Preparation method and application of immobilized lignin peroxidase

A technology of peroxidase and lignin, applied in biochemical equipment and methods, oxidoreductase, chemical instruments and methods, etc., can solve problems such as ecological environment and human hazards, glutaraldehyde toxicity, etc., and achieve good stability , The preparation process is simple and the removal efficiency is high

Inactive Publication Date: 2018-12-11
QINGDAO AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above immobilization technologies have the advantages of simple operation and high treatment efficiency, but

Method used

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  • Preparation method and application of immobilized lignin peroxidase
  • Preparation method and application of immobilized lignin peroxidase
  • Preparation method and application of immobilized lignin peroxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] This embodiment prepares immobilized lignin peroxidase, the specific steps are as follows:

[0040] (1) Preparation of Fe3O4 nanoparticles

[0041] Weigh 3g of ferric chloride, 0.8g of sodium citrate and 6g of sodium acetate, dissolve them in 100mL of ethylene glycol successively under continuous stirring, then transfer to a 100mL autoclave, and react at 200°C for 10h. After cooling, black The solid precipitate was washed several times with water and ethanol in sequence, and dried in vacuum for 24 hours to obtain ferric oxide nanoparticles.

[0042](2) Preparation of magnetic silica nanoparticles

[0043] 600mg of prepared iron ferric oxide nanoparticles were uniformly dispersed in the mixed system of water, ethanol and ammonia water. The volumes of water, ethanol and ammonia water in the mixed system are 40mL, 160mL and 2mL respectively. Then, add 3.6mL tetraethyl orthosilicate dropwise, shake at 25°C for 12h, separate the product with a magnet, and wash the obtained...

Embodiment 2

[0050] This example shows the effect of the amount of enzyme added on the enzyme activity of immobilized lignin peroxidase and the binding rate of immobilized lignin peroxidase.

[0051] This embodiment is consistent with steps (1), (2), (3) in embodiment 1,

[0052] Change "30mL tartaric acid-sodium tartrate buffer containing lignin peroxidase" into "5mL, 10mL, 20mL, 40mL and 50mL" in step (4) in Example 1, and then measure the immobilized Enzyme activity of lignin peroxidase.

[0053] figure 2 It can be seen that the amount of enzyme added directly affects the activity of immobilized enzyme. Under other conditions being the same, the immobilized enzyme activity gradually increased with the increase of enzyme amount. When the amount of enzyme added exceeds 30mL, the activity of immobilized enzyme increases slowly. On the other hand, with the increase of the amount of enzyme added, the binding rate of immobilized enzyme decreased gradually. The greater the amount of enzy...

Embodiment 3

[0056] This example shows the effect of the concentration of dopamine hydrochloride on the enzyme activity of immobilized lignin peroxidase.

[0057] This embodiment is consistent with steps (1), (2), and (4) in Embodiment 1,

[0058] In step (3), add dopamine hydrochloride according to the mass concentration of 1.2mg / mL, 1.4mg / mL, 1.6mg / mL, 1.8mg / mL and 2.0mg / mL respectively, and then measure the obtained immobilized lignin peroxide Enzyme activity.

[0059] image 3 The effect of dopamine hydrochloride concentration on the activity of immobilized lignin peroxidase was given. With the increase of dopamine hydrochloride concentration, the immobilized enzyme activity increased gradually. When the concentration of dopamine hydrochloride exceeds 1.6mg / mL, the immobilized enzyme activity hardly changes.

[0060] Considering comprehensively, the concentration of dopamine hydrochloride is 1.6mg / mL, and the activity of immobilized enzyme is 182.8U / g.

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Abstract

The invention discloses a preparation method and application of immobilized lignin peroxidase. The preparation method comprises the following steps of (1) preparing ferroferric oxide nano-particles through ferric trichloride, sodium citrate and sodium acetate; (2) preparing magnetic silicon dioxide nano-particles; (3) preparing dopamine polymerization embedded magnetic silicon dioxide nano-particles; and (4) preparing magnetic silicon dioxide immobilized lignin peroxidase through dopamine polymerization in-situ embedding. The immobilized lignin peroxidase prepared with the method and hydrogenperoxide are added in wastewater containing organic pollutants, and reaction is performed at room temperature to remove the organic pollutants. A preparation technology is simple and convenient; an adopted immobilized carrier is nanometer sized particles, is safe without toxicity, is high in stability, large in specific surface area, high in binding rate with enzyme, relatively good in dispersibility in a catalytic system and higher in removal efficiency to the pollutants without the risk of secondary pollution and can recycle immobilized enzyme through magnetic separation and recovery.

Description

technical field [0001] The invention relates to a preparation method of immobilized lignin peroxidase and its application in treating organic pollutants in waste water, belonging to the technical field of treating organic pollutants in waste water. Background technique [0002] The main organic pollutants in industrial and livestock wastewater are phenolic compounds, benzene compounds, halocarbon compounds and antibiotics. These organic pollutants are highly toxic and not easily degraded by microorganisms in the environment. At present, the methods for treating organic pollutants in wastewater mainly include chemical methods, physical methods and biological methods. Among them, the physical method generally has relatively expensive equipment and requires high treatment conditions; the chemical method has quick results and a short cycle in the process of treatment, but it is prone to secondary pollution; Harmful to the growth of microorganisms. [0003] Enzymes are green c...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/08C12N11/04C02F3/34
CPCC02F3/342C12N9/0065C12N11/04C12N11/08C12N11/14C12Y111/01014
Inventor 柴超郭瑾葛蔚张小梅柳修楚陈小宇
Owner QINGDAO AGRI UNIV
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