NDLI: Optimization of cold methanol quenching for quantitative metabolomics of Penicillium chrysogenum

Content Provider	Springer Nature Link
Author	Jonge, Lodewijk P. Douma, Rutger D. Heijnen, Joseph J. Gulik, Walter M.
Copyright Year	2011
Abstract	A sampling procedure for quantitative metabolomics in Penicillium chrysogenum based on cold aqueous methanol quenching was re-evaluated and optimized to reduce metabolite leakage during sample treatment. The optimization study included amino acids and intermediates of the glycolysis and the TCA-cycle. Metabolite leakage was found to be minimal for a methanol content of the quenching solution (QS) of 40% (v/v) while keeping the temperature of the quenched sample near −20°C. The average metabolite recovery under these conditions was 95.7% (±1.1%). Several observations support the hypothesis that metabolite leakage from quenched mycelia of P. chrysogenum occurs by diffusion over the cell membrane. First, a prolonged contact time between mycelia and the QS lead to a somewhat higher extent of leakage. Second, when suboptimal quenching liquids were used, increased metabolite leakage was found to be correlated with lower molecular weight and with lower absolute net charge. The finding that lowering the methanol content of the quenching liquid reduces metabolite leakage in P. chrysogenum contrasts with recently published quenching studies for two other eukaryotic micro-organisms. This demonstrates that it is necessary to validate and, if needed, optimize the quenching conditions for each particular micro-organism.
Starting Page	727
Ending Page	735
Page Count	9
File Format	PDF
ISSN	15733882
Journal	Metabolomics
Volume Number	8
Issue Number	4
e-ISSN	15733890
Language	English
Publisher	Springer US
Publisher Date	2011-10-07
Publisher Institution	Metabolomics Society
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Quantitative metabolomics Intracellular metabolites Cold methanol quenching Metabolite leakage Penicillium chrysogenum Biomedicine general Biochemistry Developmental Biology Cell Biology Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Endocrinology, Diabetes and Metabolism Clinical Biochemistry Biochemistry

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