Utilization of fumarate by sulfur-reducing bacteria Desulfuromonas sp.

O. Сhayka; T. Peretjatko; S. Gudz; A. Halushka

doi:10.15421/011643

O. Сhayka Ivan Franko National University of Lviv
T. Peretjatko Ivan Franko National University of Lviv
S. Gudz Ivan Franko National University of Lviv
A. Halushka Ivan Franko National University of Lviv

Keywords: reducing bacteria, succinate, fumarate reductase, hydrogenase, fumarate respiration

Abstract

The main goal of the work was to study the utilization of fumarate by sulfur-reducing bacteria Desulfuromonas sp. under different growth conditions and accumulation of hydrogen sulfide by bacteria in the media with sulfur and different electron donors. Sulfur-reducing bacteria Desulfuromonas sp., isolated from soil in Yazivske sulfur deposit, were used in the reasearch. Bacteria were grown in the medium Postgate C without sulfates. The content of hydrogen sulfide was determined by formation of methylene blue. The content of organic acids (fumarate, succinate, lactate, acetate) was determined by high performance liquid chromatography (HPLC). The biomass of cells was determined by the photoelectrocolorymetry method using KFK-3. The highest level of accumulation of hydrogen sulfide by bacteria Desulfuromonas sp. was found in media with sodium lactate and sodium pyruvate. The maximal concentration of hydrogen sulfide was 1.9 mM. Maximal accumulation of biomass was observed in the media with malate, lactate and fumarate with the presence of elemental sulfur. Sulfur-reducing bacteria Desulfuromonas sp. are able to utilize fumarate as an electron donor and acceptor in the absence of elemental sulfur in the medium. After the incubation of Desulfuromonas sp. in the medium with fumarate, chromatographic analysis of culture liquid showed that fumarate is converted to succinate and small quantities of acetate The presence of acetate is, probably, due to the particularaties of the functioning of citric acid cycle in bacteria of the genus Desulfuromonas. Consequently, the results indicate that the fumarate serves as a donor and acceptor of electrons.The simultaneous introduction of two electron donors – fumarate and elemental sulfur – was accompanied by inhibition of sulfur reduction. After an additional source of carbon (sodium lactate) and electron acceptor (elemental sulfur) was added to the medium with fumarate a fivefold increase of sulfidogenic activity was observed. Thus, regulation of respiration in bacteria Desulfuromonas sp. is directed to the primary utilization of the most energetically favorable electron acceptors.

References

Butler, J.E., Glaven, R.H., Esteve-Nunez, A., Nunez, C., Shelobolina, E.S., Bond, D.R., Lovley, D.R., Butler, J.E., 2006. Genetic characterization of a single bifunctional enzyme for fumarate reduction and succinate oxidation in Geobacter sulfurreducens and engineering of fumarate reduction in Geobacter metallireducens. J. Bacteriol. 188(2), 450–455. >> doi.org/10.1128/jb.188.2.450-455.2006

Gebhardt, N.A.,Thauer, R.K., Linder, D., Kaulfers, P.-M., Pfennig, N., 1985. Mechanism of acetate oxidation to CO2 with elemental sulfur in Desulfuromonas acetoxidans. Arch. Microbiol. 141, 392–398. >> doi.org/10.1007/bf00428855

Hedderich, R., Klimmek, O., Kroger, A., Dirmeier, R., Keller, M., Stetter, K.O., 1999. Anaerobic respiration with elemental sulfur and with disulfides. FEMS Microbiol. Rev. 22, 353–381. >> doi.org/10.1111/j.1574-6976.1998.tb00376.x

Kerem, Z., Bravdo, B., Shoseyov, O., Tugendhaft, Y., 2004. Rapid liquid chromatography – ultraviolet determination of organic acids and phenolic compounds in red wine and must. J. Cromatogr. A. 1052, 211–215. >> doi.org/10.1016/j.chroma.2004.08.105

Kozlova, I.P., Radchenko, O.S., Stepura, L.G., Kondratyuk, T.O., Pilyashenko-Novokhatnyy, A.I., 2008. Geoximichna diyalnist mikroorganizmiv ta yiyi prykladni aspekty [The geochemical activity of microorganisms and its practical aspects]. Naukova Dumka, Kyiv (in Ukrainian).

Kröger, A., Biel, S., Simon, J., Gross, R., Unden, G., Lancaster, C., Roy, D., 2002. Fumarate respiration of Wolinella succinogenes: Enzymology, energetics and coupling mechanism. Biochim. Biophys. Acta 1553, 23–38. >> doi.org/10.1016/s0005-2728(01)00234-1

Lancaster, C., Roy, D., Simon, J., 2002. Succinate: Quinone ox-ido-reductases from e-proteobacteria. Biochim. Biophys. Acta 1553, 84–101. >> doi.org/10.1016/s0005-2728(01)00230-4

Lemos, R.S., Fernandes, A.S., Pereira, M.M., Gomes, C.M., Teixeira, M., 2002. Quinol:fumarate oxidoreductases and succinate:quinone oxidoreductases: Phylogenetic relationships, metal centres and membrane attachment. Biochim. Biophys. Acta 1553, 158–170. >> doi.org/10.1016/s0005-2728(01)00239-0

Postgate, J.R., 1984. The sulfate-reducing bacteria. 2nd ed. Cambridge University, Cambridge. >> doi.org/10.1002/jobm.3620250205

Sugiyama, M., 2002. Reagent composition for measuring hydrogen sulfide and method for measuring hydrogen sulfide. United States Patent, №6340596.

Thauer, R.K., 1988. Citricacid cycle, 50 years on modifications and an alternative pathway in anaerobic bacteria. Eur. J. Biochem. 176, 497–508. >> doi.org/10.1111/j.1432-1033.1988.tb14307.x

Zaunmüller, T., Kelly, D.J., Glöckner, F.O., Unden, G., 2006. Succinate dehydrogenase functioning by a reverse redox loop mechanism and fumarate reductase in sulphate-reducing bacteria. Microbiology 152, 2443–2453. >> doi.org/10.1099/mic.0.28849-0

Сhayka, O., Peretjatko, T., Gudz, S., 2010. Sirkovidnovlyuval’ni bakteriyi vodoym Yazivs’koho sirkovoho rodovyshcha [Sulfur reducing bacteria of Yazivske sulfur deposit]. Nauk. Visn. Uzhhorod. Univ. Ser. Biol. 28, 52–55 (in Ukrainian).