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养殖石斑鱼溃疡病病原哈维氏弧菌胞外产物的研究
陈雅芳1, 王 军1, 苏永全1, 覃映雪1, 王世锋1
厦门大学海洋与环境学院
摘要:
以网箱养殖的患溃疡病青石斑鱼(Epinephlus awoara)和斜带石斑鱼(Epinephlus coioides)分离的哈维氏弧菌(Vibrio harveyi)TS-628菌株为研究对象,采用平板玻璃纸覆盖技术,获得该菌株的胞外产物(ECP),并对其成分进行分析。利用杯碟法研究表明,哈维氏弧菌TS-628菌株的胞外产物具有酪蛋白酶、明胶蛋白酶、淀粉酶、脂肪酶、卵磷脂酶等多种酶活性和溶血活性,且溶血性物质具有热不稳定性。SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)及考马斯亮蓝R-250染色分析表明,ECP蛋白条带主要集中在分子质量范围25.0~66.2 ku之间,经不同时间培养所获得的ECP在凝胶上显示出的蛋白条带几乎一致。采用复性电泳方法(G-PAGE)分析了哈维氏弧菌及其它4种弧菌属细菌的ECP蛋白酶活性,结果发现哈维氏弧菌ECP在酸性条件下,蛋白酶活性较强,在碱性条件下,蛋白酶带明显减少,活性很弱。在相同的pH条件下,5种弧菌中以副溶血弧菌的ECP蛋白酶活性最强,种类多,分子质量范围宽;而哈维氏弧菌的蛋白酶带较少,活性也稍弱。
关键词:  青石斑鱼(Epinephlus awoara)  斜带石斑鱼(Epinephlus coioides)  哈维氏弧菌(Vibrio harveyi)  胞外产物  溃疡病
DOI:
分类号:
基金项目:福建省科技重大资助项目(2001N009);国家863计划资助项目(2001AA621010)
Study on the extra cellular products(ECP) of the pathogenic Vibrio harveyi isolated from the farmed groupers
Abstract:
A pathogenic strain of Vibrio harveyi named TS-628 was isolated from the farmed Epinephlus awoara and Epinephlus coioides suffering from ulcered syndrome. The extra cellular products(ECP) of the bacteria were found to show gelatinase, amylase,lipase,phospholipase,caseinase activities and haemolytic property. ECP samples obtained from the SDS-polyacrylamide gel eletrophoresis(SDS-PAGE) after different incubation periods produced almost the same bands. The molecular weights of the proteins in these ECP mainly ranged from 25.0 ku to 66.2 ku. The extracellular proteinase(ECPase) activities of V. harveyi and the other four Vibrio species(V.fischeri, V.parahaemolyticus, V.fluvialis, V.alginolyticus) were compared by SDS-gelatin-polyacrylamide gel electrophoresis (SDS-GPAGE) and stained with coomassie brilliant blue R-250. The results from the SDS-G-PAGE of these ECPases indicated that: (1) For V. harveyi, the activity and the sorts of ECPase under acidic condition were higher and more than those under basic condition; (2) In the same case, ECPase of V.parahaemolyticus had the strongest activity and various styles as well as wide molecular weight range among the five Vibrio species, and it seemed that there are the lower proteinase activity and less sorts of the ECPase in V. harveyi.
Key words:  Epinephlus awoara  Epinephlus coioides  Vibrio harveyi  extra cellular products(ECP)  ulcer disease
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