枯草芽孢杆菌抗菌肽的分离纯化及质谱检测  

Study on purification and mass spectrometric detection of antimicrobial peptide from Bacillus subtilis

作　　者：喻钢[1] 田万红[1] 董思国[1] 王军志[1] 曾明[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2014年第10期1902-1906,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：十二五"重大新药创制"专项课题"国家疫苗生产菌种基因组测序及蛋白指纹图谱库的建设和应用(2013ZX09304101);国家高技术研究发展计划(863计划)益生菌类资源开发及应用(2014AA022210)

摘　　要：目的:研究枯草芽孢杆菌拮抗幽门螺杆菌的能力及分离纯化其抑菌活性物质。方法:以幽门螺杆菌国际标准株NCTC11637为指示菌,采用打孔法检测枯草芽孢杆菌抑制幽门螺旋杆菌能力;挑选拮抗能力最强菌株,对其活性物质进行分离纯化;质谱检测活性物质分子量。结果:枯草芽孢杆菌发酵上清液具有显著抑制幽门螺旋杆菌的能力,其中枯草芽孢杆菌KM抑制幽门螺旋杆菌的能力最强;活性物质经分离纯化后,质谱检测分子量为3402.524D,推测为Subtilosin A结构类似物;10株枯草芽孢杆菌Subtilosin A的前体基因的表达水平与抗幽门螺旋杆菌的能力呈正相关。结论:枯草芽孢杆菌具有不同程度体外拮抗幽门螺旋杆菌能力,其抑菌机制与分泌抗菌肽Subtilosin A结构类似物相关,该类物质可作为治疗幽门螺杆菌感染的候选药物。Objective ：To study the anti - Helicobacter pylori activity and its possible mechanism for Bacillus subtilis. Method ：The anti - H. pylori （ NCTC11637 ） of B. subtilis was tested through the agar diffusion method. After purifica- tion, the molecular weight of antibacterial substance of the strain with the highest anti - H. pylori activity was detected by mass spectrometry. Results：The culture supematant of B. subtilis was efficient to antagonize H. pylori and the anti - H. pylori ability differs in different B. subtilis strains. B. subtilis Km has the most significant anti - H. pylori ability in 10 B. subtilis strains. Mass spectrometry analysis showed that the molecular weight of the antibacterial substance was 3402. 524D,corresponding to the isoform of Subtilosin A. Gene expression analysis of sboA, the precursor of Subtilosin A, showed that they were positive correlated with their anti - H. pylori ability in ten B. subtilis strains. Conclusion： B. subtilisis available to antagonize H. pylori, and the ability is strain specific. The anti -H. pylori ability is relevant with Subtilosin A,an antimicrobial peptide.

关 键 词：枯草芽孢杆菌 幽门螺杆菌 抗菌肽 质谱检测 Subtilosin A 活性成分分离纯化 

分 类 号：R917[医药卫生—药物分析学]