鳗弧菌胞外产物中致病因子的分离纯化及性质研究  

Purification and Characterization of a Virulent Factor in Extracellular Products of Vibrio Anguillarum

作　　者：陈师勇[1] 莫照兰[1] 张振冬[1] 徐永立[1] 张培军[1] 

机构地区：[1]中国科学院海洋研究所海洋生物学开放室,青岛266071

出　　处：《高技术通讯》2002年第8期96-100,共5页Chinese High Technology Letters

基　　金：国家重点基础研究 (G19990 12 0 0 3)资助项目

摘　　要：鳗弧菌致病株M3的胞外产物经超滤、SephadexG10 0凝胶层析、DEAE cellulose离子交换层析和HPLC凝胶色谱层析纯化后 ,得到一个纯化的毒性蛋白酶 ,其对金鱼的半数致死量为 1 2微克蛋白 /克体重。该蛋白酶的分子量为 36 5kDa ,等电点为 5 1;与底物A zocasin作用的最适pH为 8 0 ,最适作用温度 5 5℃ ,85℃作用 30min后酶被灭活。该酶活性可被EDTA、EGTA及 1,10 菲咯啉抑制 ,表明它是金属蛋白酶。N端部分氨基酸序列经测定为AGATGTGPGGAGLTG。An extracellular protease produced by Vibrio anguillarum strain M3 was purified following the steps of ultrafiltration, gel filtration with Sephadex G100, ion-exchange chromatography with DEAE-cellulose A52, HPLC gel filtration. The purified protease was lethal for goldfish at LD 50 value of 1.2μg protein/g body weight. With an isoelectric point of 5.1 and molecular weight of 36.5kDa, it showed maximal activities at pH8.0 and 55℃. After incubated at 85℃ in 30min, the protease lost all its activity. The fact that chelating agents (EDTA?EDGA and 1,10-phenanthroline) could inhibit the protease indicated it was a metalloprotease. Partial N-terminal amino acid sequence of the protease was AGATGTGPGGAGLTG.

关 键 词：胞外产物 鳗弧菌 致病因子 蛋白酶 分离纯化 鱼类 毒性 流行性疾病 

分 类 号：S941[农业科学—水产养殖]