Amperometric glucose biosensor utilizing FAD-dependent glucose dehydrogenase immobilized on nanocomposite electrode

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• 作者：Rastislav Mono&scaron ; í ; k^a ; ^{rastislav.monosik@stuba.sk} ; ^{rasto.monosik@gmail.com} ; Miroslav Stre膹anský ; ^b ; Karol Lu&scaron ; pai^c ; Peter Magdolen^d ; Ernest &Scaron ; turdí ; k^a
• 关键词：Biosensor ; Nanocomposite ; Carbon nanotube ; FAD-glucose dehydrogenase ; Phenazine methyl sulfate ; Beverage analysis
• 刊名：Enzyme and Microbial Technology
• 出版年：2012
• 期刊代码：26_01410229
• 类别：ce
• 出版时间：5 April, 2012
• 卷：50
• 期：4-5
• 页码：227-232
• 文件大小：427 K

摘要

Amperometric glucose biosensors utilizing commercially available FAD-dependent glucose dehydrogenases from two strains of Aspergillus species are described. Enzymes were immobilized on nanocomposite electrode consisting of multi-walled carbon nanotubes by entrapment between chitosan layers. Unlike the common glucose oxidase based biosensor, the presented biosensors appeared to be O₂-independent. The optimal amount of enzymes, working potential and pH value of working media of the glucose biosensors were determined. The biosensor utilizing enzyme isolated from Aspergillus sp. showed linearity over the range from 50 to 960 渭M and from 70 to 620 渭M for enzyme from Aspergillus oryzae. The detection limits were 4.45 渭M and 4.15 渭M, respectively. The time of response was found to be 60 s. The biosensors showed excellent operational stability - no loss of sensitivity after 100 consecutive measurements and after the storage for 4 weeks at 4 掳C in phosphate buffer solution. When biosensors were held in a dessicator at room temperature without use, they kept the same response ability at least after 6 months. Finally, the results obtained from measurements of beverages and wine samples were compared with those obtained with the enzymatic-spectrophotometric and standard HPLC methods, respectively. Good correlation between results in case of analysis of real samples and good analytical performance of presented glucose biosensor allows to use presented concept for mass production and commercial use.