Global metabolite analysis of yeast:: evaluation of sample preparation methods
Global metabolite analysis of yeast:: evaluation of sample preparation methods
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DOI：
10.1002/yea.1308
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发表时间：
2005-10-30
期刊：
影响因子：
2.6
通讯作者：
Nielsen, J
中科院分区：
文献类型：
--
作者：
Villas-Bôas, SG;Hojer-Pedersen, J;Nielsen, J
样品制备被认为是微生物代谢组学分析中的限制性步骤之一。真核生物和原核生物在用于代谢物分析的经典样品制备方法的几个步骤中表现得非常不同。即使在真核生物王国中，细胞结构也有很大的多样性，这使得盲目采用为特定微生物群体设计的方案是不明智的。因此，我们审查和评估了整个样品制备程序的酵母代谢物分析。我们的重点一直是在代谢组学分析，这是一个非常不同的化学和物理性质的大量代谢物的分析目前的需求。这项工作报告了在用冷甲醇溶液淬灭酵母细胞期间观察到的细胞内代谢物的泄漏，六种不同方法提取细胞内代谢物的功效，以及通过冻干和溶剂蒸发进行样品浓缩期间注意到的损失。一个更可靠的程序，建议淬火酵母细胞与冷甲醇溶液，然后提取细胞内代谢产物的纯甲醇。该方法可以与减压溶剂蒸发相结合，因此代表了一种有吸引力的样品制备程序，用于酵母的高通量代谢组学分析。版权所有（c）2005年约翰威利父子有限公司。
Sample preparation is considered one of the limiting steps in microbial metabolome analysis. Eukaryotes and prokaryotes behave very differently during the several steps of classical sample preparation methods for analysis of metabolites. Even within the eukaryote kingdom there is a vast diversity of cell structures that make it imprudent to blindly adopt protocols that were designed for a specific group of microorganisms. We have therefore reviewed and evaluated the whole sample preparation procedures for analysis of yeast metabolites. Our focus has been on the current needs in metabolome analysis, which is the analysis of a large number of metabolites with very diverse chemical and physical properties. This work reports the leakage of intracellular metabolites observed during quenching yeast cells with cold methanol solution, the efficacy of six different methods for the extraction of intracellular metabolites, and the losses noticed during sample concentration by lyophilization and solvent evaporation. A more reliable procedure is suggested for quenching yeast cells with cold methanol solution, followed by extraction of intracellular metabolites by pure methanol. The method can be combined with reduced pressure solvent evaporation and therefore represents an attractive sample preparation procedure for high-throughput metabolome analysis of yeasts. Copyright (c) 2005 John Wiley & Sons, Ltd.