CN103642764A - Process for fermenting glucose oxidase - Google Patents
Process for fermenting glucose oxidase Download PDFInfo
- Publication number
- CN103642764A CN103642764A CN201310588356.7A CN201310588356A CN103642764A CN 103642764 A CN103642764 A CN 103642764A CN 201310588356 A CN201310588356 A CN 201310588356A CN 103642764 A CN103642764 A CN 103642764A
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- Prior art keywords
- glucose oxidase
- fermentation
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- aspergillus niger
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
- C12Y101/03004—Glucose oxidase (1.1.3.4)
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- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention discloses a process for fermenting glucose oxidase. The process comprises the following specific steps: selecting Aspergillus niger as a starting strain for fermentation production of glucose oxidase, and carrying out an infrared mutagenesis treatment on the Aspergillus niger; screening to obtain high-yield strains; continuously carrying out a passage experiment 3-5 times on the high-yield strains to obtain strains with stable genetic characters; placing into a culture medium to culture; adopting a double-layer yarn membrane to carrying out dialysis; carrying out conventional fermentation for at least 24 h; transferring into a fermentation tank, and sealing for at least 12 h; and carrying out the passage experiment 3-5 times, wherein the volume of the fermentation tank and the fermentation substance are the same, the volume can not exceed 10% of the volume of the fermentation substance, and maltose can be adopted to replace the Aspergillus niger. The process has the following beneficial effects that: the enzyme production reaction has high activity, the change is relatively stable, the metabolite generation during the fermentation process is effectively reduced, and the obstruction effect of the metabolite generation on the product synthesis is effectively reduced.
Description
Technical field
The present invention relates to the preparation method of biochemical substances, specifically, refer to a kind of technique that glucose oxidase is fermented.
Background technology
Glucose oxidase (G l u c o s eOxidase is called for short GOD) system β-D-Glucose by name: it can become gluconic acid and hydrogen peroxide by β-D-glucose oxidase oxydo-reductase (EC1.1.3.4) single-mindedly.Glucose oxidase is a kind of very important enzyme in zymotechnic field, due to the substantial connection of glucose oxidase and life-critical material glucose and oxygen, has caused its application widely on scientific research, medicine and foodstuffs industry are produced.20 beginning of the century Maximou find oxygen uptake enzyme system in aspergillus niger culture.Nineteen twenty-four, Malliard found gluconic acid.Nineteen twenty-eight, the enzymology that Muller carries out gluconic acid formation with the compacting of black-koji mould filament, verifies energy oxygen consumed when glucose exists, and called after glucose oxidase, and it is included into desaturase.Later many investigators have done a large amount of work to the character of this enzyme, especially prothetic group one flavin adenine dinucleotide (FAD) of glucose oxidase have been done to deep research, and have given detailed explanation.BentleyNeubergar application isotropic substance 18O2218O is clear and definite at the series of experiments having under catalase existence.Keilin etc. have also done more detailed research to the kinetics of the glucose oxidase of aspergillus niger and action mode thereof.Within 1961, the international biochemical society enzyme council is called the systematic name of this enzyme (β-D-glucose-oxydo-reductase (EC1.1.3.4).
Summary of the invention
For overcoming above-mentioned technical problem, we have proposed following technical scheme:
The technique that glucose oxidase is fermented, concrete steps are as follows:
Select aspergillus niger as the starting strain of fermentative production glucose oxidase, it is carried out to ultrared mutagenic treatment;
Screening, obtains superior strain;
Continuously 3-5 time to its experiment of going down to posterity, and obtains the bacterial strain of stabilization characteristics of genetics;
Be positioned in substratum and cultivate;
Use double-deck yarn film to dialyse;
Normal fermentation, at least 24 hours;
Be transferred in fermentor tank, seal at least 12 hours;
The experiment of again going down to posterity, 3-5 time.
In the present invention, the volume of described fermentor tank must be suitable with fermented product, and volume must not surpass 10 of its volume.
In the present invention, can use maltose replace black aspergillus.
The invention has the beneficial effects as follows: it is high to produce enzyme reaction activity, change more steady, the generation that has effectively reduced metabolite in fermenting process with and the inhibition synthetic to product.
Embodiment
The technique that glucose oxidase is fermented, concrete steps are as follows:
Select aspergillus niger as the starting strain of fermentative production glucose oxidase, it is carried out to ultrared mutagenic treatment;
Screening, obtains superior strain;
Continuous 5 times to its experiment of going down to posterity, obtain the bacterial strain of stabilization characteristics of genetics;
Be positioned in substratum and cultivate;
Use double-deck yarn film to dialyse;
Normal fermentation, 24 hours;
Be transferred in fermentor tank, seal 12 hours;
The experiment of again going down to posterity, 3 times.
The volume of described fermentor tank must be suitable with fermented product, and volume is its volume 10.
The above; it is only preferably embodiment of the present invention; but protection scope of the present invention is not limited to this; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; according to technical scheme of the present invention and inventive concept thereof, be equal to replacement or changed, within all should being encompassed in protection scope of the present invention.
Claims (3)
1. a technique of glucose oxidase being fermented, is characterized in that, concrete steps are as follows:
Select aspergillus niger as the starting strain of fermentative production glucose oxidase, it is carried out to ultrared mutagenic treatment;
Screening, obtains superior strain;
Continuously 3-5 time to its experiment of going down to posterity, and obtains the bacterial strain of stabilization characteristics of genetics;
Be positioned in substratum and cultivate;
Use double-deck yarn film to dialyse;
Normal fermentation, at least 24 hours;
Be transferred in fermentor tank, seal at least 12 hours;
The experiment of again going down to posterity, 3-5 time.
2. a kind of technique that glucose oxidase is fermented as claimed in claim 1, is characterized in that, the volume of described fermentor tank must be suitable with fermented product, and volume must not surpass 10 of its volume.
3. a kind of technique that glucose oxidase is fermented as claimed in claim 1, is characterized in that, can use maltose replace black aspergillus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310588356.7A CN103642764A (en) | 2013-11-21 | 2013-11-21 | Process for fermenting glucose oxidase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310588356.7A CN103642764A (en) | 2013-11-21 | 2013-11-21 | Process for fermenting glucose oxidase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103642764A true CN103642764A (en) | 2014-03-19 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310588356.7A Pending CN103642764A (en) | 2013-11-21 | 2013-11-21 | Process for fermenting glucose oxidase |
Country Status (1)
| Country | Link |
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| CN (1) | CN103642764A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385609A (en) * | 2015-12-24 | 2016-03-09 | 山东宝来利来生物工程股份有限公司 | Aspergillus niger for high-yield glucose oxidase and application thereof |
| CN109234247A (en) * | 2018-09-18 | 2019-01-18 | 四川省食品发酵工业研究设计院 | A kind of glucose oxidase and preparation method thereof |
-
2013
- 2013-11-21 CN CN201310588356.7A patent/CN103642764A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385609A (en) * | 2015-12-24 | 2016-03-09 | 山东宝来利来生物工程股份有限公司 | Aspergillus niger for high-yield glucose oxidase and application thereof |
| CN109234247A (en) * | 2018-09-18 | 2019-01-18 | 四川省食品发酵工业研究设计院 | A kind of glucose oxidase and preparation method thereof |
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140319 |