Covalent immobilization of redox protein via click chemistry and carbodiimide reaction: direct electron transfer and biocatalysis.
Covalent immobilization of redox protein via click chemistry and carbodiimide reaction: direct electron transfer and biocatalysis.
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DOI：
10.1016/j.jcis.2010.07.018
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发表时间：
2010-10
影响因子：
9.9
通讯作者：
Liwei Wang;Q. Ran;Yuan Tian;Jingjing Xu;Y. Xian;Ru Peng;L. Jin
中科院分区：
文献类型：
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作者：
Liwei Wang;Q. Ran;Yuan Tian;Jingjing Xu;Y. Xian;Ru Peng;L. Jin
本文提出了一种简便易行、通用性强的Au表面与Hb共价生物功能化的策略。在叠氮十一硫醇和稀硫醇的混合溶液中，在金表面形成了结构清晰的叠氮末端有机自组装单分子膜。通过选择性、可靠、可靠的点击反应和广泛使用的碳二亚胺反应，4-戊烯酸作为双官能团连接物用于固定化Hb。采用反射吸收红外光谱(RAIR)、电化学阻抗谱(EIS)和循环伏安法(CV)对其进行了表征。实验数据表明，所提出的方法是一种高度通用的策略，可以定量、稳定和共价地将生物大分子附着到固体界面上。在0.1mol/L pH 5.0缓冲溶液中，Hb功能化Au电极在约0.210V(vs.SCE)处出现一对清晰的准可逆峰，代表Hb Fe(III)/Fe(II)氧化还原对的还原和氧化。电子转移速率常数(Ks)为0.78s−-1。固定化Hb的最大表面覆盖率(Г)为8.3×10-−-12molcm-−-2，表明形成了单分子膜。此外，还研究了HB生物功能化电极的生物催化活性，观察到了对O2和H2O2的良好电催化还原。
In this paper, a simple, facile and general strategy was proposed for the covalent biofunctionalization of Au surface with hemoglobin (Hb). Structurally well-defined azide-terminated organic self-assembled monolayers (SAMs) were formed on Au surface from a mixture solution of azidoundecanethiol and dilute thiol. 4-Pentynoic acid was used as a bifunctional linker to immobilized Hb via a selective, reliable, robust click reaction and a widely used carbodiimide reaction. The surface modification was characterized by reflectance absorption infrared (RAIR) spectroscopy, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The experimental data indicate that the proposed methodology is a highly versatile strategy for quantitative, stable and covalent attachment of biomacromolecules onto solid interface. The Hb functionalized Au electrode shows a pair of well-defined and quasi-reversible peaks at about −0.210V (vs. SCE) in 0.1mol/L pH 5.0 buffers, which stands for the reduction and oxidation of Hb Fe(III)/Fe(II) redox couple. The electron transfer rate constant (ks) was estimated to be 0.78s−1. The maximal surface coverage (Г) of the immobilized Hb was 8.3×10−12molcm−2, indicating the formation of a monolayer. Moreover, the biocatalytic activity of the Hb biofunctonalized electrode was investigated and an excellent electrocatalytic reduction toward O2and H2O2was observed.