Majima M, Takagaki K, Igarashi S, Nakamura T, Endo M
J Biochem Biophys Methods. 1984 Dec;10(3-4):143-51. doi: 10.1016/0165-022x(84)90034-4.
The Milner-Avigad method (Milner, Y. and Avigad, G. (1967) Carbohydr. Res. 4, 359-361) has been shown to have high specificity for measuring the reducing powers of free hexuronic acids; its applicability in the quantitative determination of endouronidase activity was the purpose of this investigation. Among the constituent monosaccharides of glycosaminoglycans, hexuronic acids showed high color yield by this method, while xylose, galactose and N-acetylhexosamine recorded negligible color yield. Among the monosaccharide residues at the reducing terminals of oligosaccharides, only hexuronic acids exhibited color yield. However, the color yield was less than that of free hexuronic acids. Gel filtration chromatography of reaction products revealed that the cleavage of the oligosaccharide chains and the resultant exposure of new reducing terminals were not caused by the reaction procedures involved in this method. These data indicate that the Milner-Avigad method is useful for determining the presence of hexuronic acid residues preferentially at reducing terminals of glycosaminoglycan moieties. Thus, it supported the conclusion that the Milner-Avigad method is applicable for the quantitative determination of endouronidase activity using glycosaminoglycan as a substrate.
米尔纳-阿维加德法（米尔纳，Y. 和阿维加德，G.（1967年）《碳水化合物研究》4，359 - 361）已被证明在测定游离己糖醛酸的还原能力方面具有高度特异性；本研究的目的是探讨其在定量测定内切糖苷酶活性中的适用性。在糖胺聚糖的组成单糖中，己糖醛酸通过该方法显示出高显色率，而木糖、半乳糖和N-乙酰己糖胺的显色率可忽略不计。在寡糖还原末端的单糖残基中，只有己糖醛酸表现出显色率。然而，其显色率低于游离己糖醛酸。反应产物的凝胶过滤色谱显示，寡糖链的断裂以及由此产生的新还原末端的暴露并非由该方法所涉及的反应程序引起。这些数据表明，米尔纳-阿维加德法可用于优先确定糖胺聚糖部分还原末端己糖醛酸残基的存在。因此，这支持了米尔纳-阿维加德法适用于以糖胺聚糖为底物定量测定内切糖苷酶活性的结论。
