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Heterologous production of new lasso peptide koreensin based on genome mining

The Journal of Antibiotics volume 74, pages 42–50 (2021)Cite this article

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Abstract

Lasso peptides are a class of ribosomally biosynthesized and posttranslationally modified peptides with a knot structure as a common motif. Based on a genome search, a new biosynthetic gene cluster of lasso peptide was found in the genome of the proteobacterium Sphingomonas koreensis. Interestingly, the amino acid sequence of the precursor peptide gene includes two cell adhesion motif sequences (KGD and DGR). Heterologous production of the new lasso peptide was performed using the cryptic biosynthetic gene cluster of S. koreensis. As a result, a new lasso peptide named koreensin was produced by the gene expression system in the host strain Sphingomonas subterranea. The structure of koreensin was determined by NMR and ESI–MS analysis. The three-dimensional structure of koreensin was obtained based on an NOE experiment and the coupling constants. A variant peptide (koreensin-RGD), which had RGD instead of KGD, was produced by heterologous production with site-directed mutagenesis experiment. Koreensin and koreensin-RGD did not show cell adhesion inhibitory activity, although the molecules possessed cell adhesion motifs. The possible presence of a salt bridge between the motifs in koreensin was indicated, and it may prevent the cell adhesion motif from functioning.

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Acknowledgements

This study was supported by the Japan Society for the Promotion of Science by Grants-in-Aids (grant number 20K05848), Koyanagi Foundation, and Takahashi Industrial and Economic Research Foundation. The NMR spectra were recorded on Bruker Avance 600 and Avance III HD 800 spectrometers at Advanced Analysis Center, NARO.

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Authors and Affiliations

  1. Graduate School of Integrated Science and Technology, Shizuoka University, Shizuoka, Japan

    Hiroki Fuwa & Shinya Kodani

  2. Food Research Institute, National Agriculture and Food Research Organization (NARO), Ibaraki, Japan

    Hikaru Hemmi

  3. Graduate School of Science and Technology, Shizuoka University, Shizuoka, Japan

    Issara Kaweewan & Shinya Kodani

  4. Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Japan

    Ikko Kozaki & Hiroyuki Honda

  5. Academic Institute, Shizuoka University, Shizuoka, Japan

    Shinya Kodani

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  1. Hiroki Fuwa
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Correspondence to Shinya Kodani.

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Fuwa, H., Hemmi, H., Kaweewan, I. et al. Heterologous production of new lasso peptide koreensin based on genome mining. J Antibiot 74, 42–50 (2021). https://doi.org/10.1038/s41429-020-00363-5

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