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Kumar B. N., Vinay ; Guo, Shuxia ; Bocklitz, Thomas ; Rösch, Petra ; Popp, Jürgen:
Demonstration of carbon catabolite repression in naphthalene degrading soil bacteria via Raman spectroscopy based stable isotope probing.
In: Analytical Chemistry.
Bd. 88
(2016)
Heft 15
.
- S. 7574-7582.
ISSN 1520-6882
DOI: https://doi.org/10.1021/acs.analchem.6b01046
Abstract
Carbon catabolite repression (CCR) is a regulatory phenomenon occurring in both lower organisms like bacteria and higher organisms like yeast, which allows them to preferentially utilize a specific carbon source to achieve highest metabolic activity and cell growth. CCR has been intensely studied in the model organisms Escherichia coli and Bacillus subtilis by following diauxic growth curves, assays to estimate the utilization or depletion of carbon sources, enzyme assays, Western blotting and mass spectrometric analysis to monitor and quantify the involvement of specific enzymes and proteins involved in CCR. In this study, we have visualized this process in three species of naphthalene degrading soil bacteria at a single cell level via Raman spectroscopy based stable isotope probing (Raman-SIP) using a single and double labeling approach. This is achieved using a combination of 2H and 13C isotope labeled carbon sources like glucose, galactose, fructose, and naphthalene. Time dependent metabolic flux of 13C and 2H isotopes has been followed via semi quantification and 2D Raman correlation analysis. For this, the relative intensities of Raman marker bands corresponding to 2H and 13C incorporation in newly synthesized macromolecules like proteins and lipids have been utilized. The 2D correlation analysis of time dependent Raman spectra readily identified small sequential changes resulting from isotope incorporation. Overall, we show that Raman-SIP has the potential to be used to obtain information about regulatory processes like CCR in bacteria at a single cell level within a time span of 3 h in fast growing bacteria. We also demonstrate the potential of this approach in identifying the most efficient naphthalene degraders asserting its importance for use in bioremediation.
Weitere Angaben
| Publikationsform: | Artikel in einer Zeitschrift |
|---|---|
| Begutachteter Beitrag: | Ja |
| Institutionen der Universität: | Fakultäten > Fakultät für Mathematik, Physik und Informatik > Institut für Informatik > Lehrstuhl Künstliche Intelligenz in der Mikroskopie und Spektroskopie > Lehrstuhl Künstliche Intelligenz in der Mikroskopie und Spektroskopie - Univ.-Prof. Dr. Thomas Wilhelm Bocklitz |
| Titel an der UBT entstanden: | Nein |
| Themengebiete aus DDC: | 500 Naturwissenschaften und Mathematik > 530 Physik |
| Eingestellt am: | 16 Mai 2023 11:48 |
| Letzte Änderung: | 16 Mai 2023 11:48 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/76336 |