菲律宾蛤仔(Ruditapes philippinesis)血浆中防卫素的纯化及其抑菌功能  

Purification of a Big Defensin from Ruditapes philippinesis and Its Antibacterial Activity

作　　者：魏玉西[1] 郭道森[1] 李荣贵[1] 陈皓文[2] 陈培勋[1] 

机构地区：[1]青岛大学生物系,青岛266071 [2]国家海洋局第一海洋研究所,青岛266061

出　　处：《生物化学与生物物理学报》2003年第12期1145-1148,共4页

基　　金：国家自然科学基金资助项目 (No .40 0 860 0 1)~~

摘　　要：经硫酸铵沉淀、凝胶排阻层析和离子交换层析 ,自菲律宾蛤仔 (Ruditapesphilippinnesis)血浆中分离纯化得到一种新的蛋白质———防卫素RPD 1(Ruditapesphilippinesisdefensin 1)。经SDS PAGE分析 ,其估计分子量为 2 4 .8kD ;经ABI4 73蛋白质自动测序仪测定 ,其N端部分氨基酸序列为AVPDVAFNAYG。在NCBI和EBI EMBL两个蛋白质数据库中未发现任何与该蛋白质有同源性的已知蛋白质。该蛋白质对革兰氏阳性菌和革兰氏阴性菌具有广谱抗性。对金黄色葡萄球菌 (Staphylococcusaureus)、枯草杆菌 (Bacillussubtilis)、四联微球菌 (Micrococcustetragenus)、大肠杆菌 (Escherichiacoli)、副溶血弧菌 (Vibrioparahaemolyticus)和鳗弧菌 (Vibrioanguillarum)的最小抑菌浓度分别为 9.6mg L、76 .8mg L、38.4mg L、76 .8mg L、19.2mg L和 19.2mg L。实验结果证明RPD 1为一种新的防卫素蛋白质。A novel big defensin was isolated and characterized from the plasma of Ruditapes philippinesis. It was purified to homogeneity by means of precipitation with (NH 4) 2SO 4, gel-exclusion chromatography, two kinds of cation-exchange chromatography and named RPD-1. Its relative molecular mass was 24.8 kD by means of SDS-PAGE. By means of ABI437 amino acid sequence analyser, its 11 NH 2-terminal amino acid sequence is AVPDVAFNAYG. Two databank systems (NCBI and EBI/EMBL) was indexed, no sequence with homology to RPD-1 was found. Furthermore, it exhibited strong inhibition on the growth of Gram-negative and -positive bacteria. Minimal inhibitory concentration (MIC) of RPD-1 against Staphylococcus aureus, Bacillus subtilis, Micrococcus tetragenus, Escherichia coli, Vibrio parahaemolyticus, Vibrio anguillarum was 9.6 mg/L, 76.8 mg/L, 38.4 mg/L, 76.8 mg/L, 19.2 mg/L, 19.2 mg/L respectively.

关 键 词：菲律宾蛤仔 血浆 蛋白质 防卫素RPD-1 抑菌功能 纯化 

分 类 号：S917[农业科学—水产科学]