O. M. Chayka, T. B. Peretyatko, A. A. Halushka



Introduction. Thermophilic sulfate-reducing bacteria attract attention of scientists as the potential agents of purification of wastewater polluted by sulfur and its compounds, heavy metal ions and organic compounds. These bacteria oxidize different organic substrates using metals with variable valency as electron acceptors and transform them into non-toxic or less toxic forms for living organisms. However, wastewater contains high concentrations of different toxic xenobiotics, particularly, metal ions that have negative influence on living organisms. For this reason, it is important to use resistant strains of microorganisms for the purification of wastewater.
The aim of this work was to identify the thermophilic sulfur-reducing bacteria, isolated from “Nadiia” pit spoil heap of Chervonohrad mining region, and to study their properties.
Materials and Methods. Thermophilic sulfur-reducing bacteria were isolated from the samples of rock of “Nadiia” pit heap at 50 cm depth. Bacteria were cultivated in TF medium under the anaerobic conditions in  anaerostates. Cell biomass was measured turbidimetrically using the photoelectric colorimeter KFK-3 (λ = 340 nm, 3 mm cuvette). Hydrogen sulfide content was measured photoelectrocolorymetrically by the production of methylene blue. Organic acids content was measured by high performance liquid chromatography. Cr(VI), Fe(III), Мn(IV) and NO3 content was measured turbidimetrically.
Results. Thermophilic sulfur-reducing bacteria were isolated from the rock of “Nadiia” pit heap of Chervonohrad mining region. They were identified as Moorela thermoacetica based on the morpho-physiological and biochemical properties and on the results of phylogenetic analysis. M. thermoacetica Nadia-3 grow in the synthetic TF medium, have the shape of elongated rods, are gram-positive, endospore-forming. They form light brown colonies. Optimal growth was observed at 50–55 °C, pH 6.5–7. The bacteria utilize glucose, starch, fructose, maltose, lactose, sodium lactate, arabinose, cellulose, maltose, glycerol, fumarate, and ethanol as carbon sources. The highest sulfidogenic activity of M. thermoacetica Nadia-3 was found in media with glycerol, lactose, and glucose. M. thermoacetica Nadia-3 reduce SO42-, S2O32-, Fe(III), NO3, Cr(VI) compounds besides elemental sulfur. They accumulate biomass at K2Cr2O7 concentrations of 0.1–1 mM. Sulfur reduction is not the main way of energy accumulation.
Conclusions. Thermophilic chromium-resistant sulfur-reducing bacteria M. thermoacetica Nadia-3, that produce hydrogen sulfide during the oxidation of different organic compounds, were isolated from the rock of “Nadiia” pit heap. They reduce Fe(III), Cr(VI), NO3, SO42-, S2O32-, besides elemental sulfur.


thermophilic sulfur-reducing bacteria, elemental sulfur, glucose, starch

Full Text:



1. Balk M., van Gelder T., Weelink S. A., Stams A. J. (Per)chlorate reduction by the thermophilic bacterium Moorella perchloratireducens sp. nov., isolated from underground gas storage. Applied and Environmental Microbiology, 2008; 74(2): 403-409.
CrossrefPubMedPMCGoogle Scholar

2. Cai J., Wang Y., Liu D., Zeng Y., Xue Y., Ma Y., Feng Y. Fervidobacterium changbaicum sp. nov., a novel thermophilic anaerobic bacterium isolated from a hot spring of the Changbai Mountains, China. International Journal of Systematic and Evolutionary Microbiology, 2007; 57(10): 2333-2336.
CrossrefPubMedGoogle Scholar

3. Drake H.L., Daniel S.L. Physiology of the thermophilic acetogen Moorella thermoacetica. Research in Microbiology, 2004; 155(6): 869-883.
CrossrefPubMedGoogle Scholar

4. Drake H.L., Gossner A.S., Daniel S.L. Old acetogens, new light. Annals of the New York Academy of Sciences, 2008; 1125(1): 100-128.
CrossrefPubMedGoogle Scholar

5. Friedrich A.B., Antranikian G. Keratin Degradation by Fervidobacterium pennavorans, a Novel Thermophilic Anaerobic Species of the Order Thermotogales. Applied and environmental microbiology, 1996; 62(8): 2875-2882.
CrossrefPubMedGoogle Scholar

6. Green M.R., Sambrook J., Harbor C.S., MacCallum P. Molecular Cloning: Laboratory Manual, Fourth Edition. New York, USA: Cold Spring Harbor, 2012. 2028 pp.
Google Scholar

7. Harris D.S. Quantitative сhemical аnalysis. New York, USA: W H Freeman & Co, 2003. 125 p.

8. Islam M.A., Zengler K., Edwards E. A., Mahadevan R., Stephanopoulos G. Investigating Moorella thermoacetica metabolism with a genome-scale constraint-based metabolic model. Integrative Biology, 2015; 7(8): 869-882.
CrossrefPubMedGoogle Scholar

9. Jiang B., Henstra A.M., Paulo P.L., Balk M., Van Doesburg W., Stams A.J. Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain. Archives of Microbiology, 2009; 191(2): 123-131.
CrossrefPubMedGoogle Scholar

10. Kanoksilapatham W., Keawram Р. Diversity of Hyperthermophilic Bacteria Belonging to Order Thermotogales Thriving in Three Hot Springs in Thailand: Resources of Genes Encoding Thermostable Enzymes. Science, Engineering and Health Studies, 2013; 7(2): 17-38.
Google Scholar

11. Kerem Z., Bravdo B., Shoseyov O., Tugendhaft Y. Rapid liquid chromatography - ultraviolet determination of organic acids and phenolic compounds in red wine and must. Journal of Chromatography A, 2004; 1052(1-2): 211-215.
CrossrefPubMedGoogle Scholar

12. Pierce E., Xie G., Barabote R.D., Saunders E., Han C.S., Detter J.C., Richardson P., Brettin T.S., Das А., Ljungdahl L.G., Ragsdale S.W. The complete genome sequence of Moorella thermoacetica (f. Сlostridium thermoacetica). Environmental Microbiology, 2008; 10(10): 2550-2573.
CrossrefPubMedPMCGoogle Scholar

13. Redl S., Poehlein A., Esser C., Bengelsdorf F.R., Jensen T., Jendresen C.B., Tindall B.J., Daniel R., Dьrre P., Nielsen A.T. Genome-Based Comparison of All Species of the Genus Moorella, and Status of the Species Moorella thermoacetica and Moorella thermoautotrophica. Frontiers in Microbiology, 2020; 17(10): 3070.
CrossrefPubMedPMCGoogle Scholar

14. Sugiyama M. Inc assignee. Reagent composition for measuring hydrogen sulfide and method for measuring hydrogen. United States Patent 6,340,596 B1. 2002 Jan 22.

15. Turner S., Pryer K.M., Miao V.P.W., Palmer J.D. Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. The Journal of Eukaryotic Microbiology, 1999; 46(4): 327-338.
CrossrefPubMedGoogle Scholar


  • There are currently no refbacks.

Copyright (c) 2021 O. M. Chayka, T. B. Peretyatko, A. A. Halushka

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.