Global metabolite analysis of yeast: evaluation of sample preparation methods

Silas Granato Villas-Bôas, Jesper Højer-Pedersen, Mats Fredrik Åkesson, Jørn Smedsgaard, Jens Nielsen

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    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.
    Original languageEnglish
    JournalYeast
    Volume22
    Issue number14
    Pages (from-to)1155-1169
    ISSN0749-503X
    DOIs
    Publication statusPublished - 2005

    Fingerprint

    Dive into the research topics of 'Global metabolite analysis of yeast: evaluation of sample preparation methods'. Together they form a unique fingerprint.

    Cite this