{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T20:06:45Z","timestamp":1779912405749,"version":"3.53.1"},"reference-count":45,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T00:00:00Z","timestamp":1738800000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010132","name":"\u00d6sterreichische Nationalstiftung f\u00fcr Forschung, Technologie und Entwicklung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010132","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002428","name":"Austrian Science Fund","doi-asserted-by":"publisher","award":["P33545"],"award-info":[{"award-number":["P33545"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002428","name":"Austrian Science Fund","doi-asserted-by":"publisher","award":["W1224"],"award-info":[{"award-number":["W1224"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006012","name":"Christian Doppler Forschungsgesellschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006012","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":[[2025,5]]},"abstract":"<jats:p>\n                    Pyranose oxidase (POx) and\n                    <jats:italic>C<\/jats:italic>\n                    \u2010glycoside oxidase (CGOx) are FAD\u2010dependent oxidoreductases belonging to the glucose\u2010methanol\u2010choline oxidoreductase superfamily and share the same sequence space. Despite a shared structural fold, these two members possess homologous domains that enable (arm and head domain) or disable (insertion\u20101 domain and barrel\u2010shaped bottom) oligomerization. POxs with a higher oligomerization state (dimeric or tetrameric) exclusively catalyze the oxidation of monosaccharides (\n                    <jats:sc>d<\/jats:sc>\n                    \u2010glucose,\n                    <jats:sc>d<\/jats:sc>\n                    \u2010xylose). In contrast, the monomeric state of POxs\/CGOxs is observed to prefer glycosides (homoorientin, phlorizin) and has low activity with free monosaccharides. We aimed to engineer dimeric POx from\n                    <jats:italic>Kitasatospora aureofaciens<\/jats:italic>\n                    (\n                    <jats:italic>Ka<\/jats:italic>\n                    POx) to form a functional monomer, and monomeric POx\/CGOx from\n                    <jats:italic>Streptomyces canus<\/jats:italic>\n                    (\n                    <jats:italic>Sc<\/jats:italic>\n                    POx) to a dimeric structure. Deletion of the head and arm domains of the\n                    <jats:italic>Ka<\/jats:italic>\n                    POx subunit resulted in enzyme variants with a less hydrophobic surface, thus affecting its oligomerization. These monomeric\n                    <jats:italic>Ka<\/jats:italic>\n                    POx variants\n                    <jats:italic>Ka<\/jats:italic>\n                    POx_xal and\n                    <jats:italic>Ka<\/jats:italic>\n                    POx_xalh resembled monomeric wild\u2010type POxs\/CGOxs and preferred glycosides as substrates over monosaccharides with catalytic efficiencies for phlorizin being 24\u2009\u00d7\u200910\n                    <jats:sup>6<\/jats:sup>\n                    higher compared to those for\n                    <jats:sc>d<\/jats:sc>\n                    \u2010xylose. The wild\u2010type dimeric\n                    <jats:italic>Ka<\/jats:italic>\n                    POx showed no activity towards glycosides. We hypothesize that\n                    <jats:italic>Ka<\/jats:italic>\n                    POx_xalh is unable to react with monosaccharides because the introduced mutations alter the positions of monosaccharide\u2010binding residues. The inability of\n                    <jats:italic>Ka<\/jats:italic>\n                    POx to react with glycosides is likely caused by steric hindrance and the inaccessibility of the active site to bulky glycosides due to dimerization. The attempt to engineer\n                    <jats:italic>Sc<\/jats:italic>\n                    POx into a dimeric structure failed at the stage of soluble expression, likely due to exposed hydrophobic patches and aggregation.\n                  <\/jats:p>","DOI":"10.1111\/febs.70004","type":"journal-article","created":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T23:49:01Z","timestamp":1738885741000},"page":"2323-2337","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Shifting the substrate scope of dimeric pyranose oxidase from monosaccharide to glycoside preference through oligomeric state modification"],"prefix":"10.1111","volume":"292","author":[{"given":"Anja","family":"Kostelac","sequence":"first","affiliation":[{"name":"Department of Food Science and Technology BOKU University  Vienna Austria"},{"name":"Doctoral Programme BioToP \u2013 Biomolecular Technology of Proteins BOKU University  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enik\u0151","family":"Hermann","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology BOKU University  Vienna Austria"},{"name":"Doctoral Programme BioToP \u2013 Biomolecular Technology of Proteins BOKU University  Vienna Austria"},{"name":"Department of Material Science and Life Sciences BOKU University  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clemens","family":"Peterbauer","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology BOKU University  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chris","family":"Oostenbrink","sequence":"additional","affiliation":[{"name":"Department of Material Science and Life Sciences BOKU University  Vienna Austria"},{"name":"Christian Doppler Laboratory for Molecular Informatics in the Biosciences BOKU University  Vienna 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