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Size exclusion chromatography—An improved method to harvest Corynebacterium glutamicum cells for the analysis of cytosolic metabolites

Paper ID Volume ID Publish Year Pages File Format Full-Text
23955 43484 2011 8 PDF Available
Title
Size exclusion chromatography—An improved method to harvest Corynebacterium glutamicum cells for the analysis of cytosolic metabolites
Abstract

The efficient separation of Corynebacterium glutamicum cells from culture medium by size exclusion chromatography (SEC) is presented. Residue analysis demonstrated that this method effectively depletes extracellular compounds. For evaluation, SEC was compared with the common methods cold methanol treatment, fast centrifugation and fast filtration. For this purpose, samples of C. glutamicum cells from fermenter cultures were harvested and subjected to a metabolome analysis. In particular, the wild type strain C. glutamicum ATCC13032 and the lysine production strain C. glutamicum DM1730 were grown in a minimal or in a complex medium. Comparison of metabolite pool sizes after harvesting C. glutamicum cells by the methods mentioned above by gas chromatography coupled to mass spectrometry (GC–MS) revealed that SEC is the most suitable method when intracellular metabolite pools are to be measured during growth in complex media or in the presence of significant amounts of secreted metabolites. In contrast to the other methods tested, the SEC method turned out to be fast and able to remove extracellular compounds almost completely.

Keywords
Corynebacterium glutamicum; Cell harvesting; Quenching; Metabolites; Metabolome analysis
First Page Preview
Size exclusion chromatography—An improved method to harvest Corynebacterium glutamicum cells for the analysis of cytosolic metabolites
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 154, Issues 2–3, 10 July 2011, Pages 171–178
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering