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Orientated Immobilization of FAD-Dependent Glucose Dehydrogenase on Electrode by Carbohydrate-Binding Module Fusion for Efficient Glucose Assay.
- Source :
-
International journal of molecular sciences [Int J Mol Sci] 2021 May 24; Vol. 22 (11). Date of Electronic Publication: 2021 May 24. - Publication Year :
- 2021
-
Abstract
- The discovery or engineering of fungus-derived FAD-dependent glucose 1-dehydrogenase (FAD-GDH) is especially important in the fabrication and performance of glucose biosensors. In this study, a novel FAD-GDH gene, phylogenetically distantly with other FAD-GDHs from Aspergillus species, was identified. Additionally, the wild-type GDH enzyme, and its fusion enzyme (GDH-NL-CBM2) with a carbohydrate binding module family 2 (CBM2) tag attached by a natural linker (NL), were successfully heterogeneously expressed. In addition, while the GDH was randomly immobilized on the electrode by conventional methods, the GDH-NL-CBM2 was orientationally immobilized on the nanocellulose-modified electrode by the CBM2 affinity adsorption tag through a simple one-step approach. A comparison of the performance of the two electrodes demonstrated that both electrodes responded linearly to glucose in the range of 0.12 to 40.7 mM with a coefficient of determination R <superscript>2</superscript> > 0.999, but the sensitivity of immobilized GDH-NL-CBM2 (2.1362 × 10 <superscript>-</superscript> <superscript>2</superscript> A/(M*cm <superscript>2</superscript> )) was about 1-fold higher than that of GDH (1.2067 × 10 <superscript>-2</superscript> A/(M*cm <superscript>2</superscript> )). Moreover, a lower detection limit (51 µM), better reproducibility (<5%) and stability, and shorter response time (≈18 s) and activation time were observed for the GDH-NL-CBM2-modified electrode. This facile and easy immobilization approach used in the preparation of a GDH biosensor may open up new avenues in the development of high-performance amperometric biosensors.
- Subjects :
- Animals
Aspergillus flavus chemistry
Aspergillus flavus metabolism
Biosensing Techniques instrumentation
Blood Glucose analysis
Electrodes
Enzymes, Immobilized chemistry
Escherichia coli metabolism
Fungi chemistry
Gene Expression
Glucose 1-Dehydrogenase chemistry
Glucose 1-Dehydrogenase genetics
Hydrogen-Ion Concentration
Microscopy, Electron, Scanning
Phylogeny
Rats
Recombinant Proteins genetics
Recombinant Proteins metabolism
Reproducibility of Results
Sequence Alignment
Temperature
Biosensing Techniques methods
Enzyme Assays methods
Enzymes, Immobilized metabolism
Flavin-Adenine Dinucleotide metabolism
Glucose analysis
Glucose 1-Dehydrogenase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1422-0067
- Volume :
- 22
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- International journal of molecular sciences
- Publication Type :
- Academic Journal
- Accession number :
- 34073858
- Full Text :
- https://doi.org/10.3390/ijms22115529