Citation: WANG Wei, Hong Jun, HANG Kun, YANG Wei-Yun, XIAO Bao-Lin, ZHAO Ying-Xue, Gao Yun-Fei. Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(10): 1543-1548. doi: 10.3724/SP.J.1096.2012.20208 shu

Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy

1.
College of Life Science, Henan University, Kaifeng 475000, China

Corresponding author: Hong Jun,
Received Date: 31 March 2012
Available Online: 29 May 2012
Fund Project: 本文系河南大学特聘教授科研启动基金(No.5443D08010) (No.5443D08010)

A nano-cluster with highly efficient peroxide activity was constructed self-assembly based on nafion (NF) and cytochrome c (Cyt c), The UV-Vis spectrometry, Circular Dichroism (CD) and transmission electron microscopic (TEM) methods were utilized for additional characterization of the artificial peroxidase (AP). The nano-cluster was composed of chain-ball structure, with an average ball size of approximately 40 nm detected by TEM method. The Michaelis–Menten (Km) and catalytic rate (kcat) constants of the AP were determined to be (2.5±0.4) μmol/L and (0.069±0.001) s^-1, respectively, in 50 mmol/L PBS at pH 7.0. The catalytic efficiency of AP was evaluated to be (0.028±0.005) μmol/L^-1s^-1, which was 39%±5% as efficient as the native HRP. AP generated here can be used in place of HRP with high stability and activity. The AP was also immobilized on a functional MWNCTs-gold nano-complex modified glassy carbon electrode. The cyclic voltammetry of AP on the nano complex modified glassy carbon electrode showed a pair of well-defined quasi-reversible redox peaks with the formal potential of (E^o') of (-45±2) mV (vs. Ag/AgCl) at scan rate of 0.05 V/s. The heterogeneous electron transfer rate constant was evaluated to be 0.65 s^-1 in 50 mmol/L Na₂HPO₄-NaH₂PO₄ buffer solution at pH 7.0. The cathodic peak current of the modified electrode increased with increasing concentration of hydrogen peroxide(from 0.02 to 10 nmol/L) with a detection limit of 0.05 nmol/L. The apparent Michaelis-Menten constant (Kmapp) was 0.23 nmol/L.
- Self-assembled,
- Nano-cluster,
- Artificial peroxidase,
- Spectroscopy,
- Electrochemistry
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