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Redox competition and generation-collection modes based scanning electrochemical microscopy for the evaluation of immobilised glucose oxidase-catalysed reactions

机译:基于氧化还原竞争和生成-收集模式的扫描电化学显微镜,用于评估固定化葡萄糖氧化酶催化的反应

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摘要

Redox competition (RC-SECM) and generation-collection (GC-SECM) modes of scanning electrochemical microscopy were applied for the evaluation of a glucose oxidase (GOx)-modified non-conducting poly(methyl methacrylate) surface. The current vs. distance curves in RC-SECM mode were registered at -600 mV vs. Ag/AgCl to determinate local O-2 concentration, taking into account that the O-2 is consumed in the GOx-catalysed enzymatic reaction. This measurement was performed in phosphate-acetate buffer at pH 6.6 with 0-30 mmol L-1 of glucose using a platinum ultramicroelectrode (UME) as the moving working electrode in a three-electrode electrochemical cell. The UME current, which is related to oxygen reduction rate, decreased when glucose was added to the solution. Another part of the investigation was performed in GC-SECM mode at +600 mV vs. Ag/AgCl in order to measure local H2O2 concentration, which is formed during the GOx-catalysed enzymatic reaction. The same SECM mode was used for imaging the GOx-catalysed reaction without any redox mediator. The imaging distance was chosen based on both the RC-SECM and GC-SECM experimental results. The RC-SECM and GC-SECM modes are described, and the processes that occurred on the UME- and GOx-modified surfaces are revealed.
机译:氧化还原竞争(RC-SECM)和世代收集(GC-SECM)扫描电化学显微镜的模式用于评估葡萄糖氧化酶(GOx)改性的非导电聚甲基丙烯酸甲酯表面。考虑到O-2在GOx催化的酶促反应中被消耗,RC-SECM模式下的电流与距离的关系曲线在-600 mV对Ag / AgCl的条件下确定了局部O-2的浓度。使用三电极电化学池中的铂超微电极(UME)作为移动工作电极,在pH值为6.6的磷酸盐-乙酸盐缓冲液中使用0-30 mmol L-1的葡萄糖进行此测量。当将葡萄糖添加到溶液中时,与氧气还原速率有关的UME电流降低。研究的另一部分是在相对于Ag / AgCl的+600 mV的GC-SECM模式下进行的,目的是测量在GOx催化的酶促反应过程中形成的局部H2O2浓度。在没有任何氧化还原介体的情况下,使用相同的SECM模式对GOx催化的反应进行成像。根据RC-SECM和GC-SECM实验结果选择成像距离。描述了RC-SECM和GC-SECM模式,并揭示了在UME和GOx修改的表面上发生的过程。

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