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. 2013 Jul 9:1:e101.
doi: 10.7717/peerj.101. Print 2013.

Looking at microbial metabolism by high-resolution (2)H-NMR spectroscopy

Victor P Kutyshenko  1 , Petr M Beskaravayny, Maxim V Molchanov, Svetlana I Paskevich, Dmitry A Prokhorov, Vladimir N Uversky
Affiliations

Looking at microbial metabolism by high-resolution (2)H-NMR spectroscopy

Victor P Kutyshenko et al. PeerJ. .

Abstract

We analyzed the applicability of high-resolution (2)H-HMR spectroscopy for the analysis of microbe metabolism in samples of mitochondrion isolated from rat liver and from aqueous extracts of homogenates of rat liver and other organs and tissues in the presence of high D2O contents. Such analysis is possible due to the fast microbe adaptation to life in the heavy water. It is also shown that some enzymatic processes typical for the intact cells are preserved in the homogenized tissue preparations. The microbial and cellular metabolic processes can be differentiated via the strategic use of cell poisons and antibiotics.

Keywords: 2H-HMR spectroscopy; Heavy water; High resolution NMR; Microbe adaptation; Microbial metabolism.

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Figures

Figure 1
Figure 1. 1H-NMR spectra of biological fluids.
(A) Mitochondrion isolated from rat liver. Measurements were taken immediately after mitochondrion isolation. (B) Mitochondrion isolated from rat liver. Measurements were taken one day after isolation. (C) Aqueous extract of the rat liver homogenate.
Figure 2
Figure 2. Kinetic changes in the integral intensities of the NMR spectra.
Time courses of changes in the integral intensities of the aliphatic part of 1H-NMR (black circles) and 2H-NMR spectra (open circles).
Figure 3
Figure 3. 2H-NMR spectra of biological fluids.
(A) Mitochondrion isolated using D2O-based protocol. (B) Mitochondrion isolated using D2O-based protocol. (C) Aqueous extract of the rat liver homogenate. (D) Aqueous extract of the rat liver homogenate with sodium azide added.
Figure 4
Figure 4. Model representation of the metabolite conversion pathways.
Various pathways of the metabolite conversion in cytosol and mitochondrion of rat liver at which hydrocarbon skeleton of resulting compounds can be deuterated.
Figure 5
Figure 5. Time course of changes in the 2H-NMR spectra of mitochondrion samples with added antibiotics.
(A) Spectrum is taken on the sixth day after the sample preparation. (B) Spectrum is taken on the second day after the sample preparation. (C) Spectrum is taken 8 h after the sample preparation.
See this image and copyright information in PMC

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