期刊
FEMS YEAST RESEARCH
卷 15, 期 6, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/femsyr/fov049
关键词
metabolomics; quenching; filtration; centrifugation; metabolite loss; Pichia pastoris
资金
- Biomin Research Center
- Boehringer-Ingelheim RCV
- Lonza AG
- Biocrates Life Sciences AG
- VTU Technology GmbH
- Sandoz GmbH
- Federal Ministry of Science, Research and Economy (BMWFW)
- Federal Ministry of Traffic, Innovation and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol
- ZIT-Technology Agency of the City of Vienna through COMET
Metabolomics can be defined as the quantitative assessment of a large number of metabolites of a biological system. A prerequisite for the accurate determination of intracellular metabolite concentrations is a reliable and reproducible sample preparation method, which needs to be optimized for each organism individually. Here, we compare the performance of rapid filtration and centrifugation after quenching of Pichia pastoris cells in cold methanol. During incubation in the quenching solution, metabolites are lost from the cells with a half-life of 70-180 min. Metabolites with lower molecular weights showed lower half-lifes compared to metabolites with higher molecular weight. Rapid filtration within 2 min after quenching leads to only minor losses below 2%, and is thus the preferred method for cell separation.
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