Abstract
Adsorption and bioelectrocatalytic activity of native horseradish peroxidase (HRP) and its recombinant forms on polycrystalline gold electrodes were studied. Recombinant forms of HRP were produced by a genetic engineering approach using an E. coli expression system. According to direct mass measurements with a quartz crystal microbalance, all the forms of HRP formed monolayer coverage of the enzyme on the gold surface. However, only gold electrodes modified with the recombinant HRP forms (non-glycosylated) exhibited high and stable current response to H2O2 due to its bioelectrocatalytic reduction based on direct electron transfer (ET) between gold and the active site of the enzyme. Introduction of a six-His tag either at the C-terminus or at the N-terminus of the enzyme molecule additionally increased the strength of the enzyme binding with the gold surface and the efficiency of direct ET. Immobilization of recombinant forms of HRP containing histidine functional groups on the surface of the gold electrode was used both for the development of a P-chip, a biosensor for hydrogen peroxide determination based on direct ET, and for the development of a bienzyme biosensor electrode for the determination of L-lysine based on co-immobilized recombinant forms of HRP and L-lysine-α-oxidase.
Similar content being viewed by others
REFERENCES
Dunford, H. B. (1982) Adv. Inorg. Biochem., 4, 41-68.
Welinder, K. G. (1979) Eur. J. Biochem., 96, 483-502.
Ruzgas, T., Gorton, L., Emneus, J., and Marco-Varga, G. (1995) J. Electroanalyt. Chem., 391, 41-49.
Yaropolov, A. I., Tarasevich, M. R., and Varfolomeev, S. D. (1978) Bioelectrochem. Bioenerg., 5, 18-24.
Bogdanovskaya, V. A. (1993) Elektrokhimiya, 29, 441-447.
Razumas, V., Jasaitis, J., and Kulys, J. (1984) Bioelectrochem. Bioenerg., 12, 297-302.
Bogdanovskaya, V. A., Tarasevich, M. R., Hintsche, R., and Sheller, F. (1988) Bioelectrochem. Bioenerg., 19, 581-584.
Lindgren, A., Munteanu, F., Gazaryan, I., Ruzgas, T., and Gorton, L. (1998) J. Electroanalyt. Chem., 458, 113-120.
Lindgren, A., Tanaka, M., Ruzgas, T., Gorton, L., Gazaryan, I., Ishimori, K., and Morishima, I. (1999) Electrochem. Commun., 1, 171-175.
Zhao, J., Henkens, R. W., Stonehuerner, J., O.Daly, J. P., and Crumbliss, A. L. (1992) J. Electroanalyt. Chem., 32, 109-119.
Razumas, V. J., Gudavicius, A. V., and Kulys, J. J. (1986) J. Electroanalyt. Chem., 198, 81-87.
Presnova, G., Grigorenko, V., Egorov, A., Ruzgas, T., Lindgren, A., Gorton, L., and Borchers, T. (2000) Faraday Discuss., 116, 281-289.
Durliat, H., Courteix, A., and Comtat, M. (1989) Bioelectrochem. Bioenerg., 22, 197-209.
Ruzgas, T., Gorton, L., Emneus, J., and Marco-Varga, G. (1996) Anal. Chim. Acta, 330, 123-138.
Khudyakova, R. V., Soloshko, S. V., and Safronov, A. Yu. (1997) Elektrokhimiya, 33, 1165-1171.
Safronov, A. Yu., Tarasevich, M. R., Bogdanovskaya, V. A., and Chernyak, A. S. (1983) Elektrokhimiya, 19, 421-424.
Egorov, A. M., Gazaryan, I. G., Kim, B. B., Doseeva, V. V., Kapeliuch, J. L., Veryovkin, A. N., and Fechina, V. A. (1994) Annals NY Acad. Sci., 721, 73-81.
Grigorenko, V., Chubar, T., Kapeliuch, Yu., Burchers, T., Spener, F., and Egorov, A. (1999) Biocatalysis and Biotransformations, 17(5), 359-379.
Ward, M. D., and Buttry, D. A. (1990) Science, 249, 1000-1007.
Sauerbrey, G. Z. (1959) Z. Physik., 155, 206-210.
Gajhede, M., Schuller, D. J., Henriksen, A., Smith, A. T., and Poulos, T. L. (1997) Nat. Struct. Biol., 4(12), 1032-1038.
Csoregi, E., Jönsson-Pettersson, G., and Gorton, L. (1993) J. Biotechnol., 30, 315-337.
Kulys, J., and Schmid, R. D. (1990) Bioelectrochem. Bioenerg., 24, 305-311.
Eremin, S. A., Vlasenko, S. B., Osipov, A. P., Eremina, I. D., and Egorov, A. M. (1989) Analyt. Lett., 22(9), 2037-2050.
Rubtsova, M., Kovba, G., and Egorov, A. (1998) Biosens. Bioelectron., 13, 75-85.
Wendzinski, F., Grundig, B., Renneberg, R., and Spener, F. (1997) Biosens. Bioelectron., 12, 43-52.
Yu, P., and Wilson, G. S. (2000) Faraday Discuss., 116, 305-317.
Vrbova, E., Marek, M., and Ralys, E. (1992) Analyt. Chim. Acta, 270, 131-136.
Curulli, A., Kelly, S., O. Sullivan, C., Guilbault, G. G., and Palleschi, G. (1998) Biosens. Bioelectron., 13, 1245-1250.
Saurina, J., Hernandez-Cassou, S., Alegret, S., and Fabregas, E. (1999) Biosens. Bioelectron., 14, 211-220.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ferapontova, E.E., Grigorenko, V.G. & Egorov, A.M. P-Chip and P-Chip Bienzyme Electrodes Based on Recombinant Forms of Horseradish Peroxidase Immobilized on Gold Electrodes. Biochemistry (Moscow) 66, 832–839 (2001). https://doi.org/10.1023/A:1011992316837
Issue Date:
DOI: https://doi.org/10.1023/A:1011992316837