PL EN


Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2015 | 64 | 2 |

Tytuł artykułu

Kinetic properties of pyruvate ferredoxin oxidoreductase of intestinal sulfate-reducing bacteria Desulfovibrio piger Vib-7 and Desulfomicrobium sp. Rod-9

Autorzy

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Intestinal sulfate-reducing bacteria reduce sulfate ions to hydrogen sulfide causing inflammatory bowel diseases of humans and animals. The bacteria consume lactate as electron donor which is oxidized to acetate via pyruvate in process of the dissimilatory sulfate reduction. Pyruvate-ferredoxin oxidoreductase activity and the kinetic properties of the enzyme from intestinal sulfate-reducing bacteria Desulfovibrio piger and Desulfomicrobium sp. have never been well-characterized and have not been yet studied. In this paper we present for the first time the specific activity of pyruvate-ferredoxin oxidoreductase and the kinetic properties of the enzyme in cell-free extracts of both D. piger Vib-7 and Desulfomicrobium sp. Rod-9 intestinal bacterial strains. Microbiological, biochemical, biophysical and statistical methods were used in this work. The optimal temperature (+35°C) and pH 8.5 for enzyme reaction were determined. The spectral analysis of the purified pyruvate-ferredoxin oxidoreductase from the cell-free extracts was demonstrated. Analysis of the kinetic properties of the studied enzyme was carried out. Initial (instantaneous) reaction velocity (V₀), maximum amount of the product of reaction (Pmax), the reaction time (half saturation period) and maximum velocity of the pyruvate-ferredoxin oxidoreductase reaction (Vmax) were defined. Michaelis constants (Km) of the enzyme reaction were calculated for both intestinal bacterial strains. The studies of the kinetic enzyme properties in the intestinal sulfate-reducing bacteria strains in detail can be prospects for clarifying the etiological role of these bacteria in the development of inflammatory bowel diseases.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

64

Numer

2

Opis fizyczny

p.107-114,fig.,ref.

Twórcy

  • Institute of Animal Biology, NAAS of Ukraine, Lviv, Ukraine

Bibliografia

  • Akagi J.M. 1967. Electron carriers for the phosphoroclastic reaction of Desulfovibrio desulfuncans. J. Biol. Chem. 242: 2478–2483.
  • Bailey N.T.J. 1995. Statistical Methods in Biology. Cambridge: Cambridge University Press, p. 252.
  • Barton L.L. and W.A. Hamilton. 2010. Sulphate-Reducing Bacteria. Environmental and Engineered Systems. Cambridge: Cambridge University Press, p. 553.
  • Cummings J.H., G.T. Macfarlane and S. Macfarlane. 2003. Intestinal bacteria and ulcerative colitis. Curr. Issues Intest. Microbiol. 4: 9–20.
  • Fernandez V.M., E.C. Hatchikian and R. Cammack. 1985. Properties and reactivation of two different deactivated forms of Desulfovibrio gigas hydrogenase. Biochim. Biophys. Acta 832: 69–79.
  • Furdui C. and S.W. Ragsdale. 2000. The role of pyruvate ferredoxin oxidoreductase in pyruvate synthesis during autotrophic growth by the Wood-Ljungdahl pathway. J. Biol. Chem. 275(37): 28494–28499.
  • Garczarek F., M. Dong, D. Typke, H.E. Witkowska, T.C. Hazen, E. Nogales, M.D. Biggin and R.M. Glaeser. 2007. Octomeric pyruvate-ferredoxin oxidoreductase from Desulfovibrio vulgaris. J. Struc. Biol. 159: 9–18.
  • Gavel O.Y., S.A. Bursakov, J.J. Calvete, G.N. George, J.J. Moura and I. Moura. 1998.ATP sulfurylases from sulfate-reducing bacteria of the genus Desulfovibrio. A novel metalloprotein containing cobalt and zinc. Biochem. 37: 16225–16232.
  • Gibson G.R., J.H. Cummings and G.T. Macfarlane. 1991. Growth and activities of sulphate-reducing bacteria in gut contents of health subjects and patients with ulcerative colitis. FEMS Microbiol. Ecol. 86: 103–112.
  • Guerlesquin F., M. Bruschi, G. Bovier-Lapierre and G. Fauque. 1980. Comparative study of two ferredoxins from Desulfovibrio desulfuricans Norway. Bioch. Biophys. Acta 626: 127–135.
  • Hatchikian E.C., H.E. Jones and M. Bruschi. 1979. Isolation and characterization of a rubredoxin and two ferredoxins from Desulfovibrio africanus. Bioch. Biophys. Acta 548: 471–483.
  • Keleti T. 1988. Basic Enzyme Kinetics. Akademiai Kiado, 422 p. Kushkevych I.V. 2012a. Sulfate-reducing bacteria of the human intestine. I. Dissimilatory sulfate reduction. Sci. Int. J. Biological studies/Studia Biologica 6(1): 149–180.
  • Kushkevych I.V. 2012b. Sulfate-reducing bacteria of the human intestine. II. The role in the diseases development. Sci. Int. J. Biological Studies/Studia Biologica 6(2): 221–250.
  • Kushkevych I.V. 2013. Identification of sulfate-reducing bacteria strains of human large intestine. Sci. Int. J. Biological Studies/Studia Biologica. 7(3): 115–124.
  • Kushkevych I.V., M. Bartos and L. Bartosova. 2014. Sequence analysis of the 16S rRNA gene of sulfate-reducing bacteria isolated from human intestine. Int. J. Curr. Microbiol. Appl. Sci. 3(2): 239–248.
  • Lowry O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall. 1951. Protein determination with the Folin phenol reagent . J. Biol. Chem. 193: 265–275.
  • Ma K., A. Hutchins, S.J. Sung and M.W. Adams. 1997. Pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon, Pyrococcus furiosus, functions as a CoA-dependent pyruvate decarboxylase. Proc. Natl. Acad. Sci. 94(18): 9608–9613.
  • Meinecke B., J. Betram and G. Gottschalk. 1989. Purification and characterization of the pyruvate-ferredoxin oxidoreductase from Clostridium acetobutylicum. Arch. Microbiol. 152(3): 244–250.
  • Pieulle L., B. Guigliarelli, M. Asso, F. Dole, A. Bernadac and E.C. Hatchikian. 1995. Isolation and characterization of the pyruvate-ferredoxin oxidoreductase from the sulfate-reducing bacterium Desulfovibrio africanus. Bioch. et Bioph. Acta 1250: 49–59.
  • Pitcher M.C. and J.H. Cummings. 1996: Hydrogen sulphide: a bacterial toxin in ulcerative colitis? Gut, 39: 1–4.
  • Raeburn S. and J.C. Rabinowitz. 1971. Pyruvate: ferredoxin oxidoreductase-I. The pyruvate-CO₂ exchange reaction. Arch. Biochem. Biophys. 146: 9–20.
  • Raeburn S. and J.C. Rabinowitz. 1971. Pyruvate: ferredoxin oxidoreductase. II. Characteristics of the forward and reverse reactions and properties of the enzyme. Arch. Biochem. Biophys. 146: 21–33.
  • Segal I.H. 1975. Enzyme kinetics: behavior and analysis of rapid equilibrium and steady-state enzyme systems. John Wiley & Sons, New York.
  • Uyeda K. and J.C. Rabinowitz. 1971. Pyruvate-ferredoxin oxidoreductase. IV. Studies on the reaction mechanism. J. Biol. Chem. 246: 3120–3125.
  • Uyeda K. and J.C. Rabinowitz. 1971. Pyruvate-ferredoxin oxidoreductase. III. Purification and properties of the enzyme. J. Biol. Chem. 246: 3111–3119.
  • Zeikus J.G., G. Fuchs, W. Kenealy, R.K. Thauer. 1977. Oxidoreductases involved in cell carbon synthesis of Methanobacterium thermoautotrophicum. J. Bacteriol. 132: 604–613.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

bwmeta1.element.agro-7a7e2d96-a4f5-43c6-bec8-dd988250f46e
JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.