The direct electrochemistry of cryo-hydrogel immobilized myoglobin at a glassy carbon electrode

Abstract

A cryo-hydrogel membrane (CHM) immobilized at a glassy carbon (GC) electrode is reported for the direct electron transfer of redox proteins. The most attractive characteristics of this CHM were its hydrophilic micro-environment for incorporated proteins to retain their activities, its high ability for protection against interference of denatured and adsorbed proteins at the electrode, its potential applications for various proteins or enzymes, as well as its high mechanical strength and thermal stability. A clear well developed and stable redox wave was obtained for commercially available horse heart myoglobin without further purification, giving a peak to peak separation ΔE p = 93 mV at 5 mV s −1 and the formal electrode potential E 0′ = −0.158 V (vs. Ag|Agcl). The formal heterogeneous electron transfer rate constant was calculated as k 0′ = 5.7 × 10 −4 cm s −1 at pH 6.5, showing rapid electron transfer was achieved. The pH controlled conformational equilibria, acid state η natural state η basic I state η basic II state, of myoglobin at the CHM GC electrode in the pH range 0–13.8 were also observed and are discussed in detail.