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Tyrosinases have an additional monooxygenation ability to hydroxylate monophenol to <jats:italic>o<\/jats:italic>\u2010diphenol. It is still not clear what causes the difference in the catalytic activities. We solved a complex structure of <jats:italic>Aspergillus oryzae<\/jats:italic> catechol oxidase with resorcinol bound into the active site. Catalytic activity of <jats:italic>A.\u2005oryzae<\/jats:italic> catechol oxidase was studied, for the first time, by high\u2010resolution FT\u2010ICR mass spectrometry to shed light on the reaction mechanism. The enzyme was also found to catalyze monooxygenation of small phenolics, which provides a novel perspective for the discussion of differences in the catalytic activity between tyrosinases and catechol oxidases. According to the results, two binding modes for resorcinol are suggested and a reaction mechanism for coupled binuclear copper enzymes is discussed.<\/jats:p>","DOI":"10.1002\/cbic.201800387","type":"journal-article","created":{"date-parts":[[2018,9,11]],"date-time":"2018-09-11T13:00:47Z","timestamp":1536670847000},"page":"2348-2352","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Unraveling Substrate Specificity and Catalytic Promiscuity of <i>Aspergillus oryzae<\/i> Catechol Oxidase"],"prefix":"10.1002","volume":"19","author":[{"given":"Leena","family":"Penttinen","sequence":"first","affiliation":[{"name":"Department of Chemistry University of Eastern Finland  Yliopistokatu 7 80130 Joensuu Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Rutanen","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Eastern Finland  Yliopistokatu 7 80130 Joensuu Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8446-4704","authenticated-orcid":false,"given":"Janne","family":"J\u00e4nis","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Eastern Finland  Yliopistokatu 7 80130 Joensuu Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juha","family":"Rouvinen","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Eastern Finland  Yliopistokatu 7 80130 Joensuu Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4471-7188","authenticated-orcid":false,"given":"Nina","family":"Hakulinen","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Eastern Finland  Yliopistokatu 7 80130 Joensuu Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,10,31]]},"reference":[{"key":"e_1_2_2_1_1","first-page":"498276","author":"Solano F.","year":"2014","journal-title":"New J. 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