Purification and properties of cytochromecand two peroxidases from spinach leaves

摘要

1. A simultaneous purification procedure of cytochromec, peroxidases, ferredoxin, ferredoxin-NADP reductase and sulfite reductase from spinach leaves is described. Cytochromec, ferredoxin and ferredoxin-NADP reductase were prepared in crystalline states. The two peroxidases were obtained in homogeneous states as evidenced by their electrophoretic patterns on acrylamide gel and sedimentation analysis.2. Crystalline cytochromecshowed a molecular weight of 13,800 and an E0′of 270 mv at pH 7.0. In addition to these properties, its spectral pattern also indicated that this cytochromecwas derived from mitochondria.3. Two peroxidases were isolated in high spin forms after treatment with HgCl2. They hada-peaks at 556 mμin their reduced forms. Although both peroxidases showed small differences in chromatographic behavior on a carboxymethyl cellulose column, ' they had similar spectral properties, dissociation constants of peroxidase-cyanide complex and rate constants for peroxidase reactio