哈维氏弧菌TS-628菌株胞外产物(ECP)蛋白酶活性的研究  

Study on protease activities of the ECP of pathogenic bacterium Vibrio harveyi strain TS-628

作　　者：覃映雪[1,2] 苏永全[2] 陈雅芳[2] 王军[2] 

机构地区：[1]集美大学水产学院,厦门361021 [2]厦门大学海洋系,亚热带海洋研究所,361005

出　　处：《海洋水产研究》2008年第1期81-85,共5页Marine Fisheries Research

基　　金：福建省青年科技人才创新基金项目(2006F3096);集美大学科研基金项目共同资助

摘　　要：采用复性电泳技术,研究环境pH、温度及培养时间对哈维氏弧菌TS-628菌株胞外产物(ECP)蛋白酶活性的影响。结果表明,胞外产物蛋白酶最适反应pH在8.5左右,酸对ECP蛋白酶活性的抑制明显强于碱对它的抑制作用;最适反应温度在20～50℃之间,20℃以下或50℃以上蛋白酶活性则受到明显抑制,表明蛋白酶不仅对高温敏感,对低温也很敏感;培养12h的胞外产物蛋白酶活性最强。最适温度和pH条件下,主要出现3种蛋白酶,其中分子量为94kDa和26kDa的两种蛋白显示了很强的蛋白酶活性,而且能在比较极端的温度和pH值条件下保持活性,而分子量为35kDa蛋白酶只在最适反应条件下出现,并且该蛋白酶的最适反应条件与弧菌病暴发时的环境条件极为相似。由此可推测分子量94kDa和分子量为26kDa的两种蛋白可能是弧菌维持生存所必需,而分子量为35kDa的蛋白酶虽然只出现在特定条件下,却可能在致病过程中起重要作用。SDS-gelatin-PAGE assay was and growing time on the protease activities of used to study the effects of pH, temperatures, the extracellular products （ECP） of Vibrio barveyi TS-628 strain. The results showed that the optimal pH for ECP protease activity was about 8.5 and the protease activity was inhibited more easily by lower values of pH than by higher ones. The optimal temperature for the protease was 20-50℃, and significant decrease in activity was observed at temperatures below 20℃ or above 50℃, which indicated that the protease was sensitive to both low and high temperatures. The highest level of the extracellular product protease activity was present after 12h incubation. Under the condition of optimal temperatures and pH, there were three kinds of protease present. 94kDa, 26kDa and 35kDa. The 94kDa and 26kDa proteases displayed high activity and could remain active under extreme temperatures and pH, while the 35kDa protease was only active under the optimal condition, which was similar to the condition for the outburst of pathogenic vibrio. It is suggested that the 94kDa and 26kDa proteases were necessary in maintaining the vibrio survival and the 35kDa protease could play an important role in the pathogenicity of the bacteria.

关 键 词：哈维氏孤菌 胞外产物 蛋白酶 

分 类 号：S917.1[农业科学—水产科学]