Purification and Characterization of Catalase from Indigenous Fungi of Neurospora crassa InaCC F226

Abstract

Microbial catalase is an important industrial enzyme that catalyzes the decomposition of hydrogen peroxide into water and oxygen. This enzyme posseses great potential of application in food, textile and pharmaceutical industries. In this paper, Neurospora crassa InaCC F226 was used to produce catalase. The aim of this research was to purify the catalase producing by Neurospora crassa InaCC F226 using gel filtration column chromatography and characterization of temperature, pH, Vmax, Km and molecular weight. In this study, the Neurospora crassa was cultured on Vogel's medium for 5 days. The cells were harvested in the 50 mM buffer sodium phosphate (pH 7) containing 3% hydrogen peroxide. Supernatant containing crude enzymes of Catalase was separated from cells by centrifugation at 15 minutes on 13000 x g and 4^oC. The purification result using gel filtration was specific activity 1464.9 U/mg, 10.7 fold and 21.7% yield. Characterization of the purified enzyme toward pH and temperature optimum was 7.0 (271.6 U/mL) and 40^oC (286.1 U/mL). The K_m and V_max found to be 8.8 mM and 5.7 s.mM^-1.  The catalase activity increased by additiom of Fe⁺², Ca⁺² and inhibited by EDTA, Cu⁺² and Mn⁺². The molecular weight of  the catalase was 59.4 kDa