1种致病性鳗弧菌胞外蛋白酶的分离纯化及性质  

Purification of an extracellular protease from Vibrio anguillarum and its physicochemical properties

作　　者：陈吉祥[1] 刘霜[1] 李筠[1] 王祥红[1] 杜宗军[1] 于德华[1] 纪伟尚[1] 徐怀恕[1] 

机构地区：[1]青岛海洋大学海洋生命学院,达尔文实验室山东青岛266003

出　　处：《中国水产科学》2002年第4期318-322,共5页Journal of Fishery Sciences of China

基　　金：国家自然科学基金资助项目 (3 9870 5 81) ;英国达尔文项目(162 /8/0 65 )

摘　　要：从山东省莱州海区自然发病的花鲈 (Lateolobraxjapanicus)体内分离到 1株致病性鳗弧菌 ,经硫酸铵盐析、DEAE SepharoseFastFlow和Sephadex G10 0凝胶层析等方法从其培养液中分离纯化了 1种胞外蛋白酶。用SDS PAGE电泳测得蛋白质的分子量为 36 7kD ,酶的最适温度为 5 0℃ ,对热不稳定 ,70℃ 15min完全失去活力 ;最适pH为 7 0 ;1mmol/L的PMSF对酶活性无影响 ,部分金属离子如Cu2 + 、Fe2 + 、Fe3 + 、Zn2 + 对酶活力有抑制作用 ,而Ca2 + 对酶有一定程度的激活作用 ,1mmol/LEDTA能完全抑制酶的活性 ,表明该酶是A strain of Vibrio anguillarum W-1 was isolated from the diseased seaperch (Lateolabrax japonicus) in Laizhou Bay, Shandong Province, and an extracellular protease was partially purified from the culture solution of the V. anguillarum by ammonium sulfate, DEAE epharose fast flow and Sephadex G-100 . The results show that, the molecular weight of the purified enzyme from V. anguillarum by SDS-PAGE is 36.7 kD; the optimum temperature for the enzyme activity is 50 ℃ ,and the enzyme is heatlabile that it would entirely lose its activity in 15 min at 70℃ ; the optimum pH is 7.0 ;1 mmol/L of EDTA and some metal ions (Cu 2+ ,Fe 2+ ,Fe 3+ ,Zn 2+ ) can inhibit the enzyme activity ,but Ca 2+ is on the contrary that it can activate the enzyme to some extent. 1 mmol/L of PMSF has no effect on the enzyme ,and the enzyme is suggested a kind of melalloprotease.

关 键 词：致病性 鳗弧菌 胞外蛋白酶 分离纯化 理化性质 

分 类 号：S941.42[农业科学—水产养殖]