{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T22:06:56Z","timestamp":1788991616113,"version":"build-2803163510"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"35","license":[{"start":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T00:00:00Z","timestamp":1787702400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T00:00:00Z","timestamp":1787702400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T00:00:00Z","timestamp":1787702400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project","doi-asserted-by":"publisher","award":["TSBICIP-CXRC-056"],"award-info":[{"award-number":["TSBICIP-CXRC-056"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"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>Jujubosides are the principal bioactive constituents of the traditional Chinese medicine Ziziphi Spinosae Semen and are well-known for their sedative, anxiolytic, and sleep-promoting activities. These compounds are dammarane-type triterpenoid saponins characterized by a unique aglycone, jujubogenin, featuring a 5,6-spiroketal moiety that forms a rigid D\/E\/F tricyclic scaffold. However, the enzymatic basis underlying the formation of this distinctive spiroketal structure remains elusive. In this study, we identified and functionally characterized three cytochrome P450 enzymes from wild jujubes and Bacopa monnieri. These enzymes cooperatively catalyze an oxidative cascade in which site-selective oxidations at C-16, C-23, and C-30 drive spiroketal formation, converting dammarenediol II into jujubogenin. We further reconstituted the complete biosynthetic pathway in engineered yeast, enabling the de novo production of jujubogenin. This study establishes a foundational platform for the complete biosynthesis of jujubosides in microbial cell factories.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06727","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:00:21Z","timestamp":1788253221000},"page":"27580-27591","source":"Crossref","is-referenced-by-count":0,"title":["Identification of Three Cytochrome P450 Enzymes Responsible\nfor the Biosynthesis of Jujubogenin and Its\n                    <i>De Novo<\/i>\n                    Production in Yeast"],"prefix":"10.1021","volume":"74","author":[{"given":"Jun","family":"Tang","sequence":"first","affiliation":[{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Liu","sequence":"additional","affiliation":[{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"Tianjin University of Traditional Chinese Medicine , ,","place":["Tianjin, China, 301617"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linhua","family":"Zhao","sequence":"additional","affiliation":[{"name":"Guang\u2019anmen Hospital, China Academy of Chinese Medical Sciences , , ,","place":["Beijing, China, 100053"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7076-1273","authenticated-orcid":true,"given":"Tao","family":"Liu","sequence":"additional","affiliation":[{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolin","family":"Tong","sequence":"additional","affiliation":[{"name":"Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"Guang\u2019anmen Hospital, China Academy of Chinese Medical Sciences , , ,","place":["Beijing, China, 100053"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,26]]},"reference":[{"issue":"8","key":"2026090917080501000_cit1","doi-asserted-by":"publisher","first-page":"1349","DOI":"10.1016\/S0031-9422(00)94587-4","article-title":"The structure of jujubosides A and B, the saponins\nisolated from the seeds of Zizyphus jujuba","volume":"17","author":"Otsuka","year":"1978","journal-title":"Phytochemistry"},{"issue":"1","key":"2026090917080501000_cit2","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/j.jep.2010.03.023","article-title":"Hypnotic effect\nof jujubosides from Semen Ziziphi Spinosae","volume":"130","author":"Cao","year":"2010","journal-title":"J. Ethnopharmacol."},{"issue":"1","key":"2026090917080501000_cit3","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1080\/07391102.2008.10507223","article-title":"What is\nthe effective\ncomponent in suanzaoren decoction for curing insomnia? Discovery by\nvirtual screening and molecular dynamic simulation","volume":"26","author":"Chen","year":"2008","journal-title":"J. Biomol. Struct. Dyn."},{"issue":"2","key":"2026090917080501000_cit4","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1016\/j.jep.2010.01.034","article-title":"Effects on the expression\nof GABA receptor subunits by jujuboside A treatment in rat hippocampal\nneurons","volume":"128","author":"You","year":"2010","journal-title":"J. Ethnopharmacol."},{"issue":"15","key":"2026090917080501000_cit5","doi-asserted-by":"publisher","first-page":"4262","DOI":"10.7150\/thno.26164","article-title":"Jujuboside\nA promotes A\u03b2 clearance and ameliorates\ncognitive deficiency in Alzheimer\u2019s disease through activating\nAxl\/HSP90\/PPAR\u03b3 pathway","volume":"8","author":"Zhang","year":"2018","journal-title":"Theranostics"},{"key":"2026090917080501000_cit6","doi-asserted-by":"publisher","first-page":"4512","DOI":"10.1038\/s41598-019-41114-3","article-title":"Jujuboside A prevents\nsleep loss-induced disturbance of hippocampal\nneuronal excitability and memory impairment in young APP\/PS1 mice","volume":"9","author":"Tabassum","year":"2019","journal-title":"Sci. Rep."},{"issue":"1","key":"2026090917080501000_cit7","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1515\/BC.2000.009","article-title":"Adjuvant and\nhaemolytic activities of 47 saponins derived\nfrom medicinal and food plants","volume":"381","author":"Oda","year":"2000","journal-title":"Biol. Chem."},{"issue":"1","key":"2026090917080501000_cit8","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/S0031-9422(00)97026-2","article-title":"Bacoside\nA3-a triterpenoid saponin\nfrom Bacopa monniera","volume":"36","author":"Rastogi","year":"1994","journal-title":"Phytochemistry"},{"issue":"3","key":"2026090917080501000_cit9","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1055\/s-2007-969908","article-title":"13C-NMR\nstudy and pharmacology of two saponins from Colubrina asiatica","volume":"48","author":"Wagner","year":"1983","journal-title":"Planta Med."},{"issue":"9","key":"2026090917080501000_cit10","doi-asserted-by":"publisher","first-page":"2287","DOI":"10.1248\/cpb.40.2287","article-title":"Antisweet natural products.\nVII. Hodulosides I, II, III, IV, and V from the leaves of Hovenia\ndulcis THUNB","volume":"40","author":"Yoshikawa","year":"1992","journal-title":"Chem. Pharm. Bull."},{"issue":"15","key":"2026090917080501000_cit11","doi-asserted-by":"publisher","first-page":"5856","DOI":"10.1039\/C4AY00800F","article-title":"Analysis\nof six bioactive components in Semen Ziziphi Spinosae by UPLC-ELSD\nand UPLC-Q\/TOF-MS","volume":"6","author":"Zhao","year":"2014","journal-title":"Anal. Methods"},{"issue":"26","key":"2026090917080501000_cit12","doi-asserted-by":"publisher","first-page":"3370","DOI":"10.1016\/j.tet.2018.04.051","article-title":"Synthesis of the hexacyclic triterpene\ncore of the jujuboside saponins via tandem Wolff rearrangement-intramolecular\nketene hetero-Diels-Alder reaction","volume":"74","author":"Karimov","year":"2018","journal-title":"Tetrahedron"},{"issue":"1","key":"2026090917080501000_cit13","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1038\/s41421-018-0075-5","article-title":"Synthesizing ginsenoside\nRh2 in Saccharomyces cerevisiae cell factory at high-efficiency","volume":"5","author":"Wang","year":"2019","journal-title":"Cell Discovery"},{"issue":"7446","key":"2026090917080501000_cit14","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1038\/nature12051","article-title":"High-level semi-synthetic\nproduction of the potent\nantimalarial artemisinin","volume":"496","author":"Paddon","year":"2013","journal-title":"Nature"},{"key":"2026090917080501000_cit15","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1016\/j.ymben.2025.01.002","article-title":"De novo\nbiosynthesis of mogroside V by multiplexed engineered yeasts","volume":"88","author":"Qu","year":"2025","journal-title":"Metab. Eng."},{"issue":"8013","key":"2026090917080501000_cit16","doi-asserted-by":"publisher","first-page":"937","DOI":"10.1038\/s41586-024-07345-9","article-title":"Complete biosynthesis\nof QS-21 in engineered\nyeast","volume":"629","author":"Liu","year":"2024","journal-title":"Nature"},{"issue":"22","key":"2026090917080501000_cit17","doi-asserted-by":"publisher","first-page":"5143","DOI":"10.1016\/j.febslet.2006.08.044","article-title":"Dammarenediol-II synthase,\nthe first dedicated enzyme for ginsenoside biosynthesis, in Panax\nginseng","volume":"580","author":"Tansakul","year":"2006","journal-title":"FEBS Lett."},{"issue":"17","key":"2026090917080501000_cit18","doi-asserted-by":"publisher","first-page":"4431","DOI":"10.1021\/acs.jafc.8b01272","article-title":"Metabolic\nengineering of for high-level production of salidroside\nfrom glucose","volume":"66","author":"Jiang","year":"2018","journal-title":"J. Agr. Food Chem."},{"issue":"9","key":"2026090917080501000_cit19","doi-asserted-by":"publisher","first-page":"5083","DOI":"10.1021\/jacs.2c10838","article-title":"Post-cyclization skeletal rearrangements\nin plant triterpenoid biosynthesis by a pair of branchpoint isomerases","volume":"145","author":"Chuang","year":"2023","journal-title":"J. Am. Chem. Soc."},{"key":"2026090917080501000_cit20","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1093\/hr\/uhae071","article-title":"Haplotype-resolved\nT2T reference genomes for wild and domesticated accessions shed new\ninsights into the domestication of jujube","volume":"11","author":"Li","year":"2024","journal-title":"Hortic.\nRes."},{"key":"2026090917080501000_cit21","doi-asserted-by":"publisher","first-page":"1289","DOI":"10.3762\/bjoc.18.135","article-title":"Cytochrome P450 monooxygenase-mediated tailoring of\ntriterpenoids and steroids in plants","volume":"18","author":"Malhotra","year":"2022","journal-title":"Beilstein\nJ. Org. 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Soc.,\nB"},{"issue":"1","key":"2026090917080501000_cit24","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1111\/j.1365-313X.2011.04529.x","article-title":"A P450-centric view of plant evolution","volume":"66","author":"Nelson","year":"2011","journal-title":"Plant J."},{"issue":"3","key":"2026090917080501000_cit25","doi-asserted-by":"publisher","first-page":"1236","DOI":"10.1111\/tpj.17044","article-title":"Sequence diversity\nin the monooxygenases involved in oxime production in plant defense\nand signaling: a conservative revision in the nomenclature of the\nhighly complex CYP79 family","volume":"120","author":"Koleva","year":"2024","journal-title":"Plant J."},{"key":"2026090917080501000_cit26","doi-asserted-by":"publisher","first-page":"1886","DOI":"10.3389\/fpls.2017.01886","article-title":"Triterpene\nstructural diversification by plant cytochrome P450 enzymes","volume":"8","author":"Ghosh","year":"2017","journal-title":"Front. Plant Sci."},{"issue":"12","key":"2026090917080501000_cit27","doi-asserted-by":"publisher","first-page":"2062","DOI":"10.1093\/pcp\/pcr150","article-title":"Cyt P450 enzyme\nCYP716A47 catalyzes the formation of protopanaxadiol from dammarenediol-II\nduring ginsenoside biosynthesis in Panax ginseng","volume":"52","author":"Han","year":"2011","journal-title":"Plant Cell Physiol."},{"issue":"9","key":"2026090917080501000_cit28","doi-asserted-by":"publisher","first-page":"1535","DOI":"10.1093\/pcp\/pcs106","article-title":"Cytochrome\nP450 CYP716A53v2 catalyzes the formation of protopanaxatriol from\nprotopanaxadiol during ginsenoside biosynthesis in Panax ginseng","volume":"53","author":"Han","year":"2012","journal-title":"Plant Cell Physiol."},{"key":"2026090917080501000_cit29","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.jbiotec.2015.10.005","article-title":"Metabolic\nengineering of Pichia pastoris for the production of dammarenediol-II","volume":"216","author":"Liu","year":"2015","journal-title":"J. Biotechnol."},{"key":"2026090917080501000_cit30","doi-asserted-by":"publisher","DOI":"10.1002\/ange.202425303","article-title":"Completing the biosynthesis\nof the clinically important diterpenoid\nandrographolide in Andrographis paniculata","volume":"137","author":"Wang","year":"2025","journal-title":"Angew. Chem., Int. Ed."},{"issue":"5","key":"2026090917080501000_cit31","doi-asserted-by":"publisher","first-page":"1947","DOI":"10.1073\/pnas.1215788110","article-title":"CYP714B1 and\nCYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity\nin rice","volume":"110","author":"Magome","year":"2013","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"49","key":"2026090917080501000_cit32","doi-asserted-by":"publisher","first-page":"18662","DOI":"10.1074\/jbc.RA119.009944","article-title":"Allylic hydroxylation of triterpenoids by a plant cytochrome\nP450 triggers key chemical transformations that produce a variety\nof bitter compounds","volume":"294","author":"Takase","year":"2019","journal-title":"J. Biol. Chem."},{"issue":"37","key":"2026090917080501000_cit33","doi-asserted-by":"publisher","first-page":"14204","DOI":"10.1073\/pnas.0803876105","article-title":"Licorice beta-amyrin 11-oxidase, a cytochrome P450\nwith a key role in the biosynthesis of the triterpene sweetener glycyrrhizin","volume":"105","author":"Seki","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"26","key":"2026090917080501000_cit34","doi-asserted-by":"publisher","first-page":"7871","DOI":"10.1021\/acs.jafc.2c01905","article-title":"Wild jujube\n(Ziziphus\njujuba var. spinosa): a review of its phytonutrients, health benefits,\nmetabolism, and applications","volume":"70","author":"Hua","year":"2022","journal-title":"J. Agric. Food\nChem."},{"key":"2026090917080501000_cit35","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.ymben.2024.11.012","article-title":"High-level\nsustainable\nproduction of complex phenylethanoid glycosides from glucose through\nengineered yeast cell factories","volume":"87","author":"Bai","year":"2025","journal-title":"Metab. Eng."},{"issue":"6728","key":"2026090917080501000_cit36","doi-asserted-by":"publisher","first-page":"1366","DOI":"10.1126\/science.ado3409","article-title":"A scaffold protein manages the biosynthesis\nof steroidal\ndefense metabolites in plants","volume":"386","author":"Boccia","year":"2024","journal-title":"Science"},{"issue":"5","key":"2026090917080501000_cit37","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1038\/s41477-018-0142-9","article-title":"The scaffold proteins\nof lignin biosynthetic cytochrome P450 enzymes","volume":"4","author":"Gou","year":"2018","journal-title":"Nat. Plants"},{"issue":"1","key":"2026090917080501000_cit38","doi-asserted-by":"publisher","first-page":"7101","DOI":"10.1038\/s41467-023-42877-0","article-title":"Deciphering\ntriterpenoid saponin biosynthesis by leveraging transcriptome response\nto methyl jasmonate elicitation in Saponaria vaccaria","volume":"14","author":"Chen","year":"2023","journal-title":"Nat. Commun."},{"key":"2026090917080501000_cit39","doi-asserted-by":"publisher","first-page":"102005","DOI":"10.1016\/j.pbi.2021.102005","article-title":"Rational\ndesign strategies for functional\nreconstitution of plant cytochrome P450s in microbial systems","volume":"60","author":"Zhou","year":"2021","journal-title":"Curr. Opin. Plant Biol."},{"issue":"4","key":"2026090917080501000_cit40","doi-asserted-by":"publisher","first-page":"1540","DOI":"10.1021\/ja311339u","article-title":"Spiroketal\nformation and modification in avermectin biosynthesis involves a dual\nactivity of AveC","volume":"135","author":"Sun","year":"2013","journal-title":"J. Am. Chem. Soc."},{"issue":"52","key":"2026090917080501000_cit41","doi-asserted-by":"publisher","first-page":"26960","DOI":"10.1002\/anie.202109384","article-title":"Catalytic control of\nspiroketal formation in rubromycin polyketide biosynthesis","volume":"60","author":"Toplak","year":"2021","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090917080501000_cit42","doi-asserted-by":"publisher","first-page":"3206","DOI":"10.1038\/s41467-019-11286-7","article-title":"Repeated evolution\nof cytochrome P450-mediated spiroketal\nsteroid biosynthesis in plants","volume":"10","author":"Christ","year":"2019","journal-title":"Nat. Commun."},{"key":"2026090917080501000_cit43","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/j.fitote.2013.07.013","article-title":"New triterpene glycosides\nfrom Ziziphi Spinosae Semen","volume":"90","author":"Wang","year":"2013","journal-title":"Fitoterapia"},{"issue":"12","key":"2026090917080501000_cit44","doi-asserted-by":"publisher","first-page":"1744","DOI":"10.1248\/cpb.47.1744","article-title":"Bioactive\nsaponins and glycosides.: XIV.: Structure elucidation and immunological\nadjuvant activity of novel protojujubogenin type triterpene bisdesmosides,\nprotojujubosides A, B, and B, from the seeds of Zizyphus jujuba var.\nspinosa (Zizyphi Spinosi Semen)","volume":"47","author":"Matsuda","year":"1999","journal-title":"Chem. Pharm.\nBull."}],"container-title":["Journal of Agricultural and Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c06727","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/35\/27580\/67418335\/acs.jafc.6c06727.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/35\/27580\/67418335\/acs.jafc.6c06727.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T21:13:01Z","timestamp":1788988381000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/35\/27580\/5329654\/Identification-of-Three-Cytochrome-P450-Enzymes"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,26]]},"references-count":44,"journal-issue":{"issue":"35","published-online":{"date-parts":[[2026,8,26]]},"published-print":{"date-parts":[[2026,9,9]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c06727","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,26]]}}}