论文部分内容阅读
从厦门文昌鱼Branchiostoma belcheri(Gray)分离提纯酸性磷酯酶(EC3.1,3.2),进一步经Sephadex G-75和羧甲基纤维素(CM-52)柱层析纯化,获得聚丙烯酰胺凝胶电泳单一纯酶制剂,比活力为351μm/min mg。酶液吸收峰在280nm,320nm,500nm,表现出含铁离子蛋白的特征吸收峰,用原子吸收法和化学法(邻啡绕啉比色法)分析,证明了文昌鱼酸性磷酸酯酶(ACPase)含有铁离子;还原剂Na_2S_2O_4处理浓度提高,酶活力及荧光强度下降,280nm和500nm峰下降,320nm峰消失,以Na_2S_2O_4测定ACPase失活和变性速度常激,结果表明,Na_2S_2O_4的失活常数大于变性带数。
Acid phosphatase (EC 3.1, 3.2) was isolated and purified from Xiamen amphioxus Branchiostoma belcheri (Gray) and purified by Sephadex G-75 and CM-52 column chromatography to obtain polyacrylamide gel Gel electrophoresis single pure enzyme preparation, the specific activity of 351μm / min mg. Enzyme absorption peak at 280nm, 320nm, 500nm, showing the characteristic absorption peak of iron-containing protein, with atomic absorption and chemical methods (o -morpholine method) analysis, to prove that amphioxus acid phosphatase (ACPase ) Contained ferric ions. The concentration of reducing agent Na_2S_2O_4 increased, the enzyme activity and fluorescence intensity decreased, the peaks at 280nm and 500nm decreased, and the peak at 320nm disappeared. The deactivation and denaturation rate of ACPase were determined by Na_2S_2O_4. The results showed that the deactivation constant of Na_2S_2O_4 was greater than Degeneration with the number.