{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:17:31Z","timestamp":1788434251138,"version":"build-2803163510"},"reference-count":34,"publisher":"American Chemical Society (ACS)","issue":"17","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32270062"],"award-info":[{"award-number":["32270062"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32271358"],"award-info":[{"award-number":["32271358"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010040","name":"Taishan Scholar Project of Shandong Province","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100010040","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The cleavage of the C10\u2013C4a bond in anthraquinones is a key step in generating ring-opened quinone derivatives in filamentous fungi. GedF, a short-chain dehydrogenase\/reductase (SDR) from Aspergillus terreus, together with the dioxygenase GedK, mediates this transformation, yet the enzymatic mechanism of GedF remains unclear. Here, isotope labeling experiments confirm that reduction of Questin-to-Questin hydroquinone incorporates one proton from NADPH and one from water. Structural modeling, molecular docking, and site-directed mutagenesis reveal that GedF employs a noncanonical catalytic architecture featuring a conserved Ser\u2013Tyr catalytic core instead of the classical Asn-Ser-Tyr-Lys tetrad typical of SDRs. Notably, mutagenesis and comparative analysis indicate that a positively charged residue is required for catalysis but is not strictly position-conserved, consistent with a role in maintaining the catalytic microenvironment and facilitating proton transfer. Phylogenetic and sequence analyses show that GedF belongs to the NAD(P)H-dependent SDR clade, and that variation in the positioning of basic residues occurs among homologues while preserving the conserved Ser\u2013Tyr catalytic core. These findings elucidate the catalytic mechanism of GedF and uncover an alternative SDR catalytic strategy involved in anthraquinone ring-opening biosynthesis in filamentous fungi.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00266","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T12:01:41Z","timestamp":1786536101000},"page":"2679-2688","source":"Crossref","is-referenced-by-count":0,"title":["Mechanistic Characterization of GedF Reveals a Ser\u2013Tyr\nCatalytic Core with a Flexible Basic Residue in a Filamentous Fungal\nAnthraquinone SDR"],"prefix":"10.1021","volume":"65","author":[{"given":"Ning","family":"Xu","sequence":"first","affiliation":[{"name":"University of Jinan , , , ,","place":["Jinan, Shandong, China, 250022"]},{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongsong","family":"Wu","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dan","family":"Liu","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keke","family":"Zhang","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danni","family":"Wu","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Du","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuenian","family":"Huang","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]},{"name":"Qingdao New Energy Shandong Laboratory , ,","place":["Qingdao, China, 266101"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 101408"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0570-9196","authenticated-orcid":true,"given":"Xuefeng","family":"Lu","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]},{"name":"Qingdao New Energy Shandong Laboratory , ,","place":["Qingdao, China, 266101"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 101408"]},{"name":"Qingdao National Laboratory for Marine Science and Technology , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Honglei","family":"Ma","sequence":"additional","affiliation":[{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong Energy Institute , ,","place":["Qingdao, China, 266101"]},{"name":"Qingdao New Energy Shandong Laboratory , ,","place":["Qingdao, China, 266101"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 101408"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2937-4909","authenticated-orcid":true,"given":"Feifei","family":"Qi","sequence":"additional","affiliation":[{"name":"University of Jinan , , , ,","place":["Jinan, Shandong, China, 250022"]},{"name":"Qingdao Institute of Bioenergy and Bioprocess Technology , , ,","place":["Qingdao, China, 266101"]},{"name":"Shandong University of Science and Technology , , ,","place":["Qingdao, China, 266590"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"issue":"8","key":"2026090306405816000_cit1","doi-asserted-by":"publisher","first-page":"3498","DOI":"10.7150\/ijbs.70447","article-title":"The versatile emodin:\nA natural easily acquired anthraquinone possesses\npromising anticancer properties against a variety of cancers","volume":"18","author":"Zhang","year":"2022","journal-title":"Int. J. Biol. Sci."},{"key":"2026090306405816000_cit2","doi-asserted-by":"publisher","first-page":"112854","DOI":"10.1016\/j.phytochem.2021.112854","article-title":"Emodin - A natural\nanthraquinone derivative with diverse pharmacological activities","volume":"190","author":"Semwal","year":"2021","journal-title":"Phytochem."},{"issue":"11","key":"2026090306405816000_cit3","doi-asserted-by":"publisher","first-page":"1016","DOI":"10.1177\/004051750107101113","article-title":"Emodin and dermocybin natural anthraquinones\nas mordant dyes for wool and polyamide","volume":"71","author":"Ra\u0308isa\u0308nen","year":"2001","journal-title":"Text.\nRes. J."},{"key":"2026090306405816000_cit4","doi-asserted-by":"publisher","first-page":"124176","DOI":"10.1016\/j.jhazmat.2020.124176","article-title":"Biodegradation and\nmetabolic pathway of anthraquinone dyes by Trametes hirsuta D7 immobilized in light expanded clay aggregate and cytotoxicity\nassessment","volume":"405","author":"Alam","year":"2021","journal-title":"J. Hazard. Mater."},{"issue":"3","key":"2026090306405816000_cit5","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1039\/C8NP00021B","article-title":"Classic fungal natural products in\nthe genomic age: the molecular\nlegacy of Harold Raistrick","volume":"35","author":"Schor","year":"2018","journal-title":"Nat. Prod. Rep."},{"issue":"1","key":"2026090306405816000_cit6","doi-asserted-by":"publisher","first-page":"174","DOI":"10.1039\/D2NP00040G","article-title":"The \u2019emodin family\u2019\nof fungal natural products-amalgamating a century of research with\nrecent genomics-based advances","volume":"40","author":"De Mattos-Shipley","year":"2023","journal-title":"Nat. Prod. Rep."},{"issue":"40","key":"2026090306405816000_cit7","doi-asserted-by":"publisher","first-page":"16326","DOI":"10.1021\/jacs.1c07182","article-title":"Bienzyme-catalytic\nand dioxygenation-mediated anthraquinone\nring opening","volume":"143","author":"Qi","year":"2021","journal-title":"J. Am. Chem. Soc."},{"issue":"2","key":"2026090306405816000_cit8","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1021\/acscatal.3c04053","article-title":"Mechanistic investigation\non C\u2013C bond cleavage of anthraquinone\ncatalyzed by an atypical nonheme iron-dependent dioxygenase BTG13","volume":"14","author":"Deng","year":"2024","journal-title":"ACS Catal."},{"issue":"24","key":"2026090306405816000_cit9","doi-asserted-by":"publisher","first-page":"10968","DOI":"10.1021\/jacs.2c04080","article-title":"Characterization\nof the oxazinomycin biosynthetic pathway revealing the key role of\na nonheme iron-dependent mono-oxygenase","volume":"144","author":"Ren","year":"2022","journal-title":"J.\nAm. Chem. Soc."},{"issue":"1","key":"2026090306405816000_cit10","doi-asserted-by":"publisher","first-page":"7682","DOI":"10.1038\/s41467-024-52101-2","article-title":"Substrate specificity of a branch of aromatic dioxygenases\ndetermined by three distinct motifs","volume":"15","author":"Cui","year":"2024","journal-title":"Nat. Commun."},{"issue":"18","key":"2026090306405816000_cit11","doi-asserted-by":"publisher","first-page":"6003","DOI":"10.1021\/bi00018a001","article-title":"Short-chain\ndehydrogenases\/reductases\n(SDR)","volume":"34","author":"Jo\u0308rnvall","year":"1995","journal-title":"Biochemistry"},{"key":"2026090306405816000_cit12","doi-asserted-by":"publisher","first-page":"3895","DOI":"10.1007\/s00018-008-8588-y","article-title":"The SDR superfamily:\nfunctional and structural diversity within a family of metabolic and\nregulatory enzymes","volume":"65","author":"Kavanagh","year":"2008","journal-title":"Cell. Mol. Life Sci."},{"key":"2026090306405816000_cit13","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.cbi.2012.11.009","article-title":"Classification and\nnomenclature of the superfamily\nof short-chain dehydrogenases\/reductases (SDRs)","volume":"202","author":"Persson","year":"2013","journal-title":"Chem.-Biol. Interact."},{"issue":"28","key":"2026090306405816000_cit14","doi-asserted-by":"publisher","first-page":"25677","DOI":"10.1074\/jbc.M202160200","article-title":"Critical residues for structure and\ncatalysis in short-chain\ndehydrogenases\/reductases","volume":"277","author":"Filling","year":"2002","journal-title":"J. Biol. Chem."},{"issue":"1","key":"2026090306405816000_cit15","doi-asserted-by":"publisher","first-page":"105596","DOI":"10.1016\/j.jbc.2023.105596","article-title":"Specific residues\nand conformational plasticity define the substrate specificity of\nshort-chain dehydrogenases\/reductases","volume":"300","author":"Qian","year":"2024","journal-title":"J. Biol.\nChem."},{"issue":"7","key":"2026090306405816000_cit16","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1016\/S0969-2126(00)00064-2","article-title":"The refined\nthree-dimensional structure of 3\u03b1,20\u03b2-hydroxysteroid dehydrogenase and possible roles\nof the residues conserved in short-chain dehydrogenases","volume":"2","author":"Ghosh","year":"1994","journal-title":"Structure"},{"issue":"12","key":"2026090306405816000_cit17","doi-asserted-by":"publisher","first-page":"8716","DOI":"10.1021\/jacs.4c01284","article-title":"Discovery of a fungal\nP450 with an unusual two-step mechanism for constructing a bicyclo[3.2.2]nonane\nskeleton","volume":"146","author":"Xu","year":"2024","journal-title":"J. Am. Chem. Soc."},{"issue":"31","key":"2026090306405816000_cit18","doi-asserted-by":"publisher","first-page":"19551","DOI":"10.1021\/acs.jafc.5c03575","article-title":"Mining UDP-glucosyltransferases\nto\ndevelop high-efficiency production strategies for emodin glucosides.\nJ","volume":"73","author":"Wu","year":"2025","journal-title":"Agric. Food Chem."},{"key":"2026090306405816000_cit19","doi-asserted-by":"publisher","first-page":"615955","DOI":"10.1101\/2024.10.10.615955","article-title":"Chai-1: decoding the molecular interactions\nof life","volume":"2024","author":"Boitreaud","year":"2024","journal-title":"bioRxiv"},{"issue":"2","key":"2026090306405816000_cit20","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1093\/bioinformatics\/10.2.189","article-title":"MEGA: Molecular\nevolutionary genetics\nanalysis software for microcomputers","volume":"10","author":"Kumar","year":"1994","journal-title":"Comput.\nAppl. Biosci."},{"issue":"4","key":"2026090306405816000_cit21","doi-asserted-by":"publisher","first-page":"406","DOI":"10.1093\/oxfordjournals.molbev.a040454","article-title":"The neighbor-joining\nmethod: a new method for reconstructing phylogenetic\ntrees","volume":"4","author":"Saitou","year":"1987","journal-title":"Mol. Biol. Evol."},{"issue":"W1","key":"2026090306405816000_cit22","doi-asserted-by":"publisher","first-page":"W242","DOI":"10.1093\/nar\/gkw290","article-title":"Interactive tree of\nlife (iTOL) v3: an online tool for the display\nand annotation of phylogenetic and other trees","volume":"44","author":"Letunic","year":"2016","journal-title":"Nucleic Acids Res."},{"issue":"13","key":"2026090306405816000_cit23","doi-asserted-by":"publisher","first-page":"3962","DOI":"10.1128\/AEM.00261-14","article-title":"Deciphering\nthe effect of the different N-glycosylation sites on the secretion,\nactivity, and stability of cellobiohydrolase I from Trichoderma reesei","volume":"80","author":"Qi","year":"2014","journal-title":"Appl. Environ. Microbiol."},{"issue":"19","key":"2026090306405816000_cit24","doi-asserted-by":"publisher","first-page":"4933","DOI":"10.1093\/emboj\/cdg479","article-title":"Structure and mechanism\nof a bacterial haloalcohol dehalogenase: a new variation of the short-chain\ndehydrogenase\/reductase fold without an NAD(P)H binding site","volume":"22","author":"De Jong","year":"2003","journal-title":"EMBO J."},{"issue":"9","key":"2026090306405816000_cit25","doi-asserted-by":"publisher","first-page":"3709","DOI":"10.1105\/tpc.114.127399","article-title":"Structural studies of cinnamoyl-CoA\nreductase and cinnamyl-alcohol dehydrogenase, key enzymes of monolignol\nbiosynthesis","volume":"26","author":"Pan","year":"2014","journal-title":"Plant Cell"},{"issue":"10","key":"2026090306405816000_cit26","doi-asserted-by":"publisher","first-page":"1318","DOI":"10.1107\/s2053230x14019785","article-title":"Structure\nof a short-chain dehydrogenase\/reductase (SDR) within a genomic island\nfrom a clinical strain of Acinetobacter baumannii","volume":"70","author":"Shah","year":"2014","journal-title":"Acta Crystallogr. F Struct. Biol. Commun."},{"issue":"22","key":"2026090306405816000_cit27","doi-asserted-by":"publisher","first-page":"15582","DOI":"10.1074\/jbc.274.22.15582","article-title":"Crystal\nstructure of the Mycobacterium tuberculosis enoyl-ACP\nreductase, InhA, in complex with NAD+ and a C16 fatty acyl substrate","volume":"274","author":"Rozwarski","year":"1999","journal-title":"J. Biol. Chem."},{"issue":"2","key":"2026090306405816000_cit28","doi-asserted-by":"publisher","first-page":"527","DOI":"10.1006\/jmbi.1999.3240","article-title":"Crystallographic\nanalysis of triclosan bound to enoyl reductase","volume":"294","author":"Roujeinikova","year":"1999","journal-title":"J. Mol. Biol."},{"issue":"12","key":"2026090306405816000_cit29","doi-asserted-by":"publisher","first-page":"1115","DOI":"10.1038\/s41929-019-0382-8","article-title":"Deciphering the enzymatic mechanism of sugar ring contraction\nin UDP-apiose biosynthesis","volume":"2","author":"Savino","year":"2019","journal-title":"Nat. Catal."},{"issue":"44","key":"2026090306405816000_cit30","doi-asserted-by":"publisher","first-page":"8844","DOI":"10.1021\/bi301135b","article-title":"Human UDP-\u03b1-D-xylose synthase and Escherichia coli ArnA conserve\na conformational shunt that controls whether xylose or 4-keto-xylose\nis produced","volume":"51","author":"Polizzi","year":"2012","journal-title":"Biochemistry"},{"issue":"24","key":"2026090306405816000_cit31","doi-asserted-by":"publisher","first-page":"23000","DOI":"10.1074\/jbc.M501534200","article-title":"Structure and function\nof both domains of ArnA, a dual function decarboxylase and a formyltransferase,\ninvolved in 4-amino-4-deoxy-L-arabinose biosynthesis","volume":"280","author":"Williams","year":"2005","journal-title":"J. Biol. Chem."},{"issue":"11","key":"2026090306405816000_cit32","doi-asserted-by":"publisher","first-page":"4051","DOI":"10.1128\/JB.01866-05","article-title":"The X-ray structure of the haloalcohol dehalogenase\nHheA from Arthrobacter sp. strain AD2: insight into enantioselectivity\nand halide binding in the haloalcohol dehalogenase family","volume":"188","author":"De Jong","year":"2006","journal-title":"J. Bacteriol."},{"issue":"12","key":"2026090306405816000_cit33","doi-asserted-by":"publisher","first-page":"2230","DOI":"10.1002\/prot.24938","article-title":"Crystal structures of halohydrin hydrogen-halide-lyases from Corynebacterium sp. N-1074","volume":"83","author":"Watanabe","year":"2015","journal-title":"Proteins"},{"issue":"6","key":"2026090306405816000_cit34","doi-asserted-by":"publisher","first-page":"1188","DOI":"10.1101\/gr.849004","article-title":"WebLogo: a sequence logo generator","volume":"14","author":"Crooks","year":"2004","journal-title":"Genome Res."}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.biochem.6c00266","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/17\/2679\/66507957\/acs.biochem.6c00266.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/17\/2679\/66507957\/acs.biochem.6c00266.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:41:04Z","timestamp":1788432064000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/65\/17\/2679\/5253254\/Mechanistic-Characterization-of-GedF-Reveals-a-Ser"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,12]]},"references-count":34,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,8,12]]},"published-print":{"date-parts":[[2026,9,1]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.biochem.6c00266","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,12]]}}}