Isolation, purification and characterization of catalase from Phanerochaete Chrysosporium

Catalase is one of the antioxidant enzymes that plays a major role in the protection of the cells and the tissues from the toxic effects of hydrogen peroxide.In this study, 129.10-fold catalase enzyme purification was obtained from white rot fungus Phanerochaete chrysosporium by 60 percentage ammonium sulphate precipitation, 60 percentage ethanol precipitation, DEAE-cellulose anion exchange and by applyingsephacryl-s-200 gel filtration chromatography as a final purification step. The molecular weight of purified catalase was found about 290 kDa with gel filtration chromatography. SDS-gel electrophoresis results indicated that Phanerochaete chrysosporium catalase consists of four apparently identical subunits, with a molecular weight of around 72 kDa. Optimum pH and the temperature values of the purified catalase was found as 7.5 and 30 celcius, respectively. It was found that purified catalase was more stable in the basic region and it lost its catalitic acitivity about 25 percentage at 60 celcius. The Km and Vmax values were found as 290.69 mM and 250000 U/mg respectively for the purified catalase. It was not observed that imazolyl and procymidon had inhibition effects on purified catalase. However, it was found that benomyl inhibited purified catalase as non-competitive inhibitor and Ki value was determined as 1.158 mM.