Jung K, Becker S
Department of Experimental Organ Transplantation, University Hospital Charité, Humboldt University Berlin, FRG.
Biomed Biochim Acta. 1991;50(7):861-7.
The two major tissue forms of N-acetyl-beta-D-glucosaminidase, A and B, and a minor form M were isolated from human urine by chromatography on DEAE-Trisacryl M and Con A-Sepharose. Forms A and B have a Km-value of about 0.75 mM for 4-nitrophenyl-beta-2-acetamido-2-deoxy-D-glucopyranoside, form M a corresponding value of 1.57 mM. The pH optimum was for all forms between pH 4.0 and 4.8. Forms A and M were heat labile, while form B was relatively heat stable. Using different binding properties of forms A and B on Trisacryl, a batch method was developed to differentiate both forms in human urine. This method can be easily performed and gives reproducible results. The intra-run and between-run precision for the separation of the two forms were below 10%.
通过在DEAE-三乙醇胺甲基丙烯酸酯M和刀豆球蛋白A-琼脂糖上进行色谱分离，从人尿中分离出N-乙酰-β-D-氨基葡萄糖苷酶的两种主要组织形式A和B以及一种次要形式M。形式A和B对4-硝基苯基-β-2-乙酰氨基-2-脱氧-D-吡喃葡萄糖苷的Km值约为0.75 mM，形式M的相应值为1.57 mM。所有形式的最适pH在4.0至4.8之间。形式A和M对热不稳定，而形式B相对热稳定。利用形式A和B在三乙醇胺甲基丙烯酸酯上的不同结合特性，开发了一种分批法来区分人尿中的这两种形式。该方法易于实施且结果可重复。两种形式分离的批内和批间精密度均低于10%。
