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Improved FK506 production by the precursors and product-tolerant mutant of Streptomyces tsukubaensis based on genome shuffling and dynamic fed-batch strategies

  • Fermentation, Cell Culture and Bioengineering
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

FK506, a secondary metabolite produced by Streptomyces tsukubaensis, is well known for its immunosuppressant properties to prevent rejection of transplanted organs and treat autoimmune diseases. However, the low titer of FK506 in the original producer strain limits the further industrialization efforts and restricts its clinical applications. To address this issue, a highly efficient method combined genome shuffling and dynamic fed-batch strategies was systematically performed in this work. Firstly, after five rounds of genome shuffling based on precursors and product resistances, a higher yielding strain TJ-P325 was successfully acquired, whose production reached 365.6 mg/L, 11-fold increase compared with the original strain. Then, the possible mechanism of different production capabilities between TJ-P325 and the wild type was explored through comparative gene expression analysis of key genes. Results showed that the transcription level of key genes was altered significantly in the mutant. Moreover, precursors addition enhanced the FK506 production by 28 %, as well as reduced the by-products biosynthesis. Finally, the disodium malonate and disodium methylmalonate dynamic fed-batch strategies dramatically led to the production of 514.5 mg/L in a 7.5-L bioreactor. These results demonstrated that genome shuffling and dynamic fed-batch strategies could be successfully applied to generate high-yield strains with value-added natural products during industrial microbial fermentation.

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Acknowledgments

This work was financially supported by the National 973 Project of China (No. 2013CB733600), the Key Program of National Natural Science Foundation of China (No. 21236005), the National Natural Science Foundation of China (No. 20936002), the Fundamental Research Funds for the Central Universities (No. 65141028) and the Programme of Introducing Talents of Discipline to Universities (No. B06006).

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Authors and Affiliations

  1. Department of Biological Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China

    Wenjie Du, Menglei Xia & Jianping Wen

  2. TEDA School of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, 300457, China

    Di Huang

  3. Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin, 300072, China

    Jianping Wen

  4. Collaborative Innovation Center of Chemical Science and Engineering, Tianjin, 300072, China

    Jianping Wen

  5. CORA, Dalian University of Technology, Dalian, 116024, China

    Ming Huang

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  1. Wenjie Du
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  2. Di Huang
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  3. Menglei Xia
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  4. Jianping Wen
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  5. Ming Huang
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Correspondence to Di Huang or Jianping Wen.

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Du, W., Huang, D., Xia, M. et al. Improved FK506 production by the precursors and product-tolerant mutant of Streptomyces tsukubaensis based on genome shuffling and dynamic fed-batch strategies. J Ind Microbiol Biotechnol 41, 1131–1143 (2014). https://doi.org/10.1007/s10295-014-1450-3

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