Co-immobilization of a bi-enzymatic cascade into hierarchically porous MIL-53 for efficient 6′-sialyllactose production

• Published in: Nanoscale 2024-08, Vol.16 (31), p.14932-14939
• Main Authors: Peng, Xinlv, Pei, Caixia, Qian, Eika W, Du, Yuguang, Li, Jian-Jun
• Format: Article
• Language: English
• Subjects: beta-D-Galactoside alpha 2-6-Sialyltransferase; Bioreactors; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Humans; Hydrogen-Ion Concentration; Immobilization; Industrial applications; Lactose - analogs & derivatives; Lactose - chemistry; Lactose - metabolism; Milk, Human - chemistry; Milk, Human - metabolism; N-Acylneuraminate Cytidylyltransferase - chemistry; N-Acylneuraminate Cytidylyltransferase - metabolism; Oligosaccharides; Oligosaccharides - biosynthesis; Oligosaccharides - chemistry; Oligosaccharides - metabolism; Porosity; Sialyltransferases - metabolism; Storage stability; Synthesis
• ISSN: 2040-3364; 2040-3372; 2040-3372
• DOI: 10.1039/d4nr01775g

6′-Sialyllactose (6′-SL), the most abundant sialylated human milk oligosaccharide, has attracted attention for its potential application in supplementary infant formulas. Herein, we report a facile strategy to construct a cascade bioreactor for the enzymatic synthesis of 6-SL by co-immobilizing an enzymatic module consisting of CMP-sialic acid synthase and α-2,6-sialyltransferase into hierarchically porous MIL-53 (HP-MIL-53). The as-prepared HP-MIL-53 showed high enzyme immobilization capacity, reaching 226 mg g −1 . Furthermore, the co-immobilized enzymes exhibited higher initial catalytic efficiency, and thermal, pH and storage stability than the free ones. Finally, the 6-SL yield remained >80% after 13 cycles of use. We expect that HP-MIL-53 would have potential industrial applications in the enzymatic modular synthesis of 6-SL and other glycans. CMP-sialic acid synthase and α-2,6-sialyltransferase were co-immobilized into HP-MIL-53. The co-immobilized enzymes exhibited high initial catalytic efficiency, reusability and thermal, pH and storage stability.