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Enzymatic biotransformation provides a green and selective route for Rh2 production, but current systems suffer from inefficient cascade conversion and limited enzyme stability. Here, two \u03b2-glucosidases enabling the stepwise and selective conversion of 20(S)-ginsenoside Rd to 20(S)-ginsenoside Rh2 were identified and assembled into a fusion biocatalyst through linker engineering. Compared with the parental two-enzyme cascade under equimolar enzyme loading, the optimized fusion enzyme reduced the accumulation of the intermediate Rg3 and increased 20(S)\u2013Rh2 production rate by 40%. A Zn\u2013Ni metal\u2013organic framework was further introduced to enable simultaneous purification and immobilization. The immobilized fusion enzyme showed enhanced operational stability, retaining 82% of its initial activity after seven successive substrate-feeding intervals. This study presents a multiscale strategy integrating enzyme fusion and MOF immobilization for the efficient, operationally stable, and sustainable production of 20(S)-ginsenoside Rh2.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c09373","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T02:07:57Z","timestamp":1788314877000},"page":"27795-27807","source":"Crossref","is-referenced-by-count":0,"title":["An Engineered Fusion\nEnzyme-MOF Platform for Enhanced\nBiotransformation of Ginsenoside Rd to Rare Ginsenoside Rh2"],"prefix":"10.1021","volume":"74","author":[{"given":"Zinuo","family":"Chen","sequence":"first","affiliation":[{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Xie","sequence":"additional","affiliation":[{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fenglin","family":"Ye","sequence":"additional","affiliation":[{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingsheng","family":"Chen","sequence":"additional","affiliation":[{"name":"Guangzhou Huanya Cosmetics Technology Co., Ltd. , ,","place":["Guangzhou, China, 510663"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyu","family":"He","sequence":"additional","affiliation":[{"name":"Guangzhou Huanya Cosmetics Technology Co., Ltd. , ,","place":["Guangzhou, China, 510663"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4742-4009","authenticated-orcid":true,"given":"Yufei","family":"Cao","sequence":"additional","affiliation":[{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3474-3446","authenticated-orcid":true,"given":"Wen-Yong","family":"Lou","sequence":"additional","affiliation":[{"name":"South China University of Technology , , ,","place":["Guangzhou, China, 510640"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,28]]},"reference":[{"issue":"23","key":"2026090917072605200_cit1","doi-asserted-by":"publisher","first-page":"4367","DOI":"10.3390\/molecules24234367","article-title":"Preparation of polyethylene glycol-ginsenoside\nRh1 and Rh2 conjugates and their efficacy against lung cancer and\ninflammation","volume":"24","author":"Mathiyalagan","year":"2019","journal-title":"Molecules"},{"issue":"1","key":"2026090917072605200_cit2","doi-asserted-by":"publisher","first-page":"100018","DOI":"10.1016\/j.foodw.2025.100018","article-title":"Harnessing probiotic fermentation\nto enhance the bioavailability\nand health impact of dietary phytochemicals","volume":"1","author":"Kanimozhi","year":"2025","journal-title":"Food Wellness"},{"issue":"1","key":"2026090917072605200_cit3","doi-asserted-by":"publisher","first-page":"100006","DOI":"10.1016\/j.foodw.2025.100006","article-title":"Exploration of all-natural\naggregation-induced emission\nnanoparticles from co-assembly of bioactive herbal molecules","volume":"1","author":"Liu","year":"2025","journal-title":"Food Wellness"},{"issue":"8","key":"2026090917072605200_cit4","doi-asserted-by":"publisher","first-page":"773","DOI":"10.1007\/s11434-015-0773-4","article-title":"Anti-inflammatory\nactivity of ginsenosides\nin LPS-stimulated RAW 264.7 cells","volume":"60","author":"Ahn","year":"2015","journal-title":"Sci. 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Technol."},{"key":"2026090917072605200_cit40","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1016\/j.ijbiomac.2021.02.027","article-title":"Covalently\nimmobilize crude d-amino\nacid transaminase onto UiO-66-NH2 surface for d-Ala biosynthesis","volume":"175","author":"Wang","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"issue":"15","key":"2026090917072605200_cit41","doi-asserted-by":"publisher","first-page":"9048","DOI":"10.1021\/acs.inorgchem.8b01055","article-title":"Improvement\nin crystallinity and porosity of poorly crystalline metal\u2013organic\nframeworks (MOFs) through their induced growth on a well-crystalline\nMOF template","volume":"57","author":"Ji","year":"2018","journal-title":"Inorg. Chem."},{"issue":"8","key":"2026090917072605200_cit42","doi-asserted-by":"publisher","first-page":"1002","DOI":"10.3390\/catal11081002","article-title":"Sustainable one-pot\nimmobilization of enzymes in\/on\nmetal-organic framework materials","volume":"11","author":"Molina","year":"2021","journal-title":"Catalysts"},{"issue":"12","key":"2026090917072605200_cit43","doi-asserted-by":"publisher","first-page":"14561","DOI":"10.1021\/acsami.3c18928","article-title":"One-Step Purification and Immobilization\nof Glycosyltransferase\nwith Zn\u2013Ni MOF for the Synthesis of Rare Ginsenoside Rh2","volume":"16","author":"Li","year":"2024","journal-title":"ACS Appl. Mater. 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Biotechnol."},{"issue":"7","key":"2026090917072605200_cit46","doi-asserted-by":"publisher","first-page":"5122","DOI":"10.1021\/acs.inorgchem.1c00164","article-title":"Defective\nHierarchical\nPore Engineering of a Zn\u2013Ni MOF by Labile Coordination Bonding\nModulation","volume":"60","author":"Ding","year":"2021","journal-title":"Inorg. 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