Glucose oxidase preparation technology

A technology for glucose oxidase and glucose, which is applied in the directions of oxidoreductase, biochemical equipment and methods, enzymes, etc., can solve the problems of low enzyme activity, and achieve the effects of high enzyme activity stability, simple process and reduced production cost.

Inactive Publication Date: 2017-04-26
温县兴发生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Similarly, Aspergillus niger is also used to ferment and produce glucose oxidase, but the enzyme activity is generally low at present, mostly at 2-7U / mL

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Separating and rejuvenating the preserved Aspergillus niger to obtain a genetically stable Aspergillus niger strain;

[0017] (2) Prepare seed medium and fermentation medium, 100mL of seed medium contains: glucose 5g, KCl 0.02g; KH 2 PO 4 0.02g; MgSO 4 0.012g, (NH 4 ) 2 HPO 4 0.06g, yeast extract powder 0.3g, soybean peptone 0.2g. The fermentation medium was glucose 172g / L, corn steep liquor 11 g / L, calcium carbonate 53 g / L, beef peptone 3.5 g / L, (NH 4 ) 2 HPO 4 0.5g / L, KCl 0.15g / L, MgSO 4 0.125 g / L, FeSO4 0.125 g / L. The seed medium also contains 0.01 g / L of fructo-oligosaccharide and 0.05 g / L of corn steep liquor; the fermentation medium also contains 0.1 g / L of fructo-oligosaccharide and 0.5 g / L of corn steep liquor;

[0018] (3) Select the optimal enzyme production conditions of Aspergillus niger, programmatically add cell osmotic agent during fermentation, and obtain glucose oxidase after fermentation. During the fermentation process of Aspergill...

Embodiment 2

[0021] (1) Separating and rejuvenating the preserved Aspergillus niger to obtain a genetically stable Aspergillus niger strain;

[0022] (2) Prepare seed medium and fermentation medium, 100mL of seed medium contains: glucose 5g, KCl 0.02g, KH 2 PO 4 0.02g, MgSO 4 0.012g, (NH 4 ) 2 HPO 4 0.06g, yeast extract powder 0.3g, soybean peptone 0.2g. The fermentation medium was glucose 172g / L, corn steep liquor 11 g / L, calcium carbonate 53 g / L, beef peptone 3.5 g / L, (NH 4 ) 2 HPO 4 0.5g / L, KCl 0.15g / L, MgSO 4 0.125 g / L, FeSO4 0.125 g / L. The seed medium also contains 0.03 g / L of fructo-oligosaccharide and 0.1 g / L of corn steep liquor; the fermentation medium also contains 0.15 g / L of fructo-oligosaccharide and 0.8 g / L of corn steep liquor;

[0023] (3) Select the optimal enzyme production conditions of Aspergillus niger, programmatically add cell osmotic agent during fermentation, and obtain glucose oxidase after fermentation. During the fermentation process of Aspergill...

Embodiment 3

[0026] (1) Separating and rejuvenating the preserved Aspergillus niger to obtain a genetically stable Aspergillus niger strain;

[0027] (2) Prepare seed medium and fermentation medium, 100mL of seed medium contains: glucose 5g, KCl 0.02g, KH 2 PO 4 0.02g, MgSO 4 0.012g, (NH 4 ) 2 HPO 4 0.06g, yeast extract powder 0.3g, soybean peptone 0.2g. The fermentation medium was glucose 172g / L, corn steep liquor 11 g / L, calcium carbonate 53 g / L, beef peptone 3.5 g / L, (NH 4 ) 2 HPO 4 0.5g / L, KCl 0.15g / L, MgSO 4 0.125 g / L, FeSO4 0.125 g / L. The seed medium also contains 0.05 g / L of fructo-oligosaccharide and 0.2 g / L of corn steep liquor; the fermentation medium also contains 0.2 g / L of fructo-oligosaccharide and 1.0 g / L of corn steep liquor;

[0028] (3) Select the optimal enzyme production conditions of Aspergillus niger, programmatically add cell osmotic agent during fermentation, and obtain glucose oxidase after fermentation. During the fermentation process of Aspergillu...

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Abstract

The invention belongs to the technical field of food biology, and particularly discloses a glucose oxidase preparation technology. The preparation technology includes the following steps: (1) bacterial strain rejuvenation, wherein separation and rejuvenation are conducted for stored aspergillus niger to obtain an aspergillus niger strain with genetic stability; 2), culture medium preparation, wherein a seed culture medium and a fermentation culture medium are prepared; 3), fermentation control, wherein the optimal enzyme production condition of the aspergillus niger is selected, a cell penetrating agent is added in the fermentation process in sequence, and glucose oxidase is obtained after the fermentation is finished. The aspergillus niger is used as an enzyme production strain, enters a fermentation cylinder for enzyme production and fermentation after seed cultivation, the cell penetrating agent is added into the fermentation process to make the glucose oxidase rapidly secreted in a fermentation liquor, and the addition of the cell penetrating agent can effectively improve enzyme activity per unit in the fermentation liquor. The production cost of the glucose oxidase can be effectively reduced, the industrial production of the glucose oxidase is convenient to achieve, and at the same time, a cheap enzymic preparation is provided for preparation of sodium gluconate by an enzymic method.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a glucose oxidase preparation technology. Background technique [0002] Glucose oxidase (GOD for short) can specifically catalyze β-D-glucose to generate gluconic acid and hydrogen peroxide in the presence of oxygen. The preparation of high-purity GOD is light yellow powder, easily soluble in water, and completely insoluble in ether , chloroform, glycerin and ethylene glycol. 50% acetone, 66% methanol can make it precipitate. It is widely distributed in animals, plants and microorganisms. However, due to the characteristics of rapid growth and reproduction of microorganisms and wide sources, it has become the main source of glucose oxidase. The main production strains of microorganisms are Aspergillus niger and Penicillium. Glucose oxidase is an aerobic dehydrogenase produced by fermentation of Aspergillus niger, etc. It is non-toxic and has no side effects on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12R1/685
CPCC12N9/0006C12Y101/03004
Inventor 吴丽安民安庆堂冯斌斌
Owner 温县兴发生物科技有限公司
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