【作者】 缪晓青； 张其康； 樊少华； 吴珍红；

【Author】 Miao Xiaoqing Zhang Qikang Fan Shaohua Wu Zhenhong (Apitherapy Institute of F.A.F.U. 350002 Fuzhou)

【机构】 福建农林大学蜂疗研究所；

【摘要】 以中华蜜蜂的工蜂成体为提酶材料,经正丁醇提取、硫酸铵分级沉淀,0．15％NaCl 溶液透析,得粗酶液。用Sephadex G-150葡聚糖层析柱纯化粗酶,获得提纯的碱性磷酸酶,提纯倍数为16．79,比活力达到135．85 U／mg。以对硝基苯磷酸二钠(PNPP)为底物,测定提纯后的碱性磷酸酶的理化性质。结果表明:该酶的最适温度为45℃,最适pn值为8．6,米氏常数(Km值) 为0．97×10-3mol／L。甲醛、甲醇、乙醇和乙二醇对碱性磷酸酶均有抑制作用。更多还原

【Abstract】 The alkaline phosphatase (AKP) has been isolated from the worker honeybees of Apis cerana cerana Fabricius by n-butanol extraction, ammonium sulfate fractionation and sodium chloride dialyzation. The partially purified enzyme was gained by means of gel filtration on Sephadex G-150 column. The purification attained to 16.79 folds and the specific activity was 135.85 U/mg. Some properties of the enzyme were undertaken. The optimum temperature for the enzyme to catalyze the hydrolysis of p-nitrophenyl phosphate(pNPP) is at 45℃. If the pH value of the assay system was equal to 8.6, the enzyme would become the most active. The Michaelis-Menten constant(Km) of the hydrolysis pNPP catalyzed by the enzyme is 0.97 mmol/L. The AKP is inhibited by formaldehyde, carbinok ethanol and glycol.更多还原