色谱

色谱

• 研究论文 • 上一篇    下一篇

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

陈华,王丽,袁成凌,郑之明,余增亮

  

  1. Key Laboratory of Ion Beam Bioengineering, the Chinese Academy of Sciences, Hefei 230031, China
  • 收稿日期:2007-10-10 修回日期:2007-12-10 出版日期:2008-05-30 发布日期:1984-03-25
  • 通讯作者: 郑之明

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

CHEN Hua, WANG Li, YUAN Chengling, ZHENG Zhiming, YU Zengliang   

  1. Key Laboratory of Ion Beam Bioengineering, the Chinese Academy of Sciences, Hefei 230031, China
  • Received:2007-10-10 Revised:2007-12-10 Online:2008-05-30 Published:1984-03-25
  • Contact: ZHENG Zhiming

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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。

关键词: 高效液相色谱-电喷雾质谱法, 枯草芽孢杆菌, 脂肽类化合物

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.

Key words: Bacillus
subtilis, lipopeptides , high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS)