The potency of the broad spectrum bacteriocin, bactofencin A, against staphylococci is highly dependent on primary structure, N-terminal charge and disulphide formation

dc.contributor.authorO'Connor, Paula M.
dc.contributor.authorO'Shea, Eileen F.
dc.contributor.authorCotter, Paul D.
dc.contributor.authorHill, Colin
dc.contributor.authorRoss, R. Paul
dc.contributor.funderScience Foundation Ireland
dc.contributor.funderDepartment of Agriculture, Food and the Marine
dc.date.accessioned2018-09-27T12:08:24Z
dc.date.available2018-09-27T12:08:24Z
dc.date.issued2018
dc.description.abstractBactofencin A is a novel class IId bacteriocin, produced by the intestinal isolate Lactobacillus salivarius DPC6502, which has potent activity against medically significant pathogens including Staphylococcus aureus. This bacteriocin is unusual in that it has a highly cationic N terminus and a single disulfide bond between Cys7 and Cys22, resulting in a large C terminal loop. In this study, a library of synthetic bactofencin A variants were screened against the mastitis isolate, S. aureus DPC5246, to identify key residues responsible for activity. It was apparent that substituting either cysteine of the disulfide bond with either serine or alanine significantly reduced the activity of the bacteriocin, confirming the importance of the C terminal loop. Substituting N terminal amino acids with alanine had no effect on activity, whereas sequential removal of the N terminal positively charged residues resulted in an increasingly inactive peptide. A complete (synthetic) alanine scanning analysis revealed that the residues between Val9 and Gly17 were most affected by substitution suggesting that this area has a major influence on the potency of the bacteriocin. Substituting residues in the loop region between Cys7 and Cys22 for D-amino acid equivalents had a more detrimental effect on activity than L-alanine substitutions. Specifically Y10A, N11A, P15A and T16A are active at 4, 16, 1 and 16 μM respectively while their D equivalents were inactive at 1000 μM, the highest concentration tested. Ultimately, this study identifies the critical features in the primary structure of the bacteriocin which gives it such potent activity against pathogenic staphylococci.en
dc.description.sponsorshipDepartment of Agriculture, Fisheries and Food (Food Institutional Research Measure (04/R&D/C/232)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleid11833
dc.identifier.citationO’ Connor, P. M., O’ Shea, E. F., Cotter, P. D., Hill, C. and Ross, R. P. (2018) 'The potency of the broad spectrum bacteriocin, bactofencin A, against staphylococci is highly dependent on primary structure, N-terminal charge and disulphide formation', Scientific Reports, 8(1), 11833 (8pp). doi: 10.1038/s41598-018-30271-6en
dc.identifier.doi10.1038/s41598-018-30271-6
dc.identifier.endpage8
dc.identifier.issn2045-2322
dc.identifier.journaltitleScientific Reportsen
dc.identifier.startpage1
dc.identifier.urihttps://hdl.handle.net/10468/6946
dc.identifier.volume8
dc.language.isoenen
dc.publisherSpringer Natureen
dc.relation.projectinfo:eu-repo/grantAgreement/SFI/SFI Research Centres/12/RC/2273/IE/Alimentary Pharmabiotic Centre (APC) - Interfacing Food & Medicine/
dc.relation.urihttps://www.nature.com/articles/s41598-018-30271-6
dc.rights© 2018, the Authors. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectLactobacillus salivariusen
dc.subjectAmino aciden
dc.subjectPathogenic staphylococcien
dc.subjectN terminalen
dc.titleThe potency of the broad spectrum bacteriocin, bactofencin A, against staphylococci is highly dependent on primary structure, N-terminal charge and disulphide formationen
dc.typeArticle (peer-reviewed)en
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