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While the functions of epidermal CDases are known, roles of neutral CDases secreted by skin\u2010residing microbes are undefined. Here, we developed a one\u2010step fluorogenic substrate,\n                    <jats:bold>S\u2010B<\/jats:bold>\n                    , for specific detection of bacterial CDase activity and inhibitor screening. We identified a non\u2010hydrolyzable substrate mimic,\n                    <jats:bold>C6<\/jats:bold>\n                    , as the best hit. Based on\n                    <jats:bold>C6<\/jats:bold>\n                    , we designed a photoaffinity probe,\n                    <jats:bold>JX\u20101<\/jats:bold>\n                    , which efficiently detects bacterial CDases. Using\n                    <jats:bold>JX\u20101<\/jats:bold>\n                    , we identified endogenous low\u2010abundance PaCDase in a\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    monoculture and in a mixed skin bacteria culture. Harnessing both\n                    <jats:bold>S\u2010B<\/jats:bold>\n                    and\n                    <jats:bold>JX\u20101<\/jats:bold>\n                    , we found that CDase activity positively correlates with the relative abundance of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    and is negatively associated with wound area reduction in clinical diabetic foot ulcer patient samples. Overall, our study demonstrates that bacterial CDases are important regulators of skin ceramides and potentially play a role in wound healing.\n                  <\/jats:p>","DOI":"10.1002\/anie.202307553","type":"journal-article","created":{"date-parts":[[2023,6,21]],"date-time":"2023-06-21T02:01:07Z","timestamp":1687312867000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Detection of Bacterial Neutral Ceramidase in Diabetic Foot Ulcers with an Optimized Substrate and Chemoenzymatic Probes"],"prefix":"10.1002","volume":"62","author":[{"given":"Jiao Xia","family":"Zou","sequence":"first","affiliation":[{"name":"Department of Chemistry National University of Singapore  3 Science Drive 3 Singapore 117543 Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wisely","family":"Chua","sequence":"additional","affiliation":[{"name":"Molecular Engineering Lab, Institute of Molecular and Cell Biology 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