光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

引用本文: 王玮, 洪军, 黄琨, 肖保林, 杨卫云, 赵莹雪, 高云飞. 光谱和电化学方法研究自组装纳米簇模拟过氧化物酶[J]. 分析化学, 2012, 40(10): 1543-1548. doi: 10.3724/SP.J.1096.2012.20208 shu

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

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

基金项目:
本文系河南大学特聘教授科研启动基金(No.5443D08010) (No.5443D08010)

伊朗国家科学基金资助

摘要: 利用细胞色素c与Nafion,通过自组装方式,构建了纳米簇模拟过氧化物酶;利用电子透射显微镜、圆二色谱仪、紫外可见光分光光度仪及电化学工作站等设备测量模拟酶结构变化、酶促动力学参数以及电化学特性;在50 mmol/L Na₂HPO₄-NaH₂PO₄缓冲溶液(pH 10.0)中,测得模拟酶米氏常数K_m、催化速率及催化效率分别为2.5 μmol/L、0.069 s^-1、0.028±0.005 (μmol/L)^-1s^-1,其催化效率为天然辣根过氧化物酶的39%±5%,该模拟酶具有很好的稳定性和活性,能够替代辣根过氧化物酶。该模拟酶可以固定在功能化多壁纳米管与纳米金复合材料修饰玻碳电极上,在50 mmol/L Na₂HPO₄-NaH₂PO₄缓冲溶液(pH 7.0)中测得模拟酶与电极之间电子迁移速度常数k_s为0.65 s^-1。利用循环伏安法测量模拟酶催化浓度为0.02-10 nmol/L的H₂O₂,其阴极峰电流随H₂O₂浓度增加而增大,并且得出模拟酶催化H₂O₂检出限为0.05 nmol/L,电化学表观米氏常数K_mapp为2.3×10^-10mol/L。

关键词:

自组装
/ 纳米簇
/ 模拟酶
/ 光谱学
/ 电化学

English

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

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

Corresponding author: 洪军

Received Date: 31 March 2012
Fund Project:
本文系河南大学特聘教授科研启动基金(No.5443D08010)

伊朗国家科学基金资助

Abstract: 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.

Key words:

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

English

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

Corresponding author: 洪军

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

English

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

Corresponding author: 洪军

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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