Roth W W, Srinivasan V R
Prep Biochem. 1978;8(1):57-71. doi: 10.1080/00327487808068218.
A beta-glucosidase was isolated from Candida guilliermondii, a yeast capable of growth on cellobiose. The enzyme was partially purified by treatment with polyethyleneimine and ammonium sulfate precipitation. Further purification was achieved by affinity chromatography using a Sepharose 4B matrix to which oxidized salicin was coupled through adipic dihydrazide. The final product was a 12.5-fold purification of the crude extract with a recovery of 27% of the initial enzyme activity. Polyacrylamide disc electrophoresis of the purified enzyme gave a single band. A km of 1.25 x 10(-4)M was obtained using p-nitrophenyl beta-D-glucopyranoside as the substrate. The optimum pH for enzyme activity was 6.8. Maximum activity was observed at temperature of 37 degrees C. Enzyme activity was completely inhibited by Hg++, Pb++, and Zn++ ions. The molecular weight of the enzyme is 48,000 as estimated by sucrose density gradient centrifugation.
从季也蒙毕赤酵母（一种能够在纤维二糖上生长的酵母）中分离出一种β-葡萄糖苷酶。该酶通过用聚乙烯亚胺处理和硫酸铵沉淀进行部分纯化。进一步的纯化是通过使用经己二酸二酰肼偶联氧化水杨苷的琼脂糖4B基质进行亲和色谱来实现的。最终产物是粗提物的12.5倍纯化，初始酶活性回收率为27%。纯化酶的聚丙烯酰胺圆盘电泳产生一条带。以对硝基苯基β-D-吡喃葡萄糖苷为底物时，得到的Km值为1.25×10⁻⁴M。酶活性的最适pH为6.8。在37℃温度下观察到最大活性。Hg²⁺、Pb²⁺和Zn²⁺离子完全抑制酶活性。通过蔗糖密度梯度离心估计该酶的分子量为48,000。
