Carrier free cross-linked peroxidase aggregates: synthesis and characterization

Abstract

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 initial enzyme concentration (0.05 mg/ml), BSA (5 mg) and glutaraldehyde (GA) (4 & #956; l/ml) amounts were optimized and maximum immobilized activities were obtained. The kinetic parameters of free and immobilised enzyme have been calculated by using Lineweaver-Burk plots. The saturation constant (Km) and the maximum reaction rate (Vm) of free enzyme were found as 2.33 mM and 0.670mmol/l.sec, respectively. The apparent kinetic parameters ($Km _{app}$ and $Vm _{app}$) of cross linked enzyme aggregates have been calculated as 33.4 mM and 0.127 mmol/l.sec, respectively.