大肠杆菌代谢物组测定中抽提方法的比较分析

代谢物组学作为一种重要的功能基因组学工具,已经在微生物性状改良等方面得到了应用。实施代谢物组学研究需要定性和定量分析细胞内的全部代谢物。所以,必须有一种能够提供良好的重复性和相对抽提效率较高的细胞内代谢物抽提方法。本研究以特定厌氧条件下培养的大肠杆菌为研究对象,以电喷雾质谱直接进样测定为评价标准,比较并分析了冰甲醇、热乙醇和碱抽提3种抽提方法。结果显示,冰甲醇抽提的样品在电喷雾质谱实验中表现出良好的重复性和最高的峰强度,并能够同时检测到超过20种不同的胞内代谢物。

Comparisons of Different Extraction Methods in Escherichia Coli Metabolome Analysis

Metabolomics,as an important functional genomics tool,had been used practically in strain improvement.In order to qualify and quantify the intracellular global metabolites,there must be a reliable cultivation and extraction methodology,which can provide good reproducibility and high peak intensities.This research focused on Escherichia coli,which is under regulated anaerobic fermentation conditions.Three different extraction methods cold methanol,hot ethanol and alkali extraction were analyzed and compared using the direct infusion electrospray ionization-mass spectrometry(ESI-MS) as the evaluation standard.The results show that cold methanol extraction method has the best reproducibility;highest signal intensities and can simultaneously detect more than 20 intracellular metabolites.