Extraction, Purification and Characterization Of Peroxidase From cabbage (Brassica oleracea Var) .

Abstract

The activity of peroxidase (POD) in cabbage was evaluated using spectrophotometric method. The enzyme was extracted from the cabbage leaves with 0.1 M phosphate buffer solution pH 7. 0 . POD activity was determined using (O-dianisidine) as a substrate. The effects of the amounts of enzyme extract, substrate concentration, pH and temperature were investigated. The highest activity of POD was recored at 2 mg/ml. The highest activity of POD was optimized with 16 mM O-dianisidine, The optimum pH was 7.0 for POD , The optimum temperature was 30°C for POD. These optimum conditions were used to determined the enzyme activities in cabbage sample. Acetone fractionated peroxidase from crude extract of Brassica oleracea leaves (Cabbage) was purified on DEAE-Cellulose chromatographic columns. The specific activity of purified POD is 103.70 (U/mg) ,which is 5.37 times more than the crude extract with 28.72% recovery. Maximum pH, thermal activity and stability of this purified enzyme are also determined were 40°C .