biostudies-literatureShen KKey Technologies Research and Development ProgramScience and Technology Commission of Shanghai Municipality104https://www.ebi.ac.uk/biostudies/studies/S-EPMC12474828biostudies-literatureUnknown12(1)Fructose-6-phosphate is a key ketose monophosphate intermediate that plays a central role in the metabolic pathway of glycolysis. Sorbitol is a naturally occurring sugar alcohol with diverse biological properties, which is widely applied in the food, cosmetic, and pharmaceutical industries due to its favorable physicochemical and physiological characteristics. Although a biosynthetic pathway from starch to fructose-6-phosphate has been well established in metabolic engineering studies, the downstream enzymatic route leading to the biosynthesis of sorbitol from this intermediate has not yet been elucidated in the literature. In this study, we performed the design and implementation of an artificial multi-enzymatic cascade system for the biosynthesis of sorbitol from fructose-6-phosphate. FiBioresources and bioprocessingDevelopment of an in vitro multi-enzyme system for efficient one-pot biosynthesis of sorbitol from fructose-6-phosphate.PMC1247482825ZR1401078, 25HC2810400, 24HC2810300 and 23HC14002002021YFC2102300，2019YFA0905000Shen KChen QZhu CNZheng GWXu JHfalseDevelopment of an in vitro multi-enzyme system for efficient one-pot biosynthesis of sorbitol from fructose-6-phosphate.Fructose-6-phosphate is a key ketose monophosphate intermediate that plays a central role in the metabolic pathway of glycolysis. Sorbitol is a naturally occurring sugar alcohol with diverse biological properties, which is widely applied in the food, cosmetic, and pharmaceutical industries due to its favorable physicochemical and physiological characteristics. Although a biosynthetic pathway from starch to fructose-6-phosphate has been well established in metabolic engineering studies, the downstream enzymatic route leading to the biosynthesis of sorbitol from this intermediate has not yet been elucidated in the literature. In this study, we performed the design and implementation of an artificial multi-enzymatic cascade system for the biosynthesis of sorbitol from fructose-6-phosphate. Fi2025-01-01T00:00:00Z2025 Sep2026-06-03T21:43:44.459Z2026-05-02T03:08:13.383ZS-EPMC124748284100392110.1186/s40643-025-00943-z