元素硒的无机氧化动力学研究i双氧水的氧化.pdfVIP

※生物工程 食品科学 2011, Vol. 32, No. 03 181 H1-9b 苏 茉，高亚朋，梁建荣，黄 洁，唐云明 * (西南大学生命科学学院，重庆市甘薯工程研究中心，三峡库区生态环境教育部重点实验室，重庆 400715) 采用研磨法破碎黑曲霉 H1-9b 菌丝体，经pH5.7 磷酸缓冲液抽提获得粗酶液，粗酶液经 DEAE-Sepharose 离子交换层析和 Superdex-200 凝胶过滤层析，得到葡萄糖氧化酶纯品。该酶的比活力为 30569.7U/mg ，回收率为 30.2% ，提纯倍数为41.4 倍，分子质量为 94.1kD ，为单亚基酶。该酶最适温度为37 ℃，最适pH 值为 5.7 ，在 30～40 ℃温度范围内稳定性较好，在pH4.0 ～8.0 范围内活性稳定。以不同浓度葡萄糖为底物在最适条件下测得该 酶Km 值为 30.69mmol/L ，Vmax 为 21.88 μmol/L 。Ag+ 、Cu2+ 、Zn2+ 对该酶活性有较大抑制作用。结果表明，该酶 稳定性较好，有一定的应用价值。 黑曲霉；葡萄糖氧化酶；分离；纯化；性质 Isolation, Purification and Some Properties of Glucose Oxidase from Aspergillius niger H1-9b SU Mo，GAO Ya-peng ，LIANG Jian-rong ，HUANG Jie ，TANG Yun-ming* (Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweetpotato Engineering Research Center, School of Life Science, Southwest University, Chongqing 400715, China) Abstract ：Glucose oxidase (GOD), an H2O2-producing enzyme, was isolated and purified from Aspergillius niger H1-9b through electrophoretic homogeneity, ion-exchange chromatography on DEAE-Sepharose column and Superdex-200 gel filtra- tion chromatography. The purified GOD had a molecular weight of 94.1 kD as a monomer. The specific activity of GOD was 30569.7 U/mg with recovery rate of 30.2% and purification fold of 41.4. The optimal reaction pH and temperature of GOD were 5.7 and 37 ℃, respectively. The GOD displayed an excellent stability under the conditions of 30－40 ℃ and pH 4.0 －8.0. The kinetic parameters such as Km and Vmax were 30.69 mmol/L and 21.88 μmol/L while using glucose as the substrate. Obvious inhibitory effects of Ag+, Cu2+ and Zn2+ on the activity of GOD were observed. Therefore, glucose oxidase from Aspe