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Denitrification in a binary culture and thiocyanate metabolism in Thiohalophilus thiocyanoxidans gen. nov. sp. nov. – a moderately halophilic chemolithoautotrophic sulfur-oxidizing Gammaproteobacterium from hypersaline lakes

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Abstract

Anaerobic enrichment culture with thiocyanate as electron donor and nitrate as electron acceptor at 2 M NaCl inoculated with a mixture of sediments from hypersaline lakes in Kulunda Steppe (Altai, Russia) resulted in a selection of a binary consortium of moderately halophilic, obligately chemolithoautotrophic sulfur-oxidizing bacteria (SOB) capable of complete denitrification of nitrate with thiosulfate as the electron donor. One consortium member, strain HRhD 3sp, was isolated into pure culture with nitrate and thiosulfate using a density gradient. This strain was responsible for the reduction of nitrate to nitrite in the consortium, while a second strain, HRhD 2, isolated under microoxic conditions with thiosulfate as substrate, was capable of anaerobic growth with nitrite and thiosulfate. Nitrite, either as substrate or as product, was already toxic at very low concentrations for both strains. As a result, optimal growth under anaerobic conditions could only be achieved within the consortium. On the basis of phylogenetic analysis, both organisms were identified as new lineages within the Gammaproteobacteria. As well as thiosulfate, strain HRhD 2 can also use thiocyanate as electron donor, representing a first halophilic SOB capable of growth with thiocyanate at 2–4 M NaCl. Product and enzymatic analysis identified the “carbonyl sulfide (COS) pathway” of primary thiocyanate degradation in this new species. On the basis of phenotypic and genetic analysis, strain HRhD 2 is proposed to be assigned to a new genus and species Thiohalophilus thiocyanoxidans.

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Acknowledgements

This work was supported by NWO-RFBR grant (047.011.2004.010), RFBR (grant 04–04–48647), and by the Program on Molecular and Cell Biology RAS.

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Authors and Affiliations

  1. Winogradsky Institute of Microbiology, Russian Academy of Sciences, Prospect 60-let Octyabrya, 7/2, 117312, Moscow, Russia

    Dimitry Y. Sorokin & Tatjana P. Tourova

  2. Department of Biotechnology, Delft University of Technology, Delft, The Netherlands

    Dimitry Y. Sorokin & Gerard Muyzer

  3. Department of Microbiology, Radboud University of Nijmegen, Nijmegen, The Netherlands

    Arjan Pol

  4. A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia

    Ekatherina Y. Bezsoudnova

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  1. Dimitry Y. Sorokin
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  2. Tatjana P. Tourova
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  3. Ekatherina Y. Bezsoudnova
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  4. Arjan Pol
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  5. Gerard Muyzer
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Correspondence to Dimitry Y. Sorokin.

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Supplementary Figure.

Dialysis culture of strain HRhD 3sp grown anaerobically with thiosulfate and nitrate. The culture is contained in the 20 ml Centricon tube (1) with 10 kDa membrane (2) put inside the 1 L medium tank (3). The toxic product of nitrate reduction (nitrite) is diffusing out of (1) into (3) through (2); (4), magnetic bars (PPT 2.51 Mb)

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Sorokin, D.Y., Tourova, T.P., Bezsoudnova, E.Y. et al. Denitrification in a binary culture and thiocyanate metabolism in Thiohalophilus thiocyanoxidans gen. nov. sp. nov. – a moderately halophilic chemolithoautotrophic sulfur-oxidizing Gammaproteobacterium from hypersaline lakes. Arch Microbiol 187, 441–450 (2007). https://doi.org/10.1007/s00203-006-0208-3

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  • DOI: https://doi.org/10.1007/s00203-006-0208-3

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