LESA 质谱高通量筛查细菌表面的抗生素

Rapid Commun Mass Spectrom. 2012 Oct 30;26(20):2477-82. doi: 10.1002/rcm.6365.
 

Kai M, González I, Genilloud O, Singh SB, Svatoš A.
Research Group Mass Spectrometry, Max Planck Institute for Chemical Ecology, Jena, Germany.

德国耶拿，马克斯·普朗克化学生态学研究所，质谱研究组

RATIONALE背景：

There is a need to find new antibiotic agents to fight resistant pathogenic bacteria. To search successfully for novel antibiotics from bacteria cultivated under diverse conditions, we need a fast and cost-effective screening method.

研发新的抗生素来对抗耐药菌刻不容缓。我们需要一种快速、经济的筛选方法，以便自多种培养条件下的大量细菌中成功地寻找新型抗生素。

METHODS方法:

A combination of Liquid Extraction Surface Analysis (LESA), automated chip-based nanoelectrospray ionization, and high-resolution mass or tandem mass spectrometry using an Orbitrap XL was tested as the screening platform. Actinobacteria, known to produce well-recognized thiazolyl peptide antibiotics, were cultivated on a plate of solid medium and the antibiotics were extracted by organic solvent mixtures from the surface of colonies grown on the plate and analyzed using mass spectrometry (MS).

液滴萃取表面分析技术（LESA）结合基于芯片的自动化纳升电喷雾离子化和高分辨质谱或串联质谱 Orbitrap XL被选为为筛选平台。放线菌以产生易识别的噻唑肽类抗生素为众人所熟悉，我们把它放置在培养皿中培养，采用混合有机溶液萃取培养好后的细菌簇表面并进行质谱分析。

RESULTS结果:

LESA combined with high-resolution MS is a powerful tool with which to extract and detect thiazolyl peptide antibiotics from different Actinobacteria. Known antibiotics were correctly detected with high mass accuracy (<4 ppm) and structurally characterized using tandem mass spectra. Our method is the first step toward the development of a novel high-throughput extraction and identification tool for antibiotics in particular and natural products in general.

LESA结合高分辨质谱在萃取及检测不同放线菌产生的噻唑肽类抗生素上展示出强大的优势，通过高质量精度（小于4 ppm），我们可以准确检测出已知抗生素，并根据串联质谱图进行结构确认。我们的方法是为设计针对天然产物开发尤其对特殊抗生素的高通量萃取和鉴定工具而迈出的第一步

CONCLUSIONS结论:

The method described in this paper is suitable for (1) screening the natural products produced by bacterial colonies on cultivation plates within the first 2 min following extraction and (2) detecting antibiotics at high mass accuracy; the cost is around 2 Euro per sample.

本文所描述的方法非常适合（1）快速检测在培养皿上培养的细菌菌群产生的天然产物，每个样品在萃取完后两分钟内完成检测，和（2）利用高质量精度来检测抗生素；每个样本检测成本约为2欧元。