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However, there is a limited number of high\u2010throughput screening protocols for PET\u2010hydrolyzing activity that avoid the use of surrogate substrates. Herein, a microplate fluorescence screening assay was described. It was based on the coupled activity of ketoreductases (KREDs) and diaphorase to release resorufin in the presence of the products of PET degradation. Six KREDs were identified in a commercial panel that were able to use the PET building block, ethylene glycol, as substrate. The most efficient KRED, KRED61, was combined with the diaphorase from <jats:italic>Clostridium kluyveri<\/jats:italic> to monitor the PET degradation reaction catalyzed by the thermostable variant of the cutinase\u2010type polyesterase from <jats:italic>Saccharomonospora viridis<\/jats:italic> AHK190. The PET degradation products were measured both fluorimetrically and by HPLC, with excellent correlation between both methods.<\/jats:p>","DOI":"10.1002\/cssc.202102750","type":"journal-article","created":{"date-parts":[[2022,3,22]],"date-time":"2022-03-22T11:03:44Z","timestamp":1647947024000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["A Coupled Ketoreductase\u2010Diaphorase Assay for the Detection of Polyethylene Terephthalate\u2010Hydrolyzing Activity"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2663-7339","authenticated-orcid":false,"given":"Mar\u00eda","family":"Gimeno\u2010P\u00e9rez","sequence":"first","affiliation":[{"name":"Department of Molecular Biology Universidad Aut\u00f3noma de Madrid  Campus de Cantoblanco Madrid 28049 Spain"},{"name":"Center of Molecular Biology \u201cSevero Ochoa\u201d (UAM-CSIC)  Nicol\u00e1s Cabrera 1 Madrid 28049 Spain"},{"name":"Institute for Molecular Biology-IUBM Universidad Aut\u00f3noma de Madrid  Campus de Cantoblanco Madrid 28049 Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8514-784X","authenticated-orcid":false,"given":"James D.","family":"Finnigan","sequence":"additional","affiliation":[{"name":"Prozomix Ltd.  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