Direct voltammetry and electrochemical catalysis with horseradish peroxidase in polyacrylamide hydrogel films

This paper reports the direct voltammetry of horseradish peroxidase (HRP) incorporated in amphiphilic polyacrylamide (PAM) films modified on pyrolytic graphite (PG) electrodes. Cyclic voltammetry of HRP-PAM films showed a pair of well-defined, nearly reversible peaks at approximately -0.33 V vs. SCE in pH 7.0 buffers, characteristic of HRP heme Fe(III)/Fe(II) redox couple. The PAM films in solution contained large amounts of water and formed a hydrogel, and provided a favorable microenvironment for HRP and facilitated its direct electron transfer with underlying PG electrodes. The apparent heterogeneous electron transfer rate constant (k(s)) and formal potential (E*') were estimated by fitting the data of square wave voltammetry (SWV) with the non-linear regression analysis. UV-vis absorption spectra demonstrated that HRP in PAM films retained its secondary structure similar to its native state. The embedded HRP in PAM films showed the electrocatalytic activity to various substrates such as nitrite, oxygen and hydrogen peroxide. The possible mechanism of catalytic reaction of H(2)O(2) with HRP-PAM films was proposed.