{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T15:21:39Z","timestamp":1790263299492,"version":"4.1.0"},"reference-count":25,"publisher":"Wiley","issue":"35","license":[{"start":{"date-parts":[[2025,6,8]],"date-time":"2025-06-08T00:00:00Z","timestamp":1749340800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007601","name":"Horizon 2020","doi-asserted-by":"publisher","award":["Marie"],"award-info":[{"award-number":["Marie"]}],"id":[{"id":"10.13039\/501100007601","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007601","name":"Horizon 2020","doi-asserted-by":"publisher","award":["Sklodowska"],"award-info":[{"award-number":["Sklodowska"]}],"id":[{"id":"10.13039\/501100007601","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007601","name":"Horizon 2020","doi-asserted-by":"publisher","award":["\u2010Curie grant agreement no. 956631"],"award-info":[{"award-number":["\u2010Curie grant agreement no. 956631"]}],"id":[{"id":"10.13039\/501100007601","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Chemistry A European J"],"published-print":{"date-parts":[[2025,6,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The development of (bio)catalytic methods to chiral tertiary alcohols is in demand, yet the enantioselective synthesis of tertiary alcohols with sterically hindered moieties is problematic. JanthE is a novel thiamine diphosphate\u2010dependent enzyme that catalyzes aldehyde\u2010ketone cross\u2010coupling reactions with a wide range of donor and acceptor substrates. However, product formation is low and precludes large\u2010scale production. We investigated the conversion of 2\u2010oxobutanoate and the bulky ketone phenoxy\u20102\u2010propanone to the product 2\u2010hydroxy\u20102\u2010methyl\u20101\u2010phenoxypentan\u20103\u2010one as a model reaction for the synthetic capabilities of JanthE. As the reaction design did not significantly improve the yield, we proceeded to rational protein engineering. Docking experiments identified V121, Y268, P293, Y297, and K567 in the active site as important residues for catalysis. Remarkably, the single\u2010point variant JanthE K567S led to a fivefold increase in product formation (90%\n                    <jats:italic>ee<\/jats:italic>\n                    ) compared to the wild type enzyme (93%\n                    <jats:italic>ee<\/jats:italic>\n                    ). The reaction was performed at preparative scale proving the direct possibility of application. Additionally, when the reaction was extended to longer 2\u2010oxopentanoate, the double variant Y297P_K567S showed a 5\u2010fold increase in product formation compared to the wild type. Our results demonstrate the evolvability and versatility of JanthE for the enantioselective synthesis of sterically hindered tertiary alcohols.\n                  <\/jats:p>","DOI":"10.1002\/chem.202500890","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T10:54:41Z","timestamp":1747738481000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Engineering of the Thiamine Diphosphate\u2010Dependent JanthE for the Synthesis of Tertiary Alcohols"],"prefix":"10.1002","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9294-2932","authenticated-orcid":false,"given":"Lucrezia","family":"Lanza","sequence":"first","affiliation":[{"name":"Institute of Pharmaceutical Sciences Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Albertstrasse 25 Freiburg 79104 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UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akash","family":"Pandya","sequence":"additional","affiliation":[{"name":"Johnson Matthey 28 Cambridge Science Park  Milton Road Cambridge CB4 0FP UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8213-1132","authenticated-orcid":false,"given":"Carmen","family":"Aranda","sequence":"additional","affiliation":[{"name":"Johnson Matthey 28 Cambridge Science Park  Milton Road Cambridge CB4 0FP UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5312-8951","authenticated-orcid":false,"given":"Zvjezdana Findrik","family":"Bla\u017eevi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Chemical Engineering and Technology University of Zagreb  Trg Marka Maruli\u0107a 19 Zagreb 10000 Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2742-4992","authenticated-orcid":false,"given":"Michael","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical Sciences Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Albertstrasse 25 Freiburg 79104 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