title: Purification and some properties of thiosulfate dehydrogenase from Acidithiobacillus ferrooxidans.
abstract: Thiosulfate dehydrogenase was purified from Acidithiobacillus ferrooxidans using three purification steps. The purification procedure involved ammonium sulfate fractionation, ion-exchange chromatography, and gel permeation chromatography. Specific activity of the purified enzyme (after IEC) was 3.26 nkat/mg, and yield of the enzyme was 78%. The purity of the enzyme was checked by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme is a tetramer composed of four probably identical subunits of relative molecular weight 45,000. The pH optimum of the enzyme reaction in the direction of substrate oxidation was found to be 3.0. The isoelectric point of the enzyme was 8.3. Enzyme activity was found to be particularly sensitive to the histidine-selective reagent diethylpyrocarbonate. Reagents selective for arginine, cysteine, and tryptophane had no effect on enzyme activity.
authors: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, Brno, Czech Republic. janiczek@chemi.muni.cz
authors: O
authors: O
authors: Janiczek
authors: J
authors: J
authors: Zemanova
authors: M
authors: M
authors: Mandl
mesh_terms: D042763:Acidithiobacillus / Q000201:enzymology*; D000645:Ammonium Sulfate / Q000737:chemistry; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects*; D006639:Histidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D012967:Sodium Dodecyl Sulfate; D006639:Histidine; D010088:Oxidoreductases; C432587:thiosulfate dehydrogenase; D004047:Diethyl Pyrocarbonate; D000645:Ammonium Sulfate