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Improved production of clavulanic acid by reverse engineering and overexpression of the regulatory genes in an industrial Streptomyces clavuligerus strain

  • Genetics and Molecular Biology of Industrial Organisms -Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Genomic analysis of the clavulanic acid (CA)-high-producing Streptomyces clavuligerus strains, OL13 and OR, developed through random mutagenesis revealed a frameshift mutation in the cas1 gene-encoding clavaminate synthase 1. Overexpression of the intact cas1 in S. clavuligerus OR enhanced the CA titer by approximately 25%, producing ~ 4.95 g/L of CA, over the OR strain in the flask culture. Moreover, overexpression of the pathway-specific positive regulatory genes, ccaR and claR, in the OR strain improved CA yield by approximately 43% (~ 5.66 g/L) in the flask. However, co-expression of the intact cas1 with ccaR-claR did not further improve CA production. In the 7 L fermenter culture, maximum CA production by the OR strain expressing the wild-type cas1 and ccaR-claR reached approximately 5.52 g/L and 6.01 g/L, respectively, demonstrating that reverse engineering or simple rational metabolic engineering is an efficient method for further improvement of industrial strains.

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Acknowledgements

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korean Government (MSIT) (2015K1A1A2028365, 2019R1A2B5B03069338), Bio and Medical Technology Development Program (2018M3A9F3079662, 2018M3A9F3079664) through NRF funded by MSIT, and the Intelligent Synthetic Biology Center of the Global Frontier Project funded by MSIT (20110031961).

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Author notes

  1. Hang Soo Cho, ** Chul Jo, and Chang-Hun Shin have contributed equally to this work.

Authors and Affiliations

  1. Department of Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea

    Hang Soo Cho & Ho Jeong Kwon

  2. Institute of Nano-Bio Technology, Ewha Womans University, Seoul, 03760, Republic of Korea

    ** Chul Jo

  3. Department of Biotechnology, Korea University, Seoul, 02841, Republic of Korea

    Chang-Hun Shin & Chan-Wha Kim

  4. Fermentation Technology Team, Research Institute of CKD Bio, Ansan, 15604, Republic of Korea

    Chang-Hun Shin

  5. Department of Biological Sciences and KI for the BioCentury, Korea Advanced Institute of Science and Technology, Daejeon, 34141, Republic of Korea

    Namil Lee & Byung-Kwan Cho

  6. School of Biological Sciences, Seoul National University, Seoul, 08826, Republic of Korea

    Joon-Sun Choi & Jung-Hye Roe

  7. Intelligenet Synthetic Biology Center, Daejeon, 34141, Republic of Korea

    Byung-Kwan Cho

  8. Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, 03760, Republic of Korea

    Yeo Joon Yoon

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  1. Hang Soo Cho
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Correspondence to Ho Jeong Kwon or Yeo Joon Yoon.

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Cho, H.S., Jo, J.C., Shin, CH. et al. Improved production of clavulanic acid by reverse engineering and overexpression of the regulatory genes in an industrial Streptomyces clavuligerus strain. J Ind Microbiol Biotechnol 46, 1205–1215 (2019). https://doi.org/10.1007/s10295-019-02196-0

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  • DOI: https://doi.org/10.1007/s10295-019-02196-0

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