Carrier Free Cross-linked Peroxidase Aggregates: Synthesis and Characterization

In this study, cross-linked Horseradish Peroxidase HRP aggregates were prepared. Bovin Serum Albumin BSA ,a proteic feeder, and glutaraldehyde, a bifunctional reactive, a crosslinker agent has also been used. The initialenzyme concentration 0.05 mg/ml , BSA 5 mg and glutaraldehyde GA 4 µl/ml amounts were optimized andmaximum immobilized activities were obtained. The kinetic parameters of free and immobilised enzyme have beencalculated by using Lineweaver-Burk plots. The saturation constant Km and the maximum reaction rate Vm offree enzyme were found as 2.33 mM and 0.670mmol/l.sec, respectively. The apparent kinetic parameters Kmapp andVmapp of cross linked enzyme aggregates have been calculated as 33.4 mM and 0.127 mmol/l.sec, respectively