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