Optimization of conditions for glutaraldehyde immobilization of soyabean peroxidase

Abstract

Phenols are considered priority pollutants of wastewaters, persistently present in the environment and its conventional treatment could be overcome using enzymatic methods. Although the application of enzymes, such as peroxidases, has been investigated widely, its main disadvantage is the cost of purified enzymes. Immobilized enzymes, on the other hand, have the advantage of greater stability and easy separation from the reaction medium, which makes them reusable. The aim of this study was to optimize the conditions for soyabean peroxidase glutaraldehyde immobilization technique. Immobilization was done in small tubes, by mixing 1 ml of enzyme solution (0.05-2.5 mg/ml) with 50 mg of support matrix (macroporous glycidyl methacrylates previosly synthesized in our lab) on a rotary shaker (24h at 25°C, 250 rpm). Enzyme activity was measured using pyrogallol and H2O2 as substrates (13 mM and 10 mM, respectively), and the reaction products were measured following absorbance at 420 nm, for 3 min using a spectrophotometer. Our data demonstrate that soyabean peroxidase can be successfully immobilized on macroporous glycidyl methacrylate using glutaraldehyde activation.