Mega T, Matsushima Y
J Biochem. 1979 Feb;85(2):335-41. doi: 10.1093/oxfordjournals.jbchem.a132338.
beta-Glucosidase [beta-D-glucoside glucohydrolase EC 3.2.1.21] and beta-galactosidase [beta-D-galactoside galactohydrolase, EC 3.2.1.23] of Takadiastase were purified by acetone fractionation, DEAE-cellulose, and hydroxylapatite chromatography. Purity was confirmed by disc electrophoresis, ultracentrifugation and measurement of other glycosidase activities which coexisted in Takadiastase. Molecular weight of the beta-glucosidase was 218,000 by sedimentation equilibrium and 110,000-116,000 by SDS-disc electrophoresis. Molecular weight of the beta-galactosidase was 112,000 by sedimentation and 56,000-59,000 by SDS-disc electrophoresis. These values showed that both enzymes consisted of two subunits. Taka-beta-N-acetylglucosaminidase also consisted of two subunits. Both enzymes were glycoproteins containing glucosamine and neutral sugar. Stability, pH optima, isoelectric points, and some specificities were observed.
通过丙酮分级分离、DEAE - 纤维素柱层析和羟基磷灰石柱层析对高峰淀粉酶的β - 葡萄糖苷酶[β - D - 葡萄糖苷葡糖水解酶，EC 3.2.1.21]和β - 半乳糖苷酶[β - D - 半乳糖苷半乳糖水解酶，EC 3.2.1.23]进行了纯化。通过圆盘电泳、超速离心以及对高峰淀粉酶中共存的其他糖苷酶活性的测定来确认纯度。β - 葡萄糖苷酶通过沉降平衡法测得的分子量为218,000，通过SDS - 圆盘电泳法测得的分子量为110,000 - 116,000。β - 半乳糖苷酶通过沉降法测得的分子量为112,000，通过SDS - 圆盘电泳法测得的分子量为56,000 - 59,000。这些数值表明这两种酶均由两个亚基组成。高峰β - N - 乙酰葡糖胺酶也由两个亚基组成。这两种酶均为含有葡糖胺和中性糖的糖蛋白。观察到了它们的稳定性、最适pH值、等电点以及一些特异性。
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