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Updated: Sep 16, 2025

Hierarchical and Programmable One-Pot Oligosaccharide Synthesis
Published on: September 6, 2019
A Cellular Coupling Strategy Leveraging Pathway Modularization and Cofactor Regeneration for the Biosynthesis of
Zimeng Zhang1,2, Hongtao Zhang1,2, Zhijie Wang1,2
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
None:
3'-Sialyllactose is a major sialylated human milk oligosaccharide that plays an important role in promoting infant development and supporting various physiological functions critical for early life. In this study, N-acetylneuraminic acid and 3'-sialyllactose were successfully synthesized utilizing four genes across three modules through a whole-cell catalysis. Among these, Module III was employed to facilitate the conversion of cytidine-5'-monophosphate to cytidine 5'-triphosphate utilizing the intrinsic enzyme system of Saccharomyces cerevisiae. This study presents the inaugural report demonstrating the synergistic synthesis of 3'-sialyllactose through three modules derived from Escherichia coli and S. cerevisiae, utilizing pyruvate, N-acetylglucosamine, lactose, and cytidine-5'-monophosphate as substrates. The induction and the whole-cell catalytic substrate dosage of the engineered E. coli JM109(DE3) were optimized, leading to a 3'-sialyllactose titer of 71.62 g/L. Ultimately, the strain underwent culture and whole-cell catalysis in a 5 L bioreactor, achieving a maximum titer of 78.03 g/L for 3'-sialyllactose.
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