瘦肉型猪(PIC344)精液酸性磷酸酶部分性质研究  

Partial Characterisitcs of Acid Phosphatase from Sperm of PIC Pigs(PIC344)

作　　者：黄新河[1] 冯兴[2] 刘鑫[1] 李佳[1] 张勇侠[1] 国锦琳[1] 肖舸[1] 刘克武[1] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]四川抗菌素工业研究所

出　　处：《四川动物》2005年第1期17-21,26,共6页Sichuan Journal of Zoology

基　　金：四川省成都市大熊猫精液研究专项基金资助

摘　　要：　　 用纱网滤掉瘦肉型猪 (PIC344) 新鲜精液中胶状物得原精液, 该原精液经硫酸铵分段盐析、DEAE Sepharose F F 离子交换柱层析、Sephacryl S 200 凝胶过滤后分离纯化到酸性磷酸酶 (Acid Phosphatase, 简称ACPase)。纯化倍数为22 78, 酶液比活力为15 26U/mg蛋白。纯化酶液经非还原性SDS PAGE检测, 呈现单一蛋白着色带。测得该酶相对分子质量为52 3kD, 等电点为5 1, 米氏常数 (Km 值) 为3 08×10-3mol/L。测得该酶最适pH为3 6, 最适温度为52℃。ACPase在pH 3 5~6 0范围内稳定, 在40℃以下稳定, 50℃保温30min后酶活仍能保持59 2%。Fresh sperm is obtained from PIC344 through filtering the jelly. The acid phosphatase (ACPase) is got by ammonium sulfate fractionation, ion-exchange with DEAE-Sepharose F.F. and gel filtration on Sephacryl S-200 column. The purification fold is 22.78 and the specific activity is 15.26U/mg protein.The purified enzyme shows as a single protein band on non-reduced SDS-PAGE. Its relative molecular weight is 52.3kD determined with non-reduced SDS-PAGE.The pI of the enzyme is about 5.1. The Michaelis-Menten constant(Km) is 3.08×10^(-3)mol/L.The optimum pH value of the enzyme is 3.6. The optimum temperature of the enzyme is 52℃.ACPase is stable between pH 3.5～6.0. Its activity is stable below 40℃ and the activity remains 59.2% after incubating at 50℃for 30min.

关 键 词：部分性 新鲜精液 酸性磷酸酶 离子交换柱层析 g蛋白 纯化 凝胶过滤 瘦肉型猪 纱网 胶状物 

分 类 号：S828.1[农业科学—畜牧学]