Purification and Properties of Liver Catalase from Water Buffalo (Bubalus bubalis)

Abstract

Catalase is ubiquitous enzyme which catalyses the breakdown of hydrogen peroxide (H 2 O 2 ). The aim of present study was to purify and characterize the liver catalase from Bubalus bubalis, which is an important domestic animal in South Asia. The enzyme was purified to electrophoretic homogeneity by selective acetone precipitations and anion-exchange chromatography based on DEAE-Sephadex Colum. Purified enzyme displayed specific activity of 35753 units per mg of protein with 30.5 % recovery and 123 fold purification. Optimum activity was measured at pH 7 and 30C. The K M value of purified catalase was calculated as 58mM for H 2 O 2 . Gel-filtration chromatography experiments indicate that the purified enzyme was a tetramer with MW of about 240 kDa. There was no effect of 10 to 50mM EDTA on the activity of enzyme after incubation for 30 min at 4C. Our study reveals the first report on the purification and properties of catalase from river buffalo which is an unexplored species for its proteins. The isolated enzyme with high yield and specific activity, offers an attractive alternate for the bovine and porcine enzymes used in clinical laboratories.