title: Use of immobilized cytochrome c as a ligand for affinity chromatography of thiosulfate dehydrogenase from Acidithiobacillus ferrooxidans.
abstract: Three matrices were used for immobilizing the cytochrome c: Sepharose CL-4B, Silasorb SPH amine and a laboratory-prepared new matrix based on crosslinked triazine (2,4,6-tris(aminoethylamine)-1,3,5-triazine) (TAT). Cytochrome c was immobilized on the matrices by several procedures and the amount of incorporated cytochrome c was determined. Cytochrome c immobilized on Sepharose CL-4B with periodate activation, cytochrome c immobilized on Silasorb-amine with carbodiimide activation and cytochrome c immobilized on crosslinked triazine were suitable for purification of thiosulfate dehydrogenase from Acidithiobacillus ferrooxidans. The yield with all matrices was about 90%. The purification factor of the above matrices was about 15. A new matrix based on TAT with cytochrome c represented a suitable way for thiosulfate dehydrogenase purification.
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic. janiczek@chemi.muni.cz
authors.forename: Oldrich
authors.initials: O
authors.lastname: Janiczek
authors.forename: Blanka
authors.initials: B
authors.lastname: Pokorna
authors.forename: Jana
authors.initials: J
authors.lastname: Zemanova
authors.forename: Martin
authors.initials: M
authors.lastname: Mandl
mesh_terms: D042763:Acidithiobacillus / Q000201:enzymology*; D002846:Chromatography, Affinity; D045304:Cytochromes c / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008024:Ligands; D010088:Oxidoreductases / Q000302:isolation & purification*; D011485:Protein Binding
chemical_list: D004800:Enzymes, Immobilized; D008024:Ligands; D045304:Cytochromes c; D010088:Oxidoreductases; C432587:thiosulfate dehydrogenase