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The roots of\n                    <jats:italic>Astragalus membranaceus<\/jats:italic>\n                    are a popular herbal medicine worldwide. It contains a series of cycloartane\u2010type saponins (astragalosides) as hepatoprotective and antivirus components. However, astragalosides exhibit complex sugar substitution patterns which hindered their purification and bioactivity investigation. In this work, glycosyltransferases (GT) from\n                    <jats:italic>A. membranaceus<\/jats:italic>\n                    were studied to synthesize structurally diverse astragalosides. Three new GTs, AmGT1\/5 and AmGT9, were characterized as 3\u2010\n                    <jats:italic>O<\/jats:italic>\n                    \u2010glycosyltransferase and 25\u2010\n                    <jats:italic>O<\/jats:italic>\n                    \u2010glycosyltransferase of cycloastragenol respectively. AmGT1\n                    <jats:sub>G146V\/I<\/jats:sub>\n                    variants were obtained as specific 3\u2010\n                    <jats:italic>O<\/jats:italic>\n                    \u2010xylosyltransferases by sequence alignment, molecular modelling and site\u2010directed mutagenesis. A combinatorial synthesis system was established using AmGT1\/5\/9, AmGT1\n                    <jats:sub>G146V\/S<\/jats:sub>\n                    and the reported AmGT8 and AmGT8\n                    <jats:sub>A394F<\/jats:sub>\n                    . The system allowed the synthesis of 13 astragalosides in Astragalus root with conversion rates from 22.6% to 98.7%, covering most of the sugar\u2010substitution patterns for astragalosides. In addition, AmGT1 exhibited remarkable sugar donor promiscuity to use 10 different donors, and was used to synthesize three novel astragalosides and ginsenosides. Glycosylation remarkably improved the hepatoprotective and SARS\u2010CoV\u20102 inhibition activities for triterpenoids. This is one of the first attempts to produce a series of herbal constituents\n                    <jats:italic>via<\/jats:italic>\n                    combinatorial synthesis. The results provided new biocatalytic tools for saponin biosynthesis.\n                  <\/jats:p>","DOI":"10.1111\/pbi.13983","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T00:41:38Z","timestamp":1671410498000},"page":"698-710","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane\u2010type saponins in\n                    <i>Astragalus membranaceus<\/i>"],"prefix":"10.1111","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3030-8447","authenticated-orcid":false,"given":"Kuan","family":"Chen","sequence":"first","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"},{"name":"Beijing Institute of Clinical Pharmacy, Beijing Friendship Hospital Capital Medical University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baihan","family":"Gao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aobulikasimu","family":"Hasan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy Uppsala University  Uppsala Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang'oujie","family":"Bao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingjing","family":"Fan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Yi","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans","family":"\u00c5gren","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy Uppsala University  Uppsala Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zilong","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiyang","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Phytochemistry and Plant Resources in West China and Yunnan Key Laboratory of Natural Medicinal Chemistry, Kunming Institute of Botany Chinese Academy of Sciences  Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9952-2380","authenticated-orcid":false,"given":"Min","family":"Ye","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8771-7877","authenticated-orcid":false,"given":"Xue","family":"Qiao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences Peking University  Beijing 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