Method for preparing tyrosinase mimic enzyme

A technology of tyrosinase and imitating enzymes, which is applied in the direction of redox enzymes, etc., can solve the problems of harsh operating conditions, easy inactivation, high price, etc., and achieve the effect of high catalytic activity

Inactive Publication Date: 2009-02-11
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with ordinary chemical catalysts, enzyme catalysis has the advantages of high efficiency, specificity and mild reaction conditions, while natural enzymes are too sensitive to the environment, demanding operating conditions, easy to inactivate, and expensive.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] Embodiment 1, the preparation method of tyrosinase mimetic enzyme of the present invention, it is: A) beta-cyclodextrin is added in dry pyridine to dissolve in mass ratio 1: 15, then press beta-cyclodextrin and The molar ratio of o-iodobenzoic acid is 1:0.2, add isophthalic disulfonyl chloride dropwise; react at 25°C for 1 hour, stop the reaction, evaporate pyridine under reduced pressure below 25°C, light brown residue Under the protection of liquid nitrogen, vacuum dry below 25°C, the obtained crude product was dissolved in 40% ethanol aqueous solution with a mass ratio of 1:8, and the insoluble matter was filtered out and then added dropwise at a mass ratio of 1:8. into an aqueous solution of acetonitrile with a concentration of 83% by mass, suction-filtered to remove the white precipitate, evaporate the solvent under reduced pressure at 30°C, dissolve the residue in an appropriate amount of aqueous ethanol with a concentration of 20% by mass, stir for 1 hour, The so...

Embodiment 2

[0009] Example 2, which is: A) β-cyclodextrin is dissolved in dry pyridine at a mass ratio of 1:25, and then the molar ratio of β-cyclodextrin to o-iodobenzoic acid is 1:0.4 dropwise Add isophthalic disulfonyl chloride; react at 25°C for 1 hour, stop the reaction, evaporate pyridine under reduced pressure below 25°C, and dry the light brown residue in vacuum at below 25°C under the protection of liquid nitrogen to obtain The crude product is dissolved in the ethanol aqueous solution of 40% mass percentage concentration by mass ratio 1: 15, after filtering off insoluble matter, add dropwise in the acetonitrile aqueous solution of 83% mass percentage concentration by mass ratio 1: 15, suction filtration, remove White precipitate, evaporate the solvent under reduced pressure at 30°C, dissolve the residue in an appropriate amount of 20% by mass ethanol aqueous solution, stir for 1 hour, evaporate the solvent at 30°C, and dry under vacuum at 30°C to obtain 6 Disubstituted cyclodext...

Embodiment 3

[0010]Example 3, it is: A) Cyclodextrin is added into dry pyridine according to the mass ratio of 1:20 to dissolve, and then the molar ratio of cyclodextrin to o-iodylbenzoic acid is 1:0.3 and isophthalic acid is added dropwise. Sulfonyl chloride; react at 25°C for 1 hour, stop the reaction, evaporate pyridine under reduced pressure below 25°C, and dry the light brown residue in vacuum at below 25°C under the protection of liquid nitrogen, and obtain the crude product according to Mass ratio 1: 10 is dissolved in the ethanol aqueous solution of 40% mass percentage concentration, after filtering out insoluble matter, add dropwise in the acetonitrile aqueous solution of 83% mass percentage concentration by mass ratio 1: 12, suction filtration, remove white precipitate, in Evaporate the solvent under reduced pressure at 30°C, dissolve the residue in an appropriate amount of 20% by mass ethanol aqueous solution, stir for 1 hour, evaporate the solvent at 30°C, and dry under vacuum a...

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Abstract

The invention relates to a method for preparing a tyrosinase mimetic enzyme, the preparation method takes Beta-cyclodextrin as a model, the tyrosinase mimetic enzyme with the natural enzyme activity is obtained by the complexation with Cu2+ on the basis of cyclodextrin dual-histidine derivatives which are obtained by preparation, the tyrosinase mimetic enzyme can catalyze the oxidation of tyrosine to generate dihydroxyphenylalanine; compared with the prior art, the preparation method is characterized by high catalytic activity, no inactivation, and the like.

Description

technical field [0001] The invention relates to a method for preparing a tyrosinase mimic enzyme, which belongs to the technical field of biomimetic catalysis. Background technique [0002] Tyrosinase, a type of polyphenol oxidase, widely exists in plants, animals, and microorganisms. Tyrosinase has two activities of oxygenase and dehydrogenase, which can catalyze the oxidation of tyrosine to generate dihydroxyphenylalanine. Compared with ordinary chemical catalysts, enzyme catalysis has the advantages of high efficiency, specificity and mild reaction conditions. However, natural enzymes are too sensitive to the environment, require harsh operating conditions, are easily inactivated, and are expensive. [0003] There are many factors to consider in designing a synthetic tyrosinase model, the key being the choice of ligand. In the past research, people used pyridyl to imitate the imidazole group of the active center of the enzyme, and synthesized a series of tyrosinase mode...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02
Inventor 胡智文王秉陈文兴
Owner ZHEJIANG SCI-TECH UNIV
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