Abstract
Whole-cell suspensions of T. ferrireducens reduced Fe(III) citrate, Fe(III)-EDTA, and ferrihydrite with glycerol as an electron donor. After cell disruption, the highest activity was registered with Fe(III)-EDTA as the electron acceptor and NADH or NADPH as electron donors. About 80% of the NAD(P)H-dependent Fe(III)-EDTA reductase activities were associated with the membrane fraction of the cells. Treatment of the membranes with lauryl maltoside led to complete solubilization of the NADH-dependent and 70% solubilization of the NADPH-dependent Fe(III)-EDTA reductase activities. After purification by ion-exchange chromatography, the NADH-dependent activity was concentrated 8-fold, and the NADPH-dependent activity was concentrated 11-fold, with a yield of about 10% for both activities. The Fe(III)-EDTA-reducing enzyme complex included c-type cytochromes and a protein with a molecular mass of ca. 115 kDa, consisting of two polypeptides. This is the first description of membrane-bound Fe(III)-reducing oxidoreductase activities from a grampositive dissimilatory Fe(III)-reducing bacterium.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Lovley, D.R., Holmes, D.E., and Nevin, K.P., Dissimilatory Fe(III) and Mn(IV) Reduction, Adv. Microb. Physiol., 2004, vol. 49, pp. 219–286.
Slobodkin, A.I., Thermophilic Microbial Metal Reduction, Mikrobiologiya, 2005, vol. 74, no. 5, pp. 581–595 [Microbiology (Engl. Transl.), vol. 74, no. 5, pp. 501–514].
Leang, C., Adams, L.A., Chin, K.-J., Nevin, K.P., Met, B.A., Webster, J., Sharma, M.L., and Lovley, D.R., Adaptation to Disruption of the Electron Transfer Pathway for Fe(III) Reduction in Geobacter sulfurreducens, J. Bacteriol., 2005, vol. 187, pp. 5918–5926.
Lies, D.P., Hernandez, M.E., Kappler, A., Mielke, R.E., Gralnick, J.A., and Newman, D.K., Shewanella oneidensis MR-1 Uses Overlapping Pathways for Iron Reduction at a Distance and by Direct Contact under Conditions Relevant for Biofilms, Appl. Environ. Microbiol., 2005, vol. 71, pp. 4414–4426.
Gaspard, S., Vazquez, F., and Holliger, C., Localization and Solubilization of the Iron(III) Reductase of Geobacter sulfurreducens, Appl. Environ. Microbiol., 1998, vol. 64, pp. 3188–3194.
Magnuson, T.S., Hodges-Myerson, A.L., and Lovley, D.R., Characterization of a Membrane-Bound NADH-Dependent Fe3+ Reductase from the Dissimilatory Fe3+-Reducing Bacterium Geobacter sulfurreducens, FEMS Microbiol. Lett., 2000, vol. 185, pp. 205–211.
Slobodkin, A., Reysenbach, A.-L., Strutz, N., Dreier, M., and Wiegel, J., Thermoterrabacterium ferrireducens gen. nov., sp. nov., a Thermophilic Anaerobic Dissimilatory Fe(III)-Reducing Bacterium from a Continental Hot Spring, Int. J. Syst. Bacteriol., 1997, vol. 47, pp. 541–547.
Gavrilov, S.N., Bonch-Osmolovskaya, E.A., and Slobodkin, A.I., Physiology of Organotrophic and Lithotrophic Growth of the Thermophilic Iron-Reducing Bacteria Thermoterrabacterium ferrireducens and Thermoanaerobacter siderophilus, Mikrobiologiya, 2003, vol. 72, no. 2, pp. 161–167 [Microbiology (Engl. Transl.), vol. 72, no. 2, pp. 132–137].
Khijniak, T.V., Slobodkin, A.I., Coker, V., Renshaw, J.C., Livens, F.R., Bonch-Osmolovskaya, E.A., Birkeland, N.-K., Medvedeva-Lyalikova, N.N., and Lloyd, J.R., Reduction of Uranium(VI) Phosphate during Growth of the Thermophilic Bacterium Thermoterrabacterium ferrireducens, Appl. Environ. Microbiol., 2005, vol. 71, pp. 6423–6426.
Manual of Methods for General Bacteriology, Gerhardt, P. et al., Eds., Washington, DC: Am. Soc. Microbiol., 1981, vol. 2.
Moody, M.G. and Dailey, H.A., Aerobic Ferrisiderophore Reductase Assay and Activity Stain for Native PAGE, Anal. Biochem., 1983, vol. 134, pp. 235–239.
Thomas, P.E., Ryan, D., and Levin, W., An Improved Staining Procedure for the Detection of the Peroxidase Activity of Cytochrome P450 on Sodium Dodecyl Sulfate Polyacrylamide Gels, Anal. Biochem., 1976, vol. 75, pp. 168–76.
Dobbin, P.S., Carter, J.P., San, Juan, C.G., von Hobe, M., Powell, A.K., and Richardson, D.J., Dissimilatory Fe(III) Reduction by Clostridium beijerinckii Isolated from Freshwater Sediment Using Fe(III) Maltol Enrichment, FEMS Microbiol. Lett., 1999, vol. 176, pp. 131–138.
Vadas, A., Monbouquette, H.G., Johnson, E., and Schroeder, I., Identification and Characterization of a Novel Ferric Reductase from the Hyperthermophilic Archaeon Archaeoglobus fuldigus, J. Biol. Chem., 1999, vol. 274, pp. 36715–36721.
Childers, S.E. and Lovley, D.R., Differences in Fe(III) Reduction in the Hyperthermophilic Archaeon Pyrobaculum islandicum versus Mesophilic Fe(III)-Reducing Bacteria, FEMS Microbiol. Lett., 2001, vol. 195, pp. 253–258.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published in Russian in Mikrobiologiya, 2007, Vol. 76, No. 2, pp. 164–171.
The article was translated by the authors.
Rights and permissions
About this article
Cite this article
Gavrilov, S.N., Slobodkin, A.I., Robb, F.T. et al. Characterization of membrane-bound Fe(III)-EDTA reductase activities of the thermophilic gram-positive dissimilatory iron-reducing bacterium Thermoterrabacterium ferrireducens . Microbiology 76, 139–146 (2007). https://doi.org/10.1134/S0026261707020026
Received:
Issue Date:
DOI: https://doi.org/10.1134/S0026261707020026