Peroxidase immobilized on phospholipid bilayers supported on au (111) by DTT self-assembled monolayers: Application to dopamine determination
Peroxidase immobilized on phospholipid bilayers supported on au (111) by DTT self-assembled monolayers: Application to dopamine determination
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DOI：
10.1002/bit.24721
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发表时间：
2013-02-01
影响因子：
3.8
通讯作者：
Creczynski-Pasa, Tania B.
中科院分区：
文献类型：
--
作者：
Fritzen-Garcia, Mauricia B.;Zoldan, Vinicius C.;Creczynski-Pasa, Tania B.
本工作利用二硫苏糖醇（DTT）自组装膜将辣根过氧化物酶（HRP）固定在以Au（111）为载体的二肉豆蔻酰磷脂酰胆碱（DMPC）双层膜上，并将其作为纳米结构的电化学生物传感器用于多巴胺的测定。磷脂双层的形态和固定的HRP到这些层的特征在于原子力显微镜（AFM）。方波伏安法（SWV）实验研究了HRP修饰电极的性能。AFM图像表明，酶被吸附在脂质双层的外层，虽然没有测量表面上的电荷，但由于实验中使用的pH值，酶和磷脂表面相互作用可能通过静电力发生。有趣的是，本系统可以用作快速检测多巴胺的一次性传感器。该系统的分析性能是线性的多巴胺浓度从3.3？x？10-5到1.3？x？10比3摩尔？L-1（r =？0.9997），检出限为2.0？x？10-6？摩尔？L-1.我们的研究结果表明，HRP-DMPC双层系统的使用可能是有用的，不仅在开发新的纳米结构材料的技术目的，但可能是非常有用的基础研究，以调查不同的微观和大分子之间的相互作用，甚至与可溶性蛋白质，和脂膜。Biotechnol. Bioeng. 2013; 110：374382. (c)2012 Wiley Periodicals，Inc.
In this work, horseradish peroxidase (HRP) was immobilized on dimyristoylphosphatidylcholine (DMPC) bilayers supported on Au (111) by dithiotreitol (DTT) self-assembled monolayers and used as a nanostructured electrochemical biosensor to dopamine determination. The morphology of the phospholipid bilayers and the immobilization of HRP to these layers were characterized by atomic force microscopy (AFM). Square-wave voltammetry (SWV) experiments were done to investigate the performance of the HRP-modified electrode. The AFM images indicate that the enzyme is adsorbed at the external layer of the lipid bilayer and, although the electrical charges on the surface were not measured, the enzyme and phospholipids surface interaction occurs probably by electrostatic forces due to the pH used in the experiments. Interestingly, the present system can be used as one-shot sensor for the rapid detection of dopamine. The analytical performance of this system was linear for dopamine concentrations from 3.3?x?10-5 to 1.3?x?10-3?mol?L-1 (r?=?0.9997) with a detection limit of 2.0?x?10-6?mol?L-1. Our results indicate that the use of HRP-DMPC bilayer system may be useful not only in developing new nanostructured materials for technological purposes, but could be very useful in fundamental studies to investigate the interactions between different micro-and macromolecules, even with soluble proteins, and lipid membranes. Biotechnol. Bioeng. 2013; 110: 374382. (c) 2012 Wiley Periodicals, Inc.