Institute for Structural Biology, Drug Discovery and Development, Virginia Commonwealth University, Richmond, VA 23219, USA.
Department of Medicinal Chemistry, Virginia Commonwealth University, Richmond, VA 23298, USA.
Glycobiology. 2020 Sep 28;30(10):768-773. doi: 10.1093/glycob/cwaa027.
Although structurally diverse, longer glycosaminoglycan (GAG) oligosaccharides are critical to understand human biology, few are available. The major bottleneck has been the predominant production of oligosaccharides, primarily disaccharides, upon enzymatic depolymerization of GAGs. In this work, we employ enzyme immobilization to prepare hexasaccharide and longer sequences of chondroitin sulfate in good yields with reasonable homogeneity. Immobilized chondroitinase ABC displayed good efficiency, robust operational pH range, broad thermal stability, high recycle ability and excellent distribution of products in comparison to the free enzyme. Diverse sequences could be chromatographically resolved into well-defined peaks and characterized using LC-MS. Enzyme immobilization technology could enable easier access to diverse longer GAG sequences.
尽管结构多样，但了解人类生物学的关键是更长的糖胺聚糖 (GAG) 寡糖，而这些寡糖的供应却非常有限。主要的瓶颈是 GAG 酶解时主要产生二糖等寡糖。在这项工作中，我们采用酶固定化技术来制备硫酸软骨素的六糖和更长序列，收率高，均一性好。与游离酶相比，固定化 ABC 酶具有较高的效率、较宽的操作 pH 范围、良好的热稳定性、较强的循环使用能力和产物分布更优。多种序列可以通过色谱分离成明确的峰，并通过 LC-MS 进行表征。酶固定化技术可以更方便地获得更多样的长 GAG 序列。
