Respiratory enzymes of Thiobacillus ferrooxidans. A kinetic study of electron transfer between iron and rusticyanin in sulfate media.
Thiobacillus ferrooxidans is a chemolithotrophic bacterium capable of fulfilling all of its energy requirements from the oxidation of soluble ferrous sulfate. Rusticyanin is a soluble blue copper protein found in abundance in the periplasmic space of this bacterium. The one-electron transfer reaction between soluble iron and purified rusticyanin has been studied by stopped flow spectrophotometry in acidic solutions containing sulfate. Second order rate constants for the reduction of rusticyanin by Fe2+, FeHSO4+, and FeSO4(0) were 0.022, 0.73, and 2.30 M-1 s-1, respectively. The pseudo-first order rate constant for the reduction of rusticyanin exhibited substrate saturation when the concentration of the total ferrous ion was varied in solutions of limiting sulfate. This saturation behavior was quantitatively described using the values of the second order rate constants listed above and the distribution of the total ferrous ion into its water-, bisulfate-, and sulfate-coordinated forms. Second order rate constants for the oxidation of rusticyanin by Fe3+ and FeSO4+ were 0.73 and 0.26 M-1 s-1, respectively. The electron transfer reactions between iron and rusticyanin monitored in vitro were far too slow to support the hypothesis that rusticyanin is the primary oxidant of ferrous ions in the iron-dependent respiratory electron transport chain of T. ferrooxidans.
Blake
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RC
Department of Biochemistry, Meharry Medical College, Nashville, Tennessee 37208.
Shute
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EA
D001400:Azurin / Q000031:analogs & derivatives*; D003470:Culture Media; D004579:Electron Transport; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008667:Metalloproteins / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D013431:Sulfates; D013855:Thiobacillus / Q000201:enzymology* / Q000254:growth & development
D003470:Culture Media; D008667:Metalloproteins; D013431:Sulfates; C012543:rusticyanin; D001400:Azurin; D007501:Iron