中华蜜蜂工蜂碱性磷酸酶的纯化及部分性质研究  

Isolation, Purification and Some Properties of Alkaline Phosphatase from Apis cerana cerana Fabricius

作　　者：吴珍红[1] 樊少华[2] 张其康[1] 缪晓青[1] 

机构地区：[1]福建农林大学蜂疗研究所,福州350002 [2]徐州师范大学生命科学学院生化教研室,徐州221116

出　　处：《中国养蜂》2005年第11期12-14,共3页Apiculture of China

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。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.

关 键 词：中华蜜蜂 碱性磷酸酶 分离纯化 

分 类 号：S891[农业科学—特种经济动物饲养]