Biosensor Based on Entrapment of Glucose Oxidase within Biomimetic Silica Nanoparticles for Glucose Detection
- Biosensor Based on Entrapment of Glucose Oxidase within Biomimetic Silica Nanoparticles for Glucose Detection
- 최옥경; 김병천; 안지혜; 김용환; 엄영순; 상병인
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- Biomimetic synthesis raises a concern because it can take place under mild conditions with more facilitation (1). Biosilicification is one of biomimetic syntheses and we recently efficiently immobilized GFP (green fluorescence protein) using biosilicification by silaffin-fused proteins (GFP-Rx) (2). Glucose oxidase (GOx, EC 184.108.40.206) is one of broadly used enzymes for biosensors with high enzyme activity to detect glucose concentration such as to track blood sugar levels. We constructed R5-GOx-fused protein and it was expressed on Pichia pastoris. Secreted GOx from Pichia was immobilized by R5 (fused with GOx) in pH 5 citric buffer using 0.1 M tetramethoxysilane. To apply silaffin-GOx-silica nanoparticles for a biosensor, biosilicification occurred on a grassy carbon electrode. Electric cell consisted of Ag/AgCl reference electrode, platinum counter electrode (cathode), and working electrode (anode) modified with poly(neutral red)(PNR)/GOx/nafion. Electrochemically formed PNR was used as an electron carrier from GOx to an electrode. Scanning electron microscopy image supported a formation of PNR/GOx complex. A current was increased to 2 ㎂·cm-1 by 1 mM glucose at -0.4 V applied potential. A biosensor of GC/PNR/GOx/nafion showed a substrate selectivity to detect glucose concentration in mixed glucose samples with other carbohydrates.
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