Production and Purification of Peroxidase from Tissue Cultures of Cauliflower (Brassica oleracea L. var. Botrytis)

Abstract

The plant peroxidases are remarkable enzymes due to their widespread use in industry. Theseenzymes, which are capable of catalysing the oxidation of various organic and inorganicsubstrates, have been used in clinical diagnosis, detoxification reactions and organic synthesis.In this study, in vitro production and purification of peroxidase enzyme from cauliflower plantwas proposed. Firstly, sterile seedlings were obtained from MS/B5 nutrient medium withoutgrowth regulator from cauliflower seeds and calluses from medium containing 0.5 mg / L 2.4-D. Callus and seedlings were powdered with liquid nitrogen and then homogenized. Peroxidaseenzymes were purified from these homogenates using affinity technique. SDS-PAGEelectrophoresis was performed to determine the molecular weight of the purified enzymes andsingle bands was observed at approximately 46 kDa. In addition, KM and Vmax values of thecallus peroxidase enzyme were determined for guaiacol, pyrogallol and H2O2 substrates.