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Method for preparing glucose oxidase from microbial fermentation

A technology of glucose oxidase and microbial fermentation, which is applied in the field of enzyme separation and can solve problems such as inhibition

Active Publication Date: 2018-08-10
宁波天皓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The action temperature of glucose oxidase is 30-60°C, the enzyme is not inhibited by EDTA, KCN and NaF, but HgCl2, AgCl, p-chloromercuric benzoic acid and phenylhydrazine can inhibit the enzyme

Method used

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  • Method for preparing glucose oxidase from microbial fermentation
  • Method for preparing glucose oxidase from microbial fermentation
  • Method for preparing glucose oxidase from microbial fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A method utilizing microbial fermentation to prepare glucose oxidase, comprising the steps of:

[0035] Aspergillus niger was streak-inoculated on the slant medium and then inoculated into the seed medium for cultivation to obtain a density of 1×10 8 Aspergillus niger seed liquid of CFU / mL; The components of the slant medium are: potato 150g / L, sucrose 20g / L, agar 15g / L; the components of the seed medium are: sucrose 20g / L, yeast Cream 10g / L, ammonium sulfate 5g / L, potassium dihydrogen phosphate 1g / L, dipotassium hydrogen phosphate 1g / L, magnesium sulfate heptahydrate 0.02g / L, manganese sulfate monohydrate 0.01g / L, ferrous sulfate heptahydrate 0.01g / L;

[0036] Insert Lactobacillus casei into MRS agar medium for culture, then inoculate into MRS liquid medium for seed culture, and obtain a density of 3×10 8 Lactobacillus casei seed solution of CFU / mL;

[0037] The first stage of cultivation: Aspergillus niger seed liquid was cultivated in a fermenter containing fermen...

Embodiment 2

[0041] A method utilizing microbial fermentation to prepare glucose oxidase, comprising the steps of:

[0042] Aspergillus niger was streak-inoculated on the slant medium and then inoculated into the seed medium for cultivation to obtain a density of 2×10 8 Aspergillus niger seed liquid of CFU / mL; The components of the slant culture medium are: potato 150g / L, sucrose 20g / L, agar 15g / L; the components of the seed culture medium are: sucrose 20g / L, yeast Cream 10g / L, ammonium sulfate 5g / L, potassium dihydrogen phosphate 1g / L, dipotassium hydrogen phosphate 1g / L, magnesium sulfate heptahydrate 0.02g / L, manganese sulfate monohydrate 0.01g / L, ferrous sulfate heptahydrate 0.01g / L;

[0043] Insert Lactobacillus casei into MRS agar medium for culture, then inoculate into MRS liquid medium for seed culture, and obtain a density of 5×10 8 Lactobacillus casei seed solution of CFU / mL;

[0044] The first stage of cultivation: Aspergillus niger seed solution was cultivated in a fermenter...

Embodiment 3

[0059] Taking Example 1 and Comparative Example 1-2 as an example, after 35 hours of fermentation and after the end of fermentation, the concentration of glucose in the fermentation broth, the concentration of gluconic acid and the biomass of bacteria after the end of fermentation were detected. The specific results are shown in Table 1:

[0060] Table 1

[0061] group

[0062] As shown in table 1, embodiment 1 adopts the mode of successive inoculation, and after fermentation finishes, biomass is the highest, and the concentration of gluconic acid is lowest; An increase in concentration will produce feedback inhibition on the synthesis of glucose oxidase, and Lactobacillus casei can use gluconic acid as a carbon source to effectively release the inhibitory effect, thereby improving the enzyme production efficiency.

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Abstract

The invention belongs to the technical field of microbial fermentation, and discloses a method for preparing glucose oxidase from microbial fermentation. The method comprises the steps of first-stagecultivation and second-stage cultivation; according to the first-stage cultivation, an Aspergillus niger seed solution is cultivated for 30-40 h, and then, a Lactobacillus casei seed solution is inoculated for continuous cultivation for 20-30 h. The method is high in fermentation enzyme production activity and short in fermentation time.

Description

technical field [0001] The invention belongs to the technical field of enzyme separation, and in particular relates to a method for preparing glucose oxidase by microbial fermentation. Background technique [0002] Glucose oxidase (glucose oxidase, GOD) specifically catalyzes the oxidation of β-D-glucose into 1,5-gluconolactone and hydrogen peroxide in the presence of molecular oxygen, and then gluconolactone spontaneously combines with water to produce glucose acids and hydrogen peroxide. Glucose oxidase is a very important enzyme in the field of enzyme technology, and is widely used in food processing, fermentation industry, and medical detection. [0003] Glucose oxidase is light yellow powder, easily soluble in water, completely insoluble in ether, chloroform, butanol, pyridine, glycerol, ethylene glycol, etc. 50% acetone and 66% methanol can make it precipitate, and general products contain catalase. The molecular weight is about 150,000, and each gram of enzyme co...

Claims

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Application Information

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IPC IPC(8): C12N9/04C12N1/14C12N1/20C12R1/685C12R1/245
CPCC12N1/14C12N1/20C12N9/0006C12Y101/03004
Inventor 张宝华潘宏渊洪成蒋一南
Owner 宁波天皓生物科技有限公司
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