Synthesis of value-added uridine 5'-diphosphate-glucose from sucrose applying an engineered sucrose synthase counteracts the activity-stability trade-off

Liting Zhao; Zhongbao Ma; Linpei Zhang; Yu Shen; Lei Chen; Youran Li; Sha Xu; Guiyang Shi; Daming Fan; Zhongyang Ding

doi:10.1016/j.foodchem.2024.141765

Synthesis of value-added uridine 5'-diphosphate-glucose from sucrose applying an engineered sucrose synthase counteracts the activity-stability trade-off

Food Chem. 2024 Oct 23;464(Pt 2):141765. doi: 10.1016/j.foodchem.2024.141765. Online ahead of print.

Authors

Liting Zhao¹, Zhongbao Ma², Linpei Zhang³, Yu Shen³, Lei Chen², Youran Li², Sha Xu², Guiyang Shi², Daming Fan⁴, Zhongyang Ding⁵

Affiliations

¹ 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.
² 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.
³ School of Biotechnology and Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China.
⁴ School of Food Science and Technology, Jiangnan University, Wuxi 214122, China. Electronic address: [email protected].
⁵ 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. Electronic address: [email protected].

PMID: 39503094
DOI: 10.1016/j.foodchem.2024.141765

Abstract

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_M6) 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_M6 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_M6, 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.

Keywords: Cascade catalysis; Productivity; Sucrose synthase; UDP-Glc; UDP-gal.