枯草芽孢杆菌抗菌肽的分离纯化及质谱检测

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 purification,the molecular weight of antibacterial substance of the strain with the highest anti-H.pylori activity was detected by mass spectrometry. Results: The culture supernatant 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.

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