枯草芽孢杆菌B21抗菌蛋白的分离纯化及特性研究  

Purification and characterization of an antifungal protein from Bacillus subtilis B21

作　　者：齐爱勇[1] 魏东盛[2] 刘大群[1] 

机构地区：[1]河北农业大学科技处,河北省农作物病虫害生物防治工程技术研究中心,河北保定071001 [2]南开大学生命科学学院,天津300071

出　　处：《河北农业大学学报》2011年第3期56-59,共4页Journal of Hebei Agricultural University

基　　金：河北省科技厅攻关项目“棚室蔬菜灰霉病生物防治拮抗菌株筛选及应用”(01220139D)资助

摘　　要：枯草芽孢杆菌（Bacillus subtilis）B21对番茄灰霉病菌（Botrytis cinerea Pers.）有较强的抑制作用,其产生的抗菌物质主要是抗菌蛋白。为明确该抗菌蛋白的理化性质,利用饱和硫酸铵沉淀法得到拮抗粗提蛋白,所获粗提蛋白不耐高温,对胰蛋白酶和蛋白酶K部分敏感,对链霉蛋白酶不敏感而对氯仿敏感,作用活性pH值范围在5~11之间。经过Sephacryl S-200 HR凝胶过滤柱层析和DEAE Fast Flow离子交换层析等方法从B21的发酵液中纯化了抗菌蛋白,通过SDS-PAGE电泳分析,确定该蛋白质的分子量为43 kDa。Bacillus subtilis B21 showed strongly antibacterial activity against Botrytis cinerea Pers..Antifungal protein produced by B21 was the main antifungal substance.To study the physicochemical characteristics of antifungal proteins isolated from strain B21,crude proteins was extracted from precipitation of the cell culture of strain B21 with ammonium sulphate at 80% saturation.The determination of bioactivity of this antagonistic protein showed that it could not endure high temperature,was partially sensitive to Trypsin and Proteinase K,was insensitive to Pronase but sensitive to Chloroform.Its active pH range was wide,from 5.0 to 11.0.Through Sephacryl S-200 HR column chromatography and DEAE Fast Flow column chromatography,the antifungal protein was prepared and purified.The purified protein was demonstrated to be one single protein band by SDS-PAGE with a molecular weight of 43KD.

关 键 词：枯草芽孢杆菌B21 番茄灰霉病菌 抗菌蛋白 分离纯化 

分 类 号：S432[农业科学—植物病理学]