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.

中文翻译：

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酶固定化提供了一种强大的工具来扩大更长、多样、天然的糖胺聚糖寡糖的生产

尽管结构多样、较长的糖胺聚糖 (GAG) 寡糖对于理解人类生物学至关重要，但可用的却很少。主要瓶颈是在 GAG 的酶促解聚时主要产生寡糖，主要是二糖。在这项工作中，我们采用酶固定化制备六糖和更长序列的硫酸软骨素，产量高且均匀性合理。与游离酶相比，固定化软骨素酶 ABC 表现出良好的效率、稳健的操作 pH 范围、广泛的热稳定性、高回收能力和优异的产品分布。不同的序列可以通过色谱分离成明确的峰，并使用 LC-MS 进行表征。酶固定化技术可以更容易地获得不同的较长 GAG 序列。