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In this study, through an extensive screening of 10 sucrose synthases derived from both eukaryotic and prokaryotic microorganisms, we identified a broad\u2010spectrum, highly efficient enzymatic scaffold suitable for the synthesis of UDP\u2010sugars. A rational design integrating PROSS and ThermoMPNN algorithms with molecular dynamics (MD) analysis yielded the V353L mutant, which retained 97% of wild\u2010type activity and extended the half\u2010life at 55\u00b0C from 0.4 to 5.5 h. Tetrameric MD simulations indicated that V353L strengthens hydrophobic stacking in the GT\u2010B linker region, reducing conformational fluctuations and driving the enzyme toward a stable low\u2010energy state. In a dual\u2010enzyme cascade synthesizing UDPG and UDPGA, the mutant sustained UDP consumption and increased the accumulated UDPGA concentration from 0.60 to 2.09\u00a0mM after 8\u00a0h. Engineering sucrose synthase thermostability thus improves UDPG supply in cascade biocatalysis and offers a viable enzymatic strategy for UDPGA\u2010centered synthesis of high\u2010value carbohydrate derivatives.<\/jats:p>","DOI":"10.1002\/biot.70306","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T05:51:58Z","timestamp":1789105918000},"source":"Crossref","is-referenced-by-count":0,"title":["Engineering the Stability of Sucrose Synthase to Enable Efficient Coupled Production of UDP\u2010Glucuronic Acid"],"prefix":"10.1002","volume":"21","author":[{"given":"Qian","family":"Zhao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ya\u2010min","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi\u2010min","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology  Shanghai China"},{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei\u2010hua","family":"Xiong","sequence":"additional","affiliation":[{"name":"So.S Agriculture (Shanghai) Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1729-9593","authenticated-orcid":false,"given":"Zong\u2010lin","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology  Shanghai 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