Orientated Immobilization of FAD-Dependent Glucose Dehydrogenase on Electrode by Carbohydrate-Binding Module Fusion for Efficient Glucose Assay
Abstract
:1. Introduction
2. Results and Discussion
2.1. Heterologous Expression and Purification of GDH and GDH-NL-CBM2
2.2. Analysis of the Enzyme Activity of GDH and GDH-NL-CBM2
2.3. Morphological Characterization of the Modified Electrode Surface
2.4. Electrochemical Characterization of the Electrodes Prepared by Two Different Methods
2.5. Electrochemical Behavior of the Electrodes Prepared by Two Methods to Glucose
2.6. Specificity and Anti-Interference Studies
2.7. Stability and Reproducibility of the Electrodes
2.8. Determination of Glucose in Rat Serum Samples and Glucose Drinks
3. Materials and Methods
3.1. Sequencing and Phylogenetic Analysis
3.2. Materials and Chemical Reagents
3.3. Construction of Recombinant Plasmid
3.4. Heterologous Expression of GDH and GDH-NL-CBM2 in Pichia pastoris
3.5. Purification of GDH and GDH-NL-CBM2
3.6. Determination of Optimum Temperature and pH Value
3.7. Preparation of GA/CS/GDH/S-MWNT/GCE and GDH-NL-CBM2/NC/S-MWNT/GCE
3.8. Electrochemical Measurements
3.9. Detection of Glucose in Rat Serum and Drink Samples
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Modified Electrode | Linear Range (mM) | Sensitivity (A/(M*cm2)) | Detection Limit (µM) | Response Time (s) | RSD | Ref |
---|---|---|---|---|---|---|
GOx/PdNPs/CS-GR [a] | 0.001–1.0 | 3.12 × 10−2 | 0.2 | ≈10 | 5.1% | [33] |
GOx/Pd NPAs/GCE [b] | 0.04–22 | -- | 6.1 | -- | 5.1% | [34] |
GOx–GQD/CCE [c] | 0.005–1.27 | 8.5 × 10−2 | 1.73 | ~3 | 5% | [35] |
GOx/rGO/GCE [d] | 0.1–27 | 1.85 × 10−3 | -- | <5 | 4.9% | [36] |
GOx/AuNPs-EGr/SPCE [e] | 0.05–1.6 | 2.55 × 10−1 | 2.5 | -- | 3.8% | [37] |
TNT-GNP/[Demin]Br/ Nafion/GOx/GCE [f] | 0.01–1.2 | 5.1 × 10−3 | -- | -- | -- | [38] |
GOx/rGO-Zn-Ag/GCE | 0.1–12.0 | -- | 10.6 | -- | 6.7% | [39] |
GDH-GBP/Au [g] | 3–30 | 1.33 × 10−3 | 3410 | 5–30 | <10% | [40] |
PPF/GDH/SWCNT-SC/PPF/Au [h] | 0.05–3.2 | 1.10 × 10−1 | 0.83 | 7 | -- | [26] |
rGO/PTZ-O/GDH/GCE [i] | 0.5–12 | 4.2 × 10−2 | -- | -- | -- | [41] |
CS/GDH/MWCNT [j] | 0.07–0.62 | 2.05 × 10−2 | 4.2 | 60 | -- | [42] |
DM/GDH/GCE [k] | 5–30 | 5.3 × 10−3 | 10 | 200 | 5% | [43] |
GA/CS/GDH/S-MWNT/GCE | 0.12–40.7 | 1.2067 × 10−2 | 81 | ~40 | <10% | This work |
Nafion/GDH-NL-CBM2/S-MWNT/GCE | 0.12–40.7 | 2.1362 × 10−2 | 51 | ~18 | <5% | This work |
Concentrations of Glucose (mM) (RSD, n = 3) | ||||||||
---|---|---|---|---|---|---|---|---|
Samples | SBA Glucose Biosensor | Reducing Sugar Analyzer | GA/CS/GDH/S-MWNT/GCE | GDH-NL-CBM2/NC/S-MWNT/GCE | ||||
Average Value | RSD /% | Average Value | RSD /% | Average Value | RSD /% | Average Value | RSD /% | |
Glucose drink (≈2777.8 mM) | 2882.7 | 1.51 | 2793.6 | 1.5 | 2917.3 | 2.64 | 2832.1 | 1.59 |
Pregnant mouse serum | 9.5 | 1.95 | 9.35 | 1.62 | 9.14 | 2.93 | 9.29 | 1.85 |
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Han, Q.; Gong, W.; Zhang, Z.; Wang, L.; Wang, B.; Cai, L.; Meng, Q.; Li, Y.; Liu, Q.; Yang, Y.; et al. Orientated Immobilization of FAD-Dependent Glucose Dehydrogenase on Electrode by Carbohydrate-Binding Module Fusion for Efficient Glucose Assay. Int. J. Mol. Sci. 2021, 22, 5529. https://doi.org/10.3390/ijms22115529
Han Q, Gong W, Zhang Z, Wang L, Wang B, Cai L, Meng Q, Li Y, Liu Q, Yang Y, et al. Orientated Immobilization of FAD-Dependent Glucose Dehydrogenase on Electrode by Carbohydrate-Binding Module Fusion for Efficient Glucose Assay. International Journal of Molecular Sciences. 2021; 22(11):5529. https://doi.org/10.3390/ijms22115529
Chicago/Turabian StyleHan, Qingye, Weili Gong, Zhenyu Zhang, Lushan Wang, Binglian Wang, Lei Cai, Qingjun Meng, Yiwei Li, Qingai Liu, Yan Yang, and et al. 2021. "Orientated Immobilization of FAD-Dependent Glucose Dehydrogenase on Electrode by Carbohydrate-Binding Module Fusion for Efficient Glucose Assay" International Journal of Molecular Sciences 22, no. 11: 5529. https://doi.org/10.3390/ijms22115529