洋葱伯克氏菌Burkholderia contaminans抗菌蛋白分离纯化及生物学特性分析  

Isolation,Purification and Characterization of an Antifungal Protein from Burkholderia contaminans

作　　者：张婧婷[1] 施俊凤[2] 范三红[1] ZHANG Jingting;SHI Junfeng;FAN Sanhong(College of Life Science,Shanxi University,Taiyuan 030006,China;？Institute of Agricultural Product Storage and Fresh Keeping,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,China)

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]山西省农业科学院农产品贮藏保鲜研究所,山西太原030031

出　　处：《食品科学》2018年第14期179-184,共6页Food Science

基　　金：山西省重点研发项目(201703D221010-1);山西省农业科学院博士后基金项目(BSH-2015JJ-003);山西省农业科学院特色农业项目(YGG17001)

摘　　要：洋葱伯克氏菌(Burkholderia contaminans)从杏果实表面分离,实验证明其对果蔬采后多种病原真菌具有较强的抑制作用。为了进一步揭示其抑菌机理,通过菌体裂解、硫酸铵分级盐析、纤维素DE-52阴离子交换柱层析及Sephadex G-100凝胶过滤柱层析,得到一种具有抗菌活性的蛋白。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)检测结果表明,该蛋白呈单一条带,其分子质量约为17.6kDa。粗提蛋白溶液具有一定热稳定性,在不同pH值条件下稳定,对有机试剂、紫外照射、蛋白酶不敏感。50mg/mL的Tween-80处理粗提液,活性比对照增加了12.5%,对还原剂SDS和二硫苏糖醇敏感。Burkholderia contaminans B-1, isolated from the surface of apricots, is an antagonistic bacterium against Botrytis cinerea and other important postharvest pathogenic fungi in fruits and vegetables. In order to investigate the mechanism of action of B-1, we extracted a protein with antimicrobial activity after lysis of the cells. The protein was purified by bioassayguided fractionation using fractional ammonium sulfate precipitation, DEAE-cellulose DE-52 ion-exchange and Sephadex G-100 gel filtration column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） analysis suggested that the purified protein was homogeneous and its apparent molecular mass was 17.6 kDa. The protein was stable to heating and pH variations and was insensitive to organic reagents, protease and UV irradiation. Its inhibitory activity was increased by 12.5% in the presence 50 mg/mL Tween-80 as compare to that of the control（CK）. The protein was sensitive to SDS and dithiothreitol（DTT）.

关 键 词：洋葱伯克氏菌(Burkholderia contaminans) 抗菌蛋白 分离纯化 理化性质 

分 类 号：Q93[生物学—微生物学]