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Mutagenesis of nisin’s leader peptide proline strongly modulates export of precursor nisin

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Abstract

The lantibiotic nisin is produced by Lactococcus lactis as a precursor peptide comprising a 23 amino acid leader peptide and a 34 amino acid post-translationally modifiable core peptide. We previously demonstrated that the conserved FNLD part of the leader is essential for intracellular enzyme-catalyzed introduction of lanthionines in the core peptide and also for transporter-mediated export, whereas other positions are subject to large mutational freedom. We here demonstrate that, in the absence of the extracellular leader peptidase, NisP, export of precursor nisin via the modification and transporter enzymes, NisBTC, is strongly affected by multiple substitutions of the leader residue at position -2, but not by substitution of positions in the vicinity of this site. Export levels of precursor nisin increased by more than 70% for position -2 mutants Asp, Thr, Ser, Trp, Lys, Val and decreased more than 70% for Cys, His, Met. In a strain with leader peptidase, the Pro-2Lys and Pro-2Asp precursor nisins were less efficiently cleaved by NisP than wild type precursor nisin. Taken together, the wild type precursor nisin with a proline at position -2 allows balanced export and cleavage efficiencies by precursor nisin’s transporter and leader peptidase.

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Acknowledgements

Sander Smits and Lutz Schmitt are gratefully acknowledged for helpful discussions on mechanistic models for transport.

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Author notes

  1. Annechien Plat

    Present address: Gynecological Oncology, University Medical Center Groningen, 9700 RB, Groningen, The Netherlands

Authors and Affiliations

  1. Biomade Technology Foundation, Nijenborgh 4, 9747 AG, Groningen, The Netherlands

    Annechien Plat

  2. Lanthio Pharma, MorphoSys AG, Rozenburglaan 13B, 9727 DL, Groningen, The Netherlands

    Anneke Kuipers, Rick Rink & Gert N. Moll

  3. ImmuCell Corp., 56 Evergreen Drive, Portland, ME, 04130, USA

    Joe Crabb

  4. Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands

    Gert N. Moll

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  1. Annechien Plat
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Correspondence to Gert N. Moll.

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Plat, A., Kuipers, A., Crabb, J. et al. Mutagenesis of nisin’s leader peptide proline strongly modulates export of precursor nisin. Antonie van Leeuwenhoek 110, 321–330 (2017). https://doi.org/10.1007/s10482-016-0802-6

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