School of Biotechnology and Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi 214122, China; National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China.
School of Biotechnology and Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China; National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China.
Food Chem. 2025 Feb 1;464(Pt 2):141765. doi: 10.1016/j.foodchem.2024.141765. Epub 2024 Oct 23.
The one-step deconstruction of sucrose into uridine 5'-diphosphate-glucose (UDP-Glc), an important sugar donor for transglycosylation, employing sucrose synthase (Susy) is emerging as a valuable sucrose utilization process. The insufficient activity and stability of Susy limit the productivity of UDP-Glc from sucrose. Here, an engineered Susy (Susy) that counteracted the activity-stability trade-off was developed with the half-life time and activity being 43-fold and 1.4-fold of wild-type, respectively. Tighter hydrophobic patches and stabilization of the SSN2 domain contributed to greater activity and stability. The use of Susy in UDP-Glc production resulted in a satisfactory space-time yield of 73 g/L/h within 1 h. The cascade of different biocatalysts with Susy, focusing on utilizing two products of sucrose decomposition, fructose and UDP-Glc, expanded sucrose utilization, efficiently promoting the UDP-Glc productivity and giving a cost-effective method for UDP-galactose (UDP-Gal) synthesis. This study demonstrated promising green pathways for producing multiple value-added products from sucrose using Susy.
利用蔗糖合酶（Susy）将蔗糖一步解构为尿苷 5′-二磷酸葡萄糖（UDP-Glc），这是一种用于糖基转移反应的重要糖供体，正逐渐成为一种有价值的蔗糖利用方法。然而，Susy 的活性和稳定性不足限制了 UDP-Glc 从蔗糖中的产量。本研究通过工程化改造Susy，得到了半衰期和活性分别是野生型的 43 倍和 1.4 倍的Susy，克服了活性-稳定性权衡问题。更紧密的疏水性斑块和 SSN2 结构域的稳定有助于提高活性和稳定性。在 UDP-Glc 生产中使用 Susy，1 小时内时空产率达到了 73 g/L/h，令人满意。利用 Susy 进行蔗糖分解产物果糖和 UDP-Glc 的级联反应，拓展了蔗糖的利用途径，有效地提高了 UDP-Glc 的产量，为 UDP-半乳糖（UDP-Gal）的合成提供了一种具有成本效益的方法。本研究为利用 Susy 从蔗糖生产多种高附加值产品展示了有前景的绿色途径。
