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Characterization of modification enzyme NukM and engineering of a novel thioether bridge in lantibiotic nukacin ISK-1

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

The lantibiotic nukacin ISK-1 is an antimicrobial peptide containing unusual amino acids such as lanthionine and dehydrobutyrine. The nukacin ISK-1 prepeptide (NukA) undergoes posttranslational modifications, such as the dehydration and cyclization reactions required to form the unusual amino acids by the modification enzyme NukM. We have previously constructed a system for the introduction of unusual amino acids into NukA by coexpression of NukM in Escherichia coli. Using this system, we describe the substrate specificity of NukM by the coexpression of a series of NukA mutants. Our results revealed the following characteristics of NukM: (1) its dehydration activity is not coupled to its cyclization activity; (2) its dehydration activity is site-specific; (3) the length of the substrate is important for its dehydration activity. Furthermore, we succeeded in introducing a novel thioether bridge in NukA by replacing an unmodified Ser at position 27 with a Cys residue.

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Acknowledgments

We thank N. Futami (Toray Research Center, Tokyo, Japan) for the amino acid sequence analysis. Our work on lantibiotics nukacin ISK-1 has been partially supported by grants from the following organizations: the Japan Society for the Promotion of Science, the Japan Science Society, the Novartis Foundation (Japan) for the Promotion of Science, the Novozymes Japan Research Fund, and the Nagase Science and Technology Foundation.

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Authors and Affiliations

  1. Laboratory of Microbial Technology, Division of Microbial Science and Technology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Kouki Shioya, Yoshitaka Harada, Jiro Nakayama & Kenji Sonomoto

  2. Section of Infection Biology, Department of Functional Bioscience, Fukuoka Dental College, 2-15-1 Tamura, Sawara-ku, Fukuoka, 814-0913, Japan

    Jun-ichi Nagao

  3. Laboratory of Functional Food Design, Department of Functional Metabolic Design, Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Kenji Sonomoto

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  1. Kouki Shioya
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  2. Yoshitaka Harada
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  3. Jun-ichi Nagao
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  4. Jiro Nakayama
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  5. Kenji Sonomoto
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Correspondence to Kenji Sonomoto.

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Shioya, K., Harada, Y., Nagao, Ji. et al. Characterization of modification enzyme NukM and engineering of a novel thioether bridge in lantibiotic nukacin ISK-1. Appl Microbiol Biotechnol 86, 891–899 (2010). https://doi.org/10.1007/s00253-009-2334-8

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  • DOI: https://doi.org/10.1007/s00253-009-2334-8

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