Determination of Kinetics of Peroxidase Enzyme Isolated From Brassica oleracea

Abstract

Peroxidases are extensively found in plants, microorganisms and animals and are resourceful biocatalysts with a wide number of applications in biomedicine as well as biotechnology. The enzyme peroxidase was isolated from the leaves of Brassica oleracea using 0.2 M phosphate buffer at pH 7.0. Different dilutions of this crude enzyme were then examined for peroxidase activity assay. The substrate 4-aminoantipyrine-phenol was used to determine peroxidase activity. The effect of enzyme concentration, substrate concentration as well as temperature and pH on enzyme activity was then determined. The kinetic constants K m and V max were also determined. Based on Lineweaver-Burk plot with substrate concentration in the range of 4.25 × 10-4 M to 16.15 × 10-4 M the K m was found to be 7.14 mM and V max was found to be 0.1 mole/min. The optimum temperature was 50°C and the optimum pH was 8.0 for the peroxidase enzyme. These optimum conditions were used to determine enzyme activities in the cabbage sample.