Intensive mutagenesis of the nisin hinge leads to the rational design of enhanced derivatives

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dc.contributor.authorHealy, Brian
dc.contributor.authorField, Des
dc.contributor.authorO'Connor, Paula M.
dc.contributor.authorHill, Colin
dc.contributor.authorCotter, Paul D.
dc.contributor.authorRoss, R. Paul
dc.contributor.funderDepartment of Agriculture, Food and the Marine, Ireland
dc.contributor.funderScience Foundation Ireland
dc.date.accessioned2016-02-17T11:45:31Z
dc.date.available2016-02-17T11:45:31Z
dc.date.issued2013
dc.description.abstractNisin A is the most extensively studied lantibiotic and has been used as a preservative by the food industry since 1953. This 34 amino acid peptide contains three dehydrated amino acids and five thioether rings. These rings, resulting from one lanthionine and four methyllanthionine bridges, confer the peptide with its unique structure. Nisin A has two mechanisms of action, with the N-terminal domain of the peptide inhibiting cell wall synthesis through lipid II binding and the C-terminal domain responsible for pore-formation. The focus of this study is the three amino acid 'hinge' region (N 20, M 21 and K 22) which separates these two domains and allows for conformational flexibility. As all lantibiotics are gene encoded, novel variants can be generated through manipulation of the corresponding gene. A number of derivatives in which the hinge region was altered have previously been shown to possess enhanced antimicrobial activity. Here we take this approach further by employing simultaneous, indiscriminate site-saturation mutagenesis of all three hinge residues to create a novel bank of nisin derivative producers. Screening of this bank revealed that producers of peptides with hinge regions consisting of AAK, NAI and SLS displayed enhanced bioactivity against a variety of targets. These and other results suggested a preference for small, chiral amino acids within the hinge region, leading to the design and creation of producers of peptides with hinges consisting of AAA and SAA. These producers, and the corresponding peptides, exhibited enhanced bioactivity against Lactococcus lactis HP, Streptococcus agalactiae ATCC 13813, Mycobacterium smegmatis MC2155 and Staphylococcus aureus RF122 and thus represent the first example of nisin derivatives that possess enhanced activity as a consequence of rational design.en
dc.description.sponsorshipDepartment of Agriculture, Food and the Marine, Ireland (Food Institutional Research Measure 08/RD/C/691); Science Foundation Ireland (Investigator Award 10/IN.1/B3027)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleide79563
dc.identifier.citationHealy B, Field D, O’Connor PM, Hill C, Cotter PD, Ross RP (2013) Intensive Mutagenesis of the Nisin Hinge Leads to the Rational Design of Enhanced Derivatives. PLoS ONE 8(11): e79563. doi:10.1371/journal.pone.0079563
dc.identifier.doi10.1371/journal.pone.0079563
dc.identifier.issn1932-6203
dc.identifier.issued11en
dc.identifier.journaltitlePLOS ONEen
dc.identifier.urihttps://hdl.handle.net/10468/2358
dc.identifier.volume8en
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.rights© 2015 Healy et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are crediteden
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectPrecursor lipid IIen
dc.subjectLantibiotic nisinen
dc.subjectBiosynthesisen
dc.subjectModeen
dc.subjectPeptidesen
dc.subjectVarianten
dc.subjectIdentificationen
dc.subjectBacteriocinsen
dc.subjectAntibioticsen
dc.subjectImmunityen
dc.titleIntensive mutagenesis of the nisin hinge leads to the rational design of enhanced derivativesen
dc.typeArticle (peer-reviewed)en
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