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Purification of Peptide Antimicrobials and Thioether-Stabilized Molecules Produced In Vivo by Lantibiotic Modification Machineries

  • Manuel Montalban-Lopez
  • Andrius Buivydas
  • Oscar P. Kuipers
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

The presence of posttranslational modifications is a key feature for the biological activity and stability of many natural products, including lanthipeptides. Lanthipeptides are a group of posttranslationally modified peptides containing lanthionine residues in their structures. Among lanthipeptides, the subgroup of lantibiotics is receiving a renewed interest in the last years for their high antimicrobial activity. The development of production platforms for novel lantibiotics and peptides containing posttranslational modifications is taking place in parallel to the understanding of the mechanistic aspects of the enzymes responsible for these modifications. It is now possible to combine pieces (i.e., enzymes and regulatory elements) of different biosynthesis routes of natural products in order to modify peptides of interest in a predictable manner. In this chapter we detail the production of lanthipeptides and thioether-stabilized peptides using two well-established production platforms for Escherichia coli and Lactococcus lactis. These are based on the dehydration and cyclization reactions carried out by the enzymes NisB and NisC, respectively, on peptides fused to the nisin leader peptide. Furthermore, we provide appropriate protocols for the purification and characterization of the peptides produced and modified with these biosynthetic tools.

Keywords

Lanthipeptide Lantibiotic Nisin Posttranslational modification Synthetic biology 

Notes

Acknowledgments

Andrius Buivydas and Manuel Montalban-Lopez are supported by the Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO) projects ALW 821.02.018 and 855.01.162, respectively. M. Montalban-Lopez is also part of the EU Project Synpeptide.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Manuel Montalban-Lopez
    • 1
  • Andrius Buivydas
    • 1
  • Oscar P. Kuipers
    • 1
  1. 1.Department of Molecular GeneticsUniversity of GroningenGroningenThe Netherlands

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