Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

• Published in: Applied surface science Vol. 320; pp. 448 - 454
• Main Authors: Lee, Taek, Chung, Yong-Ho, Yoon, Jinho, Min, Junhong, Choi, Jeong-Woo
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 30.11.2014; Elsevier
• Subjects: Atomic force microscopy; Azurin–myoglobin fusion protein; Biofilms; Condensed matter: electronic structure, electrical, magnetic, and optical properties; Condensed matter: structure, mechanical and thermal properties; Cross-disciplinary physics: materials science; rheology; Devices; Dual-level biomemory; Electrochemical analysis; Exact sciences and technology; Immobilization; Multi-potential chronoamperometry; Myoglobin; Physics; Proteins; Raman spectroscopy; Recombinant; Myoglobin; Atomic force microscopy; Multi-potential chronoamperometry; Dual-level biomemory; Azurin–myoglobin fusion protein; Azurin-myoglobin fusion protein
• ISSN: 0169-4332; 1873-5584
• DOI: 10.1016/j.apsusc.2014.09.020

•We developed the fusion protein-based biofilm on the inorganic surface.•For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method.•The developed fusion protein shows unique electrochemical property.•The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development.