Detection of volatile metabolites of <Emphasis Type="Italic">Escherichia coli</Emphasis> by multi capillary column coupled ion mobility spectrometry |
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Authors: | Sasidhar Maddula Lars Mathias Blank Andreas Schmid Jörg Ingo Baumbach |
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Institution: | 1.TU Dortmund, Faculty of Biochemical and Chemical Engineering,Laboratory of Chemical Biotechnology,Dortmund,Germany;2.ISAS-Institute for Analytical Sciences,Dortmund,Germany |
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Abstract: | Detection and immediate quantification of microbial metabolic activities is of high interest in fields as diverse as biotechnology
and infection biology. Interestingly, the most direct signals of microbial metabolism, the evolution of volatile metabolites,
is largely ignored in the literature, and rather, metabolite concentrations in the microbial surrounding or even disruptive
methods for intracellular metabolite measurements (i.e., metabolome analysis) are favored. Here, the development of a multi
capillary column coupled ion mobility spectrometer (MCC-IMS) was described for the detection of volatile organic compounds
from microbes and the MCC-IMS was used for characterization of metabolic activity of growing Escherichia coli. The MCC-IMS chromatogram of the microbial culture off-gas of the acetone-producing E. coli strain BL21 pLB4 revealed four analytes that positively correlated with growth, which were identified as ethanol, propanone
(acetone), heptan-2-one, and nonan-2-one. The occurrence of these analytes was cross-validated by solid-phase micro-extraction
coupled with gas chromatography mass spectrometry analysis. With this information in hand, the dynamic relationship between
the E. coli biomass concentration and the metabolite concentrations in the headspace was measured. The results suggest that the metabolic
pathways of heptan-2-one and nonan-2-one synthesis are regulated independent of each other. It is shown that the MCC-IMS in-line
off-gas analysis is a simple method for real-time detection of microbial metabolic activity and discussed its potential for
application in metabolic engineering, bioprocess control, and health care. |
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Keywords: | Volatile metabolites MVOC Escherichia coli Acetone Heptan-2-one Nonan-2-one Metabolism Metabolic network |
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