Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
Shandong Provincial Key Laboratory of Biochemical Engineering, College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao 266042, China.
Mar Drugs. 2019 Mar 20;17(3):185. doi: 10.3390/md17030185.
Chondroitinase (ChSase), a type of glycosaminoglycan (GAG) lyase, can degrade chondroitin sulfate (CS) to unsaturate oligosaccharides, with various functional activities. In this study, ChSase AC II from a newly isolated marine bacterium sp. CS01 was cloned, expressed in X33, purified, and characterized. ChSase AC II, with a molecular weight of approximately 100 kDa and a specific activity of 18.7 U/mg, showed the highest activity at 37 °C and pH 6.5 and maintained stability at a broad range of pH (5⁻7.5) and temperature (below 35 °C). The enzyme activity was increased in the presence of Mn and was strongly inhibited by Hg. Moreover, the kinetic parameters of ChSase AC II against CS-A, CS-C, and HA were determined. TLC and ESI-MS analysis of the degradation products indicated that ChSase AC II displayed an exolytic action mode and completely hydrolyzed three substrates into oligosaccharides with low degrees of polymerization (DPs). All these features make ChSase AC II a promising candidate for the full use of GAG to produce oligosaccharides.
软骨素酶（ChSase）是一种糖胺聚糖（GAG）裂解酶，可将硫酸软骨素（CS）降解为不饱和低聚糖，具有多种功能活性。本研究克隆并表达了一株新分离的海洋细菌 sp. CS01 中的 ChSase AC II，经 X33 培养、纯化和表征。ChSase AC II 的分子量约为 100 kDa，比活为 18.7 U/mg，最适反应温度和 pH 分别为 37°C 和 6.5，在 pH5⁻7.5 和低于 35°C 的较宽温度范围内稳定。该酶在 Mn 存在时活性增加，Hg 强烈抑制其活性。此外，还测定了 ChSase AC II 对 CS-A、CS-C 和 HA 的动力学参数。CS 降解产物的 TLC 和 ESI-MS 分析表明，ChSase AC II 表现出外切酶作用模式，可将三种底物完全水解成低聚合度（DP）的寡糖。所有这些特性使 ChSase AC II 成为充分利用 GAG 生产寡糖的有前途的候选酶。
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