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Preparation method of immobilized glucose oxidase

A technology of glucose oxidase and seeds, applied in the directions of oxidoreductase, immobilized on/in organic carriers, etc., can solve the problems of non-reusability, poor mechanical strength of glucose oxidase, low enzymatic activity, etc., and achieve a way of obtaining Simple, easy to obtain, enhance the effect of enzyme activity

Inactive Publication Date: 2017-12-22
蒋文明
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention: the object of the present invention is to solve the problem that general glucose oxidase has poor mechanical strength, low enzymatic activity, and cannot be reused, and utilizes an environment-friendly acquisition method of glucose oxidase, through the embedding method Combined with the cross-linking method to obtain a preparation method of immobilized glucose oxidase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Raw material: Isolate, screen and identify from pastoral soil, take the colony with the largest diameter as the seed strain, and use it for fermentation to obtain glucose oxidase.

[0035] Preparation of inoculated slant medium: by mass parts, take 20-30 parts of sucrose, 15 parts of tryptone, NaNO 3 2 copies, K 2 HPO 4 0.6 parts, FeSO 4 0.01 parts, 10 parts of agar, sterilized at 121℃ for 20min, pH5.4±0.2.

[0036] Preparation of screening plate medium: by mass parts, take 50 parts of glucose, 1 part of peptone, (NH4) 2 HPO 4 0.2 parts, KH 2 PO 4 0.1 part, MgSO4·7H 2 0.04 parts of O, 6 parts of potato flour, 0.8 parts of KI, 0.07 parts of sodium deoxycholate, 10 parts of agar, 0.05 parts of phosphate buffer, sterilized at 121°C for 20 minutes, pH 5.6±0.2.

[0037] Preparation of fungal plate culture medium: by mass parts, take 2 parts of peptone, 8 parts of glucose, KH 2 PO 4 0.8 parts, MgSO4 0.2 parts, Bengal red 0.01 parts, chloramphenicol 0.1 parts, st...

Embodiment 2

[0049] Raw material: Isolate, screen and identify from pastoral soil, take the colony with the largest diameter as the seed strain, and use it for fermentation to obtain glucose oxidase.

[0050] Preparation of inoculated slant medium: by mass parts, take 25 parts of sucrose, 17 parts of tryptone, NaNO 3 3 copies, K 2 HPO 4 0.8 parts, FeSO 4 0.02 parts, 15 parts of agar, sterilized at 121°C for 20 minutes, pH5.4±0.2.

[0051] Preparation of screening plate culture medium: by mass parts, get 65 parts of glucose, 2 parts of peptone, (NH4) 2 HPO 4 0.2~0.4 parts, KH 2 PO 4 0.3 parts, MgSO4·7H 2 0.08 parts of O, 8 parts of potato flour, 1.6 parts of KI, 0.15 parts of sodium deoxycholate, 15 parts of agar, 0.07 parts of phosphate buffer, sterilized at 121°C for 20 minutes, pH 5.6±0.2.

[0052] The preparation of the fungus plate medium: by mass parts, take 3 parts of peptone, 9 parts of glucose, KH 2 PO 4 1.0 parts, MgSO4 0.4 parts, Bengal red 0.25 parts, chloramphenicol...

Embodiment 3

[0064] Raw material: Isolate, screen and identify from pastoral soil, take the colony with the largest diameter as the seed strain, and use it for fermentation to obtain glucose oxidase.

[0065] Preparation of inoculated slant medium: by mass parts, take 30 parts of sucrose, 20 parts of tryptone, NaNO 3 5 copies, K 2 HPO 4 1 part FeSO 4 0.03 parts, 20 parts of agar, sterilized at 121°C for 20 minutes, pH5.4±0.2.

[0066] Preparation of screening plate culture medium: by mass parts, get 80 parts of glucose, 3 parts of peptone, (NH4) 2 HPO 4 0.4 parts, KH 2 PO 4 0.4 parts, MgSO4·7H 2 0.1 part of O, 10 parts of potato flour, 1.7 parts of KI, 0.2 part of sodium deoxycholate, 20 parts of agar, 0.1 part of phosphate buffer, sterilized at 121°C for 20 minutes, pH 5.6±0.2.

[0067] The preparation of the fungus plate medium: by mass parts, take 5 parts of peptone, 10 parts of glucose, KH 2 PO 4 1.2 parts, MgSO4 0.5 parts, Bengal red 0.35 parts, chloramphenicol 0.4 parts,...

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Abstract

The invention belongs to the field of a food preservation additive, and specifically relates to a preparation method of immobilized glucose oxidase. A fungi strain is separated and screened from soil and is used for fermenting glucose oxidase. The fungi strain is simple in acquiring approach, is green and eco-friendly, and is super easy to obtain. Through co-action of an entrapping method and a crosslinking method, polyvinyl alcohol is taken as a carrier, and bis-diazotized benzidine with the concentration being 10% is added to immobilize glucose oxidase. The immobilized glucose oxidase can be repeatedly used. The method is saved in cost and reduced in breakage rate of glucose oxidase cells. The immobilized glucose oxidase is improved in mechanical strength and enzyme activity and prolonged in enzyme activity maintaining time.

Description

technical field [0001] The invention belongs to the field of food fresh-keeping food additives, and in particular relates to a preparation method of immobilized glucose oxidase. Background technique [0002] Glucose oxidase is an aerobic dehydrogenase. Glucose oxidase is an enzyme found in molds such as Penicillium insolens and in honey. Glucose oxidase uses oxygen as the electron acceptor, and can specifically oxidize β-D-glucose to gluconic acid and hydrogen peroxide. Glucose oxidase is a very important enzyme in the field of enzyme application technology. As one of the enzyme preparations allowed by the country, it has been widely used in food, medicine, feed and other industries. [0003] Glucose oxidase is widely distributed in the biological world, and its industrial production is produced by microbial fermentation. The most common source of glucose oxidase is obtained from the fermentation of Aspergillus niger, Penicillium and Saccharomyces. Most commercial glucose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N11/02C12N11/08
Inventor 蒋文明李璐尹凯欣
Owner 蒋文明
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