A high catalytic efficient soft-template nanostructured peroxidase (SP) was constructed when cytochrome c (Cyt c, 10 μM) was mixed with sodium decyl sulfate (SDeS, 40 mM) in 100 mM sodium phosphate buffer solution (PBS). The electrochemical investigations were carried out for the SP immobilized on hydroxyl fullerenes modified glassy carbon electrode (GCE). The cyclic voltammetry of the SP-modified GCE showed a pair of well-defined redox peaks with a formal potential (\( E^{{\circ^{\prime}}} \)) of −220 ± 2 mV (vs. Ag/AgCl) in 100 mM, pH 8.0 PBS at a scan rate of 0.05 Vs−1. The heterogeneous electron transfer rate constant (ks) was calculated to be 14.3 ± 1 s−1. The apparent Michaelis–Menten constant (\( K_{m}^{app} \)) was 1.79 ± 0.03 µM. SDeS and Cyt c may be important for the structure and catalytic function of SP, respectively. The SP-modified GCE could be used as an H2O2 biosensor instead of horseradish peroxidase (HRP), with good sensitivity and stability.

A nano-micelle with highly efficient peroxide activity was constructed by self-assembly of sodium dodecyl sulfate micellar, histidine and hematin in 50 mM phosphate buffer at 25 °C. UV–Vis spectrometry methods were utilized for characterization of the nanostructured material or artificial peroxidase (AP). The Michaelis–Menten (Km) and catalytic rate (kcat) constants of the AP were obtained to be 5.5 μM and 0.06 s−1, respectively, in 50 mM phosphate buffer solution at pH 8.0. The catalytic efficiency of AP was evaluated to be 0.011 μM−1 s−1. The AP was also immobilized on a functional multi-wall carbon nanotubes-gold nanoparticles (AuNPs) nano-complex modified glassy carbon electrode (GCE). The transmission electron microscopy method was utilized for the characterization of the nano-materials. The electron-transfer rate constant (ks) and the apparent Michaelis–Menten constant Kmapp of the AP modified GCE were evaluated to be 1.36 s−1 and 0.19 μM, respectively. For a biosensor without a redox protein, the properties of the AP modified GCE were significant and will further benefit from additional studies and improvement.

By immobilizing catalase on a nanocomposite containing functionalized multi-walled carbon nanotubes and l-cysteine modified gold nanoparticles, a third generation biosensor was developed for determination of the hydrogen peroxide. The cyclic voltammograms of catalase on the nanocomposite modified glassy carbon electrode showed a pair of well-defined quasi-reversible redox peaks with the formal potential of −441 ± 2 mV versus Ag/AgCl at a scan rate of 0.05 V/s. The heterogeneous electron transfer constant was calculated to be 8.72 s−1. The enzyme electrode response toward hydrogen peroxide was linear in the concentrations ranging from 1 nM to 1 μM, with a detection limit of 0.5 nM. The apparent Michaelis–Menten constant was calculated to be 0.34 μM.

A homogeneous nanostructured enzyme (artificial peroxidase, AP) with suitable catalytic efficiency was generated using bovine heart cytochrome c (Cyt c) and sodium dodecyl sulfate nano-micelles in 50 mM phosphate buffer pH 10.5 at 25 °C. The Michaelis–Menten (Km) and catalytic rate (kcat) of the AP were determined to be 21.6 ± 1.2 μM and 0.474 ± 0.013 s−1, respectively. The catalytic efficiency of the AP was 0.0219 ± 0.002 μM−1s−1, which was 30 ± 1.5 % as efficient as the native horseradish peroxidase (HRP). The mean diameter of AP was measured to be 6.4 nm using dynamic light scattering technique. The UV–Vis spectrometry, circular dichroism, surface tension, isothermal titration calorimetric and electrochemistry methods were utilized for additional characterization of the AP. Together our results suggest that the AP generated here can be used in place of HRP in industrial and commercial fields under some extreme conditions.