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

A glutaraldehyde cross-linking, chitosanase technology, applied in the direction of immobilization on/in organic carriers, hydrolase, etc., can solve the problems of short half-life, easy inactivation, easy desorption and desorption, and achieve half-life long, low impact

Inactive Publication Date: 2011-02-02
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. Refining 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 overnight in distilled water at room temperature. Suction filtration the next day, add 4mol / L HCl to soak for 18hrs, suction filtration, add 0.5mol / L NaOH, boil for 2hrs, rinse with distilled water until neutral, add 0.005mol / L NaOH solution, soak at 50℃ for 6hrs, suction filtration; Add 0.005mol / L acetic acid solution, soak at 50°C for 4hrs, filter with suction; rinse with distilled water until neutral, dry at 70°C to obtain refined chitin.

[0027] 2. Cross-linking of Chitin

[0028] Weigh 0.2g of refined chitin as a carrier, add 1ml of cross-linking reagent, soak and stir at room temperature for 2hr, let it stand overnight, and centrifuge to remove the supernatant. Wash the carrier repeatedly with distilled water to remove residual glutaraldehyde, and filter with suction to obtain cross-link...

Embodiment 2

[0032] 1. Refining 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 overnight in distilled water at room temperature. Suction filtration the next day, add 5mol / L HCl to soak for 30hrs, suction filtration, add 1mol / L NaOH, boil for 2hrs, rinse with distilled water until neutral, add 0.01mol / L NaOH solution, soak at 40℃ for 5hrs, suction filtration; add Soak in 0.01mol / L acetic acid solution at 40°C for 4 hours, filter with suction; rinse with distilled water until neutral, dry at 90°C to obtain refined chitin.

[0034] 2. Cross-linking of Chitin

[0035] Weigh 0.5g of refined chitin, add 5ml of cross-linking reagent, soak and stir at room temperature for 5hr, let it stand overnight, and centrifuge to remove the supernatant. Wash the carrier repeatedly with distilled water to remove residual glutaraldehyde, and filter with suction to obtain cross-linked chitin. ...

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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 traditionalmethods 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 cross-linking adsorption method using chitin as an enzyme immobilization carrier and glutaraldehyde as a crosslinking agent to immobilize free chitosan enzymes. 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 uses a bifunctional or multifunctional group reagent to cross-link and bridge between the enzyme molecules The method; the above-mentioned method alone has the shortcomings of weak adsorption of the carrier produced, easy desorption and easy inactivation of the enzyme, and has short half-life at the same time, and the immob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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