{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T13:47:15Z","timestamp":1791899235780,"version":"4.3.4"},"reference-count":31,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T00:00:00Z","timestamp":1769558400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T00:00:00Z","timestamp":1769558400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100013589","name":"Research England","doi-asserted-by":"publisher","award":["E3 funding"],"award-info":[{"award-number":["E3 funding"]}],"id":[{"id":"10.13039\/501100013589","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:p>\n                    Thiamine\u2010dependent actinobacterial 2\u2010hydroxyacyl\u2010CoA lyase (AcHACL) catalyzes the reversible cleavage of 2\u2010hydroxyacyl\u2010CoAs to formyl\u2010CoA and carbonyl compounds. To exploit the enzyme's biotechnological potential, a deeper understanding of the catalysis is required. Previously, AcHACL E493 was identified as an important acid\/base catalyst. Here, wild\u2010type and E493 mutant crystal structures representing Michaelis complexes with 2\u2010hydroxyisobutyryl\u2010CoA and (\n                    <jats:italic>S<\/jats:italic>\n                    )\u20102\u2010methylglyceryl\u2010CoA are provided. Although E493 guarantees high rates of essential proton transfers in AcAHCL\u2010catalyzed on\u2010pathway cleavage of 2\u2010hydroxyacyl\u2010CoAs and off\u2010pathway carboligations with short\u2010chain aldehydes and ketones, wild\u2010type substrate accommodation is suboptimal. Not E493D, but E493A and E493S mutations improved\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>M<\/jats:sub>\n                    . However,\n                    <jats:italic>k<\/jats:italic>\n                    <jats:sub>cat<\/jats:sub>\n                    is substantially reduced in the mutants. These tradeoffs are discussed by comparing active sites of AcHACL and related enzymes either lacking or possessing an E493 homolog.\n                  <\/jats:p>","DOI":"10.1002\/2211-5463.70199","type":"journal-article","created":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T04:04:43Z","timestamp":1769659483000},"page":"1314-1327","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["C2\u03b1\u2010carbanion\u2010protonating glutamate discloses tradeoffs between substrate accommodation and reaction rate in actinobacterial 2\u2010hydroxyacyl\u2010\n                    <scp>CoA<\/scp>\n                    lyase"],"prefix":"10.1002","volume":"16","author":[{"given":"Michael","family":"Zahn","sequence":"first","affiliation":[{"name":"Centre for Enzyme Innovation, School of the Environment and Life Sciences University of Portsmouth  UK"},{"name":"Core Facility Protein Production, Biozentrum, Martin\u2010Luther\u2010University\u2010Halle\u2010Wittenberg  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Seroka","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry University of Bialystok  Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryszard","family":"Lazny","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry University of Bialystok  Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zenon","family":"Lotowski","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry University of Bialystok  Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7877-8953","authenticated-orcid":false,"given":"Thore","family":"Rohwerder","sequence":"additional","affiliation":[{"name":"Department of Microbial Biotechnology, Helmholtz Centre for Environmental Research \u2013 UFZ  Leipzig 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