Method for preparing block macroporous composite immobilized laccase constructed by CS/SiO2 ultrathin membrane

A technology for immobilizing laccase and composite materials, which is applied in the field of preparation of immobilized laccase in block macroporous composite materials, can solve the problems of low reuse rate, complicated preparation process, and poor separation effect, and achieve high recovery rate, The preparation process is simple and the effect of repeated use is convenient

Inactive Publication Date: 2016-01-27
NINGBO UNIV
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AI Technical Summary

Problems solved by technology

However, the existing immobilized laccases generally have the disadvantages of high prod

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (1) take by weighing 0.4g chitosan, add mass fraction and be in the acetic acid solution of 2%, be mixed with the chitosan solution that mass fraction is 1%;

[0016] (2) Weigh 2g block SiO 2 The macroporous material was dried at 80°C for 2 hours, placed in a ground-mouthed Erlenmeyer flask containing 1% chitosan solution, at 25°C, with a rotation speed of 50r min -1 Shake in a shaker for 6 hours, filter out the macroporous material, dry at 50°C for 2h, first wash repeatedly with 0.1M sodium carbonate solution, then wash repeatedly with distilled water, and dry at 60°C for 2h;

[0017] (3) Immerse the above-mentioned dried macroporous material in a solution containing ethylene glycol diglycidyl ether with a mass fraction of 3% and vibrate for 6 hours, filter out the macroporous material, rinse repeatedly with distilled water, and dry in vacuum to obtain immobilization Carriers for enzymes;

[0018] (4) Weigh 1g of carrier, immerse in 10ml and use pH=4.5 buffer solutio...

Embodiment 2

[0020] (1) taking by weighing the chitosan of 0.8g, adding mass fraction is in the acetic acid solution of 2%, is made into the chitosan solution that mass fraction is 2%;

[0021] (2) Weigh 2g block SiO 2 The macroporous material was dried at 80°C for 2 hours, placed in a ground-mouthed Erlenmeyer flask containing 2% chitosan solution, at 30°C, with a rotation speed of 50r min -1 Shake in a shaker for 10 hours, filter out the macroporous material, dry at 50°C for 3h, first wash repeatedly with 0.1M sodium carbonate solution, then wash repeatedly with distilled water, and dry at 60°C for 2h;

[0022] (3) Immerse the above-mentioned macroporous material dried at 60°C in a solution containing 1% glutaraldehyde mass fraction and vibrate for 12 hours, filter out the macroporous material, rinse repeatedly with ethanol and distilled water, and dry in vacuum to obtain a fixed Carriers for enzymes;

[0023] (4) Weigh 2g of carrier, immerse in 20ml and use pH=4.5 buffer solution to p...

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Abstract

The present invention relates to a method for preparing a block macroporous composite immobilized laccase, belonging to the technical field of preparing of an immobilized enzyme. The immobilized laccase is prepared by fixing laccase onto a carrier by physical adsorption method, wherein the block macroporous composite constructed by a silica / chitosan ultrathin membrane is used as the carrier. The preparation method comprises the steps preparation of the carrier and fixing of the laccase. The preparation of the carrier is as follows: chitosan is adsorbed on SiO2 macroporous material pore wall surface, and then the finished carrier, constructed by the silica / chitosan ultrathin membrane, is prepared by crosslinking of chitosan macromolecules by use of a bifunctional crosslinking agent; the fixing of the laccase is as follows: the carrier is immersed in a laccase solution and shaken for reaction at room temperature, and finished immobilized laccase can be obtained by washing and drying. The present invention provides a novel immobilized laccase and a preparation method thereof, and the novel immobilized laccase has the characteristics of simple preparation process, low cost, high immobilized laccase activity, and stable property and the like.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzyme preparation, in particular to a CS / SiO 2 A preparation method of immobilized laccase in a block macroporous composite material constructed by an ultra-thin film. Background technique [0002] Laccase is a copper-containing polyphenol oxidase widely present in many organisms. Laccase has broad substrate specificity and good stability, and can catalyze the oxidation of a variety of refractory organic pollutants, including phenolic compounds, polycyclic aromatic hydrocarbons, polychlorinated biphenyls and their derivatives, herbicides, It has important application value in wastewater treatment, biosensor construction, biopulping, food processing and organic synthesis. Therefore, scientists at home and abroad have carried out extensive research work on laccase in many fields such as biology, chemistry, physics, and medicine. [0003] The application of free laccase is limited because it ...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10
Inventor 郑璐梦梁云霄刘晓贞张瑞丰李云江思
Owner NINGBO UNIV
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