NDLI: Fast sampling for quantitative microbial metabolomics: new aspects on cold methanol quenching: metabolite co-precipitation

Content Provider	Springer Nature Link
Author	Zakhartsev, Maksim Vielhauer, Oliver Horn, Thomas Yang, Xuelian Reuss, Matthias
Copyright Year	2014
Abstract	The intra- and extracellular concentrations of 16 metabolites were measured in chemostat (D = 0.1 h$^{−1}$) anaerobic cultures of the yeast Saccharomyces cerevisiae CEN.PK-113-7D growing on minimal medium. Two independent sampling workflows were employed: (i) conventional cold methanol quenching and (ii) a differential approach. Metabolites were quantified in different sample fractions (total, extracellular, quenching supernatant, methanol/water extract and pellet) in order to derive their mass balance. The differential method in combination with absolute metabolite quantification by gas-chromatography with isotope dilution mass spectrometry (GC–IDMS) was used as a benchmark to assess quality of the cold methanol quenching procedure. Quantitative comparison of metabolite concentrations in all fractions collected by different quenching techniques indicates asystematic loss of the total mass of various metabolites in course of the cold methanol quenching. Pellet resulting from the cold methanol quenching besides biomass contains considerable amounts of precipitated inorganic salts from the fermentation media. Quantitative analysis has revealed significant co-precipitation of polar extracellular metabolites together with these salts. This phenomenon is especially significant for metabolites with large extracellular mass-fraction. We report that the co-precipitation is a hitherto neglected phenomenon and concluded that its degree strongly linked to culturing conditions (i.e. media composition) and chemical properties of the particular metabolite. Thus, intracellular metabolite levels measured from samples collected by cold methanol quenching might be uncertain and variably biased due to corruption by described phenomena.
Starting Page	286
Ending Page	301
Page Count	16
File Format	PDF
ISSN	15733882
Journal	Metabolomics
Volume Number	11
Issue Number	2
e-ISSN	15733890
Language	English
Publisher	Springer US
Publisher Date	2014-07-22
Publisher Institution	Metabolomics Society
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Yeast Sampling techniques Quantitative metabolite measurements Biochemistry Molecular Medicine Cell Biology Developmental Biology Biomedicine general
Content Type	Text
Resource Type	Article
Subject	Endocrinology, Diabetes and Metabolism Clinical Biochemistry Biochemistry

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