【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SephadexG-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135.85U/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pH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、carbinol、ethanol and glycol.更多还原