Immobilized laccase microballoon carrier and its preparation method and use

A technology for immobilizing laccase and carrier, applied in the direction of fixing on/in organic carrier, chemical instrument and method, water/sludge/sewage treatment, etc., can solve the problem of poor biocompatibility, glass bead surface Difficult functionalization and other problems, to achieve the effect of low cost, mild conditions and high enzyme activity

Inactive Publication Date: 2013-12-18
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the surface functionalization of glass

Method used

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  • Immobilized laccase microballoon carrier and its preparation method and use
  • Immobilized laccase microballoon carrier and its preparation method and use
  • Immobilized laccase microballoon carrier and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the preparation of immobilized laccase

[0036] 1) Preparation of immobilized laccase carrier:

[0037] Dissolve 3g of PVA (1799) in 30ml of water, stir mechanically in a boiling water bath for 1 hour, remove from the boiling water bath, wait until the reactant is cooled to 50°C, add 60ml of liquid paraffin. At room temperature, stir the mixture at 500rpm (the radius of the stirring blade is 2cm) for 2 hours to form microspheres, then add 1ml of 1mol L -1 hydrochloric acid solution and 2ml glutaraldehyde aqueous solution with a concentration of 50% by mass, at a rotation speed of 500rpm (the radius of the stirring paddle is 2cm), after stirring for 30 minutes, the microspheres are centrifuged at a rotation speed of 4000rpm (the radius of the centrifuge is 13.5cm) , washed three times with petroleum ether, and then vacuum-dried for 48 hours to obtain polyvinyl alcohol microspheres, the polyvinyl alcohol microspheres such as figure 2 shown. Depend on f...

Embodiment 2

[0042] Embodiment 2, the preparation of immobilized laccase

[0043] 1) Preparation of immobilized laccase carrier:

[0044]Dissolve 3g of PVA (1799) in 40ml of water, stir mechanically in a boiling water bath for 2 hours, remove from the boiling water bath, wait until the reactant is cooled to 80°C, add 80ml of liquid paraffin. At room temperature, the mixture was stirred at 1000rpm (the radius of the stirring blade was 2cm) for 24 hours to form microspheres, and then 2ml1mol L was added to a three-neck round bottom flask -1 hydrochloric acid solution and 3ml glutaraldehyde aqueous solution with a concentration of 50% by mass, at a rotation speed of 1000rpm (the radius of the stirring paddle is 2cm), after stirring for 60 minutes, the microspheres were separated by centrifugation at a rotation speed of 10000rpm (the radius of the centrifuge is 13.5cm) , washed three times with petroleum ether, and then vacuum-dried for 48 hours to obtain polyvinyl alcohol microspheres, the p...

Embodiment 3

[0049] Embodiment 3, the thermal stability of immobilized laccase

[0050] The immobilized laccase and free laccase prepared in Example 1 were respectively placed in a water bath at 30°C-70°C for 6 hours, and samples were taken to detect their enzyme activity. The enzyme activity detection was the same as in Example 1.

[0051] The results showed that the optimum temperature of immobilized laccase was 40°C, which was higher than the optimum temperature of free enzyme at 30°C, and within 30°C-70°C, the enzyme activity of immobilized laccase was higher than that of free enzyme. The activity of laccase, especially at 70°C, the retained activity of immobilized laccase can still reach 40.7%, while the activity of free laccase is only 7.4%, showing that the thermal stability of laccase is significantly improved after immobilization.

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Abstract

The invention discloses an immobilized laccase microballoon carrier and its preparation method and use. The preparation method comprises carrying out phosphorylation modification on polyvinyl alcohol microballoons to obtain the immobilized laccase microballoon carrier. The immobilized laccase microballoon carrier can undergo chemical coupling and chelating reactions with laccase. The immobilized laccase microballoon carrier can be used for industrial wastewater treatment, can be used in the field of environment monitoring, and has the characteristics of large specific surface area, small particle size, good biocompatibility, stable physical and chemical properties and low cost.

Description

technical field [0001] The invention belongs to the field of environmental monitoring and relates to an immobilized enzyme microsphere carrier and a preparation method and application thereof. Background technique [0002] Laccase, a protein that binds multiple copper ions, is widely found in plants, fungi and even insects. Laccase has quite broad substrate specificity and good stability, and can catalyze many phenols, aromatic amines, carboxylic acids, steroid hormones, biochromes, metal organic chemicals, etc., and the reaction products are Water, green and environmental protection. Therefore, it has great application potential in wastewater treatment, pulp bleaching, biosensor and biomonitoring, etc. However, laccase has disadvantages such as high price, susceptible to denaturation and inactivation by various environmental factors, and non-reusable. [0003] The immobilization of laccase can realize the repeated use and stable use of laccase. There are mainly four met...

Claims

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Application Information

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IPC IPC(8): C08J7/14C08J3/24C08L29/04C12N11/08C02F3/00
Inventor 施汉昌白雪王青李冰
Owner TSINGHUA UNIV
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