Enhancing the peroxidase activity and decreasing the protease activity of ficin with rational modification and its application to one-step colorimetric detection of glucose
| Autor: | Yijuan Long, Huzhi Zheng, Yadi Pan, Danyang Yi, Wen Zheng |
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| Rok vydání: | 2020 |
| Předmět: |
Tris
Hydrochloride medicine.medical_treatment 02 engineering and technology 010402 general chemistry 01 natural sciences Analytical Chemistry chemistry.chemical_compound medicine Humans Instrumentation Spectroscopy Peroxidase Detection limit Chromatography Protease biology Hydrogen Peroxide 021001 nanoscience & nanotechnology Atomic and Molecular Physics and Optics Enzyme assay 0104 chemical sciences Ficain Glucose chemistry Peroxidases TCEP biology.protein Colorimetry 0210 nano-technology Selectivity Oxidation-Reduction |
| Zdroj: | Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy. 247 |
| ISSN: | 1873-3557 |
| Popis: | Ficin has dual enzyme activity, i.e., protease and peroxidase-like activity. In some respects, its application is limited by the protease activity of ficin. Herein, we used tris (2-carboxyethyl) phosphine hydrochloride (TCEP) to break the three pairs of disulfide bonds of ficin, and then blocked the free thiol groups with N-ethylmaleimide (NEM) to synthesize ficin-TN. The results showed that ficin-TN had increased peroxidase-like activity and reduced protease activity. According to this phenomenon, we have exploited a colorimetric method with high sensitivity and selectivity for the one-step detection of glucose. Comparing with ficin, ficin-TN has wider detection range (0.1–300 μM) and lower detection limit (88 nM), and our method is simpler and more timesaving than other two-step methods. Furthermore, the actual appliances of ficin-TN for glucose detection in human serum have been illustrated with satisfied result, suggesting that its promising utilization in various fields. |
| Databáze: | OpenAIRE |
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