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Preparation method and applications of biological inorganic hybrid membrane based on immobilized laccase adopting copper foil carrier

A copper foil carrier and hybrid membrane technology, which is applied in biochemical equipment and methods, chemical instruments and methods, fixed on or in inorganic carriers, etc., can solve the problem of insufficient structural stability, low laccase loading, and leakage. Many problems, to achieve the effect of low cost, enhanced laccase activity, and improved use efficiency

Active Publication Date: 2017-08-11
江阴智产汇知识产权运营有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the laccase immobilized by this method has low loading capacity, unstable structure, and many leaks, and the enzyme activity is greatly affected by factors such as the temperature and pH of the surrounding environment.

Method used

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  • Preparation method and applications of biological inorganic hybrid membrane based on immobilized laccase adopting copper foil carrier
  • Preparation method and applications of biological inorganic hybrid membrane based on immobilized laccase adopting copper foil carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] a) Copper foil carrier preparation: Cut the copper foil (thickness 0.1 mm) into blocks with a length and width of 30 mm and 20 mm respectively, clean it with 2 mol / L HCl under ultrasonic for 15 min, and then wash it with deionized water and ethanol Each was cleaned for 3 min; then the treated copper foil was soaked in 2 mL of (NH 4 ) 2 S 2 o 8 (0.1 mol / L) and Na 2 HPO 4 (0.4 mol / L) in 20 mL of aqueous solution for 1 h, removed, washed with deionized water, and dried at 40 °C for 2 h under nitrogen protection to prepare the immobilized laccase carrier.

[0026] b) Fixation of laccase: prepare 20 mL of laccase solution with a concentration of 0.1 mg / mL in phosphate buffer solution (0.005 mol / L, pH 5), and stir magnetically at room temperature for 6 min; The prepared carrier was soaked in the above laccase solution for 6 h, the surface of the carrier was washed with deionized water without immobilized laccase, and the hybrid enzyme finished film was obtained by vacuum...

Embodiment 2

[0029] a) Copper foil carrier preparation: Cut the copper foil (thickness 0.1 mm) into blocks with a length and a width of 30 mm and 20 mm respectively, clean it with 5 mol / L HCl under ultrasonic for 15 min, and then wash it with deionized water and ethanol Each was cleaned for 3 min; then the treated copper foil was soaked in 3 mL of (NH 4 ) 2 S 2 o 8 (0.3 mol / L) and Na 2 HPO 4 (0.6 mol / L) in 20 mL of aqueous solution for 3 h, removed, washed with deionized water, and dried at 40 °C for 2 h under nitrogen protection to prepare the immobilized laccase carrier;

[0030] b) Fixation of laccase: prepare 20 mL of laccase solution with a concentration of 0.2 mg / mL in phosphate buffer solution (0.05 mol / L, pH 6), stir magnetically at room temperature for 7 minutes, and then mix the The prepared carrier was soaked in the above-mentioned laccase solution for 18 h, the surface of the carrier was washed with deionized water without immobilized laccase, and dried in vacuum at 30°C t...

Embodiment 3

[0033] a) Preparation of copper foil carrier: Cut the copper foil (thickness 0.1 mm) into blocks with the length and width of 30 mm and 20 mm respectively, clean them with 6 mol / L HCl under ultrasonic for 15 min, and then wash them with deionized water and ethanol respectively. Wash for 4 min; then soak the treated copper foil in 4.5 mL of (NH 4 ) 2 S 2 o 8 (0.6 mol / L) and Na 2 HPO 4 (0.7 mol / L) in 20 mL of aqueous solution for 5 h, removed, washed with deionized water, and dried at 40 °C for 2 h under nitrogen protection to prepare the immobilized laccase carrier.

[0034] b) Laccase immobilization: Prepare 20 mL of laccase solution with a concentration of 0.4 mg / mL in phosphate buffer solution (0.08 mol / L, pH 6.5), and stir magnetically at room temperature for 8 min; The prepared carrier was soaked in the above laccase solution for 16 h, the surface of the carrier was washed with deionized water without immobilized laccase, and dried in vacuum at 30°C to prepare the fin...

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Abstract

The invention belongs to the technical field of biological inorganic hybridization, and in particular relates to a preparation method and applications of a biological inorganic hybrid membrane based on immobilized laccase adopting a copper foil carrier. The preparation method comprises the following steps: cutting copper foil into blocks, washing the copper foil blocks by adopting HCl solution, deionized water and alcohol, then soaking the washed copper foil blocks in (NH4)2S2O8 aqueous solution and Na2HPO4 aqueous solution in sequence, carrying out drying for 2h at 40 DEG C under the protection of inert gas, thus preparing an immobilized laccase carrier; soaking the carrier in laccase solution for 6-24 h, washing the surface by adopting deionized water, and carrying out vacuum drying at 30 DEG C, thus obtaining the biological inorganic hybrid membrane. The invention further discloses applications of the biological inorganic hybrid membrane based on immobilized laccase adopting the copper foil carrier in the environmental pollution control including dye wastewater decoloration and the like. The preparation method is simple, the prepared product has the activity enhanced than the activity of free laccase, has the enhanced stability in resisting the pH and temperate change influences, is good in reusability, and is strong in operability, a novel thought is provided for the immobilization of laccase and other proteins, and a promoting effect is achieved in the applications including protein immobilization, environment control, medical treatment and the like.

Description

technical field [0001] The invention belongs to the technical field of bio-inorganic hybridization, and relates to a bio-inorganic hybrid film, in particular to a preparation method and application of a bio-inorganic hybrid film based on a copper foil carrier immobilizing laccase. Background technique [0002] Laccase is a protein that binds multiple copper ions. It exists widely in plants and fungi. It can use molecular oxygen as an electron acceptor to reduce it to water, so it is an environmentally friendly enzyme in nature. Laccase has low substrate specificity and high catalytic activity, and is widely used in wastewater treatment, aromatic compound conversion, biosensors, papermaking, environmental monitoring, pharmaceuticals, food, etc. However, when laccase leaves the relatively stable environment of organisms, it becomes extremely fragile, which greatly shortens its storage time at room temperature. At the same time, it is easy to inactivate and denature under high...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N9/02C02F3/34C02F101/30
CPCC02F3/342C02F2101/308C12N9/0061C12N11/14C12Y110/03002
Inventor 邱凤仙荣坚张涛朱瑶杨冬亚徐吉成
Owner 江阴智产汇知识产权运营有限公司
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