Fabrication and Characterization of a Thermostable Quinoprotein Aldose Sugar Dehydrogenase Immobilized Electrode

• Published in: Analytical Sciences Vol. 29; no. 1; pp. 79 - 83
• Main Authors: YAMADA, Yohei, HAYASHI, Tetsuya, SAKURABA, Haruhiko, YABUTANI, Tomoki, TAKAYANAGI, Toshio
• Format: Journal Article
• Language: English
• Published: Singapore The Japan Society for Analytical Chemistry 01.01.2013; Springer Nature Singapore; Japan Science and Technology Agency
• Subjects: Analytical Chemistry; Benzoquinones - chemistry; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Buffers; Carbon - chemistry; Chemistry; Enzyme Stability; Enzymes, Immobilized - chemistry; Glucose - chemistry; Glucose Dehydrogenases - chemistry; Hydrogen-Ion Concentration; Nanofibers - chemistry; Pyrobaculum - enzymology; Substrate Specificity; Surface Properties
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.29.79

We fabricated a thermostable quinoprotein aldose sugar dehydrogenase (tPQQ-ASD derived from a hyperthermophilic archaeon Pyrobaculum aerophilum) immobilized electrode. The electrode was prepared by immobilizing agarose gel mixed with the enzyme and carbon nanofiber (CNF) on a carbon paste (CP) electrode containing p-benzoquinone (BQ) as an electron mediator. The electrocatalytic response was clearly observed by the addition of D-glucose at the electrode. The electrode properties such as pH, temperature dependency and substrate selectivity basically followed the enzyme properties. The current response against D-glucose increased with measurement temperatures up to 70°C, and response perturbation caused by dissolved oxygen level was not observed at the electrode. As for the results of long-term stability evaluation, the current response was stable for 30 days when the electrode was stored in HEPES buffer solution (pH 7.0) at 4°C.