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A kind of immobilized horseradish peroxidase carrier and its preparation and application method

A horseradish peroxidase and carrier technology, applied in directions such as being fixed on/in an organic carrier, can solve the problems of unreliable binding, low affinity, environmental pollution, etc., so as to improve operational stability, The effect of stable chemical properties and low cost

Active Publication Date: 2018-03-06
ANHUI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the first three are physical methods for immobilization with weaker interaction between the enzyme and the carrier. The affinity is not high, the combination is not reliable, the enzyme is easy to fall off from the carrier, and the loss of the enzyme is relatively serious, resulting in environmental pollution. Increased costs

Method used

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  • A kind of immobilized horseradish peroxidase carrier and its preparation and application method
  • A kind of immobilized horseradish peroxidase carrier and its preparation and application method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Embodiment 1, the preparation of immobilized horseradish peroxidase

[0028] 1) Preparation of immobilized horseradish peroxidase carrier:

[0029] Dissolve 8 grams of glucose in 80 ml of distilled water and stir until clear, then transfer to a 100 ml autoclave, react at 180°C for 8 hours, cool to room temperature, centrifuge (8000 rpm, 10 minutes), and distilled water for precipitation Alternately washed with ethanol for several times and then freeze-dried to obtain carbon nanospheres;

[0030] figure 1 Fourier transform infrared spectrum (FT-IR) photos of carbon nanospheres prepared for hydrothermal carbonization of glucose, it can be seen that carbon nanospheres have obvious absorption peaks, 3417cm -1 is the stretching vibration of O-H, 1625cm- 1 is the C=O stretching vibration from the carboxyl group. It shows that nano-carbons are rich in functional groups such as carboxyl and hydroxyl groups.

[0031] Take 100 mg of the above nano-carbon spheres and add them...

Embodiment 2

[0036] Embodiment 2, the stability in the extreme environment of immobilized horseradish peroxidase

[0037] 1) The immobilized horseradish peroxidase prepared in Example 1 and the free horseradish peroxidase were respectively placed in a water bath at 5-55° C. for 2 hours, and samples were taken to detect their enzyme activity.

[0038] The results showed that the optimum temperature of immobilized horseradish peroxidase was 25°C, which was equivalent to the optimum temperature of free enzyme, but in extreme temperature environments, especially at 5°C and 55°C, the Enzyme retained enzyme activity can still reach 45.1% and 11.2%, respectively, while free horseradish peroxidase enzyme activity is only 16.9% and 1.5%, indicating that immobilized horseradish peroxidase has a wider temperature adaptation range. Therefore, compared with free horseradish peroxidase, the immobilized horseradish peroxidase prepared in this example is more conducive to adapting to the complex environme...

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Abstract

The invention belongs to the field of biotechnology, and specifically relates to an immobilized horseradish peroxidase carrier and its preparation and application method, wherein the immobilized horseradish peroxidase carrier is nano-carbon spheres prepared by hydrothermal carbonization of glucose , and then it is obtained by amination modification and glutaraldehyde activation; the immobilized horseradish peroxidase carrier of the present invention is mixed and shaken with the horseradish peroxidase solution, centrifuged, washed and freeze-dried to obtain the immobilized horseradish peroxidase root peroxidase. The invention provides a green, economical and efficient method for preparing immobilized horseradish peroxidase, the immobilized horseradish peroxidase obtained by the method is low in cost, simple in operation, stable in physical and chemical properties, especially in extreme environments High catalytic activity can still be maintained under these conditions.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a preparation method of an immobilized horseradish peroxidase carrier. Background technique [0002] Horseradish peroxidase is an enzyme composed of glycoprotein and iron porphyrin prosthetic group, which can catalyze the degradation of phenol, aniline and their substitutes, and has a wide range of substrate specificity. Therefore, it has far-reaching application potential in the fields of wastewater treatment, biosensors, environmental monitoring, immunoassays, and clinical chemistry. But because the stability of natural horseradish peroxidase comprises thermal stability, acid-base stability, organic phase stability and storage stability etc. are all relatively poor. Its half-life is about 10 hours at 37°C, and its active half-life is only about 5 minutes at 65°C. The stability is worse in some medium-strong polar organic solvents mixed with water solvents, and free enzymes in water ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/10
Inventor 陆永明杨乔月陈彦
Owner ANHUI UNIVERSITY
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