Copper-based metal organic framework material immobilized laccase as well as preparation method and application thereof

A technology for immobilizing laccase and organic framework, applied in biochemical equipment and methods, immobilizing on/in organic carrier, oxidoreductase, etc., can solve the problem of poor stability, high cost and complicated process of immobilizing laccase problem, to achieve the effect of good bisphenol A removal ability, good activity retention, and high efficiency removal rate

Active Publication Date: 2021-03-26
SOUTH CHINA UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the existing immobilization methods still have problems such as complicated process, high cost, and poor stability of immobilized laccase.

Method used

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  • Copper-based metal organic framework material immobilized laccase as well as preparation method and application thereof
  • Copper-based metal organic framework material immobilized laccase as well as preparation method and application thereof
  • Copper-based metal organic framework material immobilized laccase as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The synthesis method of Cu-PABA@Lac: first dissolve copper acetate (50mM), laccase (0.5mg / mL, purchased from sigma company) in water; PABA (12.5mM) is dissolved in acetate buffer (ABS, pH=7.0 ,50mM). The two solutions were then mixed in equal volumes and stirred at room temperature for more than 4 hours. The solid was collected by centrifugation and washed 3 times with deionized water.

Embodiment 2

[0037] The synthesis method of Cu-PABA: First, copper acetate (50 mM) was dissolved in water; PABA (12.5 mM) was dissolved in acetate buffer (ABS, pH=7.0, 50 mM). The two solutions were then combined and stirred at room temperature for 8 hours. The solid was collected by centrifugation and washed 3 times with deionized water.

[0038] From figure 1 According to the XRD spectra of Cu-PABA and Cu-PABA@lac biocomposites, there is no obvious difference in crystal structure. The intensity of the diffraction peak of Cu-PABA@lac decreased, and the results showed that the successful loading of laccase and the introduction of laccase had little effect on the structure of Cu-PABA.

[0039] figure 2 The TGA curve can also demonstrate the success of laccase loading. From figure 2 As can be seen in the Cu-PABA curve, the mass loss (about 10%) below 160 °C is due to water evaporation, and the structure is destroyed above 260 °C. However, the TGA curves of Cu-PABA@Lac showed obvious ...

Embodiment 3

[0040] Example 3: Determination of immobilized enzyme retention activity

[0041] 50mmol / L sodium succinate was used as buffer solution (pH=4.5), and 0.04mmol guaiacol and laccase / immobilized enzyme were contained in 10mL reaction mixture. After reacting at 30°C for 30min, the absorbance was measured at 465nm. Taking the free enzyme activity as 100%, calculate the retained enzyme activity of the immobilized enzyme. The maximum retained enzyme activity of the product obtained in Example 1 is 41.7%.

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Abstract

The invention relates to the field of environmental protection, and particularly discloses copper-based metal organic framework material immobilized laccase as well as a preparation method and application thereof. The preparation method comprises the following steps of dissolving copper acetate and laccase in water to prepare a solution A; dissolving a p-aminobenzoic acid solution into an acetic acid buffer solution to prepare a solution B, then mixing the two solutions, and performing stirring for 4 hours or above; and centrifugally collecting solid, performing washing with water, and performing freeze-drying to obtain the copper-based metal organic framework material immobilized laccase. According to the method, the copper-based MOF (Cu-PABA) immobilized laccase is adopted for the firsttime, a coprecipitation method is used for one-step synthesis, the method is faster and more environmentally friendly, the stability of the laccase is greatly improved, and the laccase can be recycledfor eight times or above after bisphenol A is degraded. After the laccase is immobilized, 41.7% of the activity can be retained. The copper-based metal organic framework material immobilized laccaseprovided by the invention has good bisphenol A removal capability, and the bisphenol A removal rate in 12 hours can reach about 80% in the presence of ABTS.

Description

technical field [0001] The invention relates to the field of environmental protection, in particular to a preparation method of immobilized enzyme and its application in the field of removing bisphenol A. Background technique [0002] Bisphenol A is a typical endocrine disruptor. It is widely produced in baby bottles, toys and medical equipment. In addition to being detected in surface water and groundwater, BPA has also been detected in soil, landfill leachate, air and food. come out. Common treatment methods include physical, chemical and biological methods. Physical methods generally include adsorption method and membrane separation method. Although they are relatively fast, they are not thorough and difficult to reuse, which limits their application. Chemical methods mainly include photocatalysis, electrochemistry, Fenton method, etc. Generally speaking, they need to consume more energy, and the intermediate products generated may be more toxic, causing secondary pollu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02A62D3/02A62D101/28
CPCC12N11/02C12N9/0061A62D3/02C12Y110/03002A62D2101/28
Inventor 程建华袁宇航杜克斯周心慧
Owner SOUTH CHINA UNIV OF TECH
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