{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T15:09:34Z","timestamp":1776092974134,"version":"3.50.1"},"reference-count":44,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2023,1,20]],"date-time":"2023-01-20T00:00:00Z","timestamp":1674172800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Clinical isolates of <jats:italic>Clostridioides difficile<\/jats:italic> sometimes exhibit multidrug resistance and cause diarrhea after antibiotic administration. Metronidazole and vancomycin are often used as therapeutic agents, but resistance to these antibiotics has been found clinically. Therefore, the development of alternative antimicrobial agents is needed. Nisin A, produced by <jats:italic>Lactococcus lactis<\/jats:italic>, has been demonstrated to be effective against <jats:italic>C<\/jats:italic>. <jats:italic>difficile<\/jats:italic> infection. In this study, we evaluated the susceptibility of 11 <jats:italic>C<\/jats:italic>. <jats:italic>difficile<\/jats:italic> clinical isolates to nisin A and found that they could be divided into 2 groups: high and low susceptibility. Since CprABC and DltDABC, which are responsible for nisin A efflux and cell surface charge, respectively, have been reported to be related to nisin A susceptibility, we investigated the expression of <jats:italic>cprA<\/jats:italic> and <jats:italic>dltA<\/jats:italic> among the 11 strains. <jats:italic>cprA<\/jats:italic> expression in all strains was induced by nisin A, but <jats:italic>dltA<\/jats:italic> expression was not. The expression levels of both genes did not correlate with nisin A susceptibility in these clinical isolates. To evaluate cell surface charge, we performed a cytochrome C binding assay and found no relationship between charge and nisin A susceptibility. Then, we determined the whole genome sequence of each clinical isolate and carried out phylogenetic analysis. The 11 isolates separated into two major clusters, which were consistent with the differences in nisin A susceptibility. Furthermore, we found common differences in several amino acids in the sequences of CprA, CprB, and CprC between the two clusters. Therefore, we speculated that the different amino acid sequences of CprABC might be related to nisin A susceptibility. In addition, <jats:italic>C<\/jats:italic>. <jats:italic>difficile<\/jats:italic> strains could be divided in the same two groups based on susceptibility to epidermin and mutacin III, which are structurally similar to nisin A. These results suggest that genotypic variations in <jats:italic>C<\/jats:italic>. <jats:italic>difficile<\/jats:italic> strains confer different susceptibilities to bacteriocins.<\/jats:p>","DOI":"10.1371\/journal.pone.0280676","type":"journal-article","created":{"date-parts":[[2023,1,20]],"date-time":"2023-01-20T18:48:53Z","timestamp":1674240533000},"page":"e0280676","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Different CprABC amino acid sequences affect nisin A susceptibility in Clostridioides difficile isolates"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9429-4438","authenticated-orcid":true,"given":"Noriaki","family":"Ide","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miki","family":"Kawada-Matsuo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9394-6107","authenticated-orcid":true,"given":"Mi Nguyen-Tra","family":"Le","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junzo","family":"Hisatsune","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiromi","family":"Nishi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshinori","family":"Hara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Norikazu","family":"Kitamura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seiya","family":"Kashiyama","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michiya","family":"Yokozaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroyuki","family":"Kawaguchi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroki","family":"Ohge","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Motoyuki","family":"Sugai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9094-7649","authenticated-orcid":true,"given":"Hitoshi","family":"Komatsuzawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2023,1,20]]},"reference":[{"key":"pone.0280676.ref001","first-page":"1","article-title":"Large clostridial toxins: mechanisms and roles in disease","volume":"85","author":"KE Orrell","year":"2011","journal-title":"Microbiol Mol Biol Rev"},{"key":"pone.0280676.ref002","doi-asserted-by":"crossref","first-page":"15","DOI":"10.4161\/gmic.26854","article-title":"Clostridium difficile binary toxin CDT","volume":"5","author":"DN Gerding","year":"2013","journal-title":"Gut Microbes"},{"issue":"4","key":"pone.0280676.ref003","doi-asserted-by":"crossref","first-page":"234","DOI":"10.4161\/gmic.1.4.12706","article-title":"Clostridium difficile infection: An overview of the disease and its pathogenesis, epidemiology and interventions","volume":"1","author":"VK Viswanathan","year":"2010","journal-title":"Gut Microbes"},{"issue":"12","key":"pone.0280676.ref004","doi-asserted-by":"crossref","first-page":"2459","DOI":"10.1007\/s10096-021-04311-5","article-title":"Antimicrobial resistance in Clostridioides difficile","volume":"40","author":"K O\u2019Grady","year":"2021","journal-title":"Eur J Clin Microbiol Infect Dis"},{"issue":"1","key":"pone.0280676.ref005","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1186\/s13756-020-00815-5","article-title":"Antimicrobial resistance in Clostridioides (Clostridium) difficile derived from humans: a systematic review and meta-analysis","volume":"9","author":"M Sholeh","year":"2020","journal-title":"Antimicrob Resist Infect Control"},{"key":"pone.0280676.ref006","unstructured":"O\u2019Neill J. 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