| Simple Preparation of Diaphorase/Polysiloxane Viologen Polymer Modified Electrode for Sensing NAD and NADH |
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Ji-Eun Song, Zhenyu Hong, Rajaram Krishna Nagarale, Woon-Sup Shin |
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Department of Chemistry and Interdisciplinary Program of Integrated Biotechnology, Sogang University;Department of Chemistry and Interdisciplinary Program of Integrated Biotechnology, Sogang University;Department of Chemistry and Interdisciplinary Program of Integrated Biotechnology, Sogang University;Department of Chemistry and Interdisciplinary Program of Integrated Biotechnology, Sogang University; |
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| Abstract |
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Nicotinamide adenine dinucleotide, $NAD^+$, and its reduced form, NADH, play important roles as coenzymes in many enzymatic reactions. Electrochemical methods for $NAD^+$ or NADH detection or generation are drawn attention because it can provide the simple and low cost platform with fairly good sensitivity. In this study, the polysiloxane viologen polymer/diaphorase/hydrophilic polyurethane (PSV/DI/HPU) modified electrodes were simply prepared and demonstrated for bio-electrocatalytic $NAD^+$ sensors. The electrodes were co-immobilized with diaphorase and polysiloxane viologen polymer as an electron mediator followed by the overcoating with HPU membrane. The mixture of the enzyme and the electron mediator was well stabilized within HPU membrane and exhibited good reversibility and stability. The sensitivity was 0.2 $nA{cdot}{mu}M^{-1}$ and the detection limit was 28 ${mu}M$ with a response time of 50 s ($t_{90%}$). The capability for NADH sensor was also observed on the PSV/DI/HPU electrode. |
| Key Words:
$NAD^+$ reduction, NADH generation, Electrochemical sensor, Diaphorase, Polysiloxane viologen polymer, Co-immobilization |
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