A high-throughput method for microbial metabolome analysis using gas chromatography/mass spectrometry.
An analytical high-throughput method based on gas chromatography/mass spectrometry (GC/MS) was developed for fast metabolome investigation. By parallelization and partial automation the time needed for the preanalytical steps could be reduced. In addition a strong decrease of the relative standard deviation of metabolite concentrations from independent samples on the same microtiter plate from 25 to 13% was achieved. Between different plates the relative standard deviation is comparable to the one observed in standard experiments with shaking flasks. Using a fast GC the time need for the full GC/MS-based metabolome analysis could be decreased from 60 to 18 min per run, allowing the measurement of 72 single samples per day and GC/MS machine. In samples of the model organism Corynebacterium glutamicum more than 1000 peaks in the total ion current could be detected in a single fast GC/MS run of which 650 were strong enough to be quantified. Approximately 150 compounds of these were identified using our metabolite MS-library. Correlation analysis of the concentration vectors of independent wild-type samples raised under the same conditions show very high correlations of 0.99+/-0.01 (logs). In conclusion this method allows screenings of large mutant libraries for genetically induced metabolic perturbations.
Börner
Jana
J
Institute of Biochemistry, University of Cologne, Zülpicher Str. 47, D-50674 Cologne, Germany.
Buchinger
Sebastian
S
Schomburg
Dietmar
D
D001419:Bacteria / Q000254:growth & development / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*