Preparation method of alginate lyase immobilized enzyme

A technology of alginate lyase and immobilized enzyme, which is applied in the field of preparation of alginate lyase immobilized enzyme, can solve the problems of limited development and poor stability, and achieve good stability, low cost, and excellent acid and alkali resistance environment Effect

Inactive Publication Date: 2019-03-29
青岛海大生物集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, enzyme catalysis is affected by ambient temperature and pH, and its stability is poor, which limits its development as an industrial enzyme preparation.

Method used

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  • Preparation method of alginate lyase immobilized enzyme
  • Preparation method of alginate lyase immobilized enzyme
  • Preparation method of alginate lyase immobilized enzyme

Examples

Experimental program
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Effect test

Embodiment 1

[0027] A preparation method for alginate lyase immobilized enzyme, specifically comprising the following steps:

[0028] (1) Chitosan and tamarind gum are mixed and ground into powder, passed through a 60-mesh sieve, soaked at room temperature for 4 hours, washed away impurities, and dried after suction filtration; the molecular weight of the chitosan is 30000-40000Da; The mass ratio of chitosan and tamarind gum is 4:1;

[0029] (2) Dissolve the chitosan-tamarind gum mixture with 3% acetic acid solution, the final concentration is 2%-5%, ultrasonically until the bubbles disappear and become a transparent gel, take the gel and add it to the coagulation solution, and harden overnight , washed with water until neutral, and stored at 4°C; the condensate consists of a mixture of 15% NaOH solution and 95% ethanol, with a volume ratio of 4:1;

[0030] (3) Take chitosan-tamarind gum pellets and place them in glutaraldehyde solution, the mass concentration of glutaraldehyde is 3%, the...

Embodiment 2

[0033] A preparation method for alginate lyase immobilized enzyme, specifically comprising the following steps:

[0034] (1) Chitosan and tamarind gum are mixed and ground into powder, passed through a 60-mesh sieve, soaked at room temperature for 4 hours, washed away impurities, and dried after suction filtration; the molecular weight of the chitosan is 30000-40000Da; The mass ratio of chitosan and tamarind gum is 3:1;

[0035] (2) Dissolve the chitosan-tamarind gum mixture with 3% acetic acid solution, the final concentration is 2%-5%, ultrasonically until the bubbles disappear and become a transparent gel, take the gel and add it to the coagulation solution, and harden overnight , washed with water until neutral, and stored at 4°C; the condensate consists of a mixture of 15% NaOH solution and 95% ethanol, with a volume ratio of 4:1;

[0036] (3) Take chitosan-tamarind gum pellets and place them in glutaraldehyde solution, the mass concentration of glutaraldehyde is 3%, the...

Embodiment 3

[0039] A preparation method for alginate lyase immobilized enzyme, specifically comprising the following steps:

[0040] (1) Chitosan and tamarind gum are mixed and ground into powder, passed through a 60-mesh sieve, soaked at room temperature for 4 hours, washed away impurities, and dried after suction filtration; the molecular weight of the chitosan is 30000-40000Da; The mass ratio of chitosan and tamarind gum is 5:1;

[0041] (2) Dissolve the chitosan-tamarind gum mixture with 3% acetic acid solution, the final concentration is 2%-5%, ultrasonically until the bubbles disappear and become a transparent gel, take the gel and add it to the coagulation solution, and harden overnight , washed with water until neutral, and stored at 4°C; the condensate consists of a mixture of 15% NaOH solution and 95% ethanol, with a volume ratio of 4:1;

[0042] (3) Take chitosan-tamarind gum pellets and place them in glutaraldehyde solution, the mass concentration of glutaraldehyde is 3%, the...

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Abstract

The invention discloses a preparation method of an alginate lyase immobilized enzyme. A chitosan-tamarind gum-glutaraldehyde cross-linking adsorption method is adopted, and alginate lyase is fixed toa medium. The preparation method comprises the following steps of refining a chitosan carrier, preparing a cross-linking reagent, performing chitosan- tamarind gum-glutaraldehyde cross-linking, performing extraction to obtain alginate lyase, performing fixing and the like. According to the prepared alginate lyase immobilized enzyme, the catalytic capability and stability to different environment (temperature, pH and storing time) are obviously improved, the immobilized enzyme is used for treating sodium alginate, an ion resin adsorption degradation product is adopted, and a method for filtering with a screen mesh is adopted for recovering the immobilized enzyme. An ultra filtration membrane filter method is adopted for recovering free enzymes, and repeated utilization can be realized, so that the yield of alginate oligosaccharide is increased, the production course is simple, the production cost is greatly reduced, and industrial production requirements of the alginate oligosaccharideproducts can be met.

Description

technical field [0001] The invention relates to a method for preparing an alginate lyase immobilized enzyme, and relates to the technical field of enzyme immobilization, in particular to a method for preparing an alginate lyase immobilized by using biological polysaccharides such as chitosan as a medium. Background technique [0002] Fucoidan oligosaccharides have the characteristics of good hydrophilicity, stability, high viscosity, and non-toxicity, and have unique biological activities, such as promoting plant growth, improving stress resistance, etc., and have broad application potential in the fields of medical treatment and fertilizer. Alginate oligosaccharides are degraded from alginate. Traditional chemical methods use strong acid and alkali or strong oxidants, and the products are mainly saturated oligosaccharides. The enzymatic hydrolysis method has a mild effect and mainly breaks the chemical bonds between uronic acids to obtain unsaturated oligosaccharides with b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10
CPCC12N11/10C12N9/90C12Y402/02003C12Y402/02011
Inventor 宋海妹单俊伟张术聪
Owner 青岛海大生物集团股份有限公司
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