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H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub> exhibit greater nucleophilicity than H<jats:sub>2<\/jats:sub>S while also having electrophilic characteristics, enabling unique activities such as protein S\u2010persulfidation. Despite their physiological importance, mechanisms and reactivities of H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub> remain inadequately explored due to their inherent instability in aqueous environments. Consequently, there is a need to develop biocompatible methods for controlled H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub> generation to elucidate their behaviors in biological contexts. Herein, we present a dual enzyme system (containing glucose oxidase (GOx) and chloroperoxidase (CPO)) with thioglucose as the substrate to facilitate the controlled release of H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub>. Fluorescence measurements with SSP4 and the trapping studies allowed us to confirm the production of H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub>. Such a method may be useful in elucidating the reactivity of hydrogen polysulfides in biological systems as well as provide a potential delivery of H<jats:sub>2<\/jats:sub>S<jats:sub>n<\/jats:sub> to target sites for biological applications.<\/jats:p>","DOI":"10.1002\/cbic.202400390","type":"journal-article","created":{"date-parts":[[2024,6,20]],"date-time":"2024-06-20T22:19:05Z","timestamp":1718921945000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["In\u2010Situ Generation of Hydrogen Polysulfides (H<sub>2<\/sub>S<sub>n<\/sub>) with Thioglucose, Glucose Oxidase, and Chloroperoxidase"],"prefix":"10.1002","volume":"25","author":[{"given":"Zannatun","family":"Noor","sequence":"first","affiliation":[{"name":"Department of Chemistry Brown University  Providence RI 02912 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiang","family":"Ni","sequence":"additional","affiliation":[{"name":"Department of Chemistry Brown University  Providence RI 02912 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7902-2987","authenticated-orcid":false,"given":"Ming","family":"Xian","sequence":"additional","affiliation":[{"name":"Department of Chemistry Brown University  Providence RI 02912 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