Metabolic quenching of <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis>: efficiency of methods and impact of cold shock |
| |
Authors: | Max Wellerdiek Dajana Winterhoff Waldemar Reule Jürgen Brandner Marco Oldiges |
| |
Institution: | (1) Institute of Biotechnology, Forschungszentrum Jülich GmbH, Leo-Brandt Strasse 20, 52428 Jülich, Germany;(2) Institute of Micro Process Engineering, Forschungszentrum Karlsruhe GmbH, Hermann von Helmholz Platz1, 76344 Egenstein Leopoldshafen, Germany;(3) Hochschule Furthwangen, Maschinenbau und Verfahrenstechnik, Robert-Gerwig-Platz 1, 78120 Furtwangen, Germany |
| |
Abstract: | Representative and valid cytoplasmic concentrations are essential for ensuring the significance of results in the field of
metabolome analysis. One of the most crucial points in this respect is the sampling itself. A rapid and sudden stopping of
the metabolism on a timescale that is much faster than the conversion rates of investigated metabolites is worthwhile. This
can be achieved by applying of cold methanol quenching combined with reproducible, fast, and automated sampling. Unfortunately,
quenching the metabolism by a sharp temperature shift leads to what is known as cold shock or the cell-leakage effect. In
the present work, we applied a microstructure heat exchanger to analyze the cold shock effect using Corynebacterium glutamicum as a model microorganism. Using this apparatus together with a silicon pipe, it was possible to assay the leakage effect
on a timescale starting at 1 s after cooling cell suspension. The high turnover rates not only require a rapid quenching technique,
but also the correct application. Moreover, we succeeded in showing that even when the required appropriate setup of methanol
quenching is not used, the metabolism is not stopped within the required timescale. By applying robust techniques like rapid
sampling in combination with reproducible sample processing, we ensured fast and reliable metabolic inactivation during all
steps. |
| |
Keywords: | Methanol quenching Metabolome analysis Metabolomics Microstructure heat exchanger Rapid sampling |
本文献已被 SpringerLink 等数据库收录! |
|