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高效液相色谱-电喷雾质谱法分离和鉴别枯草芽孢杆菌产生的脂肽类化合物

Isolation and identification of lipopeptides produced by Bacillus subtilis using high performance liquid chromatography and electrospray ionization mass spectrometry

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【作者】 陈华王丽袁成凌郑之明余增亮

【Author】 CHEN Hua, WANG Li, YUAN Chengling, ZHENG Zhiming, YU Zengliang (Key Laboratory of Ion Beam Bioengineering, the Chinese Academy of Sciences, Hefei 230031, China)

【机构】 中国科学院离子束生物工程学重点实验室中国科学院离子束生物工程学重点实验室 安徽合肥230031安徽合肥230031

【摘要】 枯草芽孢杆菌JA因产生多种脂肽类化合物而具有广阔的开发前景。JA发酵液经过离心、酸沉淀、甲醇抽提等步骤得到脂肽类化合物的粗提物。将粗提物溶于流动相,采用反相高效液相色谱分离,对收集的洗脱峰组分进行电喷雾质谱(ESI-MS)分析。根据质荷比推断JA菌株产生的脂肽类化合物属于3个家族,分别为surfactin,iturin和fengycin,是枯草芽孢杆菌合成的重要生物表面活性素。对一级质谱中的主成分进行串联质谱分析,进一步确定了3种脂肽类化合物的分子结构。实验证明ESI-MS是一种鉴定脂肽类化合物及其同系物的可靠方法。

【Abstract】 Bacillus subtilis JA produces a broad spectrum of bioactive lipopeptides with great potential applications in agriculture and industry. Crude lipopeptides were extracted with 100% methanol from the precipitate which was obtained by adding 6 mol/L HCl to the cell-free culture broth. The crude extract was separated on a Diamonsil C18 column (5 μm, 250 mm×4.6 mm) in reversed-phase high performance liquid chromatography (HPLC) system to separate the lipopeptide homologues. Five peaks were eluted from HPLC. Electrospray ionization mass spectrometry (ESI-MS) was used to analyze each HPLC fraction. The results showed three series of ion peaks. According to the values of m/z, the three series of ions were classified into surfactin, iturin and fengycin homologues, which were well-known biosurfactants produced by B. subtilis strains. The major ions were structurally characterized using tandem mass spectrometry. This study suggested a reasonable method for the isolation and identification of lipopeptides produced by B. subtilis strains.

【基金】 国家自然科学基金资助项目(No20576132)
  • 【文献出处】 色谱 ,Chinese Journal of Chromatography , 编辑部邮箱 ,2008年03期
  • 【分类号】Q93
  • 【被引频次】37
  • 【下载频次】933
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