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Sortase enzymes are critical for bacterial pathogenesis, and their inhibitors have become promising therapeutic targets for infection management. Filifactor alocis, a Gram-positive anaerobic bacterium, is now proposed as a diagnostic indicator of periodontal disease. Unlike other bacteria, F. alocis encodes a single putative sortase, FA1364. In this study, we functionally characterized the putative sortase FA1364 and found that it belongs to the class A family (SrtA). The SrtA-anchored surface proteins (FA1006, FA1336, FA1424, and FA1750) were identified, and MS\/MS analysis confirmed that SrtA is required for their cell-surface localization. The recombinant SrtA protein could recognize and cleave the LPKTG sorting motif with cysteine 191 and arginine 200 as essential catalytic residues. F. alocis FLL101 (\u0394FA1364::ermF) showed reduced ability to coaggregate and form biofilm, along with decreased collagen binding and survival in epithelial cells. Additionally, the FA1364-defective mutant exhibited increased sensitivity to air exposure. Collectively, these results suggest that the F. alocis SrtA protein is an important virulence factor and may represent a novel therapeutic target for the control of periodontal diseases.<\/jats:p>","DOI":"10.3390\/ijms27114783","type":"journal-article","created":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T11:02:51Z","timestamp":1779793371000},"page":"4783","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Functional Characterization of a Putative Sortase FA1364 in Filifactor alocis"],"prefix":"10.3390","volume":"27","author":[{"given":"Arunima","family":"Mishra","sequence":"first","affiliation":[{"name":"Division of Microbiology and Molecular Genetics, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2745-2062","authenticated-orcid":false,"given":"Nana Y.","family":"Sakyi Opoku","sequence":"additional","affiliation":[{"name":"Division of Microbiology and Molecular Genetics, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1057-7746","authenticated-orcid":false,"given":"Guangyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Mass Spectrometry Core Facility, Loma Linda University, Loma Linda, CA 92350, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard J.","family":"Lamont","sequence":"additional","affiliation":[{"name":"Department of Oral Immunology and Infectious Diseases, School of Dentistry, University of Louisville, Louisville, KY 40202, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7165-2159","authenticated-orcid":false,"given":"Hansel M.","family":"Fletcher","sequence":"additional","affiliation":[{"name":"Division of Microbiology and Molecular Genetics, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1099\/00207713-35-4-475","article-title":"Fusobacterium alocis sp. nov. and Fusobacterium sulci sp. nov. from the Human Gingival Sulcus","volume":"35","author":"Cato","year":"1985","journal-title":"Int. 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