{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T16:12:45Z","timestamp":1788970365739,"version":"build-2803163510"},"reference-count":34,"publisher":"American Chemical Society (ACS)","issue":"35","license":[{"start":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T00:00:00Z","timestamp":1787875200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","award":["202208350014"],"award-info":[{"award-number":["202208350014"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","award":["202406210070"],"award-info":[{"award-number":["202406210070"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]},{"name":"German Research Council (DFG)","award":["INST 160\/620-1"],"award-info":[{"award-number":["INST 160\/620-1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Lactone-containing natural products are important candidates for drug discovery. Drimane-type sesquiterpenes (DTSs), characterized by a bicyclic trans-decalin scaffold, can bear both \u03b2- and \u03b3-lactone moieties. While \u03b3-lactone-containing DTSs have frequently been reported, \u03b2-lactone-containing derivatives are rare, and their biosynthesis remains unexplored. Here, we identified a biosynthetic gene cluster (dri) in Aspergillus ustus and confirmed ustidrimane A (1), a \u03b2- and \u03b3-lactone-containing DTS, as its product. Heterologous gene expression, precursor feeding, and enzymatic investigation provided evidence for the formation of both lactone rings. In both cases, the reaction cascade is initiated by regiospecific (and stereoselective) methyl hydroxylation, followed by regiospecific and stereoselective oxidation of one hydroxymethyl group to an aldehyde. The resulting hemiacetal was proven to be subsequently oxidized to a lactone. The \u03b2-lactone formation is catalyzed by two cytochrome P450 enzymes (DriE and DriF), followed by two oxidation steps catalyzed by two dehydrogenases (DriG and DriH). These findings differ entirely from the known \u03b2-lactone formation in fatty acid-, PKS-, and NRPS-derived metabolites. The subsequent \u03b3-lactone formation is catalyzed by a P450 (DriJ) and a dehydrogenase (DriD). DriJ has been shown to be involved in both methyl hydroxylation and hemiacetal formation, while DriD is responsible for the hemiacetal oxidation and also contributes moderately to its formation. Collectively, these findings establish a sequential P450\/dehydrogenase-mediated oxidative cascade for the construction of two distinct lactone motifs within a single DTS scaffold. Moreover, they provide the first insight into the \u03b2-lactone formation in terpenes, thus unveiling a new strategy for the construction of this structural motif.<\/jats:p>","DOI":"10.1021\/jacs.6c13983","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T01:31:19Z","timestamp":1788312679000},"page":"38483-38490","source":"Crossref","is-referenced-by-count":0,"title":["Cytochrome\nP450- and Dehydrogenase-Mediated Regiospecific\nand Stereoselective Formation of \u03b2- and \u03b3-Lactones\nin Drimane-Type Sesquiterpenoid Biosynthesis"],"prefix":"10.1021","volume":"148","author":[{"given":"Meiting","family":"Wu","sequence":"first","affiliation":[{"name":"Philipps-Universita\u0308t Marburg , , Robert-Koch-Stra\u00dfe 4 , ,","place":["Marburg, Germany, 35037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingjing","family":"Wu","sequence":"additional","affiliation":[{"name":"Philipps-Universita\u0308t Marburg , , Robert-Koch-Stra\u00dfe 4 , ,","place":["Marburg, Germany, 35037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1324-9580","authenticated-orcid":true,"given":"Daniel J.","family":"Janzen","sequence":"additional","affiliation":[{"name":"Philipps-Universita\u0308t Marburg , , Robert-Koch-Stra\u00dfe 4 , ,","place":["Marburg, Germany, 35037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9184-3198","authenticated-orcid":true,"given":"Wen-Bing","family":"Yin","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Chinese Academy of Sciences , , ,","place":["Beijing, China, 100101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4583-2655","authenticated-orcid":true,"given":"Shu-Ming","family":"Li","sequence":"additional","affiliation":[{"name":"Philipps-Universita\u0308t Marburg , , Robert-Koch-Stra\u00dfe 4 , ,","place":["Marburg, Germany, 35037"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,28]]},"reference":[{"key":"2026090910102185000_cit1","doi-asserted-by":"publisher","first-page":"132001","DOI":"10.1016\/j.tet.2021.132001","article-title":"Lactones: Classification, synthesis,\nbiological activities, and industrial applications","volume":"84","author":"Sartori","year":"2021","journal-title":"Tetrahedron"},{"key":"2026090910102185000_cit2","doi-asserted-by":"publisher","first-page":"2769","DOI":"10.3390\/ijms22052769","article-title":"A review of the pharmacological activities\nand recent synthetic advances of \u03b3-butyrolactones","volume":"22","author":"Hur","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"2026090910102185000_cit3","doi-asserted-by":"publisher","first-page":"84","DOI":"10.7164\/antibiotics.42.84","article-title":"Lactivicin,\na naturally occurring non-beta-lactam antibiotic having beta-lactam-like\naction: Biological activities and mode of action","volume":"42","author":"Nozaki","year":"1989","journal-title":"J. Antibiot."},{"key":"2026090910102185000_cit4","doi-asserted-by":"publisher","first-page":"15935","DOI":"10.1038\/ncomms15935","article-title":"An L-threonine\ntransaldolase is required for L-threo-\u03b2-hydroxy-\u03b1-amino\nacid assembly during obafluorin biosynthesis","volume":"8","author":"Scott","year":"2017","journal-title":"Nat. Commun."},{"key":"2026090910102185000_cit5","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1039\/C8NP00052B","article-title":"Biosynthesis and chemical diversity\nof \u03b2-lactone natural products","volume":"36","author":"Robinson","year":"2019","journal-title":"Nat. Prod.\nRep."},{"key":"2026090910102185000_cit6","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.7164\/antibiotics.40.1081","article-title":"Lipstatin,\nan inhibitor of pancreatic lipase, produced by Streptomyces toxytricini. I. Producing organism, fermentation, isolation\nand biological activity","volume":"40","author":"Weibel","year":"1987","journal-title":"J. Antibiot."},{"key":"2026090910102185000_cit7","doi-asserted-by":"publisher","first-page":"1594","DOI":"10.7164\/antibiotics.33.1594","article-title":"Ebelactone,\nan inhibitor of esterase, produced by actinomycetes","volume":"33","author":"Umezawa","year":"1980","journal-title":"J. Antibiot."},{"key":"2026090910102185000_cit8","doi-asserted-by":"publisher","first-page":"7488","DOI":"10.1073\/pnas.84.21.7488","article-title":"Inhibition of hydroxymethylglutaryl-coenzyme\nA synthase\nby L-659,699","volume":"84","author":"Greenspan","year":"1987","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090910102185000_cit9","doi-asserted-by":"publisher","first-page":"5749","DOI":"10.1021\/ol062307u","article-title":"Vibralactone:\nA lipase inhibitor\nwith an unusual fused beta-lactone produced by cultures of the basidiomycete Boreostereum vibrans","volume":"8","author":"Liu","year":"2006","journal-title":"Org. Lett."},{"key":"2026090910102185000_cit10","doi-asserted-by":"publisher","first-page":"355","DOI":"10.1002\/anie.200390115","article-title":"Salinosporamide A:\na highly cytotoxic proteasome inhibitor\nfrom a novel microbial source, a marine bacterium of the new genus Salinospora","volume":"42","author":"Feling","year":"2003","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090910102185000_cit11","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1016\/0304-3940(81)90105-1","article-title":"Anisatin,\na potent GABA antagonist,\nisolated from Illicium anisatum","volume":"25","author":"Kudo","year":"1981","journal-title":"Neurosci. Lett."},{"key":"2026090910102185000_cit12","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1016\/0005-2760(88)90260-3","article-title":"Mode of action\nof tetrahydrolipstatin: A derivative of the naturally occurring lipase\ninhibitor lipstatin","volume":"962","author":"Borgstro\u0308m","year":"1988","journal-title":"Biochim. Biophys. Acta"},{"key":"2026090910102185000_cit13","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1021\/acs.biochem.6b01199","article-title":"\u03b2-Lactone\nsynthetase found in the olefin biosynthesis\npathway","volume":"56","author":"Christenson","year":"2017","journal-title":"Biochemistry"},{"key":"2026090910102185000_cit14","doi-asserted-by":"publisher","first-page":"3432","DOI":"10.1038\/s41467-019-11383-7","article-title":"The structural basis of N-acyl-\u03b1-amino-\u03b2-lactone\nformation catalyzed by a nonribosomal peptide synthetase","volume":"10","author":"Kreitler","year":"2019","journal-title":"Nat. Commun."},{"key":"2026090910102185000_cit15","doi-asserted-by":"publisher","first-page":"421","DOI":"10.1038\/ja.2013.48","article-title":"Biosynthesis\nof ebelactone A: Isotopic tracer, advanced\nprecursor and genetic studies reveal a thioesterase-independent cyclization\nto give a polyketide \u03b2-lactone","volume":"66","author":"Wyatt","year":"2013","journal-title":"J. Antibiot."},{"key":"2026090910102185000_cit16","doi-asserted-by":"publisher","first-page":"7209","DOI":"10.1002\/anie.202000710","article-title":"A hydrolase-catalyzed\ncyclization forms the fused bicyclic \u03b2-lactone\nin vibralactone","volume":"59","author":"Feng","year":"2020","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090910102185000_cit17","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1093\/plcell\/koaa023","article-title":"Interdependent evolution\nof biosynthetic gene clusters for momilactone production in rice","volume":"33","author":"Kitaoka","year":"2021","journal-title":"Plant Cell"},{"key":"2026090910102185000_cit18","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202502766","article-title":"Cooperative redox reactions encoded\nby two gene clusters enable intermolecular cycloaddition cascade for\nthe formation of meroaspochalasins","volume":"64","author":"Li","year":"2025","journal-title":"Angew. Chem.,\nInt. Ed."},{"key":"2026090910102185000_cit19","doi-asserted-by":"publisher","first-page":"815","DOI":"10.3762\/bjoc.20.73","article-title":"Discovery and biosynthesis\nof bacterial drimane-type sesquiterpenoids from Streptomyces clavuligerus","volume":"20","author":"Zhang","year":"2024","journal-title":"Beilstein J. Org. Chem."},{"key":"2026090910102185000_cit20","doi-asserted-by":"publisher","first-page":"737","DOI":"10.1016\/S1875-5364(22)60190-0","article-title":"Drimane-type sesquiterpenoids\nfrom fungi","volume":"20","author":"Du","year":"2022","journal-title":"Chin. J. Nat. Med."},{"key":"2026090910102185000_cit21","doi-asserted-by":"publisher","first-page":"1026","DOI":"10.3390\/jof7121026","article-title":"Sesquiterpenoids\nspecially produced\nby fungi: Structures, biological activities, chemical and biosynthesis\n(2015\u20132020)","volume":"7","author":"Dai","year":"2021","journal-title":"J. Fungi."},{"key":"2026090910102185000_cit22","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202300518","article-title":"Terpene synthases\nin the biosynthesis of drimane-type\nsesquiterpenes across diverse organisms","volume":"24","author":"Chen","year":"2023","journal-title":"ChemBiochem"},{"issue":"4","key":"2026090910102185000_cit23","doi-asserted-by":"publisher","first-page":"645","DOI":"10.3390\/molecules31040645","article-title":"Drimane sesquiterpenes\nisolated from the fruiting body of the clavarioid fungus Ramaria botrytoides","volume":"31","author":"Peng","year":"2026","journal-title":"Molecules"},{"key":"2026090910102185000_cit24","doi-asserted-by":"publisher","first-page":"1491","DOI":"10.1021\/acs.jnatprod.5c00431","article-title":"Spinulactones A-F:\nBioactive drimane-type sesquiterpenes\nfrom the endophytic Penicillium spinulosum 9\u20131","volume":"88","author":"Zhang","year":"2025","journal-title":"J. Nat. Prod."},{"key":"2026090910102185000_cit25","doi-asserted-by":"publisher","first-page":"113391","DOI":"10.1016\/j.phytochem.2022.113391","article-title":"The induced cryptic metabolites and\nantifungal activities\nfrom culture of Penicillium chrysogenum by supplementing\nwith host Ziziphus jujuba extract","volume":"203","author":"Li","year":"2022","journal-title":"Phytochemistry"},{"key":"2026090910102185000_cit26","doi-asserted-by":"publisher","first-page":"1052","DOI":"10.1111\/tpj.13527","article-title":"Identification of a drimenol synthase and drimenol oxidase from Persicaria hydropiper, involved in the biosynthesis of insect\ndeterrent drimanes","volume":"90","author":"Henquet","year":"2017","journal-title":"Plant J."},{"key":"2026090910102185000_cit27","doi-asserted-by":"publisher","first-page":"4597","DOI":"10.1016\/j.febslet.2014.10.031","article-title":"Molecular cloning\nand characterization of drimenol synthase from valerian plant (Valeriana officinalis)","volume":"588","author":"Kwon","year":"2014","journal-title":"FEBS Lett."},{"key":"2026090910102185000_cit28","doi-asserted-by":"publisher","first-page":"32865","DOI":"10.1038\/srep32865","article-title":"Identification of a\nnovel sesquiterpene biosynthetic machinery involved in astellolide\nbiosynthesis","volume":"6","author":"Shinohara","year":"2016","journal-title":"Sci. Rep."},{"key":"2026090910102185000_cit29","doi-asserted-by":"publisher","first-page":"23763","DOI":"10.1002\/anie.202108970","article-title":"Biosynthesis of fungal drimane-type\nsesquiterpene esters","volume":"60","author":"Huang","year":"2021","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090910102185000_cit30","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202215566","article-title":"The biosynthetic\ngene cluster of mushroom-derived antrocin encodes two dual-functional\nhaloacid dehalogenase-like terpene cyclases","volume":"62","author":"Chen","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090910102185000_cit31","doi-asserted-by":"publisher","first-page":"408","DOI":"10.3390\/md20070408","article-title":"Sesquiterpenoids\nfrom the mangrove-derived Aspergillus ustus 094102","volume":"20","author":"Gui","year":"2022","journal-title":"Mar. Drugs"},{"key":"2026090910102185000_cit32","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1021\/acs.jnatprod.0c00910","article-title":"Antibacterial drimane\nsesquiterpenes from Aspergillus ustus","volume":"84","author":"Neuhaus","year":"2021","journal-title":"J. Nat. Prod."},{"key":"2026090910102185000_cit33","doi-asserted-by":"publisher","first-page":"762","DOI":"10.1248\/cpb.59.762","article-title":"Drimane sesquiterpenoids\nfrom the mangrove-derived fungus Aspergillus ustus","volume":"59","author":"Zhou","year":"2011","journal-title":"Chem. Pharm. Bull."},{"key":"2026090910102185000_cit34","doi-asserted-by":"publisher","first-page":"1585","DOI":"10.1021\/np900220r","article-title":"Drimane sesquiterpenoids\nfrom the fungus Aspergillus ustus isolated from the marine sponge Suberites domuncula","volume":"72","author":"Liu","year":"2009","journal-title":"J. Nat. Prod."}],"container-title":["Journal of the American Chemical Society"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacs.6c13983","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/35\/38483\/67577593\/jacs.6c13983.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/35\/38483\/67577593\/jacs.6c13983.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T15:28:15Z","timestamp":1788967695000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jacsat\/article\/148\/35\/38483\/5346687\/Cytochrome-P450-and-Dehydrogenase-Mediated"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,28]]},"references-count":34,"journal-issue":{"issue":"35","published-online":{"date-parts":[[2026,8,28]]},"published-print":{"date-parts":[[2026,9,9]]}},"URL":"https:\/\/doi.org\/10.1021\/jacs.6c13983","relation":{},"ISSN":["0002-7863","1520-5126"],"issn-type":[{"value":"0002-7863","type":"print"},{"value":"1520-5126","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,28]]}}}