2010CB126102), the National Key Technology R&D Program (No. 2012BAD19B06), the Program for New Century Excellent Talents in University (NCET-11-0953), the National Natural Science Foundation of China (Nos. 31372006, 31171913, and 31071750), the Outstanding Youth Foundation of Heilongjiang Province (JC201201), and Chang Jiang Scholar Candidates Program for Provincial Universities in Heilongjiang (CSCP).

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Authors and Affiliations

  1. Key Laboratory of Agriculture Biological Functional Gene of Heilongjiang Provincial Education Committee, Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Hai-Yan Wang, Ji Zhang, Yue-Jing Zhang, Chong-Xi Liu, Xiang-Jing Wang & Wen-Sheng Xiang

  2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, People’s Republic of China

    Bo Zhang, Hai-Rong He & Wen-Sheng Xiang

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  1. Hai-Yan Wang
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  2. Ji Zhang
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  3. Yue-Jing Zhang
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  4. Bo Zhang
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Correspondence to Xiang-Jing Wang or Wen-Sheng Xiang.

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Wang, HY., Zhang, J., Zhang, YJ. et al. Combined application of plasma mutagenesis and gene engineering leads to 5-oxomilbemycins A3/A4 as main components from Streptomyces bingchenggensis . Appl Microbiol Biotechnol 98, 9703–9712 (2014). https://doi.org/10.1007/s00253-014-5970-6

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  • DOI: https://doi.org/10.1007/s00253-014-5970-6

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