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Purification and characterization of thermo-alkali stable ca | 16620
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International Research Journal of Biotechnology

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Purification and characterization of thermo-alkali stable catalase from Bacillus sp

Abstract

Asmaa Ali Hussein

Thermo-alkali stable catalases are promising enzymes in biotechnological applications as H2O2detoxifying systems. Catalase has been purified and characterized in this study from Bacillus sp. The purification was performed with a three step procedure consisting of ammonium sulfate precipitation, ion exchange chromatography on DEAE- cellulose and gel filtration chromatography on Sephacryl-S300 and finally achieved a 16.6-fold-purifying over the crude extract with 50% overall yield. The purified catalase evidenced an estimated molecular weight of 65 kD. The enzyme also exhibited a broad optimal pH (6.0-10.0), and remained stable (100%) in alkaline pH (8.0-10.0),also it was most active at 70°C and stable within a range of 50-80°C. 2- mercaptoethanol, sodium azide, and potassium cyanide are known protein inhibitors, inhibited catalase activity by 70, 50 and 35% respectively.

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