Electrochemical and spectroelectrochemical characterization of different mesoporous TiO₂ film electrodes for the immobilization of Cytochrome c

Abstract

In this work three different mesoporous TiO₂ film electrodes were prepared and used for the immobilization of Cytochrome c (Cyt-c). Films prepared via a standard sol–gel route (SG-films) were compared with commercially available benchmark nanotitania materials, namely P25 Degussa (P25-films) and Dyesol nanopaste (Dyesol films). Their properties, film deposition characteristics and their abilities to adsorb protein molecules in a stable and functional way were examined. We investigated whether it is possible, rather than preparing TiO₂ films using multistep, lengthy and not always reproducible sol–gel procedures, to use commercially available nanotitania materials and produce reproducible films faster that exhibit all the properties that make TiO₂ films ideal for protein immobilization. Although these materials are formulated primarily for dye-sensitized solar cell applications, in this study we found out that protein immobilization is facile and remarkably stable on all of them. We also investigated their electrochemical properties by using cyclic voltammetry and spectroelectrochemistry and found out that not only direct reduction of Fe(III)-heme to Fe(II)-heme of immobilized Cyt-c was possible on all films but that the adsorbed protein remained electroactive.

Keywords

TiO₂ films / spectroelectrochemistry / cyclic voltammetry / Cytochrome c

References

[1] Solanki P R, Kaushik A, Agrawal V V, . Nanostructured metal oxide-based biosensors. NPG Asia Materials, 2011, 3(1): 17–24

[2] Bisquert J, Fabregat-Santiago F, Mora-Sero I, . A review of recent results on electrochemical determination of the density of electronic states of nanostructured metal-oxide semiconductors and organic hole conductors. Inorganica Chimica Acta, 2008, 361(3): 684–698

[3] Obare S O, Ito T, Balfour M H, . Ferrous hemin oxidation by organic halides at nanocrystalline TiO₂ interfaces. Nano Letters, 2003, 3(8): 1151–1153

[4] Topoglidis E, Campbell C J, Cass A E G, . Factors that affect protein adsorption on nanostructured titania films. A novel spectroelectrochemical application to sensing. Langmuir, 2001, 17(25): 7899–7906

[5] Topoglidis E, Cass A E G, O’Regan B, . Immobilization and bioelectrochemistry of proteins on nanoporous TiO₂ and ZnO films. Journal of Electroanalytical Chemistry, 2001, 517(1‒2): 20–27

[6] Topoglidis E, Astuti Y, Duriaux F, . Direct electrochemistry and nitric oxide interaction of heme proteins adsorbed on nanocrystalline tin oxide electrodes. Langmuir, 2003, 19(17): 6894–6900

[7] Li Q, Luo G, Feng J. Direct electron transfer for heme proteins assembled on nanocrystalline TiO₂ film. Electroanalysis, 2001, 13(5): 359–363

[8] Jiang G, Tang H, Zhu L, . Improving electrochemical properties of liquid phase deposited TiO₂ thin films by doping sodium dodecylsulfonate and its application as bioelectrocatalytic sensor for hydrogen peroxide. Sensors and Actuators B: Chemical, 2009, 138(2): 607–612

[9] Renault C, Balland V, Martinez-Ferrero E, . Highly ordered transparent mesoporous TiO₂ thin films: an attractive matrix for efficient immobilization and spectroelectrochemical characterization of cytochrome c. Chemical Communications, 2009, 48(48): 7494–7496

[10] Yang D H, Takahara N, Mizutani N, . Fabrication of TiO₂ and cytochrome  c alternate ultrathin films via a gas-phase surface sol‒gel process. Chemistry Letters, 2006, 35(9): 990–991

[11] McKenzie K J, Marken F. Accumulation and reactivity of the redox protein cytochrome c in mesoporous films of phytate. Langmuir, 2003, 19(10): 4327–4331

[12] Kim H, Park S S, Seo J, . Stable protein device platform based on pyridine dicarboxylic acid-bound cubic-nanostructured mesoporous titania films. ACS Applied Materials & Interfaces, 2013, 5(15): 6873–6878

[13] Yang D H, Shin M J, Choi S M, . Cytochrome c assembly on fullerene nanohybrid metal oxide ultrathin films. RSC Advances, 2016, 6(23): 19173–19181

[14] Liu L, Wang N, Cao X, . Direct electrochemistry of cytochrome c at a hierarchically nanostructured TiO₂ quantum electrode. Nano Research, 2010, 3(5): 369–378

[15] McKenzie K J, Marken F, Opallo M. TiO₂ phytate films as hosts and conduits for cytochrome c electrochemistry. Bioelectrochemistry, 2005, 66(1‒2): 41–47

[16] Topoglidis E, Lutz T, Durrant J R, . Interfacial electron transfer on cytochrome-c sensitised conformally coated mesoporous TiO₂ films. Bioelectrochemistry, 2008, 74(1): 142–148

[17] Rafiee-Pour H A, Hamadanian M, Koushali S K. Nanocrystalline TiO₂ films containing sulfur and gold: Synthesis, characterization and application to immobilize and direct electrochemistry of cytochrome c. Applied Surface Science, 2016, 363: 604–612

[18] Qiu J, Zhang S, Zhao H. Recent applications of TiO₂ nanomaterials in chemical sensing in aqueous media. Sensors and Actuators B: Chemical, 2011, 160(1): 875–890

[19] Wang R, Zhang J, Hu Y. Liquid phase deposition of hemoglobin/SDS/TiO₂ hybrid film preserving photoelectrochemical activity. Bioelectrochemistry, 2011, 81(1): 34–38

[20] Luo Y, Tian Y, Zhu A, . pH-dependent electrochemical behavior of proteins with different isoelectric points of the nanostructured TiO₂ surface. Journal of Electroanalytical Chemistry, 2010, 642(2): 109–114

[21] Renault C, Nicole L, Sanchez C, . Unraveling the charge transfer/electron transport in mesoporous semiconductive TiO₂ films by voltabsorptometry. Physical Chemistry Chemical Physics, 2015, 17(16): 10592–10607

[22] Kityakarn S, Pooarporn Y, Songsiriritthigul P, . (Photo)Electrochemical characterization of nanoporous TiO₂ and Ce-doped TiO₂ sol‒gel film electrodes. Electrochimica Acta, 2012, 83: 113–124

[23] Dai G, Zhao L, Wang S, . Double-layer composite film based on sponge-like TiO₂ and P25 as photoelectrode for enhanced efficiency in dye-sensitized solar cells. Journal of Alloys and Compounds, 2012, 539: 264–270

[24] Lee M S, Cheon I C, Kim Y I. Photoelectrochemical studies on nanocrystalline TiO₂ film electrodes. Bulletin of the Korean Chemical Society, 2003, 24(8): 1155–1162

[25] Moore G R, Pettigrew G W. Cytochrome c: Evolution, Structure, and Physicochemical Aspects. Berlin: Springer-Verlag, 1990

[26] Stellwagen E. Haem exposure as the determinate of oxidation‒reduction potential of haem proteins. Nature, 1978, 275(5675): 73–74

Acknowledgement

This work was partially supported by a small initiative grant from the University of Patras.

RIGHTS & PERMISSIONS

2018 Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature