小鼠肾脏海藻糖酶：纯化与性质

Mouse kidney trehalase: purification and properties.

作者信息

Tenan M N, de Oliveira C P, Panek A D

出版信息

An Acad Bras Cienc. 1979 Mar;51(1):151-8.

Abstract

Trehalase (E.C.3.2.1.28) was isolated from mouse kidney and purified to homogeneity. The enzyme was solubilized with n-octanol and activated by freezing and thawing before precipitation with ammonium sulfate. A 1700-fold purification was achieved by chromatography on DEAE-cellulose, SP-Sephadex, followed by gel filtration on Sephadex G-200. Only a single form of enzyme activity was shown throughout the fractionation as confirmed by gel electrophoresis of the final preparation. The enzyme was specific for trehalose and its estimated molecular weight by filtration on Sephadex G-200 was 73,000. The Km for trehalose was shown to be 2.67 x 10(-3)M and the optimum pH was in the range of 5.5-5.6. We have also shown that the optimum temperature of the enzyme is 60 degrees C, but in the absence of substrate, thermal inactivation occurred at 55 degrees C.

摘要

海藻糖酶（E.C.3.2.1.28）从小鼠肾脏中分离出来并纯化至同质。该酶用正辛醇溶解，在硫酸铵沉淀之前通过冻融激活。通过DEAE - 纤维素、SP - 葡聚糖凝胶色谱，随后在葡聚糖凝胶G - 200上进行凝胶过滤，实现了1700倍的纯化。通过最终制剂的凝胶电泳证实，在整个分级分离过程中仅显示出单一形式的酶活性。该酶对海藻糖具有特异性，通过在葡聚糖凝胶G - 200上过滤估计其分子量为73,000。海藻糖的Km值为2.67×10（-3）M，最适pH值在5.5 - 5.6范围内。我们还表明该酶的最适温度为60℃，但在没有底物的情况下，在55℃发生热失活。