A Direct Electrode-Driven Immobilized Catalase for Biocatalysis of Peroxides

Abstract

The direct electrochemistry of catalase (Cat) immobilized in agarose hydrogel film on glassy carbon (GC) electrode was investigated. A pair of well-defined and quasi-reversible redox peaks both in aqueous buffer and ethanol-aqueous mixture was obtained. The formal potential (E°') indicates the reactions origin from the direct electron transfer between the CatFe(III)/(II) couple and the electrode surface. The E°' is linearly dependent on solution pH (redox Bohr effect), indicating the electron transfer of the CatFe(III)/(II) redox couple accompanied with the transfer of a proton. The electron transfer can be facilitated in ethanol-aqueous mixture for the reason of partial dehydration of the distal heme pocket. The water pools in agarose hydrogel play an important role in preventing either the change of conformation or the inactivity of Cat from polar organic solvent. Cat retains its original conformation in the agarose film that was confirmed by UV-Vis spectra and electrochemical behaviors in comparison to the controlled experiments using free hemin. The catalytic reactions to peroxides (hydrogen peroxide, cumene hydroperoxide, t-butyl hydroperoxide, 2-butanone peroxide) using Cat-agarose modified electrode were also explored both in aqueous buffer and ethanol-aqueous mixture. The method could be application to the preparation of the hydrogel based third generation biosensors or bioreactors in organic media.