Nanomaterials as Redox Mediators in Laccase-Based Amperometric Biosensors for Catechol Assay

Abstract

Laccase is a copper-containing enzyme that does not require hydrogen peroxide as a co-substrate or additional cofactors for an enzymatic reaction. Nanomaterials of various chemical structures are usually applied to the construction of enzyme-based biosensors. Metals, metal oxides, semiconductors, and composite NPs perform various functions in electrochemical transformation schemes as a platform for the enzyme immobilization, a mediator of an electron transfer, and a signal amplifier. We describe here the development of amperometric biosensors (ABSs) based on laccase and redox-active micro/nanoparticles (hereafter—NPs), which were immobilized on a graphite electrode (GE). For this purpose, we isolated a highly purified enzyme from the fungus Trametes zonatus, and then synthesized bi- and trimetallic NPs of noble and transition metals, as well as hexacyanoferrates (HCF) of noble metals; these were layered onto the surfaces of GEs. The electroactivity of many of the NPs immobilized on the GEs was characterized by cyclic voltammetry (CV) experiments. The most effective mediators of electron transfer were selected as the platform for the development of laccase-based ABSs. As a result, a number of catechol-sensitive ABSs were constructed and characterized. The laccase/CuCo/GE was demonstrated to possess the highest sensitivity to catechol (4523 A·M−1·m−2) among the tested ABSs. The proposed ABSs may be promising for the analysis of phenolic derivatives in real samples of drinking water, wastewater, and food products.

The Peroxidase-like Nanocomposites as Hydrogen Peroxide-Sensitive Elements in Cholesterol Oxidase-Based Biosensors for Cholesterol Assay

“…The XRM images (nCuFe and nCeAu) demonstrate the peaks for metals of the composites. The morphology of nAuHCF was presented in our earlier paper [ 56 ].…”

“…The sensitivity of the ChOx/nCuFe/nPt/GCE sensor has been compared with the literature data, as shown in Table 2 . To date, a variety of materials such as CNTs, polymers, metal NPs and graphene are used for the construction of biosensors for CHOL detection [ 24 , 26 , 37 , 46 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 ]. The developed bionanosensor ChOx/nCuFe/nPt/GCE has several advantages such as lower cost, high sensitivity and selectivity.…”

“…All NPs were obtained by the chemical reduction method [ 56 , 57 ]. nCeAu were obtained by the following procedure: 5 mL 10 mM HAuCl4 and 5 mL 10 mM Ce(HCO 3 ) 4 were mixed with 0.1 mL 10 mM CTAB and 1.5 mL 50 mM ascorbic acid.…”

“…To determine the HRP-like activity of the synthesized NZs in solution, the spectrophotometric method with o-dianisidine was applied [ 55 ]. The specific activity (U/mg) was calculated according to the formula [ 56 ]: where…”

“…The constructed sensors were characterized using amperometry in a three-electrode configuration with an Ag/AgCl/KCl (3 M) reference electrode, a Pt-wire counter electrode and a working GCE [ 56 ].…”