Method for biomimetic synthesis of nano zirconium dioxide immobilized laccase

A technology of nano-zirconium dioxide and immobilized laccase, which is applied to biochemical equipment and methods, redox enzymes, immobilized on or in inorganic carriers, etc., can solve difficult problems, improve utilization rate, operate Simple process and good biocompatibility

Inactive Publication Date: 2018-03-30
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to synthesize zirconia by biomimetic method, and there are still few related reports

Method used

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  • Method for biomimetic synthesis of nano zirconium dioxide immobilized laccase
  • Method for biomimetic synthesis of nano zirconium dioxide immobilized laccase
  • Method for biomimetic synthesis of nano zirconium dioxide immobilized laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] At room temperature, prepare 5mg / mL lysozyme solution and 0.1U / mL laccase in phosphate buffered saline (PBS) with pH=7.0 and concentrations of 30mmol / L, 40mmol / L, 50mmol / L, and 60mmol / L respectively solution, take 1mL of each of these two solutions and mix well, then add to 8mL 0.1mmol / L K 2 ZrF 6 In the solution, shake gently for 10min, 4000rpm, and centrifuge for 5min to obtain a white precipitate. The resulting precipitate is washed three times with PBS buffer to remove unreacted precursor and unembedded laccase, collect the precipitate, and freeze-dry to obtain nanometer 2 Zirconia immobilized laccase particles.

[0033] The enzyme activity recovery rate of gained immobilized laccase is as follows figure 1 shown. Depend on figure 1 It can be seen that when the concentration of phosphate buffer solution is 50mmol / L, the recovery rate of enzyme activity is the highest.

Embodiment 2

[0035] At room temperature, prepare 5mg / mL, 10mg / mL, 15mg / mL, 20mg / mL lysozyme solutions and 0.1U / mL laccase solutions with pH=7.0, 50mmol / L phosphate buffered saline (PBS), respectively, Take 1mL of lysozyme solutions with different concentrations, mix them with 1mL laccase solution, and add to 8mL 0.1mmol / L K 2 ZrF 6 In the solution, shake gently for 10 minutes, and centrifuge at 4000rpm for 5 minutes to obtain a white precipitate. The obtained precipitate is washed three times with PBS buffer to remove unreacted precursors and unembedded laccase. The precipitate is collected and freeze-dried to obtain nano-dioxide Zirconium-immobilized laccase particles.

[0036] The enzyme activity recovery rate of gained immobilized laccase is as follows figure 2 shown. Depend on figure 2 It can be seen that when the concentration of lysozyme is 5mg / mL, the recovery rate of enzyme activity is the highest.

Embodiment 3

[0038] At room temperature, prepare 5mg / mL lysozyme solution and 0.1U / mL laccase solution with pH=7.0, 50mmol / L phosphate buffered saline (PBS), mix 1mL of each of these two solutions, add To 8mL K 2 ZrF 6In the solution, shake gently for 10 minutes, and centrifuge at 4000rpm for 5 minutes to obtain a white precipitate. The obtained precipitate is washed three times with PBS buffer to remove unreacted precursors and unembedded laccase. The precipitate is collected and freeze-dried to obtain nano-dioxide Zirconium-immobilized laccase particles.

[0039] The enzyme activity recovery rate of gained immobilized laccase is as follows image 3 shown. Depend on image 3 It can be seen that when K 2 ZrF 6 When the concentration of the solution is 0.1mmol / L, the recovery rate of enzyme activity is the highest.

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Abstract

The invention discloses a method for biomimetic synthesis of nano zirconium dioxide immobilized laccase, wherein the method comprises the following steps: proportionally mixing a lysozyme solution anda laccase solution of certain concentration; adding the mixture into a zirconium precursor solution; shaking to obtain white precipitate; centrifuging, washing and freeze drying to obtain immobilizedlaccase particles. According to the method disclosed by the invention, the operation process is simple and easy to implement, and the reaction conditions are mild; the obtained immobilized laccase has high stability and can be used repeatedly; the defects of traditional immobilization method such as complicated immobilization process, violent reaction conditions, weak combination between enzyme and carrier and low stability of immobilized enzyme are overcome.

Description

technical field [0001] The invention belongs to the field of immobilized enzyme preparation, in particular to a method for biomimetic synthesis of nano zirconium dioxide immobilized laccase. Background technique [0002] Laccase is a polyphenol oxidase containing four copper ions, and its catalyzed reaction process uses molecular oxygen to produce a water-only by-product, which is an "eco-friendly" enzyme. Due to its wide range of substrate activity and no secondary pollution caused by oxidizing substrates, laccase has broad application value in the fields of industry, food, environmental protection, and cosmetics. Its development and application have always been the focus of experts and scholars at home and abroad. Research hotspots. However, laccase in the free state is water-soluble, sensitive to the external environment, poor in stability, and cannot be reused, which greatly limits its application in biocatalytic processes. Enzyme immobilization technology can effectiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
CPCC12N9/0061C12N11/14C12Y110/03002
Inventor 张新颖王美银吴慧敏
Owner FUZHOU UNIV
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