title: Purification and properties of thiosulfate reductase from Desulfovibrio gigas.
abstract: Thiosulfate reductase of the dissimilatory sulfate-reducing bacterium Desulfovibrio gigas has been purified 415-fold and its properties investigated. The enzyme was unstable during the different steps of purification as well as during storage at - 15 degrees C. The molecular weight of thiosulfate reductase estimated from the chromatographic behaviour of the enzyme on Sephadex G-200 was close to 220000. The absorption spectrum of the purified enzyme exhibited a protein peak at 278 nm without characteristic features in the visible region. Thiosulfate reductase catalyzed the stoichiometric production of hydrogen sulfide and sulfite from thiosulfate, and exhibited tetrathionate reductase activity. It did not show sulfite reductase activity. The optimum pH of thiosulfate reduction occurred between pH 7.4 and 8.0 and its Km value for thiosulfate was calculated to be 5 - 10(-4)M. The sensitivity of thiosulfate reductase to sulfhydryl reagent and the reversal of the inhibition by cysteine indicated that one or more sulfhydryl groups were involved in the catalytic activity. The study of electron transport between hydrogenase and thiosulfate reductase showed that the most efficient coupling was obtained with a system containing cytochromes c3 (Mr = 13000) and c3 (Mr = 26000).
mesh_terms: D003080:Cold Temperature; D003574:Cytochrome c Group; D003901:Desulfovibrio / Q000201:enzymology*; D004227:Dithionite; D004355:Drug Stability; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010088:Oxidoreductases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013776:Tetrathionic Acid; D013885:Thiosulfates; D014755:Viologens
chemical_list: D003574:Cytochrome c Group; D013439:Sulfhydryl Reagents; D013885:Thiosulfates; D014755:Viologens; D004227:Dithionite; D013776:Tetrathionic Acid; D010088:Oxidoreductases