{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T14:25:26Z","timestamp":1787235926904,"version":"build-2736575974"},"reference-count":54,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2023,1,19]],"date-time":"2023-01-19T00:00:00Z","timestamp":1674086400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR\u201017\u2010CE07\u20100047"],"award-info":[{"award-number":["ANR\u201017\u2010CE07\u20100047"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002790","name":"Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["STPGP 493781\u201016"],"award-info":[{"award-number":["STPGP 493781\u201016"]}],"id":[{"id":"10.13039\/501100002790","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004794","name":"Centre National de la Recherche Scientifique","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004794","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2023,5]]},"abstract":"<jats:p>\n                    Fungal copper radical oxidases (CROs) from the Auxiliary Activity family 5 (AA5) constitute a group of metalloenzymes that oxidize a wide panel of natural compounds, such as galactose\u2010containing saccharides or primary alcohols, into product derivatives exhibiting promising biotechnological interests. Despite a well\u2010conserved first copper\u2010coordination sphere and overall fold, some members of the AA5_2 subfamily are incapable of oxidizing galactose and galactosides but conversely efficiently catalyse the oxidation of diverse aliphatic alcohols. The objective of this study was to understand which residues dictate the substrate preferences between alcohol oxidases and galactose oxidases within the AA5_2 subfamily. Based on structural differences and molecular modelling predictions between the alcohol oxidase from\n                    <jats:italic>Colletotrichum graminicola<\/jats:italic>\n                    (\n                    <jats:italic>Cgr<\/jats:italic>\n                    AlcOx) and the archetypal galactose oxidase from\n                    <jats:italic>Fusarium graminearum<\/jats:italic>\n                    (\n                    <jats:italic>Fgr<\/jats:italic>\n                    GalOx), a rational mutagenesis approach was developed to target regions or residues potentially driving the substrate specificity of these enzymes. A set of 21 single and multiple\n                    <jats:italic>Cgr<\/jats:italic>\n                    AlcOx variants was produced and characterized leading to the identification of six residues (W39, F138, M173, F174, T246, L302), in the vicinity of the active site, crucial for substrate recognition. Two multiple\n                    <jats:italic>Cgr<\/jats:italic>\n                    AlcOx variants, i.e. M4F (W39F, F138W, M173R and T246Q) and M6 (W39F, F138W, M173R, F174Y, T246Q and L302P), exhibited a similar affinity for carbohydrate substrates when compared to\n                    <jats:italic>Fgr<\/jats:italic>\n                    GalOx. In conclusion, using a rational site\u2010directed mutagenesis approach, we identified key residues involved in the substrate selectivity of AA5_2 enzymes towards galactose\u2010containing saccharides.\n                  <\/jats:p>","DOI":"10.1111\/febs.16713","type":"journal-article","created":{"date-parts":[[2023,1,20]],"date-time":"2023-01-20T02:49:34Z","timestamp":1674182974000},"page":"2658-2672","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Rational engineering of AA5_2 copper radical oxidases to probe the molecular determinants governing their substrate selectivity"],"prefix":"10.1111","volume":"290","author":[{"given":"Radka","family":"Koncitikova","sequence":"first","affiliation":[{"name":"Centrale Marseille, iSm2 Aix Marseille Universit\u00e9, CNRS  France"},{"name":"UMR1163 Biodiversit\u00e9 et Biotechnologie Fongiques INRAE, Aix Marseille Universit\u00e9  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