水稻内生枯草芽孢杆菌G87抗菌蛋白的分离纯化及理化特性  

Isolation,purification and characterization of antifungal protein from rice endophytic bacterim Bacillus subtilis G87

作　　者：陈夕军[1] 李娟[1] 孙启利[1] 童蕴慧[1] 徐敬友[1] 

机构地区：[1]扬州大学园艺与植物保护学院,扬州225009

出　　处：《微生物学报》2010年第10期1353-1357,共5页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30571243)~~

摘　　要：【目的】为得到枯草芽孢杆菌(Bacillus subtilis)G87的抗菌蛋白,明确其蛋白理化特性。【方法】采用硫酸铵沉淀和柱层析法进行分离纯化。【结果】获得单一抗菌活性蛋白(峰6-2-1),此抗菌蛋白分子量为50.8 kDa,等电点为5.90。经初步分析,抗菌蛋白不含脂,而含有少量(0.62%)糖;其蛋白部分具有脯氨酸或羟脯氨酸,但不含芳香族氨基酸。抗菌蛋白在高温(≥60℃)和较碱(pH>8)环境下活性明显下降,但较抗紫外线、氯仿和胰蛋白酶、蛋白酶K、胃蛋白酶。【结论】枯草芽孢杆菌G87的抗菌蛋白为不含芳烃的糖蛋白,对高温和碱性条件敏感,而对蛋白酶类和紫外线等不敏感。[ Objective ] In order to obtain the antagonistic protein of Bacillus subtilis G87 and definitude its characterization. [ Methods] Methods of ammonium sulfate precipitating and column chromatography analyzing were used to isolate and purify the protein. [ Results] A purified protein （peak 6-2-1 ） was obtained which molecular weight was 50.8 kD by SDS-PAGE and isoelectric point was 5.90 by IEF-PAGE. The antifungal protein contained 0.62% saccharide and some proline or hydroxyproline, but no lipid and aromatic amino acid. The inhibitory activity of the antifungal protein would decreased distinctly at the higher temperature （ ≥60℃） and in the condition of alkalinity （pH 〉 8 ） , but tolerant to ultraviolet radiation, chloroform, trypsin, proteinase K and pepsin. [ Conclusion] Antifungal protein of Bacillus subtilis G87 was a kind of glycoprotein without aromatic hydrocarbon. It was sensitive to higher temperature and tight alkalinity but not to proteinase analog and ultraviolet radiation et al.

关 键 词：枯草芽孢杆菌 抗菌蛋白 分离纯化 理化特性 水稻内生细菌 

分 类 号：S476[农业科学—农业昆虫与害虫防治]