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Preparation method of graphene immobilized laccase

A technology of graphene and laccase, which is applied to biochemical equipment and methods, and enzymes immobilized on or in inorganic carriers, can solve the problems of expensive synthesis, complicated processing technology, and poor separation effect, and achieve enzyme retention Live, improve stability, avoid chemical damage

Pending Publication Date: 2019-07-05
NANJING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these carrier materials generally have disadvantages such as expensive synthesis, complicated processing technology, and poor separation effect.

Method used

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  • Preparation method of graphene immobilized laccase
  • Preparation method of graphene immobilized laccase
  • Preparation method of graphene immobilized laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Carrier preparation, its steps are:

[0027] (1) Accurately weigh 5g of dodecylamine powder, dissolve it in 20ml of absolute ethanol, weigh 1.590g of ferrous chloride tetrahydrate and 0.900g of ammonium dihydrogen phosphate and dissolve it in 80ml of ultrapure water (dihydrogen phosphate The molar ratio of ammonium to ferrous chloride tetrahydrate is 1:1.02, and the molar ratio of ammonium dihydrogen phosphate to dodecylamine is 1:3.45). In a water bath at 45°C, mix the anhydrous ethanol solution of dodecylamine with the mixed solution of ferrous chloride and ammonium dihydrogen phosphate, collect the solid product by centrifugation, and dry it in vacuum at 40°C for 36 hours. The dried solid product was ground into a powder and placed in a crucible. After drying, put the crucible into a tube furnace, and calcine at 600°C for 24 hours under a low-flow argon protective atmosphere;

[0028] (2) After the calcination process is over, after the product is cooled to room te...

Embodiment 2

[0033] Carrier preparation, its steps are:

[0034] (1) Accurately weigh 5g of dodecylamine powder, dissolve it in 20ml of absolute ethanol, weigh 1.590g of ferrous chloride tetrahydrate and 0.900g of ammonium dihydrogen phosphate and dissolve it in 80ml of ultrapure water (dihydrogen phosphate The molar ratio of ammonium to ferrous chloride tetrahydrate is 1:1.02, and the molar ratio of ammonium dihydrogen phosphate to dodecylamine is 1:3.45). In a water bath at 45°C, mix the anhydrous ethanol solution of dodecylamine with the mixed solution of ferrous chloride and ammonium dihydrogen phosphate, collect the solid product by centrifugation, and dry it in vacuum at 40°C for 36 hours. The dried solid product was ground into a powder and placed in a crucible. After drying, put the crucible into a tube furnace, and calcine at 600°C for 24 hours under a low-flow argon protective atmosphere;

[0035](2) After the calcination process is over, after the product is cooled to room tem...

Embodiment 3

[0040] Carrier preparation, its steps are:

[0041] (1) Accurately weigh 5g of dodecylamine powder, dissolve it in 20ml of absolute ethanol, weigh 1.590g of ferrous chloride tetrahydrate and 0.900g of ammonium dihydrogen phosphate and dissolve it in 80ml of ultrapure water (dihydrogen phosphate The molar ratio of ammonium to ferrous chloride tetrahydrate is 1:1.02, and the molar ratio of ammonium dihydrogen phosphate to dodecylamine is 1:3.45). In a water bath at 45°C, mix the anhydrous ethanol solution of dodecylamine with the mixed solution of ferrous chloride and ammonium dihydrogen phosphate, collect the solid product by centrifugation, and dry it in vacuum at 40°C for 36 hours. The dried solid product was ground into a powder and placed in a crucible. After drying, put the crucible into a tube furnace, and calcine at 600°C for 24 hours under a low-flow argon protective atmosphere;

[0042] (2) After the calcination process is over, after the product is cooled to room te...

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Abstract

The invention provides a preparation method of graphene immobilized laccase. The graphene immobilized laccase is prepared by immobilizing laccase to a carrier of two-dimensional crystal material having excellent conductivity through nonspecific adsorbing action. The preparation method comprises the steps of preparing the carrier and immobilizing laccase; particularly, the carrier, graphene, is prepared by using a phenol as a raw material to carry out graphitization on sandwiched FePO4 / dodecylamine nanolayer composite, and stripping before synthesis; the immobilized laccase is immobilized by adsorbing laccase to the surface of the evenly dispersed graphene in nonspecific manner, centrifuging, washing, and drying to obtain the finished immobilized laccase. The carrier of graphene herein helps increase electron transfer rate of enzymatic catalytic reaction process and improve reaction efficiency; the immobilized laccase prepared via the preparation method has high fixing quantity, high activity and good stability, can be recycled from the reaction system, and is reusable; the preparation process is simple and feasible.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzyme preparation, in particular to a method for preparing graphene-immobilized laccase, more specifically to prepare a carbon nanomaterial graphene-immobilized lacquer with excellent electrical conductivity and excellent mechanical stability enzyme. Background technique [0002] Laccase is a copper-containing polyphenol oxidase that exists widely in various organisms. Laccase can catalyze oxygen in the air to directly oxidize and decompose various phenolic dyes, substituted phenols, chlorophenols, thiophenols, bisphenol A, aromatic amines, etc. In the presence of laccase mediators (ABTS, HBT, etc.), laccase It can also catalyze the degradation of diphenylmethane, N-substituted p-phenylenediamine, organophosphorus compounds (pesticides) and dioxins related to lignin. Due to its wide substrate specificity and good stability, laccase has important application value in paper industry, wastewat...

Claims

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Application Information

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IPC IPC(8): C12N11/14
CPCC12N11/14C12Y110/03002C12N9/0061
Inventor 毛亮董仕鹏杨广平
Owner NANJING UNIV
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