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Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same

A polycarbonate membrane, alcohol oxidase technology, applied in biochemical equipment and methods, immobilized on/in organic carriers, determination/inspection of microorganisms, etc., can solve the problem of increasing mass transfer resistance, complex enzyme membranes , adverse reactant diffusion and other problems, to achieve the effect of short response time and high sensitivity

Inactive Publication Date: 2011-11-02
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pretreatment of polycarbonate membrane by plasma polymerization technology makes the preparation of enzyme membrane too complicated. In addition, the use of polycarbonate membrane with smaller pore size increases the mass transfer resistance, which is not conducive to the diffusion of reactants.

Method used

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  • Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same
  • Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same
  • Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same

Examples

Experimental program
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Effect test

Embodiment 1

[0022] (1) Cut the polycarbonate membrane with an average pore size of 0.45 μm and a thickness of 10 μm into discs with a diameter of 10 mm, then stick the cut polycarbonate membrane on the O-shaped rubber ring, and dry it for use;

[0023] (2) Immerse the polycarbonate membrane with the O-shaped rubber ring glued to a phosphate buffer solution with a pH value of 7.5 for 6 hours, then take it out and dry it with high-purity nitrogen;

[0024] (3) Drop-coat 5 μL of alcohol oxidase solution with a concentration of 1.02 U / μL on the treated polycarbonate membrane to form a round spot of enzyme layer with a diameter of 5 mm, and dry the enzyme membrane in a refrigerator at 4°C for 1 hour. Hour;

[0025] (4) Suspend the enzyme membrane in a closed container filled with a 50% glutaraldehyde aqueous solution, and cross-link in a refrigerator at 4°C for 12 hours;

[0026] (5) Immerse the enzyme membrane in a phosphate buffer solution with a pH value of 7.5 to wash away the alcohol ox...

Embodiment 2

[0029](1) Cut the polycarbonate membrane with an average pore size of 0.10 μm and a thickness of 30 μm into discs with a diameter of 10 mm, then stick the cut polycarbonate membrane on the O-shaped rubber ring, and dry it for use;

[0030] (2) Immerse the polycarbonate membrane with the O-shaped rubber ring glued to a phosphate buffer solution with a pH value of 6.0 for 12 hours, then take it out, and dry it with high-purity nitrogen;

[0031] (3) Drop-coat 8 μL of alcohol oxidase solution with a concentration of 1.1 U / μL on the treated polycarbonate membrane to form a round spot of enzyme layer with a diameter of 6 mm, and dry the enzyme membrane in a refrigerator at 0°C for 2 Hour;

[0032] (4) Suspend the enzyme membrane in a closed container filled with a 60% glutaraldehyde aqueous solution, and cross-link in a refrigerator at 0° C. for 18 hours;

[0033] (5) Immerse the enzyme membrane in a phosphate buffer solution with a pH value of 6.0 to wash away the weakly bound a...

Embodiment 3

[0036] (1) Cut the polycarbonate membrane with an average pore size of 0.80 μm and a thickness of 20 μm into discs with a diameter of 10 mm, then stick the cut polycarbonate membrane on the O-shaped rubber ring, and dry it for use;

[0037] (2) Immerse the polycarbonate membrane with the O-shaped rubber ring glued to a phosphate buffer solution with a pH value of 9.0 for 10 hours, then take it out, and dry it with high-purity nitrogen;

[0038] (3) Drop-coat 2 μL of alcohol oxidase solution with a concentration of 0.8 U / μL on the treated polycarbonate membrane to form a round spot of enzyme layer with a diameter of 4 mm, and dry the enzyme membrane in a refrigerator at 0°C for 0.5 Hour;

[0039] (4) Suspend the enzyme membrane in a closed container filled with a 40% glutaraldehyde aqueous solution, and cross-link in a refrigerator at 2° C. for 6 hours;

[0040] (5) Immerse the enzyme membrane in a phosphate buffer solution with a pH value of 9.0 to wash away the weakly bound...

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Abstract

The invention provides an ethanol oxidase membrane which takes a polycarbonate membrane as a matrix and a preparation method thereof, pertaining to the technical field of electrochemical biosensors and the preparation thereof. The ethanol oxidase membrane consists of a polycarbonate matrix membrane, the glutaraldehyde of ethanol oxidase and immobilized enzyme. The preparation method of the ethanol oxidase membrane comprises the following steps: the polycarbonate membrane is pasted on an O-shaped rubber ring and soaked into phosphate buffer solution after drying; the polycarbonate membrane is blow-dried by nitrogen and then the ethanol oxidase is dropped on the membrane; and glutaraldehyde steam is utilized to fix the ethanol oxidase by crosslinking. The ethanol oxidase has the advantages of shorter response time, higher sensitivity, wider linear response range, good operation stability and storage stability; in addition, the preparation method of the ethanol oxidase is simple and has low cost.

Description

technical field [0001] The invention belongs to the technical field of electrochemical biosensor and its preparation, in particular to an alcohol oxidase film with polycarbonate film as the substrate and its preparation method. Background technique [0002] Accurate and rapid detection of ethanol content has important application value in food detection, fermentation control and clinical diagnosis. There are many methods for the analysis and detection of ethanol, among which the most commonly used methods are gas chromatography, high performance liquid chromatography and spectrophotometry, etc., but most of these methods require more expensive instruments and equipment, and the detection steps are also more complicated. In recent years, people have begun to use electrochemical biosensors to detect ethanol. This method is simple, fast and has high selectivity. [0003] The electrochemical biosensor prepared with alcohol oxidase as the biologically active substance can be use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327C12N11/08C12Q1/26
Inventor 杨文胜杨利刘长霞
Owner BEIJING UNIV OF CHEM TECH
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