The influence of ferric (III) citrate on ATP-hydrolases of Desulfuromonas acetoxidans ІМV В-7384

  • O. Maslovska Ivan Franko Lviv National University
  • S. O. Hnatush Ivan Franko National University of Lviv


Desulfuromonas acetoxidans obtains energy for growth by the anaerobic oxidation of organic compounds with the carbon dioxide formation. It was found that ferrum and manganese are used as terminal electron acceptors in the processes of anaerobic respiration, such as dissimilative Fe3+- and Mn4+-reduction, carried out by these bacteria (Lovely, 1991). D. acetoxidans ІМV B-7384 can be used as anode biocatalyst in microbial fuel cell with high electron recovery through acetate oxidation to the electric current as a result of electron transfer to the anode or 3d-type transition metals, such as ferrum and manganese, in the process of their reduction. Investigation of biochemical changes of D. acetoxidans ІМV B-7384 under the influence of Fe (III) compounds is important for optimization of the process of bacterial electricity generation. ATP-hydrolase is located in cytoplasmic membrane, and its subunits are exposed to both the cytoplasm and the external environment. Therefore, the changes of that enzyme activity can be used as an indicator of various stress exposure. Presence of ferric iron ions in the bacterial growth medium could catalyze generation of organic reactive oxygen species, such as peroxyl (ROO-) and alkoxyl (RO-) radicals. Lipid peroxidation is one of the main reasons of cell damage and it’s following death under the influence of reactive oxygen metabolites. It is known that lipid peroxidation and membrane transport processes are somehow interrelated, but mechanisms of such interaction are still unidentified. In our previous researche we have shown the influence of ferric (III) citrate on the intensity of lipid peroxidation of D. аcetoxidans ІМV В-7384. Significant increase of the content of lipid peroxidation products (lipid hydroperoxides, conjugated dienes and malondialdehyde) in bacterial cells has been observed under the addition of ferric (III) citrate into the cultural medium. The increase of the concentration of lipid peroxidation products in bacterial cells confirms free radical mechanism of oxidation of polyunsaturated fatty acids. Thus, for fulfiling complete analyses of cell response against oxidative stress it was reasonable to investigate the influence of ferric (III) citrate on specific ATP-hydrolase activity, Na+, K+-ATP-hydrolase activity and Mg2+-ATP-hydrolase activity of D. acetoxidans ІМV В-7384. Bacteria were cultivated in the modified Postgaite C medium during four days under the anaerobic conditions and temperature +27°С with addition from 10 to 20 mM of ferric (III) citrate into the growth medium. Control samples didn’t contain investigated metal salt. Chosen concentrations of metal salt caused inhibition of bacterial growth by 20–50%. Activities of ATP-hydrolases were investigated as described. It was shown, that specific ATP-hydrolase activity of D. acetoxidans ІМV В-7384 is changing in dependance on duration of ferric (III) citrate exposure and concentration of the metal salt. Addition of the ferric (III) citrate in relatively low concentrations (10–12 mM) causes increasing of specific ATP-hydrolase activity of D. acetoxidans IMV B-7384 in comparison with control. Activity of investigated enzymes was inhibited under the increasing of metal salt concentration in bacterial growth medium. Increase of duration of D. acetoxidans IMV B-7384 cultivation causes decrease of ATP-hydrolase activity. Addition of ferric (III) citrate causes simultaneous increasing of Na+, K+-ATP-hydrolase activity and inhibition of Mg2+-ATP-hydrolase activity during four days of bacterial cultivation.


Danilovich, G.V., Gruzina, T.G., Ulberg, Z.R., Kosterin, S.O., 2004. Identification and catalytic properties of Mg2+-dependent ATP-hydrolase of plasmic membrane of Bacillus sp. B4253 capable to gold accumulation [Іdentifіkacіja ta katalіtichnі vlastivostі Mg2+-zalezhnoi ATF-gіdrolazi citoplazmatichnoi membrani Bacillus sp. B4253, zdatnih do nakopichennja zolota]. Ukr. Biochem. J. 76(5), 45–51 (in Ukrainian).

Danilovich, G.V., Gruzina, T.G., Ulberg, Z.R., Kosterin, S.O., 2007. Effect of ionic and colloid gold on ATP-hydrolase fermentative systems of Bacillus sp. В4253 and Bacillus sp. В4851 [Vpliv іonnogo ta koloіdnogo zolota na ATF-gіdrolaznі fermentnі sistemi v membranі mіkroorganіzmіv Bacillus sp. B4253 ta Bacillus sp. B4851]. Ukr. Biochem. J. 79(4), 46–53 (in Ukrainian).

Barton, L., 2007. Sulfate-reducing bacteria: Environmental and engineered systems. New York, USA, Cambridge University Press.

Fiske, C., Subbarow, Y., 1925. The colorimetric determination of phosphorus. J. Biol. Chem. 66, 375–400.

Gebhardt, N., Thauer, R., Linder, D., Kaulfers, P., Pfenning, N., 1985. Mechanism of acetate oxidation to CO2 with elemental sulfur in Desulfuromonas acetoxidans. Arch. Microbiol. 141, 392–398. >> doi:10.1007/BF00428855

Golovchak, N.P., Tarnovs’ka, A.V., Kocjumbas, G.І., Sanagurs’kij, D.І., 2012. Lipid peroxidation in living organisms [Procesey perekisnogo okisnennja lіpіdіv u zhivih organіzmah]. Lviv, Ivan Franko Lviv National University (in Ukrainian).

Hasan, S., Rosen, B., 1979. Properties and function of the proton-translocating adenosine triphosphatase of Clostridium perfringens. J. Bacteriol. 140(2), 745–747.

Maeda, M., Hamano, K., Hirano, Y., 1998. Structures of P-type transporting ATP-ases and chromosomal locations. Cell Structure and Function 23, 315–323. >> doi:10.1247/csf.23.315

Maslovska, O., Hnatush, S., 2013. The influence of ferric (III) citrate on the intensity of lipid peroxidation and activity of antioxidative system of Desulfuromonas acetoxidans (Proc. 5th Ukrainian–Polish Weigl Conference). Chernivtsi, Ukraine, 46.

Kuhlbrandt, W., 2004. Biology, structure and mechanism of P-type of ATP-ases. Nat. Rev. Mol. Cell Biol. 5, 282–295. >> doi.:10.1038/nrm1354

Lakin, G.F., 1990. Biometrija [Biometrics]. Moscow, Vysshaja Shkola (in Russian).

Lovely, D., Holmes, D., Nevin, K., 2004. Dissimilatory Fe (III) and Mn (IV) reduction. Adv. Microb. Physiol. 49, 246–259. >> doi:10.1016/S0065-2911(04)49005-5

Lovely, D., 1991. Dissimilatory Fe (III) and Mn (IV) reduction. Мicrobiol. Rev. 55(2), 259–287.

Lowry, O., Rosenbrough, N., Farr, L., Randall, R., 1951. Protein measurement, with the folin phenol reagent. J. Biol. Chem. 193, 265–275.

Papanikolaou, G., Pantopoulos, K., 2005. Iron metabolism and toxicity. Toxicol. Appl. Pharmacol. 202, 199–211. >> doi:10.1016/j.taap.2004.06.021

Semchyshyn, H.M., Lushchak, V.I., 2004. Oxidative stress and control of catalase activity in Escherichia colі [Oksidativnij stres і reguljacіja aktivnostі katalaz u Escherichia colі]. Ukr. Biochem. J. 76(2), 31–42 (in Ukrainian).

Vasyliv, O., Hnatush, S., 2011. Effect of transition metal compounds on catalase activity of sulfur-reducing bacterial Desulfuromonas acetoxidans cells. Visn. Lviv Univ. Biol. 57, 207–215.

Vasyliv, O., Bilyy, O., Ferensovich, J., Hnatush, S., 2012. Electric current generation by sulfur-reducing bacteria in microbial-anode fuel cell (Proc. SPIE). San-Diego, USA, 2012. 8472, 84720Z-1-7.

Zhang, E., Xu, W., Diao, G., Shuang, C., 2006. Electricity generation from acetate and glucose by sedimentary bacterium attached to elecrode in microbial-anode fuel cells. J. Power Sourses 161, 820–825. >> doi:10.1016/j.jpowsour.2006.05.004

Yoshimura, F., 1978. Properties of membrane adenosine triphosphatase of the obligately anaerobic bacteria Veillonella alcalescenes. J. Biochem. 83, 1231–1238.

Zyn, A., 2012. Prooxidant-antioxidant homeostasis and membrane transport in living organisms [Prooksidantno-antioksidantnij gomeostaz і membrannij transport u zhivih organіzmah]. Visn. Lviv Univ. Biol. 60, 20–39 (in Ukrainian).