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Method for fixing chitosan enzyme by using cross-linking adsorption of chitosan - glutaraldehyde

A technology of glutaraldehyde cross-linking and chitosan enzyme, which is applied to the direction of hydrolytic enzyme fixed on/in it, can solve the problems of short half-life, easy inactivation, easy desorption and falling off enzyme, and achieve half-life Long-lasting, low-impact effects

Inactive Publication Date: 2007-10-10
王艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The adsorption method refers to the method of immobilizing the enzyme through the secondary bond interaction between the surface of the carrier and the surface of the enzyme; the cross-linking method uses a bifunctional or multifunctional group reagent to cross-link and bridge between the enzyme molecules method; the above method alone has the disadvantages of weak adsorption of the carrier, easy desorption and easy inactivation of the enzyme, and short half-life, and the immobilization system is greatly affected by the external environment.

Method used

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  • Method for fixing chitosan enzyme by using cross-linking adsorption of chitosan - glutaraldehyde
  • Method for fixing chitosan enzyme by using cross-linking adsorption of chitosan - glutaraldehyde
  • Method for fixing chitosan enzyme by using cross-linking adsorption of chitosan - glutaraldehyde

Examples

Experimental program
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Effect test

Embodiment 1

[0025] 1. Purification of immobilized carrier chitin

[0026] Set the molecular weight to 2.0 x 10 4 The flaky chitin was ground into granules, passed through a 30-mesh sieve, and soaked in distilled water overnight at room temperature. The next day, suction filtration, add 4 mol / L HCl, soak for 18 hr, suction filtration, add 0.5 mol / L NaOH, boil for 2 hr, rinse with distilled water until neutral, add 0.005 mol / L NaOH solution, soak for 6 hr at 50 °C, and suction filtration; Add 0.005mol / L acetic acid solution, soak for 4 hours at 50°C, suction filtration; rinse with distilled water until neutral, and dry at 70°C to obtain refined chitin.

[0027] 2. Cross-linking of chitin

[0028] Weigh 0.2 g of purified chitin as a carrier, add 1 ml of a cross-linking reagent, soak at room temperature and stir for 2 hr, stand overnight, and centrifuge to remove the supernatant. The carrier was repeatedly washed with distilled water to remove residual glutaraldehyde, and filtered with suc...

Embodiment 2

[0032] 1. Purification of immobilized carrier chitin

[0033] Set the molecular weight to 2.5 x 10 4 The flaky chitin was ground into granules, passed through a 50-mesh sieve, and soaked in distilled water overnight at room temperature. The next day, suction filtration, add 5 mol / L HCl, soak for 30 hr, suction filtration, add 1 mol / L NaOH, boil for 2 hr, rinse with distilled water until neutral, add 0.01 mol / L NaOH solution, soak at 40 °C for 5 hr, suction filtration; add The 0.01mol / L acetic acid solution was soaked at 40°C for 4 hours, filtered with suction; rinsed with distilled water until neutral, and dried at 90°C to obtain refined chitin.

[0034] 2. Cross-linking of chitin

[0035] Weigh 0.5 g of purified chitin, add 5 ml of cross-linking reagent, soak at room temperature and stir for 5 hr, let stand overnight, and centrifuge to remove the supernatant. The carrier was repeatedly washed with distilled water to remove residual glutaraldehyde, and filtered with suction...

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Abstract

This invention discloses a method for preparing immobilized chitosanase by chitin-glutaraldehyde crosslinking and adsorption. The method comprises: refining chitin carrier, crosslinking with glutaraldehyde, extracting chitosanase, and immobilizing. The method overcomes such problems as low adsorption strength of the carrier, easy desorption and easy deactivation of the enzyme faced by traditional methods that utilize crosslinking or adsorption only. Besides, the also has such advantages as long half-life, and little influence of environment to the immobilized system.

Description

technical field [0001] The invention relates to an enzyme immobilization method, in particular to an enzyme immobilization crosslinking adsorption method for immobilizing free chitosanase by using chitin as an enzyme immobilization carrier and glutaraldehyde as a crosslinking agent. Background technique [0002] Enzyme immobilization methods can be divided into four types: adsorption method, covalent coupling method, cross-linking method and embedding method. The adsorption method refers to the method of immobilizing the enzyme through the secondary bond interaction between the surface of the carrier and the surface of the enzyme; the cross-linking method is to use a bifunctional or multifunctional group reagent to cross-link and bridge the immobilized enzyme between the enzyme molecules. The above method has the disadvantages of weak carrier adsorption, easy desorption and detachment, and easy inactivation of the enzyme when used alone. [0003] Chitin, also known as chiti...

Claims

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Application Information

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IPC IPC(8): C12N11/12C12N11/06C12N9/24
Inventor 王艳潘晓榕周培根王锡昌汪之和陈舜胜
Owner 王艳
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