Analysis of Intracellular Metabolites from Microorganisms: Quenching and Extraction Protocols

Metabolites

Raphael Aggio¹  Silas G. Villas-Boas²  Farhana R. Pinu³ 

[1] Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool L693BX, UK;School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland 1010, New Zealand;The New Zealand Institute for Plant & Food Research Limited, Private Bag 92169, Auckland 1142, New Zealand;

关键词: metabolite footprinting;    exometabolome;    culture medium;    metabolic modelling;    sample preparation;    analytical instruments;    data integration;    extracellular metabolites;    intracellular metabolites;   

DOI  :  10.3390/metabo7040053

来源: DOAJ

【 摘 要 】

Sample preparation is one of the most important steps in metabolome analysis. The challenges of determining microbial metabolome have been well discussed within the research community and many improvements have already been achieved in last decade. The analysis of intracellular metabolites is particularly challenging. Environmental perturbations may considerably affect microbial metabolism, which results in intracellular metabolites being rapidly degraded or metabolized by enzymatic reactions. Therefore, quenching or the complete stop of cell metabolism is a pre-requisite for accurate intracellular metabolite analysis. After quenching, metabolites need to be extracted from the intracellular compartment. The choice of the most suitable metabolite extraction method/s is another crucial step. The literature indicates that specific classes of metabolites are better extracted by different extraction protocols. In this review, we discuss the technical aspects and advancements of quenching and extraction of intracellular metabolite analysis from microbial cells.

【 授权许可】

Unknown