Chondroitin sulfate (CS)and dermatan sulfate (DS) are negatively charged polysaccharides found abundantly in animal tissue and have been extensively described to play key roles in health and disease. The most common method to analyze their structure is by digestion into disaccharides with bacterial chondroitinases, followed by chromatography and/or mass spectrometry. While studying the structure of oncofetal CS, we noted a large variation in the activity and specificity of commercially available chondroitinases. Here studied the kinetics of the enzymes and used high-performance liquid chromatography–mass spectrometry to determine the di- and oligosaccharide products resulting from the digestion of commercially available bovine CS A, shark CS C and porcine DS, focusing on chondroitinases ABC, AC and B from different vendors. Application of a standardized assay setup demonstrated large variations in the enzyme-specific activity compared to the values provided by vendors, large variation in enzyme specific activity of similar enzymes from different vendors and differences in the extent of cleavage of the substrates and the generated products. The high variability of different chondroitinases highlights the importance of testing enzyme activity and monitoring product formation in assessing the content and composition of chondroitin and DSs in cells and tissues.

中文翻译：

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当心，商业软骨素酶的活性和底物特异性各不相同

硫酸软骨素 (CS) 和硫酸皮肤素 (DS) 是在动物组织中大量发现的带负电荷的多糖，并已被广泛描述在健康和疾病中起关键作用。分析其结构的最常用方法是用细菌软骨素酶消化成二糖，然后进行色谱和/或质谱分析。在研究癌胎儿 CS 的结构时，我们注意到市售软骨素酶的活性和特异性存在很大差异。在此研究了酶的动力学，并使用高效液相色谱-质谱法来确定由市售的牛 CS A、鲨鱼 CS C 和猪 DS 消化产生的二糖和寡糖产物，重点是软骨素酶 ABC、AC 和B 来自不同的供应商。与供应商提供的值相比，标准化测定设置的应用表明酶比活性存在很大差异，来自不同供应商的类似酶的酶比活性存在很大差异，并且底物和生成的产物的裂解程度存在差异。不同软骨素酶的高度可变性突出了测试酶活性和监​​测产物形成在评估细胞和组织中软骨素和 DSs 的含量和组成方面的重要性。