Preparation method for magnetic sodium alginate-immobilized laccase

A technology of sodium alginate and curing paint, applied in biochemical equipment and methods, oxidoreductase, immobilized on or in inorganic carriers, etc., can solve the problems of difficult to maintain stability, can not be maintained for a long time, complex processing technology, etc. , to achieve a wide range of industrial use value, easy separation, high activity recovery effect

Inactive Publication Date: 2016-04-20
JIANGSU SHIKONG COATING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem mainly solved by the present invention: the current traditional laccase is easily soluble in water, cannot be recycled in water and cannot be maintained for a long time, and its stability is not easy to maintain under high temperature or other extreme environments, and it is easy to denature and inactivate, thus limiting its application , and the commonly used immobilization technology can maintain the catalytic activity of free laccase and improve the utilization rate and stability, but the immobilization materials used have low immobilization efficiency, complex processing technology, difficult separation and low operability. Provide A laccase enzyme solution prepared by fermentation of laccase-producing bacteria, mixed with magnetic particles, supplemented with sodium alginate to immobilize it, oscillating and adsorbing, and then drying to obtain magnetic sodium alginate-immobilized laccase

Method used

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Examples

Experimental program
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Effect test

example 1

[0022]First, according to the inoculum amount being 5% of the mass of the medium substrate, inoculate Amanita albicans into a 300mL Erlenmeyer flask containing 50mL of basal liquid medium, place it on a shaker at 25°C and shake it at a rate of 100rpm for 3 days. Rinse the surface of the mycelia with sterile water and collect the eluent to obtain a suspension of mycelia and store it at 4°C; then add 1% (w / v) tea leaves and 1% (w / v) a mixture of soybean meal and 1% (w / v) bagasse and 5mL copper sulfate solution with a concentration of 0.5mol / L. Ferment at ℃ for 7 days, then transfer the fermentation liquid into a horizontal centrifuge, and centrifuge at 7000r / min to obtain the supernatant liquid, which is the laccase enzyme liquid, and set it aside; / L ferric chloride and 0.2mol / L ferrous chloride solution are mixed evenly, take 1000mL of the mixed solution and put it into a 300mL three-necked flask. Continuously inject nitrogen gas and move it into a water bath, raise the tempe...

example 2

[0025] First, the inoculum amount was 6% of the mass of the medium substrate, inoculated Amanita albidoides into a 300mL Erlenmeyer flask containing 80mL of basal liquid medium, and placed it on a shaker at a constant temperature of 150rpm at 28°C for 4 days. Rinse the surface of the mycelia with sterile water and collect the eluate to obtain a mycelial suspension, which was stored at 5°C; then, 1.5% (w / v) tea leaves and 1.5% (w / v) tea leaves and 1.5% (w / v / v) A mixture of soybean meal and 1.5% (w / v) bagasse and 8mL copper sulfate solution with a concentration of 0.5mol / L, mix well, then add 23mL of the mycelia suspension prepared above, and put it in an incubator at 30°C Ferment for 7.5 days, then transfer the fermented liquid into a horizontal centrifuge, and centrifuge at 8000r / min to obtain the supernatant liquid, which is the laccase enzyme liquid, and set it aside; Mix ferric chloride and 0.2mol / L ferrous chloride solution evenly, take 150mL of the mixed solution and put...

example 3

[0028] First, the inoculum amount was 7% of the mass of the medium substrate, inoculated Amanita albicans into a 300mL Erlenmeyer flask containing 100mL of basal liquid medium, placed on a shaker, and cultured at a constant temperature of 200rpm at 30°C for 5 days. Rinse the surface of mycelia with sterile water and collect the eluent to obtain a suspension of mycelium, which was stored at 6°C; then, 2% (w / v) tea leaves and 2% (w / v / v) A mixture of soybean meal and 2% (w / v) bagasse and 10mL copper sulfate solution with a concentration of 0.5mol / L, after mixing evenly, add 25mL of the mycelia suspension prepared above, and put it in an incubator at 35°C Ferment for 8 days, then transfer the fermented liquid into a horizontal centrifuge, and centrifuge at a speed of 9000r / min to obtain a supernatant that is the laccase enzyme liquid for use; then the concentration is 0.3 mol / L at a molar ratio of 3:2 Mix ferric chloride and 0.2mol / L ferrous chloride solution evenly, take 200mL o...

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Abstract

The invention discloses a preparation method for magnetic sodium alginate-immobilized laccase, and belongs to the technical field of immobilized enzyme preparation. According to the method, a laccase enzyme liquid is prepared by fermenting a laccase production strain, magnetic particles are mixed to immobilize the laccase enzyme liquid in combination of sodium alginate in an auxiliary manner, and drying is carried out after oscillation adsorption, so as to obtain the magnetic sodium alginate-immobilized laccase. The method is simple in process and low in cost, the activity of the prepared magnetic sodium alginate-immobilized laccase is 250 IU/mL or above, the total output is increased to 85-90%, and both the activity and the total output reach a higher level; moreover, through mixing of the nanoscale magnetic particles, the adsorption is more easy to realize, and separation is easy, so that the recovery rate of the immobilized laccase is improved, the stability is remarkably enhanced, and the laccase has excellent performance.

Description

technical field [0001] The invention discloses a preparation method of magnetic sodium alginate immobilized laccase, which belongs to the technical field of immobilized enzyme preparation. Background technique [0002] Laccase is a copper-containing polyphenol oxidase widely found in fungi, bacteria and plants. Laccase has a wide range of substrate specificity and strong stability, and can catalyze many phenols and aromatic amines, such as phenol, p-chlorophenol, aniline, and polyphenols and polyamines. , biofuel cells, biotransformation and wastewater treatment and other fields have a wide range of applications. However, since laccase is easily soluble in water, the free laccase in water cannot be recovered or stored for a long time, and the stability of laccase is not easy to maintain under high temperature or other extreme environments, thus limiting the practical application of laccase. [0003] The application of free laccase is limited due to its easy denaturation an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10
CPCC12N11/14C12N9/0061C12N11/10C12Y110/03002
Inventor 陈明生张帆林大伟
Owner JIANGSU SHIKONG COATING
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