鸭肝碱性磷酸酶的分离纯化及其部分性质研究  

Isolation and Purification of Alkalina Phosphatase from Anas platyrhynchos Liver and Some of Its Properties

作　　者：张丽丽[1] 阙瑞琦[1] 徐榕敏[1] 唐云明[1] 

机构地区：[1]西南大学生命科学学院,重庆400715

出　　处：《西南大学学报（自然科学版）》2007年第10期109-113,共5页Journal of Southwest University(Natural Science Edition)

基　　金：重庆市科委资助项目(CSTC;2004AC1012)

摘　　要：经过匀浆、正丁醇除脂、硫酸铵分级分离、DEAE-Sepharose Fast Flow离子交换柱层析、Sephacryl S-200凝胶过滤等步骤,从鸭肝中分离纯化出电泳纯的碱性磷酸酶,该酶纯化倍数达58.3倍,比活为337.4 U/mg.经SDS-PAGE和凝胶过滤测得该酶的全分子量为121.0 kD,亚基分子量为60.5 kD.以磷酸苯二钠为底物,测得该酶的最适pH为9.0,最适温度为40℃.Na+、K+、Li+等一价离子对该酶活性基本无影响,Mg2+、Mn2+对该酶活性有激活作用,Cd2+对该酶活性有抑制作用.米氏常数Km为1.28 mmol/L.Alkaline phosphatase was isolated and purified from the duck （Anas platyrhynchos） liver by means of homogenation, n-butanol disposal, ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose Fast Flow column and gel filtration chromatography on Sephacryl S-200. The preparation was shown to be homogenous on polyacrylamide gel electrophoresis （PAGE）. Its specific ac- tivity was 337.4 uint/mg protein, and the purification fold was 58.3. Sephacryl S-200 chromatography and SDS-PAGE showed that the molecular weight of this AKP was 121.0 kD, and the molecular weight of subunit was 60.5 kD. With phenylphosphoric acid disodium salt being the substrate of alkaline phosphatase, its optimum pH and temperature was 9.0 and 35 ℃, respectively. Na^＋ , K^＋ and Li^＋ had no effect on the activity of the enzyme. Mg^2＋ and Mn^2＋ activated the enzyme while Cd^2＋ inhibited its actiivity. The Michaelis constant （Km） was 1.28 mmol/L.

关 键 词：鸭 肝脏 碱性磷酸酶 分离纯化 性质 

分 类 号：Q814.1[生物学—生物工程]