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Production method of glucose oxidase

A technology of glucose oxidase and production method, which is applied in the production field of glucose oxidase, can solve the problems of high production cost, low yield, low fermentation activity and the like, and achieves the effects of high production efficiency, easy storage and simple process

Active Publication Date: 2014-12-17
SUZHOU KUNLAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Glucose oxidase is widely distributed in the biological world. Its industrial production is produced by microbial fermentation, but this method has limiting factors such as low fermentation activity, low yield, and high production cost. Therefore, in order to overcome the above problems, technicians are using various methods. A lot of work has been done to improve the production performance of microorganisms by means of genetics, but there is still a lack of work done on the improvement of fermentation performance through the improvement of culture methods

Method used

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  • Production method of glucose oxidase

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Embodiment 1

[0039] A production method of glucose oxidase, comprising the following steps:

[0040] (1) Get the Aspergillus niger strain and inoculate it in the first-grade seed culture medium according to the inoculum amount of 10%, cultivate it at 32° C. for 24 hours, feed sterile air to control the dissolved oxygen above 50%, and cultivate to obtain the first-grade seed liquid.

[0041]Among them, the ingredients of the primary seed medium are: glucose 60g / L, yeast extract 15g / L, ammonium sulfate 3g / L, potassium dihydrogen phosphate 3g / L, magnesium sulfate 1g / L, manganese chloride 0.2g / L , Calcium carbonate 20g / L.

[0042] (2) Inoculate the primary seed liquid in the secondary seed medium with an inoculum amount of 15%, cultivate at 28° C. for 24 hours, pass through sterile air to control dissolved oxygen above 50%, and cultivate to obtain the secondary seed liquid.

[0043] Among them, the composition of the secondary seed medium is: glucose 60g / L, yeast extract 15g / L, ammonium sulfa...

Embodiment 2

[0048] A production method of glucose oxidase, comprising the following steps:

[0049] (1) Get the Aspergillus niger strain and inoculate it in the first-class seed culture medium with an inoculum amount of 15%, cultivate it at 30° C. for 20 hours, feed sterile air to control the dissolved oxygen above 50%, and cultivate to obtain the first-class seed liquid.

[0050] Among them, the ingredients of the primary seed medium are: glucose 80g / L, yeast extract 10g / L, ammonium sulfate 5g / L, potassium dihydrogen phosphate 3g / L, magnesium sulfate 0.5g / L, manganese chloride 0.2g / L L, calcium carbonate 15g / L.

[0051] (2) Inoculate the primary seed liquid into the secondary seed medium with an inoculation amount of 12%, cultivate at 32° C. for 24 hours, pass through sterile air to control dissolved oxygen above 50%, and cultivate to obtain the secondary seed liquid.

[0052] Among them, the composition of the secondary seed medium is: glucose 80g / L, yeast extract 10g / L, ammonium sulfa...

Embodiment 3

[0057] A production method of glucose oxidase, comprising the following steps:

[0058] (1) Get the Aspergillus niger strain and inoculate it in the first-grade seed culture medium according to the inoculum amount of 12%, cultivate it at 28° C. for 22 hours, adjust the stirring speed to control the dissolved oxygen above 50%, and cultivate to obtain the first-grade seed liquid.

[0059] Among them, the ingredients of the primary seed medium are: glucose 60g / L, yeast extract 15g / L, ammonium sulfate 4g / L, potassium dihydrogen phosphate 2g / L, magnesium sulfate 1g / L, manganese chloride 0.5g / L , Calcium carbonate 20g / L.

[0060] (2) Inoculate the primary seed liquid into the secondary seed medium with an inoculum amount of 10%, cultivate at 32° C. for 20 h, adjust the stirring speed to control dissolved oxygen above 50%, and cultivate to obtain the secondary seed liquid.

[0061] Among them, the composition of the secondary seed medium is: glucose 60g / L, yeast extract 15g / L, ammon...

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Abstract

The invention provides a production method of glucose oxidase, and the production method comprises the following steps: (1) taking aspergillus niger seed to inoculate into a first stage seed culture medium for culturing to obtain a first stage seed solution; (2) inoculating the first stage seed solution into a second stage seed culture medium for culturing to obtain a second stage seed solution; (3) inoculating the second stage seed solution into a fermentation culture medium to obtain a mixed culture medium, then adding fructose, feeding in batch glucose to obtain a fermented liquid with the enzyme activity reached above 200U / mL; (4) filtering the fermented liquid, concentrating a filter liquid by ultrafiltration to obtain a glucose oxidase enzyme solution with the enzyme activity reached above 800U / mL; (5) using starch as a carrier to perform spray drying of the glucose oxidase enzyme solution to obtain the solid glucose oxidase with the enzyme activity reached above 2500U / mL. The production method of the glucose oxidase has the advantages of simple process, easy operation and high production efficiency, and the produced glucose oxidase has a high activity and good stability, and is easy to store.

Description

technical field [0001] The invention relates to the field of glucose oxidase, in particular to a production method of glucose oxidase. Background technique [0002] Glucose oxidase (GOD) 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. Glucose oxidase is widely distributed in the biological world. Its industrial production is produced by microbial fermentation, but this method has limiting factors such as low fermentation activity, low yield, and high production cost. Therefore, in order to overcome the above problems, technicians are using various methods. A lot of work has been done to improve the production performance of microorganisms by means of genetics, but there is still a lack of wo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/04C12R1/685
Inventor 赵路宋宵因吴鹏刘宝龙郭峰
Owner SUZHOU KUNLAN BIOTECH
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