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.
出处
《西南大学学报（自然科学版）》
CAS
CSCD
北大核心
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)
关键词
鸭
肝脏
碱性磷酸酶
分离纯化
性质
duck
liver
alkaline phosphatase
isolation and purification
properties
