鳜嗜水气单胞菌GYK1株胞外产物提取、纯化及生物学活性分析  

Extraction, purification and biological activity analysis of extracellular products of Aeromonas hydrophila GYK1 strain isolated from mandarinfish(Siniperca chuatsi)

作　　者：潘厚军[1] 石存斌[1] 刘瑞明[1] 胡学峰[1] 张敏[1] 吴淑勤[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所,广东广州510380

出　　处：《南方水产》2006年第3期1-6,共6页South China Fisheries Science

基　　金：国家科技攻关计划(2004BA526B0501);农业部农业结构调整重大技术专项(041105B);广东省攻关项目(2002A20510)

摘　　要：以鳜分离的嗜水气单胞菌GYK1株为代表，采用玻璃纸覆盖技术提取了8株嗜水气单胞菌和1株温和气单胞菌的胞外产物，SDS-PAGE电泳分析显示，8株嗜水气单胞菌的胞外产物均有35kDa的共同蛋白带。酶活性和溶血性分析表明，GYK1株胞外产物具有酪蛋白酶、淀粉酶、脂肪酶、明胶酶活性，但不具备脲酶活性；溶血性较强，蛋白浓度为365 μg·mL^-1的胞外产物对鳜血细胞的溶血价为2^13，对加州鲈、银鲫、小白鼠、兔、绵羊、人O型血的红细胞的溶血价在2^11～2^14之间。应用聚丙烯酰胺葡聚糖凝胶柱层析，对GYK1株的胞外产物进行了初步纯化，得到分子量为35 kDa的蛋白带，该纯化产物具溶血性，但溶血性比粗胞外产物降低，对鳜和小鼠血细胞的溶血价均为2^5。Extracellular products （ECP） of Aeromonas hydrophila （Ah） GYK1, isolated from mandarin-fish, Siniperca chuatsi and other 7 strains of Ah and 1 strain of A. sobria were extracted by cellophane cover technique. Some characteristics of the ECP of GYK1 strain were analyzed. The ECP of GYK1 strain had enzyme activity of caseinase, amylase, lipase and gelatinase, but had not activity of urease. The ECP also had strong hemolytic activity. The hemolytic titer of crude ECP, with protein concentration of 365 μg·mL^-1, to erythrocyte of mandarinfish was 2^13, to that of other two kinds of fishes and five kinds of mammals were between 2^ll - 2^14. The result of SDS-PAGE showed that the band with molecular weight about 35 kDa is the common protein band of the ECP of the tested 8 Ah strains. ECP of GYK1 strain was purified and the band of 35 kDa was obtained by Hiprep^R Sephacryl S-200 high resolution chromatography. And the purified product had hemolytic activity, but was weaker than that of the crude ECP. The hemolytic titer of＇crude ECP to mandarinfish and mouse was 2^13 and 2^12, while that of the purified ECP was both 2^5.

关 键 词：鳜 嗜水气单胞菌 胞外产物 酶活性 

分 类 号：S917.1[农业科学—水产科学]