P-CHIP AND P-CHIP BIENZYME ELECTRODES BASED ON RECOMBINANT FORMS OF HORSERADISH PEROXIDASE IMMOBILIZED ON GOLD ELECTRODESстатья
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Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
Аннотация:Adsorption and bioelectrocatalytical activity of native horseradish peroxidase (HRP) and its recombinant forms were studied on polycrystalline gold electrodes. Recombinant forms of HRP were produced by genetic engineering approach using an E. coli expression system. According to the direct mass measurements with quartz crystal microbalance all the forms of HRP formed a monolayer coverage of the enzyme on the gold surface. However, only gold electrodes modified with the recombinant HRP forms (nonglycosilated) 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 at the C-terminus or at the N-terminus of the enzyme molecule additionally increased the strength of the enzyme binding with gold surface and efficiency of direct ET. Iimmobilisation of the recombinant forms of HRP, containing histidine functional groups, on the surface of the gold electrode was used both for the development of P-chip - a biosensor for hydrogen peroxide determination based on direct ET, and for the development of a bienzyme biosensor electrode for the detremination of L-lysine, based on co-immobilised recombinant forms of HRP and L-lysine-*-oxidase.