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Constitutive and high gene expression in the diaminopimelate pathway accelerates ε-poly-l-lysine production in Streptomyces albulus

The Journal of Antibiotics volume 76, pages 522–531 (2023)Cite this article

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Abstract

Streptomyces albulus NBRC14147 produces a homopoly(amino acid), ε-poly-l-lysine (ε-PL). Due to its antibiotic activity, thermostability, biodegradability, and non-toxicity to humans, ε-PL is used as a food preservative. In this study, homology searches of diaminopimelate (DAP) pathway genes (dapB and dapE), in an S. albulus genome database, were shown to encode predicted enzymes using dapB or dapE in Escherichia coli strain complementation assays. We observed that dapB and dapE transcriptional levels were weak during ε-PL production stages. Therefore, we strengthened this expression using an ermE constitutive promoter. Engineered strains generated faster growth and ε-PL production rates when compared with the control strain. Moreover, maximum ε-PL yields in S. albulus, where dapB was constitutively expressed, were approximately 14% higher when compared with the control strain. These findings showed that enhanced lysine biosynthetic gene expression generated faster and higher ε-PL production levels.

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Acknowledgements

The authors would like to thank Enago (www.enago.jp) for the English language review.

Funding

This research was supported by JSPS KAKENHI grants 20K21284 (Y.H.), 20H02918 (Y.H.), 20K15437 (F.H.), and 22K05390 (F.H.), and by a JSPS KAKENHI Grant-in-Aid for Transformative Research Areas (A) 22H05122 (C.M.).

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Authors and Affiliations

  1. Department of Bioscience, Fukui Prefectural University, Fukui, Japan

    Fumihito Hasebe, Kazuya Adachi, Chitose Maruyama & Yoshimitsu Hamano

  2. Fukui Bio Incubation Center (FBIC), Fukui Prefectural University, Eiheiji-cho, Fukui, Japan

    Fumihito Hasebe, Chitose Maruyama & Yoshimitsu Hamano

  3. Department of Life Science & Technology, Kansai University, Osaka, Japan

    Kazuya Yamanaka & Tadao Oikawa

  4. Graduate School of Science and Engineering, Kansai University, Osaka, Japan

    Kazuya Yamanaka & Tadao Oikawa

Authors
  1. Fumihito Hasebe
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  2. Kazuya Adachi
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  3. Kazuya Yamanaka
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  4. Tadao Oikawa
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  5. Chitose Maruyama
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  6. Yoshimitsu Hamano
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Contributions

FH, KY, TO, CM, and YH designed experiments. CM and YH performed the experiments for RT-PCR and the construction of S. albulus transformants. FH and KA conducted complementation assays and fermentation experiments on S. albulus transformants. KY performed RNA-Seq analysis. FH, KY and YH wrote the manuscript. YH led the study.

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Correspondence to Fumihito Hasebe or Yoshimitsu Hamano.

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Hasebe, F., Adachi, K., Yamanaka, K. et al. Constitutive and high gene expression in the diaminopimelate pathway accelerates ε-poly-l-lysine production in Streptomyces albulus. J Antibiot 76, 522–531 (2023). https://doi.org/10.1038/s41429-023-00636-9

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  • DOI: https://doi.org/10.1038/s41429-023-00636-9

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