Capture‐Release Strategy Facilitates Rapid Enzymatic Assembly of Oligosaccharides

Comprehensive Summary A convenient and highly efficient strategy was developed for the simplification of enzymatic synthesis and purification of oligosaccharides. Glycosyl acceptors terminated with a thiol tag were extended by enzymatic modular assembly (EMA) in the aqueous phase, and the desired products were captured during solid‐phase workup (SPW) using commercially available thiopropyl Sepharose 6B resin through a reversible disulfide linkage. Finally, the products were released from solid‐phase resin in the presence of dithiothreitol (DTT) as reducing agent and the resin could be recovered and reused. This strategy overcomes the uncertain influence of polymer‐ or ionic‐supports commonly used in enzymatic glycosylation, and showed perfect compatibility with all 10 enzyme modules for the rapid assembly of a series of complex oligosaccharides. A highly efficient and convenient liquid‐phase enzymatic synthesis and solid‐phase capture‐release purification strategy was developed for the rapid preparation of diverse oligosaccharides.