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Method for preparing mixed immobilized glucose oxidase/catalase microspheres

A technology of glucose oxidase and catalase, which is applied in the direction of immobilized on/in organic carriers, multi-enzyme systems, etc., can solve problems such as the difficulty in realizing the full utilization of amino groups, and achieve sustainable development and eliminate damage , The effect of improving industrial production efficiency

Inactive Publication Date: 2013-04-03
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage of this method is that only one enzyme is immobilized on the carrier, and under the same conditions, the same kind of enzyme has the same charge and repels each other, making it difficult to fully utilize the amino group.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for preparing co-immobilized glucose oxidase / catalase microspheres, the specific method steps are as follows:

[0025] (1) Preparation of chitosan-arginine anionic microspheres

[0026] According to the article "Chitosan-Arginine Resin Immobilization of Chymotrypsin and Its Properties" published in "Applied Chemistry" Volume 26 Issue 7 in July 2009, chitosan-arginine anionic microspheres were prepared .

[0027] (2) Prepare GOD and CAT mixed solution

[0028] After the first step (1) is completed, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 3~8 according to the ratio of GOD:CAT activity ratio of 1:1~6, The lysates containing GOD and CAT were collected. For the collected mixed solution containing GOD and CAT, it was used to prepare co-immobilized GOD / CAT.

[0029] (3) Co-immobilization of GOD and CAT

[0030] After the (2) step was completed, the mass (g) of the chitosan-arginine anion microspheres...

Embodiment 2

[0034] A method for preparing co-immobilized glucose oxidase / catalase microspheres, same as in Example 1, wherein:

[0035] In the (2) step, the activity ratio of GOD:CAT is 1: 4, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 5;

[0036] In the (3) step, the quality of the chitosan-arginine anion microsphere prepared in the (1) step: the volume ratio of the mixed solution containing GOD and CAT collected in the (2) step is 1g: 5mL, After 45 minutes of stirring and adsorption, the glutaraldehyde volume concentration is 2.5% glutaraldehyde solution: the volume ratio of the mixed solution containing GOD and CAT is 1: 7, and the reaction is 2.5 hours at 7 ° C;

[0037] In step (4), wash 4 times with 0.2 mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 5, freeze-dry for 27 hours at a temperature of -45° C. and a vacuum of 35 Pa.

Embodiment 3

[0039] A method for preparing co-immobilized glucose oxidase / catalase microspheres, same as in Example 1, wherein:

[0040] In the (2) step, the activity ratio of GOD:CAT is 1: 6, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 8;

[0041] In the (3) step, the quality of the chitosan-arginine anion microsphere prepared in the (1) step: the volume ratio of the mixed solution containing GOD and CAT collected in the (2) step is 1g: 8mL, After 60 minutes of stirring and adsorption, the glutaraldehyde volume concentration was 2.5% glutaraldehyde solution: the volume ratio of the mixed solution containing GOD and CAT was 1: 10, and the reaction was carried out at 10° C. for 3 hours;

[0042] In step (4), wash 5 times with 0.2 mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 8, freeze-dry for 30 hours at a temperature of -50° C. and a vacuum of 50 Pa in a freeze dryer.

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PUM

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Abstract

The invention provides a method for preparing mixed immobilized glucose oxidase (GOD) / catalase (CAT) microspheres and belongs to the technical field of immobilized enzyme. The method comprises the following steps: on the basis of using chitosan-arginine anionic microspheres as carrier, adding a mixed solution of GOD and CAT and glutaraldehyde, performing crosslinking reaction, filtering, washing,and performing freeze drying to prepare mixed immobilized GOD / CAT microspheres. The method has the characteristics of mild reaction conditions, higher recovery rate of enzyme activity, long half-lifeof immobilized enzyme, simplicity in operation, low cost, cleanness, safety, and the like. The product prepared by the method can be widely used in the industries of medicines, food, water treatment,cement and the like. The product can also be used as a glucose-removing reagent, an anti-oxidation reagent, a bactericide, a glucose quantitative analysis reagent and the like.

Description

1. Technical field [0001] The invention belongs to the technical field of immobilized enzymes, and in particular relates to a technology for preparing co-immobilized glucose oxidase (GOD) / catalase (CAT) microspheres by using arginine-modified cross-linked chitosan microspheres as a carrier. 2. Background technology [0002] Immobilized enzymes are new biocatalysts that bind soluble enzymes to water-insoluble carriers. Compared with free enzymes that can only be used once, immobilized enzymes not only have the catalytic properties of free enzymes, but also have good stability and can be used repeatedly. It can also realize the continuous and automatic production process, which is the main development direction of biocatalyst. [0003] Glucose oxidase (GOD) specifically catalyzes β-D-glucose to generate gluconic acid and hydrogen peroxide. In the food industry, it is used for flour improvement, seafood preservation, and the determination of sugar content in fermentation liqui...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/18C12N11/10
Inventor 周小华王颖利康宏宽付雪梅周兴
Owner CHONGQING UNIV
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