Banerjee D K, Basu D
Biochem J. 1975 Jan;145(1):113-8. doi: 10.1042/bj1450113.
N-Acetyl-beta-hexosaminidase A was purified 1000-fold from human urine by chromatography on DEAE-Sephadex followed by concanavalin A--Sepharose affinity chromatography. The optimal pH range was 4.4--4.5 for both the N-acetylglucosamine and N-acetylgalactosamine derivatives. The Km values were 0.51 mM and 0.28 mM respectively for the N-acetylglucosamine and N-acetylgalactosamine derivatives. The glycoprotein nature of the urinary enzyme was established by its affinity towards concanavalin A as well as by the presence of sialic acid, galactose, glucose, mannose and hexosamines in the molecule.
通过在DEAE-葡聚糖凝胶上进行色谱分离,随后进行伴刀豆球蛋白A-琼脂糖亲和色谱,从人尿中纯化出1000倍的N-乙酰-β-己糖胺酶A。N-乙酰葡糖胺和N-乙酰半乳糖胺衍生物的最佳pH范围均为4.4 - 4.5。N-乙酰葡糖胺和N-乙酰半乳糖胺衍生物的Km值分别为0.51 mM和0.28 mM。尿酶的糖蛋白性质通过其对伴刀豆球蛋白A的亲和力以及分子中存在的唾液酸、半乳糖、葡萄糖、甘露糖和己糖胺得以确定。