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一种将过氧化氢酶固定在多壁碳纳米管/聚(L-赖氨酸)生物复合材料上的简单策略,用于检测 H2O2 和碘酸盐。

A simple strategy for the immobilization of catalase on multi-walled carbon nanotube/poly (L-lysine) biocomposite for the detection of H2O2 and iodate.

机构信息

Electroanalysis and Bioelectrochemistry Lab, Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, No. 1, Section 3, Chung-Hsiao East Road, Taipei 106, Taiwan, ROC.

Electroanalysis and Bioelectrochemistry Lab, Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, No. 1, Section 3, Chung-Hsiao East Road, Taipei 106, Taiwan, ROC.

出版信息

Biosens Bioelectron. 2014 Nov 15;61:639-47. doi: 10.1016/j.bios.2014.05.023. Epub 2014 Jun 2.

Abstract

Herein, we report a novel third-generation H2O2 and IO3- biosensor, which was fabricated by loading catalase (CAT) onto l-lysine/multiwalled carbon nanotube (PLL/f-MWCNT) film modified glassy carbon electrode (GCE). The UV-visible (UV-vis) and Fourier-transform infrared (FTIR) spectra show that the catalase encapsulated in the PLL/f-MWCNT film can effectively retain its bioactivity. The immobilized CAT retained its bioactivity with a high protein loading of 4.072 × 10(-10) mol cm(-2), thus exhibiting a surface-controlled reversible redox reaction, with a fast heterogeneous electron transfer rate of 5.48 s(-1). The immobilized CAT shows a couple of reversible and well-defined cyclic voltammetry peaks with a formal potential (E(0)) of -0.471 V (vs. Ag/AgCl) in a pH 6.5 phosphate buffer solution (PBS). Moreover, the modified film exhibited high electrocatalytic activity for the reduction of hydrogen peroxide (H2O2). It exhibited a wide linear response to H2O2 in the concentration range of 1 × 10(-6) - 3.6 × 10(-3), with higher sensitivity (392 mA cm(-2) M(-1)) and a lower Michaelis-Menten constant (0.224 mM). It provided high-catalytic activity towards H2O2 in a shorter time (5s), with a detection limit of 8 nM. These results indicate great improvement in the electrochemical and electrocatalytic properties of the CAT/PLL/f-MWCNT biosensor, offering a new idea for the design of third-generation electrochemical biosensors.

摘要

在此,我们报告了一种新型的第三代 H2O2 和 IO3- 生物传感器,它是通过将过氧化氢酶 (CAT) 加载到赖氨酸/多壁碳纳米管 (PLL/f-MWCNT) 修饰的玻碳电极 (GCE) 上来制备的。紫外-可见 (UV-vis) 和傅里叶变换红外 (FTIR) 光谱表明,封装在 PLL/f-MWCNT 薄膜中的过氧化氢酶可以有效地保持其生物活性。固定化 CAT 具有较高的蛋白质负载量(4.072×10(-10) mol cm(-2)),保留了其生物活性,从而表现出表面控制的可逆氧化还原反应,具有 5.48 s(-1)的快速非均相电子转移速率。固定化 CAT 在 pH 6.5 磷酸盐缓冲溶液 (PBS) 中显示出一对可逆且定义明确的循环伏安峰,具有 -0.471 V(相对于 Ag/AgCl)的形式电位 (E(0))。此外,修饰后的薄膜对过氧化氢 (H2O2) 的还原表现出高电催化活性。它在 1×10(-6) - 3.6×10(-3) 的浓度范围内对 H2O2 表现出宽线性响应,具有更高的灵敏度(392 mA cm(-2) M(-1))和更低的米氏常数 (0.224 mM)。它在更短的时间(5s)内对 H2O2 表现出高催化活性,检测限为 8 nM。这些结果表明 CAT/PLL/f-MWCNT 生物传感器的电化学和电催化性能得到了极大的改善,为第三代电化学生物传感器的设计提供了新的思路。

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