{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T05:52:58Z","timestamp":1764913978638,"version":"3.46.0"},"reference-count":51,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T00:00:00Z","timestamp":1759449600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Basic Microbiology"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Clostridium perfringens<\/jats:italic>\n                    epsilon toxin (ETX) is a potent pore\u2010forming exotoxin responsible for severe enterotoxemia and necrotizing enterocolitis in ruminants. To elucidate the molecular mechanisms underlying ETX pathogenicity and attenuation, several site\u2010directed mutants (R25A, F92A, Y133A, F206A, D210A, and G221A) were constructed based on structural analysis. Cytotoxicity assays revealed reduced virulence in ETX\u2010Y133A (hereafter referred to as Y133A), F92A, and F206A, with Y133A exhibiting the most significant attenuation. To further investigate the role of residue Y133, additional mutants (Y133E, Y133F, Y133S, Y133W, and Y133G) were generated. Selected mutants were evaluated for cytotoxicity, pathogenicity in BALB\/c mice, and in vivo safety through histopathological analysis. Furthermore, their pore\u2010forming ability, binding affinity to MDCK cells, and oligomerization properties were assessed. Results demonstrated that residue Y133 is critical for ETX activity, likely due to the necessity of its aromatic side chain for pore formation. In contrast, F92 and F206 appear to be involved in host\u2013cell interactions via distinct mechanisms. These findings provide insights into ETX structure\u2013function relationships and offer potential strategies for rational attenuation in vaccine development.\n                  <\/jats:p>","DOI":"10.1002\/jobm.70107","type":"journal-article","created":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T07:41:02Z","timestamp":1759477262000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Attenuated\n                    <i>Clostridium perfringens<\/i>\n                    Epsilon Toxin Mutants and Insights Into the Mechanism of Attenuation"],"prefix":"10.1002","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-3194-2636","authenticated-orcid":false,"given":"Zibei","family":"Huang","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine Yangzhou University Yangzhou Jiangsu Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haiyan","family":"Wang","sequence":"additional","affiliation":[{"name":"Jiangsu Vocational College of Agricultural and Animal Husbandry Taizhou Jiangsu Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinlin","family":"Huang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine Yangzhou University Yangzhou Jiangsu Province China"},{"name":"Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality Ministry of Agriculture and Rural Affairs of China Yangzhou Jiangsu Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenbo","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine Yangzhou University Yangzhou Jiangsu Province China"},{"name":"Jiangsu Co\u2010innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses Yangzhou University Yangzhou Jiangsu Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,10,3]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2014.03.079"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2014.06.012"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0163-7258(80)90031-5"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anaerobe.2014.08.016"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.2001.tb00503.x"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/toxins5112138"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2007.04.025"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.60.1.102-110.1992"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.64.1.230-237.1996"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.1997.tb01888.x"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2009.09.020"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0022053"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M206731200"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/antib7040037"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-10645-8"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2020.183364"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1177\/104063870802000301"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.2250"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2011.08145.x"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi300690a"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291x(77)90506-x"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2010.05.014"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0017787"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2011.08140.x"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-144-2-333"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.021782"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M010412200"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.20.6480-6487.1997"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1191\/0960327103ht397oa"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1369\/jhc.4A6254.2004"},{"key":"e_1_2_12_32_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01672-06"},{"key":"e_1_2_12_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2007.09.013"},{"key":"e_1_2_12_34_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00562-07"},{"key":"e_1_2_12_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9975(84)90031-8"},{"key":"e_1_2_12_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9975(84)90024-0"},{"key":"e_1_2_12_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/smtd.202300028"},{"key":"e_1_2_12_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2011.12.020"},{"key":"e_1_2_12_39_1","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12373"},{"key":"e_1_2_12_40_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02513-14"},{"key":"e_1_2_12_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2015.3"},{"key":"e_1_2_12_42_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01217.x"},{"key":"e_1_2_12_43_1","doi-asserted-by":"publisher","DOI":"10.4161\/hv.25649"},{"key":"e_1_2_12_44_1","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2017.1303022"},{"key":"e_1_2_12_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2023.06.044"},{"key":"e_1_2_12_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2795(81)90165-3"},{"key":"e_1_2_12_47_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12275-015-4523-8"},{"key":"e_1_2_12_48_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.00199.2007"},{"key":"e_1_2_12_49_1","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12373"},{"key":"e_1_2_12_50_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-5822.2003.00262.x"},{"key":"e_1_2_12_51_1","doi-asserted-by":"publisher","DOI":"10.3390\/toxins16040180"},{"key":"e_1_2_12_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)50294-0"}],"container-title":["Journal of Basic Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jobm.70107","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T05:49:18Z","timestamp":1764913758000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jobm.70107"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,3]]},"references-count":51,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2025,12]]}},"alternative-id":["10.1002\/jobm.70107"],"URL":"https:\/\/doi.org\/10.1002\/jobm.70107","archive":["Portico"],"relation":{},"ISSN":["0233-111X","1521-4028"],"issn-type":[{"type":"print","value":"0233-111X"},{"type":"electronic","value":"1521-4028"}],"subject":[],"published":{"date-parts":[[2025,10,3]]},"assertion":[{"value":"2025-07-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-03","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70107"}}