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Investigation of halogenation during the biosynthesis of ramoplanin in Actinoplanes sp. ATCC33076

  • Applied genetics and molecular biotechnology
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Abstract

Ramoplanin and enduracidin are lipopeptide antibiotics effective against Gram-positive pathogens, which share close similarity in structure and biosynthetic pathway. Both compounds have chlorine atoms attached to 4-hydroxyphenylglycine (Hpg) but with different chlorinating sites and levels. Here, to probe the factor affecting the site and level of halogenation, gene inactivation and heterologous expression were carried out in Actinoplanes sp. ATCC33076 by homologous recombination. Metabolite analysis confirmed that ram20 encodes the only halogenase in ramoplanin biosynthetic pathway, and enduracidin halogenase End30 could heterologously complement the ram20-deficient mutant. Additionally, the mannosyltransferase-deficient mutant produces a dichlorinated ramoplanin aglycone with the halogenation site at Hpg13. This study has refined our understanding of how halogenation occurs in ramoplanin biosynthetic pathway, and lays the foundation for further exploitation of ramoplanin and enduracidin halogenase in combinatorial biosynthesis.

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Authors and Affiliations

  1. School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Rd., Minhang District, Shanghai, 200240, China

    Jun-Sheng Chen

  2. State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai, 200040, China

    Jun-Sheng Chen, Min Su, Lei Shao, Yuan-Xi Wang, Hui-Min Lin & Dai-Jie Chen

  3. Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, SJTU, 800 Dongchuan Rd., Minhang District, Shanghai, 200240, China

    Jun-Sheng Chen

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  1. Jun-Sheng Chen
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  2. Min Su
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  3. Lei Shao
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  4. Yuan-Xi Wang
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  5. Hui-Min Lin
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Correspondence to Lei Shao or Dai-Jie Chen.

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Funding

This work was funded in part by grants from the National Natural Science Foundation of China (81373310) and Ministry of Science and Technology of China (Grant No. 2012ZX09301002-003).

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The authors declare that they have no competing interests.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Chen, JS., Su, M., Shao, L. et al. Investigation of halogenation during the biosynthesis of ramoplanin in Actinoplanes sp. ATCC33076. Appl Microbiol Biotechnol 100, 289–298 (2016). https://doi.org/10.1007/s00253-015-7014-2

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  • DOI: https://doi.org/10.1007/s00253-015-7014-2

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