Fabrication of fusion protein-based heterolayers composed of redox protein/myoglobin for bioelectronic device

Abstract

In the present study, fusion protein heterolayers, consisting of metalloprotein conjugates were fabricated for bioelectronic device. A recombinant azurin-myoglobin (Azu-Myo) fusion protein and a cytochrome c-myoglobin (Cyt-Myo) fusion protein were conjugated through S-HyNic (succinimidyl-6-hydrazino- nicotinamide) linker and S-4FB (succinimidyl-4- formylbenzamide) linker, respectively. SDS-PAGE and UV-VIS spectroscopy were carried out to confirm the conjugation of each fusion protein. Each fusion protein conjugate was self-assembled onto an Au substrate, and then atomic force microscopy (AFM) was used to observe the surface morphology and surface roughness of each fusion protein. Moreover, the redox properties of the fusion protein heterolayers were measured via cyclic voltammetry (CV). The results indicated that the Azu-Myo and Cyt-Myo fusion protein heterolayers maintained their original electrochemical properties. The present study therefore showed that the fabricated fusion protein heterolayers can be used in electrochemical- based bioelectronic device application.