Purification and Characterization of Catalase Enzymes from Chicken Liver and Sheep Erythrocytes

Abstract

Catalase enzyme (H₂O₂: H₂O₂ oxidoreductase; E.C. 1.11.1.6) was purified from chicken liver and sheep erythrocytes, using DEAESephadex A50 ion exchange chromatography and some characteristics of the enzymes were investigated. The purification procedure was composed of 3 steps i.e., homogenate/hemolisate preparation, ammonium sulfate precipitation and DEAE-Sephadex A50 ion exchange chromatography. Chicken liver and sheep erythrocytes enzymes, having the specific activity of 560.46 and 1017.5 EU/mg proteins were purified with a yield of 30.06 and 22.23 %; 190.63 and 643.9-fold, respectively. In order to control the purification of enzymes were done, sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE). SDS-PAGE showed a single band for each enzyme. Optimal pH, stable pH, optimal temperature, KM and V_max values for H₂O₂ were also determined for the each enzyme. In addition, molecular weight and subunit molecular weights were found by SDS-PAGE and gel filtration chromatography respectively.