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Characterization of Extra Cellular Thermostable Endoglucanase from Rhizopus oryzae using Response Surface Methodology

Moumita Karmakar and Rina Rani Ray

High activity extracellular endoglucanase (E.C.3.2.1.4) produced by Rhizopus oryzae PR7MTCC 9642 was partially purified and characterized. The central composite experimental design (CCD) and response surface methodology (RSM) were employed to derive a statistical model for the effects of different physio-chemical conditions on the activity of the enzyme, which indicated that a pH 6.0 and an incubation temperature of 33ï‚°C along with 1%(w/v) substrate (carboxy methyl cellulose) concentration was required to maximize the activity of the enzyme. The enzyme was extremely thermo stable and 68% activitywas restored even after the exposure of the enzyme protein at 60ï‚°Cfor 60minutes. The enzymewas 60% stable in a broad range of pH of 4 to 8. The endoglucanase activity was found to increase in presence of thiol compounds indicating the presence of thiol groups at the active site.