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Immobilized xanthine oxidase, its prepn. and uses

A technology of xanthine oxidase and carbon nanotubes, which is applied in the field of immobilized xanthine oxidase and its preparation, can solve the problems that limit the practical application of carbon nanotubes, and achieve easy control of operating parameters, mild conditions, and simple operation Effect

Inactive Publication Date: 2006-09-20
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, carbon nanotubes are almost insoluble in all solvents, which greatly limits the practical application of carbon nanotubes.

Method used

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  • Immobilized xanthine oxidase, its prepn. and uses
  • Immobilized xanthine oxidase, its prepn. and uses

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1: immobilized xanthine oxidase and its preparation: Commercialized single-walled carbon nanotubes were refluxed with 68% concentrated nitric acid for 16 hours to carry out open-tube oxidation (Nature 372 (1994) 159-162), and washed with water to medium and dry it, take 1 mg of the obtained single-walled carbon nanotubes and 1 mg of dihexadecyl phosphoric acid, add them to 1 ml of distilled water, and ultrasonically disperse them for 10 hours to obtain stable single-walled carbon nanotubes - dihexadecane phosphoric acid dispersion. Add 2 microliters of the obtained dispersion solution dropwise on the polished glassy carbon electrode, place it under an infrared lamp, and wait for the solvent to volatilize completely to obtain a glassy carbon electrode modified by a single-walled carbon nanotube-dicetyl phosphoric acid mixed film. In the potential range of -1.0 volts to 1.5 volts, the electrochemical workstation was scanned cyclically at a scan rate of 500 mV / s...

Embodiment 2

[0020] Example 2: immobilized xanthine oxidase and its preparation: commercialized multi-walled carbon nanotubes were refluxed with 68% concentrated nitric acid for 16 hours to carry out open-tube oxidation (Nature 372 (1994) 159-162), and washed with water to medium properties and drying, take 0.5 mg of the resulting multi-walled carbon nanotubes and 0.6 mg of dihexadecyl phosphoric acid, add to 0.5 ml of distilled water, and ultrasonically disperse for 10 hours to obtain stable multi-walled carbon nanotubes - dihexadecane phosphoric acid dispersion. Add 2 microliters of the obtained dispersion solution dropwise on the polished glassy carbon electrode, place it under an infrared lamp, and wait for the solvent to volatilize completely to obtain a glassy carbon electrode modified by a multi-walled carbon nanotube-dicetyl phosphate mixed film. In the potential range of -1.0 volts to 1.5 volts, the electrochemical workstation was scanned 30 times at a scan rate of 100 mV / s to obt...

Embodiment 3

[0023]Example 3: immobilized xanthine oxidase and its preparation: Commercialized single-walled carbon nanotubes were refluxed with 68% concentrated nitric acid for 16 hours to carry out open-tube oxidation (Nature 372 (1994) 159-162), and washed with water to medium and dried, take 2 mg of the obtained single-walled carbon nanotubes and 1.7 mg of dihexadecyl phosphoric acid, add them to 1 ml of distilled water, and ultrasonically disperse them for 10 hours to obtain stable single-walled carbon nanotubes - dihexadecane phosphoric acid dispersion. Add 2 microliters of the obtained dispersion solution dropwise on the polished glassy carbon electrode, place it under an infrared lamp, and wait for the solvent to volatilize completely to obtain a glassy carbon electrode modified by a single-walled carbon nanotube-dicetyl phosphoric acid mixed film. In the potential range of -1.0 volts to 1.5 volts, the electrochemical workstation was cycle-scanned at a scan rate of 500 mV / s for 10 ...

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Abstract

This invention relates to a method for manufacturing fixed xanthine oxidase, which is composed of carbon nanotubes 40-49.5 wt.%, dihexadecyl phosphorous acid 40-49.5 wt.% and xanthine oxidase 1-20 wt.%. The method comprises the steps of: oxidizing the carbon nanotubs, washing with water to neutrality and drying, adding dried carbon nanotubes and dihexadecyl phosphorous acid into distilled water, ultrasonicating to obtain a dispersion, applying the dispersion to the substrate, drying to obtain a mixed membrane, and placing the mixed membrane in the xanthine oxidase solution for adsorption to obtain fixed xanthine oxidase. This invention can obtain three pairs of oxidation-reduction electrochemical signals of xanthine oxidase simultaneously, and the fixed xanthine oxidase can maintain its bioactivity and can catalyze the reduction of nitrate ions. The xanthine oxidase has good stability and reproducibility, and can be used to manufacture biosensors for detecting nitrate ions.

Description

technical field [0001] The invention relates to an immobilized xanthine oxidase and its preparation method and application, which are suitable for the direct electrochemical research of the xanthine oxidase and the preparation of biosensors. Background technique [0002] Xanthine oxidase is a large molecular weight oxidoreductase that contains multiple electron transfer centers, including one molybdenum, one flavin adenine dinucleotide and two iron chalcogenides. For a long time, xanthine oxidase has been used to prepare biosensors for detecting xanthine or hypoxanthine, but both require the use of electronic mediators, which belong to the second generation of biosensors. Now people are interested in the research of third-generation biosensors, that is, the direct electrochemical research of enzymes. Since the 1970s, people have studied the direct electrochemistry of xanthine oxidase, but they have only achieved partial success, and they can only get the signal of one or tw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
Inventor 胡胜水吴云华
Owner WUHAN UNIV
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