A method and application of immobilizing tyrosinase with polyacrylonitrile microspheres as carrier

Abstract

The invention provides a method for obtaining immobilized tyrosinase by using polyacrylonitrile microspheres as carriers, and an application. According to the method, the modified polyacrylonitrile microspheres are used as the carriers, tyrosinase is used as an immobilized enzyme, through glutaraldehyde covalent cross-linking combination, the tyrosinase is immobilized on the polyacrylonitrile microspheres, and the purpose of degrading phenolic compounds can be achieved. The method disclosed by the invention is simple, the cost of raw materials is low, the reaction condition is easy to realize, the cyclic utilization effect of the synthetized immobilized tyrosinase is good, the pH tolerance is high, the temperature tolerance is high, the immobilized tyrosinase is heated stably, and the tyrosinase combined to the polyacrylonitrile microspheres can effectively remove phenolic substances in waste water under the effect of oxygen. According to the method disclosed by the invention, the phenolic compounds are degraded by a biological method, and the method conforms to the current international development trend of green decontamination and environmental-friendly decontamination.

Description

technical field

[0001] The invention relates to the field of biochemical industry, in particular to a method and application of immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier.

[0002] technical background

[0003] Phenolic compounds belong to the class of aromatic compounds. The common phenol is a raw material and intermediate in the production of dyestuffs, papermaking, medicine, pesticides and other industries, but its toxic effect on the human body cannot be ignored. Phenol has a significant corrosive effect on any tissue in the human body. When in contact with the protein in the cell protoplasm, a chemical reaction can occur to form insoluble protein, which will cause the cell to lose its vitality. In recent years, with the rapid development of my country's chemical industry, the demand for phenol is increasing, and the discharge of phenol-containing wastewater is also increasing. In the United States, the National Security Agency has listed p...

Examples

Embodiment 1

[0037] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0038] 1) 2g polyacrylonitrile is dissolved in 8g N-N dimethylformamide to form a homogeneous solution, and a phase inversion method is utilized to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0039] 2) Soak the polyacrylonitrile microspheres in 10wt% sodium hydroxide solution for 1 hour at 50°C, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak the carboxylated polyacrylonitrile microspheres in 0.1M hydrochloric acid for 2 hours at room temperature. Acrylonitrile microspheres, then soak the partially carboxylated polyacrylonitrile microspheres in 10wt% ethylenediamine solution for 1h to make them amidated (reaction temperature is 25°C), to obtain modified polyacrylonitrile microspheres;

[0040] 3) Contacting the modified polyacrylonitrile microspheres with 10 wt% glutaraldehyde solut...

Embodiment 2

[0044] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0045] 1) dissolving polyacrylonitrile in N-N dimethylformamide to form a homogeneous solution, and using a phase inversion method to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0046]2) At 45°C, soak the polyacrylonitrile microspheres in 8wt% sodium hydroxide solution for 1.5h, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak them in 0.1M hydrochloric acid at room temperature for 2.5h Then the polyacrylonitrile microspheres of partial carboxylation were immersed in 8% ethylenediamine solution for 1.5h to make it amidate (the reaction temperature was 20°C), and the modified polyacrylonitrile was obtained. Microspheres;

[0047] 3) Contacting the modified polyacrylonitrile microspheres with 8 wt% glutaraldehyde solution at 4° C. for 1 h in the dark;

[0048] 4) Finally, contact the ...

Embodiment 3

[0051] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0052] 1) dissolving polyacrylonitrile in N-N dimethylformamide to form a homogeneous solution, and using a phase inversion method to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0053] 2) At 50°C, soak the polyacrylonitrile microspheres in 10wt% sodium hydroxide solution for 1.2h, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak them in 0.1M hydrochloric acid at room temperature for 2.2h Polyacrylonitrile microspheres, then soak the partially carboxylated polyacrylonitrile microspheres in 8wt% ethylenediamine solution for 1.2h to make them amidate (reaction temperature is 40°C), to obtain modified polyacrylonitrile Microspheres;

[0054] 3) Contacting the modified polyacrylonitrile microspheres with 10 wt% glutaraldehyde solution at 4° C. for 1 h in the dark;

[0055] 4) Finally, c...

technical field

[0001] The invention relates to the field of biochemical industry, in particular to a method and application of immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier.

[0002] technical background

[0003] Phenolic compounds belong to the class of aromatic compounds. The common phenol is a raw material and intermediate in the production of dyestuffs, papermaking, medicine, pesticides and other industries, but its toxic effect on the human body cannot be ignored. Phenol has a significant corrosive effect on any tissue in the human body. When in contact with the protein in the cell protoplasm, a chemical reaction can occur to form insoluble protein, which will cause the cell to lose its vitality. In recent years, with the rapid development of my country's chemical industry, the demand for phenol is increasing, and the discharge of phenol-containing wastewater is also increasing. In the United States, the National Security Agency has listed p...

Examples

Embodiment 1

[0037] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0038] 1) 2g polyacrylonitrile is dissolved in 8g N-N dimethylformamide to form a homogeneous solution, and a phase inversion method is utilized to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0039] 2) Soak the polyacrylonitrile microspheres in 10wt% sodium hydroxide solution for 1 hour at 50°C, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak the carboxylated polyacrylonitrile microspheres in 0.1M hydrochloric acid for 2 hours at room temperature. Acrylonitrile microspheres, then soak the partially carboxylated polyacrylonitrile microspheres in 10wt% ethylenediamine solution for 1h to make them amidated (reaction temperature is 25°C), to obtain modified polyacrylonitrile microspheres;

[0040] 3) Contacting the modified polyacrylonitrile microspheres with 10 wt% glutaraldehyde solut...

Embodiment 2

[0044] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0045] 1) dissolving polyacrylonitrile in N-N dimethylformamide to form a homogeneous solution, and using a phase inversion method to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0046]2) At 45°C, soak the polyacrylonitrile microspheres in 8wt% sodium hydroxide solution for 1.5h, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak them in 0.1M hydrochloric acid at room temperature for 2.5h Then the polyacrylonitrile microspheres of partial carboxylation were immersed in 8% ethylenediamine solution for 1.5h to make it amidate (the reaction temperature was 20°C), and the modified polyacrylonitrile was obtained. Microspheres;

[0047] 3) Contacting the modified polyacrylonitrile microspheres with 8 wt% glutaraldehyde solution at 4° C. for 1 h in the dark;

[0048] 4) Finally, contact the ...

Embodiment 3

[0051] The method for immobilizing tyrosinase with polyacrylonitrile microspheres as a carrier comprises the following steps:

[0052] 1) dissolving polyacrylonitrile in N-N dimethylformamide to form a homogeneous solution, and using a phase inversion method to prepare polyacrylonitrile microspheres using a peristaltic pump;

[0053] 2) At 50°C, soak the polyacrylonitrile microspheres in 10wt% sodium hydroxide solution for 1.2h, then take out the polyacrylonitrile microspheres and wash them with distilled water, then soak them in 0.1M hydrochloric acid at room temperature for 2.2h Polyacrylonitrile microspheres, then soak the partially carboxylated polyacrylonitrile microspheres in 8wt% ethylenediamine solution for 1.2h to make them amidate (reaction temperature is 40°C), to obtain modified polyacrylonitrile Microspheres;

[0054] 3) Contacting the modified polyacrylonitrile microspheres with 10 wt% glutaraldehyde solution at 4° C. for 1 h in the dark;

[0055] 4) Finally, c...

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