Peroxide-Dependent Analyte Conversion by the Heme Prosthetic Group, the Heme Peptide “Microperoxidase-11” and Cytochrome c on Chitosan Capped Gold Nanoparticles Modified Electrodes
In view of the role ascribed to the peroxidatic activity of degradation products of cytochrome c (cyt c) in the processes of apoptosis, we investigate the catalytic potential of heme and of the cyt c derived heme peptide MP-11 to catalyse the cathodic reduction of hydrogen peroxide and to oxidize aromatic compounds. In order to check whether cyt c has an enzymatic activity in the native state where the protein matrix should suppress the inherent peroxidatic activity of its heme prosthetic group, we applied a biocompatible immobilization matrix and very low concentrations of the co-substrate H 2O 2. The biocatalysts were entrapped on the surface of a glassy carbon electrode in a biocompatible chitosan layer which contained gold nanoparticles. The electrochemical signal for the peroxide reduction is generated by the redox conversion of the heme group, whilst a reaction product of the substrate oxidation is cathodically reduced in the substrate indication. The catalytic efficiency of microperoxidase-11 is sufficient for sensors indicating HRP substrates, e.g., p-aminophenol, paracetamol and catechol , but also the hydroxylation of aniline and dehalogenation of 4-fluoroaniline. The lower limit of detection for p-aminophenol is comparable to previously published papers with different enzyme systems. The peroxidatic activity of cyt c immobilized in the chitosan layer for catechol was found to be below 1 per mill and for p-aminophenol about 3% as compared with that of heme or MP-11.
References
[1]
Diederix, R.E.M.; Ubbink, M.; Canters, G.W. Effect of the protein matrix of cytochrome c in suppressing the inherent peroxidase activity of its heme prosthetic group. ChemBioChem 2002, 3, 110–112.
[2]
Lohmann, W.; Karst, U. Biomimetic modelling of oxidative drug metabolism. Anal. Bioanal. Chem. 2011, 391, 79–96.
[3]
Ahluwalia, U.; Nayeem, S.M.; Deep, S. The non-native conformations of cytochrome c in sodium dodecyl sulfate and their modulation by ATP. Eur. Biophys. J. 2011, 40, 259–271.
[4]
Vazquez-Duhalt, R. Cytochrome c as a biocatalyst. J. Mol. Catal. B Enzym. 1999, 7, 241–249.
[5]
Laszlo, J.A.; Compton, D.L. Comparison of peroxidase activities of hemin, cytochrome c, and microperoxidase-11 in molecular solvents and imidazolium-based ionic liquids. J. Mol. Catal. B Enzym. 2002, 18, 109–120.
[6]
Everse, J.; Liu, C.-J.J.; Coates, P.W. Physical and catalytic properties of a peroxidase derived from cytochrome c. Biochim. Biophys. Acta 2011, 1812, 1138–1145.
Cai, J.; Yang, J.; Jones, D.P. Mitochondrial control of Apoptosis: The role of the cytochrome c. Biochim. Biophys. Acta 1998, 1366, 139–149.
[9]
Yarman, A.; Badalyan, A.; Gajovic-Eichelmann, N.; Wollenberger, U.; Scheller, F.W. Enzyme electrode for aromatic compounds exploiting the catalytic activities of microperoxidase-11. Biosens. Bioelectron. 2011, 30, 320–323.
[10]
Bonnard, C.; Papermaster, D.S.; Kiraehenbuhl, J.-P. The streptavidin-biotin bridge technique: Applications in light and electron microscope immunocytochemistry. In Immunolabeling for Electron Microscopy; Elsevier: New York, NY, USA, 1984.
Marcus, R.A.; Sutin, N. Electron transfer in chemistry and biology. Biochim. Biophys. Acta 1985, 811, 265–322.
[13]
L？tzbeyer, T.; Schuhmann, W.; Schmidt, H.-L. Minienzymes: A review for the development of reagentless amperometric biosensors based on direct electron-transfer process. Bioelectrochem. Bioenerg. 1997, 42, 1–6.
[14]
Yarman, A.; Nagel, T.; Gajovic-Eichelmann, N.; Fischer, A.; Wollenberger, U.; Scheller, F.W. Bioelectrocatalysis by microperoxidase-11 in a multilayer architecture of chitosan embedded gold nanoparticles. Electroanalysis 2011, 23, 611–618.
[15]
Liu, J.; Qiu, J.; Sun, K.; Chen, J.; Miao, Y. Electrochemistry of hemin self-assembled from aqueoushexadecyltrimethylammonium bromide (CTAB) solution on single-wall-carbon-nanotube-modified glassy carbon electrodes. Helv. Chim. Acta 2009, 92, 462–469.
[16]
Feng, Z.Q.; Sagara, T.; Niki, K. Electroreflectance study of hemin adsorbed on pyrolytic graphite electrode surface and its coadsorption with with methylene blue. J. Electroanal. Chem. 1993, 349, 159–171.
[17]
Chen, J.; Wollenberger, U.; Lisdat, F.; Ge, B.; Scheller, F.W. Superoxide sensor based on hemin modified electrode. Sens. Actuat. B 2000, 70, 115–120.
[18]
Zheng, N.; Zeng, Y.; Osborne, P.G.; Li, Y.; Chang, W.; Wang, Z. Electrocatalytic reduction of dioxygen on hemin based carbon paste electrode. J. Appl. Electrochem. 2002, 32, 129–133.
[19]
Feng, J.-J.; Li, Z.-H.; Li, Y.-F.; Wang, A.-J.; Zhang, P.-P. Electrochemical determination of dioxygen and hydrogen peroxide using Fe3O4@SiO2@hemin microparticles. Microchim. Acta 2002, 32, 129–133.
[20]
Laviron, E. General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems. J. Electroanal. Chem. 1979, 101, 19–28.
[21]
Gorton, L.; Lindgren, A.; Larsson, T.; Munteanu, F.D.; Ruzgas, T.; Gazaryan, I. Direct electron transfer between heme-containing enzymes and electrodes as basis for third generation biosensors. Anal. Chim. Acta 1999, 400, 91–108.
[22]
Marques, M.H. Insights into porphyrin chemistry provided by the microperoxidases, the haempeptides derived from cytochrome c. Dalton Trans. 2007, 39, 4371–4385.
[23]
Ruzgas, T.; Gaigalas, A.; Gorton, L. Diffusionless electron transfer of microperoxidase-11 on gold electrodes. J. Electroanal. Chem. 1999, 469, 123–131.
[24]
Zhou, Y.; Liu, S.; Jiang, H.-J.; Yang, H.; Chen, H.Y. Direct electrochemistry and bioelectrocatalysis of microperoxidase-11 immobilized on chitosan-graphene nanocomposite. Electroanalysis 2010, 22, 1323–1328.
[25]
Feng, J.-J.; Zhao, G.; Xu, J.-J.; Chen, H.-Y. Direct electrochemistry and electrocatalysis of heme proteins immobilized on gold nanoparticles stabilized by chitosan. Anal. Biochem. 2005, 342, 280–286, doi:10.1016/j.ab.2005.04.040.
[26]
Dallacosta, C.; Casella, L.; Monzani, E. Modified microperoxidases exhibit different reactivity towards phenolic substrates. ChemBioChem 2004, 5, 1692–1699.
[27]
Cs？regi, E.; J？nsson-Petterson, G.; Gorton, L. Mediatorless electrocatalytic reduction of hydrogen peroxide at graphite electrodes chemically modified with peroxidases. J. Biotechnol. 1993, 30, 315–337.
[28]
Ranieri, A.; Battistuzzi, G.; Borsari, M.; Bortolotti, C.A.; Di Rocco, G.; Monari, S.; Sola, M. A bis-histidine-ligated unfolded cytochrome c immobilized on anionic SAM shows pseudo-peroxidase activity. Electrochem. Commun. 2012, 14, 29–31.
[29]
Dunford, H.B. Oxidations of iron(II)/(III) by hydrogen peroxide: From aquo to enzyme. Coord. Chem. Rev. 2002, 311, 233–234.
[30]
Astuti, Y.; Topoglidis, E.; Durrant, J.R. Use of microperoxidase-11 to functionalize tin dioxide electrodes for the optical and electrochemical sensing of hydrogen peroxide. Anal. Chim. Acta 2011, 686, 126–132.
[31]
Razumas, V.; Kazlauskait？, J.; Vid？iūnait？, R. Electrocatalytic reduction of hydrogen peroxide on the microperoxidase-11 modified carbon paste and graphite electrodes. Bioelectrochem. Bioenerg. 1996, 39, 139–143.
[32]
Youssoufi-Korri, H.; Desbenoit, N.; Ricoux, R.; Mahy, J.P.; Lecomte, S. Eleboration of a new hydrogen peroxide biosensor using microperoxidase 8 (MP8) immobilised on a polypyyrole coated electrode. Mater. Sci. Eng. C 2008, 28, 855–860.
[33]
Wang, M.; Shen, Y.; Liu, Y.; Wang, T.; Zhao, F.; Liu, F.; Dong, S. Direct electrochemistry of microperoxidase-11 using carbon nanotube modified electrodes. J. Electroanal. Chem. 2005, 578, 121–127.
[34]
Liu, Y.; Wang, M.; Zhao, F.; Guo, Z.; Chen, H.; Dong, S. Direct electron transfer and electrocatalysis of microperoxidase immobilised on nanohybrid film. J. Electroanal. Chem. 2005, 581, 1–10.
[35]
Zhu, X.; Yuri, I.; Gan, X.; Suzuki, I.; Li, G. Electrochemical study of the effect of nano-zinc oxide on microperoxidase and its application to more sensitive hydrogen peroxide biosensor preparation. Biosens. Bioelectron. 2007, 22, 1600–1604.
[36]
Huang, W.; Jia, J.; Zhang, Z.; Han, X.; Tang, J.; Wang, J.; Dong, S.; Wang, E. Hydrogen peroxide biosensor based on microperoxidase-11 entrapped in lipid membrane. Biosens. Bioelectron. 2003, 18, 1225–1230.
[37]
Cipriano, T.C.; Takahashi, P.M.; de Lima, D.; Oliveira, V.X.; Souza, J.A.; Martinho, H.; Alves, W.A. Spatial organization of peptide nanotubes for electrochemical devices. J. Mater. Sci. 2010, 45, 5101–5108.
[38]
Wan, J.; Bi, J.; Du, P.; Zhang, S. Biosensor based on the biocatalysis of microperoxidase-11 in nanocomposite material of multiwalled carbon nanotubes/room temperature ionic liquid for amperometric determination of hydrogen peroxide. Anal. Biochem. 2009, 386, 256–261.
[39]
Chen, Z.; Sun, D.; Zhou, Y.; Zhao, J.; Lu, T.; Huang, X.; Cai, C.; Shen, J. Nano polyurethane-assisted ultrasensitive biodetection of hydrogen peroxide over immobilized Microperoxidase-11. Biosens.Bioelectron. 2011, 29, 53–59.
[40]
Ju, H.; Liu, S.; Ge, B.; Lisdat, F.; Scheller, F.W. Electrochemistry of cytochrome c immobilized on colloidal gold modified carbon paste electrodes and its electrocatalytic activity. Electroanalysis 2002, 14, 141–147, doi:10.1002/1521-4109(200201)14:2<141::AID-ELAN141>3.0.CO;2-Y.
[41]
Wang, L.; Waldeck, D.H. Denaturation of cytochrome c and its peroxidase activity when immobilized on SAM films. J.Phys.Chem.C 2008, 112, 1351–1356.
[42]
J？nchen, M.; Scheller, F.; Prümke, H.-J.; Mohr, P. Polarographische untersuchungen zur peroxidaseaktivit？t von ligandiertem deuteroh？min. Acta Biol. Medica Ger. 1975, 34, 319–324.
[43]
Yang, S.; Li, Y.; Jiang, X.; Chen, Z.; Lin, X. Horseradish peroxidase biosensor based on layer-by-layer technique for the determination of phenolic compounds. Sens. Actuat. B 2006, 114, 774–780.
[44]
Veeger, C. Does P450-type catalysis proceed through a peroxo-iron intermediate? A review of studies with microperoxidase. J. Inorg. Biochem. 2002, 91, 35–45.
[45]
Lin, M.S.; Jan, B.I.; Chen, P.Y.; Cheng, W.C.; Chen, C.H. New determination scheme of p-aminophenol by MnO2 modified electrode coupled with flow injection analysis. Electroanalysis 2010, 22, 1278–1281.
[46]
Wollenberger, U.; Lisdat, F.; Rose, A.; Streffer, K. Bioelectrochemistry: Fundamentals, Experimental Techniques and Applications; Bartlett, P., Ed.; John Wiley & Sons: Hoboken, NJ, USA, 2008.
[47]
Wollenberger, U.; Neumann, B. Quinoprotein glucose dehydrogenase modified carbon paste electrode for the detection of phenolic compounds. Electroanalysis 1997, 9, 366–371.
[48]
Mie, Y.; Kowata, K.; Hirano, Y.; Niwa, O.; Mizutani, F. Comparison of enzymatic recycling electrodes for measuring aminophenol: Development of a highly sensitive natriuretic peptide assay system. Anal. Sci. 2008, 24, 577–582.
[49]
Solná, R.; Skládal, P. Amperometric flow-injection determination of phenolic compounds using a biosensor with immobilized laccase, peroxidase and tyrosinase. Electroanalysis 2005, 17, 2137–2146.
[50]
Zhu, Y.; Li, J.; Dong, S. Dimerization of hydroxylated species of m-aminophenolby cytochrome c with hydrogen peroxide. J. Mol. Catal. B Enzym. 1998, 5, 475–482.
[51]
Kawakami, M.; Akamatsu, N.; Koya, H.; Amada, K. Amperometric detection of phenol with cytochrome c-modified gold electrode using dual working electrode system. Anal. Lett. 2005, 38, 549–561.
