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Use of Raman spectroscopy for identification of compatible solutes in halophilic bacteria

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Abstract

We explored the use of Raman spectroscopy to detect organic osmotic solutes as biomarkers in the moderately halophilic heterotrophic bacterium Halomonas elongata grown in complex medium (accumulation of glycine betaine) and in defined medium with glucose as carbon source (biosynthesis of ectoine), and in the anoxygenic phototrophic Ectothiorhodospira marismortui known to synthesize glycine betaine in combination with minor amounts of trehalose and N-α-carbamoyl glutamineamide. We tested different methods of preparation of the material: lyophilization, two-phase extraction of water-soluble molecules, and perchlorate extraction. Raman signals of glycine betaine and ectoine were detected; perchlorate extraction followed by desalting the extract on an ion retardation column gave the best results. Lyophilized cells of E. marismortui showed strong signals of carotenoid pigments, and glycine betaine could be detected only after perchlorate extraction and desalting. The data presented show that Raman spectroscopy is a suitable tool to assess the mode of osmotic adaptation used by halophilic microorganisms.

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Acknowledgments

This work was supported by grant no. P210/10/0467 from the Grant Agency of the Czech Republic and by institutional support MSM0021620855 from the Ministry of Education of the Czech Republic (to JJ) and by grant no. 1103/10 from the Israel Science Foundation (to AO).

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Authors and Affiliations

  1. Institute of Geochemistry, Mineralogy and Mineral Resources, Charles University in Prague, Prague, Czech Republic

    Jan Jehlička & Petr Vítek

  2. Department of Plant and Environmental Microbiology, The Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel

    Aharon Oren

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  1. Jan Jehlička
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  2. Aharon Oren
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  3. Petr Vítek
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Correspondence to Aharon Oren.

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Communicated by M. da Costa.

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Jehlička, J., Oren, A. & Vítek, P. Use of Raman spectroscopy for identification of compatible solutes in halophilic bacteria. Extremophiles 16, 507–514 (2012). https://doi.org/10.1007/s00792-012-0450-3

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  • DOI: https://doi.org/10.1007/s00792-012-0450-3

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