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Among them, acyl homoserine lactones (AHSLs) play a crucial role in quorum sensing, influencing bacterial pathogenicity and biofilm formation in Gram\u2010negative bacteria. Paraoxonases (PONs), calcium\u2010containing enzymes known for their lactonase activity, have been shown to hydrolyze AHSLs and reduce the biofilm formation of several pathogenic bacteria. In this study, we explored the potential lactonase activity of a class of zinc(II) enzymes, the carbonic anhydrases (CAs), aiming to uncover new insights into their catalytic versatility. Using LC\u2010MS and MS\/MS analyses, we investigated the lactonase activity of CAs and assessed several lactones through a stopped\u2010flow kinetic assay as substrates\/inhibitors. Our findings reveal that lactones are novel \u201cprodrug\u201d inhibitors of CAs, with lactones <jats:bold>DHC<\/jats:bold> and <jats:bold>6<\/jats:bold> showing the most promising inhibition constants (<jats:italic>K<\/jats:italic><jats:sub>I<\/jats:sub>s) in the low micromolar range against both human and bacterial isozymes.<\/jats:p>","DOI":"10.1002\/ardp.202400705","type":"journal-article","created":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T13:21:58Z","timestamp":1733750518000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Lactonase activity of \u03b1\u2010carbonic anhydrases allows identification of novel inhibitors"],"prefix":"10.1002","volume":"358","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0669-7481","authenticated-orcid":false,"given":"Simone","family":"Giovannuzzi","sequence":"first","affiliation":[{"name":"NEUROFARBA Department, Pharmaceutical and Nutraceutical Section University of 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