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 o C. 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 o C (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.

Cold‐active catalase from the psychrotolerant fungus Penicillium griseofulvum

“…The specific activity was improved more than 12-fold compared with the cell-free extract while total protein content reduced to 1.96 mg in comparison with the initial protein content of 398.4 mg. Similar results concerning purification degree and enzyme yield of mesophilic CAT have been reported for S. marcescens FZSF01 [58], T. claveryi [20], N. crassa [17], and so on.…”

“…From Aspergillus terreus MTCC 6324, stable CAT with a molecular mass of 368 kDa has been isolated [16]. Purified enzyme has been also obtained from Neurospora crassa [17], Agaricus bisporus [18], opportunistic fungus Scedosporium boydii [19], and mycorrhizal fungus Terfezia claveryi TcCAT-1 [20]. Fiedurek and Gromada [21] developed a method for improvement of CAT production by increasing dissolved oxygen (DO) concentration in culture media.…”

A Novel Extracellular Catalase Produced by the Antarctic Filamentous Fungus Penicillium Rubens III11-2

“…MTCC 6324, a monofunctional enzyme with a molecular mass of 368 kDa, was isolated from A. terreus [19]. A purified enzyme was also obtained from Neurospora crassa [20], Agaricus bisporus [21], the opportunistic fungus Scedosporium boydii [22], and the mycorrhizal fungus Terfezia claveryi TcCAT-1 [23]. Fiedurek and Gromada (2000) developed a method to increase the CAT level by increasing the dissolved oxygen (DO) concentration in the culture filtrate [24].…”