DISSIMILATORY SULFITE REDUCTASE IN CELL-FREE EXTRACTS OF INTESTINAL SULFATE-REDUCING BACTERIA
DOI: http://dx.doi.org/10.30970/sbi.0802.353
Abstract
Dissimilatory sulfite reductase activity in different fractions of the sulfate-reducing bacteria Desulfovibrio piger Vib-7 and Desulfomicrobium sp. Rod-9 isolated from human intestine was studied. Sulfite reductase, an important enzyme in the process of sulfur metabolism in these bacteria, was solubilized from the membrane fraction. The highest activity of the enzyme in the cell-free extract of the bacterial strains was measured (0.032±0.0026 and 0.028±0.0022 U×mg-1 protein for D. piger Vib-7 and Desulfomicrobium sp. Rod-9, respectively) compared to other fractions. The optimal temperature (+30…35 °C) and pH 7.0 for sulfite reductase reaction in the extracts of both bacterial strains was determined. The spectral analysis of purified sulfite reductase from cell-free extracts was carried out. The absorption maxima were 284, 391, 412, 583, and 630 nm, as well as 287, 393, 545, and 581 nm for sulfite reductase of D. piger Vib-7 and Desulfomicrobium sp. Rod-9, respectively. Analysis of the kinetic properties of the bacterial sulfite reductase has been carried out. The sulfite reductase activity, initial (instantaneous) reaction rate (V0) and maximum rate of the sulfite reductase reaction (Vmax) were higher in the D. piger Vib-7 cells than in the Desulfomicrobium sp. Rod-9. However, Michaelis constants (Km) of the enzyme activity were similar for both bacterial strains. The studies of the sulfite reductase activity, the kinetic properties of this enzyme in the intestinal sulfate-reducing bacteria strains, and their production of hydrogen sulfide in detail can be useful for clarification of the etiological role of these bacteria in the development of inflammatory bowel diseases in humans and animals.
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1. Bailey N.T.J. Statistical Methods in Biology. Cambridge University Press, 1995; 255 p. | |
| |
2. Barton L.L., Hamilton W.A. Sulphate-reducing Bacteria. Environmental and Engineered. Cambridge University Press, 2007; 553 p. | |
| |
3. Cummings J.H., Macfarlane G.T., Macfarlane S. Intestinal Bacteria and Ulcerative Colitis. Curr. Issues Intest. Microbiol, 2003; 4: 9-20. | |
| |
4. Gibson G.R., Cummings J.H., Macfarlane G.T. Growth and activities of sulphate-reducing bacteria in gut contents of health subjects and patients with ulcerative colitis. FEMS Microbiol. Ecol, 1991; 86: 103-112. | |
| |
5. Keleti T. Basic Enzyme Kinetics. Akademiai Kiado, 1988; 422 p. | |
| |
6. Kobayashi K., Takahashi E., Ishimoto M. Biochemical studies on sulfate-reducing bacteria. XI. Purification and some properties of sulfite reductase, desulfoviridin. J. Biochem, 1972; 72(4): 879-887. | |
| |
7. Kobayashi K., Seki Y. and Ishimoto M. Biochemical Studies on Sulfate-reducing Bacteria XIII. Sulfite Reductase from Desulfovibrio vulgaris - Mechanism of Trithionate, Thiosulfate, and Sulfide Formation and Enzymatic Properties. J. Biochem, 1974. 75(3): 519-529. | |
| |
8. Kosterin S.A., Burchynskaya N.F. The method for determining the characteristics kinetics of the Ca2+-transport systems subcells structures of smooth muscles. Ukrainian Biochem. J, 1987; 59(2): 66-69. | |
| |
9. Kushkevych I.V. Sulfate-reducing bacteria of the human intestine. I. Dissimilatory sulfate re-duction. Sci. Int. J. Biological Studies/Studia Biologica. 2012; 6(1): 149-180. | |
| |
10. Kushkevych I.V. Sulfate-reducing bacteria of the human intestine. II. The role in the diseases development. Sci. Int. J. Biological Studies/Studia Biologica. 2012; 6(2): 221-250. | |
| |
11. Kushkevych I.V. Identification of sulfate-reducing bacteria strains of human large intestine. Sci. Int. J. Biological Studies/Studia Biologica, 2013; 7(3): 115-124. | |
| |
12. Kushkevych I.V., Bartos M., Bartosova L. Sequence analysis of the 16S rRNA gene of sulfate-reducing bacteria isolated from human intestine. Int. J. Curr. Microbiol. Appl. Sci, 2014; 3(2): 239-248. | |
| |
13. Lee J.P., Yi C.S., LeGall J., and Peck H.D.Jr. Isolation of a New Pigment, Desulforubidin, from Desulfovibrio desulfuricans (Norway Strain) and Its Role in Sulfite Reduction. J. Bacteriol, 1973; 115(1): 453-455. | |
| |
14. Lee J.P., LeGall J., Peck H.D.Jr. Isolation of Assimilatory- and Dissimilatory-Type Sulfite Reductases from Desulfovibrio vulgaris. J. Bacteriol, 1973; 115(2): 529-542. | |
| |
15. Lowry O.H., Rosebrough N.J, Farr A.L., Randall R.J. Protein determination with the Folin phenol reagent . J. Biol. Chem, 1951; 193: 265-275. | |
| |
16. Morse R., Gibson G.R., Collins M.D. Secondary structure analysis of the dissimilatory sulphite reductase in Desulfovibrio desulfuricans. Lett. Appl. Microbiol, 2000; 30: 375-378. | |
| |
17. Peck H.D., Van Beeumen J., and LeGall J. Biochemistry of Dissimilatory Sulphate Reduction and Discussion. Phil. Trans. R. Soc. Lond. B, 1982; 298: 443-466. | |
| |
18. Steuber J. and Kroneck P.M.H. Desulfoviridin, the dissimilatory sul.te reductase from Desulfovibrio desulfuricans (Essex): new structural and functional aspects of the membranous enzyme. Inorganica Chimica Acta, 1998; 276: 52-57. | |
| |
19. Steuber J., Arendsen A.F., Hagen W.R. and Kroneck P.M.H. Molecular properties of the dissimilatory sulfite reductase from Desulfovibrio desulfuricans (Essex) and comparison with the enzyme from Desulfovibrio vulgaris (Hildenborough). Eur. J. Bioch, 1995; 233: 873-879. | |
| |
20. Steuber J., Cypionka H. and Kroneck P.M.H. Mechanism of dissimilatory sulfite reduction by Desulfovibrio desulfuricans. Purification of a membrane-bound sulfite reductase and coupling with cytochrome c3 and hydrogenase. Arch. Microbiol, 1994; 162: 255-260. | |
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