Capture-Release Strategy Facilitates Rapid Enzymatic Assembly of Oligosaccharides
Capture-Release Strategy Facilitates Rapid Enzymatic Assembly of Oligosaccharides
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捕获-释放策略促进低聚糖的快速酶促组装
DOI：
10.1002/cjoc.202100744
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发表时间：
2021
影响因子：
5.4
通讯作者：
Cao Hongzhi
中科院分区：
文献类型：
--
作者：
Fang Wenyuan;Zhong Kan;Cheng Jiansong;Liu Xian-Wei;Liu Chang-Cheng;Wang Zhongfu;Cao Hongzhi
为简化寡糖的酶法合成和纯化过程提供了一种简便、高效的方法。以巯基标签封端的糖基受体在水相中通过酶模块组装（EMA）进行延伸，并在固相处理（SPW）过程中使用市售硫代丙基琼脂糖6 B树脂通过可逆二硫键捕获所需产物。最后，在二硫苏糖醇（DTT）作为还原剂的存在下，产物从固相树脂中释放出来，树脂可以回收和重复使用。这种策略克服了酶促糖基化中常用的聚合物或离子支持物的不确定影响，并与所有10种酶模块显示出完美的相容性，用于快速组装一系列复杂的寡糖。
Comprehensive SummaryA 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.