title: Metabolomics sampling of Pichia pastoris revisited: rapid filtration prevents metabolite loss during quenching.
abstract: 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.
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Hannes
authors.initials: H
authors.lastname: Russmayer
authors.affiliation: Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria Department of Chemistry, Division of Analytical Chemistry, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Christina
authors.initials: C
authors.lastname: Troyer
authors.affiliation: Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria Department of Chemistry, Division of Analytical Chemistry, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Neubauer
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Matthias G
authors.initials: MG
authors.lastname: Steiger
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gasser
authors.affiliation: Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria Department of Chemistry, Division of Analytical Chemistry, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Stephan
authors.initials: S
authors.lastname: Hann
authors.affiliation: Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria Institute of Analytical Chemistry, University of Vienna, Waehringer Str. 42, 1090 Vienna, Austria.
authors.forename: Gunda
authors.initials: G
authors.lastname: Koellensperger
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria michael.sauer@boku.ac.at.
authors.forename: Michael
authors.initials: M
authors.lastname: Sauer
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Diethard
authors.initials: D
authors.lastname: Mattanovich
mesh_terms: D002498:Centrifugation / Q000379:methods; D005374:Filtration / Q000379:methods*; D005615:Freezing; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D008828:Microbiological Techniques / Q000379:methods*; D010843:Pichia / Q000737:chemistry*; D013997:Time Factors
keywords: Pichia pastoris; centrifugation; filtration; metabolite loss; metabolomics; quenching