{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T01:58:31Z","timestamp":1785463111548,"version":"3.56.0"},"reference-count":33,"publisher":"American Chemical Society (ACS)","issue":"23","license":[{"start":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T00:00:00Z","timestamp":1780876800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T00:00:00Z","timestamp":1780876800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T00:00:00Z","timestamp":1780876800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["25TS1401500"],"award-info":[{"award-number":["25TS1401500"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004001","name":"Guizhou Provincial Science and Technology Department","doi-asserted-by":"publisher","award":["CXPTXM (2025) 010"],"award-info":[{"award-number":["CXPTXM (2025) 010"]}],"id":[{"id":"10.13039\/501100004001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004001","name":"Guizhou Provincial Science and Technology Department","doi-asserted-by":"publisher","award":["CXPTXM (2025) 021"],"award-info":[{"award-number":["CXPTXM (2025) 021"]}],"id":[{"id":"10.13039\/501100004001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004001","name":"Guizhou Provincial Science and Technology Department","doi-asserted-by":"publisher","award":["KXJZ (2024) 015"],"award-info":[{"award-number":["KXJZ (2024) 015"]}],"id":[{"id":"10.13039\/501100004001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004001","name":"Guizhou Provincial Science and Technology Department","doi-asserted-by":"publisher","award":["KXJZ (2025) 014"],"award-info":[{"award-number":["KXJZ (2025) 014"]}],"id":[{"id":"10.13039\/501100004001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010876","name":"Shanghai University of Traditional Chinese Medicine","doi-asserted-by":"publisher","award":["2023YZZ01"],"award-info":[{"award-number":["2023YZZ01"]}],"id":[{"id":"10.13039\/501100010876","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,6,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Ginsenoside Rf, a rare protopanaxatriol-type ginsenoside derived from Panax ginseng, exhibits notable neuroprotective, anti-inflammatory, and antitumor properties. However, its low natural abundance has impeded large-scale production. Here, we reprogrammed the sugar donor specificity of PnUGT94Q39 from UDP-xylose to UDP-glucose, thereby enabling efficient ginsenoside Rf biosynthesis. A single-point mutation (I142T) within a flexible loop not only switched the sugar donor preference to UDP-glucose but also established efficient catalysis (kcat\/Km, 1.18 \u03bcM\u20131\u00b7min\u20131) toward this donor. Coupling the I142T mutant with sucrose synthase GmSUS in Escherichia coli enabled the production of ginsenoside Rf (Rf) from ginsenoside Rh1 (Rh1) without exogenous UDP-glucose addition. Under optimized whole-cell biocatalysis conditions, a ginsenoside Rf titer of 1219.49 mg\u00b7L\u20131 was achieved, corresponding to a 97.26% conversion rate from Rh1 to Rf. Collectively, this work generated an efficient PnUGT94Q39-I142T mutant and established a sustainable, high-efficiency route for ginsenoside Rf production.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c04510","type":"journal-article","created":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T08:54:20Z","timestamp":1780908860000},"page":"18212-18221","source":"Crossref","is-referenced-by-count":0,"title":["Protein Engineering\nto Change the Sugar Donor and\nImprove the Activity of PnUGT94Q39 for Efficient Ginsenoside Rf Biosynthesis"],"prefix":"10.1021","volume":"74","author":[{"given":"Maoqi","family":"Hou","sequence":"first","affiliation":[{"name":"Shanghai University of Traditional Chinese Medicine , , ,","place":["Shanghai, P. R. China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sijie","family":"Chen","sequence":"additional","affiliation":[{"name":"Shanghai University of Traditional Chinese Medicine , , ,","place":["Shanghai, P. R. China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Yang","sequence":"additional","affiliation":[{"name":"Shanghai University of Traditional Chinese Medicine , , ,","place":["Shanghai, P. R. China, 201203"]},{"name":"Guizhou Medical University , , , ,","place":["Guiyang, Guizhou Province, P. R. China, 550025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2269-925X","authenticated-orcid":true,"given":"Shujuan","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shanghai University of Traditional Chinese Medicine , , ,","place":["Shanghai, P. R. China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2797-4625","authenticated-orcid":true,"given":"Zhengtao","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai University of Traditional Chinese Medicine , , ,","place":["Shanghai, P. R. China, 201203"]},{"name":"Guizhou Medical University , , , ,","place":["Guiyang, Guizhou Province, P. R. China, 550025"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,6,8]]},"reference":[{"issue":"7","key":"2026073021134248400_cit1","doi-asserted-by":"publisher","first-page":"1813","DOI":"10.1016\/j.apsb.2020.12.017","article-title":"Ginsenosides in Panax genus and their biosynthesis","volume":"11","author":"Hou","year":"2021","journal-title":"Acta Pharm. Sin. B"},{"key":"2026073021134248400_cit2","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1016\/j.jare.2024.01.003","article-title":"Rare ginsenosides: a unique perspective\nof ginseng research","volume":"66","author":"Fan","year":"2024","journal-title":"J. Adv. Res."},{"key":"2026073021134248400_cit3","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jgr.2019.12.009","article-title":"Phytochemical\nanalysis of Panax species: a review","volume":"45","author":"Yang","year":"2021","journal-title":"J. Ginseng Res."},{"key":"2026073021134248400_cit4","doi-asserted-by":"publisher","first-page":"1281","DOI":"10.1021\/ac990819z","article-title":"Differentiation\nand Authentication of Panax ginseng, Panax quinquefolius, and Ginseng\nProducts by Using HPLC\/MS.pdf","volume":"72","author":"Chan","year":"2000","journal-title":"Anal. Chem."},{"issue":"1","key":"2026073021134248400_cit5","doi-asserted-by":"publisher","first-page":"6463770","DOI":"10.1155\/2022\/6463770","article-title":"Rapid Identification\nof Characteristic Chemical Constituents of Panax ginseng, Panax quinquefolius,\nand Panax japonicus Using UPLC-Q-TOF\/MS","volume":"2022","author":"Jinbiao","year":"2022","journal-title":"J.\nAnal. Methods Chem."},{"issue":"1","key":"2026073021134248400_cit6","doi-asserted-by":"publisher","first-page":"309","DOI":"10.3233\/jad-180251","article-title":"Neuroprotective\nEffects of Ginsenoside Rf on Amyloid-\u03b2-Induced Neurotoxicity in vitro and in vivo","volume":"64","author":"Du","year":"2018","journal-title":"J. Alzheimers Dis."},{"issue":"2","key":"2026073021134248400_cit7","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/j.jgr.2018.11.007","article-title":"Ginsenoside Rf inhibits cyclooxygenase-2 induction via peroxisome\nproliferator\u2013activated receptor gamma in A549 cells","volume":"43","author":"Song","year":"2019","journal-title":"J. Ginseng Res."},{"issue":"2","key":"2026073021134248400_cit8","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/j.jgr.2017.02.005","article-title":"Antinociceptive\nand anti-inflammatory effects of ginsenoside Rf in a rat model of\nincisional pain","volume":"42","author":"Kim","year":"2018","journal-title":"J. Ginseng Res."},{"key":"2026073021134248400_cit9","doi-asserted-by":"publisher","first-page":"157403","DOI":"10.1016\/j.phymed.2025.157403","article-title":"Systematic review and\npharmacological insights of ginsenoside Rf: Current status and future\nperspectives","volume":"148","author":"Li","year":"2025","journal-title":"Phytomedicine"},{"key":"2026073021134248400_cit10","doi-asserted-by":"publisher","first-page":"1132","DOI":"10.1016\/j.biopha.2018.12.026","article-title":"The modulatory properties of Si Jun Zi Tang enhancing anticancer\nof gefitinib by an integrating approach","volume":"111","author":"Li","year":"2019","journal-title":"Biomed.\nPharmacother."},{"issue":"1","key":"2026073021134248400_cit11","doi-asserted-by":"publisher","first-page":"305","DOI":"10.3892\/or.2013.2815","article-title":"Induction\nof G2\/M phase cell cycle\narrest and apoptosis by ginsenoside Rf in human osteosarcoma MG-63\ncells through the mitochondrial pathway","volume":"31","author":"Shangguan","year":"2014","journal-title":"Oncol.\nRep."},{"key":"2026073021134248400_cit12","doi-asserted-by":"publisher","first-page":"117757","DOI":"10.1016\/j.ejmech.2025.117757","article-title":"Design, synthesis,\nand biological evaluation of C-12 modified ocotillol-type\nderivatives as novel P-glycoprotein modulators for overcoming multidrug\nresistance","volume":"294","author":"Chen","year":"2025","journal-title":"Eur. J. Med. Chem."},{"key":"2026073021134248400_cit13","doi-asserted-by":"publisher","first-page":"112507","DOI":"10.1016\/j.ejmech.2020.112507","article-title":"Design, synthesis\nand anti-inflammatory\nactivity of 3-amino acid derivatives of ocotillol-type sapogenins","volume":"202","author":"Yang","year":"2020","journal-title":"Eur. J. Med. Chem."},{"issue":"21","key":"2026073021134248400_cit14","doi-asserted-by":"publisher","first-page":"10279","DOI":"10.1021\/acs.joc.6b01265","article-title":"Synthesis of Ocotillol-Type\nGinsenosides","volume":"81","author":"Shen","year":"2016","journal-title":"J. Org. Chem."},{"issue":"4","key":"2026073021134248400_cit15","doi-asserted-by":"publisher","first-page":"278","DOI":"10.6023\/A17120544","article-title":"Total Synthesis of\nDammarane-Type Saponins Ginsenoside\nRe and Notoginsenoside R1","volume":"76","author":"Shen","year":"2018","journal-title":"Acta Chim. 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Eng."},{"key":"2026073021134248400_cit17","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.ymben.2020.05.007","article-title":"Elucidation of the\ncomplete biosynthetic pathway of\nthe main triterpene glycosylation products of Panax notoginseng using a synthetic biology platform","volume":"61","author":"Wang","year":"2020","journal-title":"Metab.\nEng."},{"key":"2026073021134248400_cit18","doi-asserted-by":"publisher","first-page":"1191102","DOI":"10.3389\/fmicb.2023.1191102","article-title":"Reconstruction\nof engineered yeast factory for high\nyield production of ginsenosides Rg3 and Rd","volume":"14","author":"Lin","year":"2023","journal-title":"Front. Microbiol."},{"key":"2026073021134248400_cit19","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1016\/j.ijbiomac.2020.10.184","article-title":"Broaden the sugar donor selectivity of blackberry glycosyltransferase\nUGT78H2 through residual substitutions","volume":"166","author":"Chen","year":"2021","journal-title":"Int.\nJ. Biol. Macromol."},{"issue":"3","key":"2026073021134248400_cit20","doi-asserted-by":"publisher","first-page":"1295","DOI":"10.1104\/pp.108.128256","article-title":"Catalytic\nkey amino acids and UDP-sugar donor specificity of a plant glucuronosyltransferase,\nUGT94B1: molecular modeling substantiated by site-specific mutagenesis\nand biochemical analyses","volume":"148","author":"Osmani","year":"2008","journal-title":"Plant Physiol."},{"issue":"2","key":"2026073021134248400_cit21","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1111\/tpj.13271","article-title":"Identification of a\nresidue responsible for UDP-sugar donor selectivity\nof a dihydroxybenzoic acid glycosyltransferase from Arabidopsis natural accessions","volume":"89","author":"Chen","year":"2017","journal-title":"Plant J."},{"issue":"8","key":"2026073021134248400_cit22","doi-asserted-by":"publisher","first-page":"3535","DOI":"10.1016\/j.apsb.2023.05.011","article-title":"A highly selective C-rhamnosyltransferase from Viola tricolor and insights into its mechanisms","volume":"13","author":"Han","year":"2023","journal-title":"Acta Pharm. Sin. B"},{"issue":"1","key":"2026073021134248400_cit23","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1021\/acs.jafc.2c05601","article-title":"New glycosyltransferases\nin Panax notoginseng perfect main ginsenosides biosynthetic\npathways","volume":"71","author":"Hou","year":"2023","journal-title":"J. Agric. Food Chem."},{"issue":"7","key":"2026073021134248400_cit24","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1021\/sb300016b","article-title":"ePathBrick: A\nSynthetic Biology Platform for Engineering Metabolic Pathways in E.\ncoli","volume":"1","author":"Xu","year":"2012","journal-title":"ACS Synth. Biol."},{"key":"2026073021134248400_cit25","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.ymben.2015.03.003","article-title":"Production of bioactive ginsenosides Rh2 and Rg3 by metabolically\nengineered yeasts","volume":"29","author":"Wang","year":"2015","journal-title":"Metab. Eng."},{"key":"2026073021134248400_cit26","doi-asserted-by":"publisher","first-page":"2177","DOI":"10.1093\/pcp\/pcu147","article-title":"Two ginseng\nUDP-glycosyltransferases synthesize ginsenoside Rg3 and Rd","volume":"55","author":"Jung","year":"2014","journal-title":"Plant Cell Physiol."},{"key":"2026073021134248400_cit27","doi-asserted-by":"publisher","first-page":"1412","DOI":"10.1016\/j.molp.2015.05.010","article-title":"Characterization\nof Panax ginseng UDP-glycosyltransferases catalyzing\nprotopanaxatriol and biosyntheses of bioactive ginsenosides F1 and\nRh1 in metabolically engineered yeasts","volume":"8","author":"Wei","year":"2015","journal-title":"Mol.\nPlant"},{"issue":"3","key":"2026073021134248400_cit28","doi-asserted-by":"publisher","first-page":"73","DOI":"10.3390\/biotech14030073","article-title":"Structure-Based Pipeline\nfor Plant Enzymes: Pilot Study Identifying Novel Ginsenoside Biosynthetic\nUGTs","volume":"14","author":"Jung","year":"2025","journal-title":"BioTech"},{"issue":"11","key":"2026073021134248400_cit29","doi-asserted-by":"publisher","first-page":"2413185","DOI":"10.1002\/advs.202413185","article-title":"Structural Insights\ninto the Substrate Recognition of Ginsenoside Glycosyltransferase\nPq3-O-UGT2","volume":"12","author":"Ji","year":"2025","journal-title":"Adv. Sci."},{"issue":"W1","key":"2026073021134248400_cit30","doi-asserted-by":"publisher","first-page":"W320","DOI":"10.1093\/nar\/gku316","article-title":"Deciphering key features in protein structures with the new ENDscript\nserver","volume":"42","author":"Robert","year":"2014","journal-title":"Nucleic Acids Res."},{"issue":"8016","key":"2026073021134248400_cit31","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of\nbiomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"issue":"2","key":"2026073021134248400_cit32","doi-asserted-by":"publisher","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: improving\nthe speed and accuracy of\ndocking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"issue":"5","key":"2026073021134248400_cit33","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1007\/s10822-010-9352-6","article-title":"Ligand docking and binding site analysis with PyMOL\nand Autodock\/Vina","volume":"24","author":"Seeliger","year":"2010","journal-title":"J. Comput.-Aided Mol. Des."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c04510","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/23\/18212\/66262650\/acs.jafc.6c04510.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/23\/18212\/66262650\/acs.jafc.6c04510.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T01:13:51Z","timestamp":1785460431000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/23\/18212\/5232902\/Protein-Engineering-to-Change-the-Sugar-Donor-and"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,8]]},"references-count":33,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2026,6,8]]},"published-print":{"date-parts":[[2026,6,17]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c04510","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,8]]}}}