Department of Chemical & Biochemical Engineering, Rutgers, The State University of New Jersey, 98 Brett Road, Piscataway, NJ 08854, USA.
Department of Chemical & Biochemical Engineering, Rutgers, The State University of New Jersey, 98 Brett Road, Piscataway, NJ 08854, USA.
Curr Opin Biotechnol. 2020 Dec;66:283-291. doi: 10.1016/j.copbio.2020.09.006. Epub 2020 Nov 8.
One of the stumbling blocks to advance the field of glycobiology has been the difficulty in synthesis of bespoke carbohydrate-based molecules like glycopolymers (e.g. human milk oligosaccharides) and glycoconjugates (e.g. glycosylated monoclonal antibodies). Recent strides towards using engineered Carbohydrate-Active enZymes (CAZymes) like glycosyl transferases, transglycosidases, and glycosynthases for glycans synthesis has allowed production of diverse glycans. Here, we discuss enzymatic routes for glycans biosynthesis and recent advances in protein engineering strategies that enable improvement of CAZyme specificity and catalytic turnover. We focus on rational and directed evolution methods that have been developed to engineer CAZymes. Finally, we discuss how improved CAZymes have been used in recent years to remodel and synthesize glycans for biotherapeutics and biotechnology related applications.
糖生物学领域的发展障碍之一一直是合成定制的基于碳水化合物的分子（如糖聚合物（例如人乳寡糖）和糖缀合物（例如糖基化单克隆抗体））的困难。最近在使用工程化的碳水化合物活性酶（CAZymes），如糖基转移酶、转糖苷酶和糖基合成酶进行聚糖合成方面取得了进展，从而可以生产出多种聚糖。在这里，我们讨论了糖生物合成的酶促途径以及最近在蛋白质工程策略方面的进展，这些策略可以提高 CAZyme 的特异性和催化周转率。我们重点介绍了为工程 CAZymes 而开发的合理和定向进化方法。最后，我们讨论了近年来如何改进 CAZymes 用于生物治疗和生物技术相关应用中的糖链重塑和合成。
