CN110129293A - A kind of industrial fermentation extracting method of glucose oxidase - Google Patents
A kind of industrial fermentation extracting method of glucose oxidase Download PDFInfo
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- CN110129293A CN110129293A CN201910476418.2A CN201910476418A CN110129293A CN 110129293 A CN110129293 A CN 110129293A CN 201910476418 A CN201910476418 A CN 201910476418A CN 110129293 A CN110129293 A CN 110129293A
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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- 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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Abstract
The invention discloses a kind of industrial fermentation extracting methods of glucose oxidase, are related to glucose oxidase extractive technique field.The present invention includes that strain is put into constant temperature incubation in slant medium to generate spore and spore is fitted into Liquid Culture bottle, and bacterium solution is made in culture after mixing well；Bacterium solution is seeded in plating medium and is cultivated until growing bacterium colony；Bacterium colony is placed in fermentor and expands fermented and cultured acquisition fermentation liquid；It is glucose oxidase liquid that supernatant is obtained after fermentation liquid is centrifuged；It is concentrated and dried glucose oxidase liquid to obtain glucose oxidase.The present invention is put into slant medium culture by strain and bacterium solution is made, and bacterium solution is seeded to plating medium and grows bacterium colony, by bacterium colony, is placed in fermentor and expands fermentation acquisition fermentation liquid；Fermentation liquid is centrifuged to obtain supernatant and be concentrated and dried supernatant and obtains glucose oxidase；Realize high-volume high efficiency extraction glucose oxidase.
Description
Technical field
The invention belongs to glucose oxidase extractive technique field, the industry more particularly to a kind of glucose oxidase is sent out
Ferment extracting method.
Background technique
Glucose oxidase can be catalyzed glucose consumption oxygen, gluconic acid and hydrogen peroxide be generated, so it is eating
It is very widely used in conduct industry, in terms of being mainly manifested in following four: remaining glucose in removal food, deoxidation, sterilization,
Measure Determination of Glucose in Food content.
The somatotrophic mechanism of action of glucose oxidase is: the glucose oxidase added in animal feed can have anti-
The function of oxidation can dispose a large amount of free radicals generated under livestock stress situation in intestinal epithelial cell, to protect enteron aisle
The integrality of epithelial cell.
Medically Tes-Tape is used to measure the glucose content in diabetes patient's urine；Glucose oxidase oxygen performance rate method
The accuracy of measurement glucose is high, the range of linearity is wide, atopic is strong, repeatability is good.
Glucose oxidase (GOD) is distributed widely in animal, plant and microbial body.But due to being included in animal and plant body
It measures less, and extracts and have certain limitation, therefore less use.
In microorganism, the production of glucose oxidase mainly uses mold fermentation method, is generally done using aspergillus and mould
It is industrial production GOD to produce strain such as aspergillus niger, aspergillus oryzae, mould, Kluyveromyces lactis and Kluyveromyces fragilis
Universal proenzyme.Currently, being all made of Production by Microorganism Fermentation glucose oxidase both at home and abroad.Under certain condition due to mould
The ability for generating glucose oxidase is strong, and therefore, the industrialized production of glucose oxidase mainly uses aspergillus niger
(Aspergillu niger) and Penicillium notatum (Penicilliunnotation).But glucose oxidase almost all depends on
Import, production cost also increase accordingly.Therefore, the higher bacterial strain of malaga carbohydrate oxidase enzyme activity is screened, there is great reality
Application value.
The present invention provides a kind of industrial fermentation extracting method of glucose oxidase, can be realized high-volume high efficiency extraction Portugal
Grape carbohydrate oxidase.
Summary of the invention
The purpose of the present invention is to provide a kind of industrial fermentation extracting methods of glucose oxidase, are put into tiltedly by strain
Simultaneously bacterium solution is made in face culture medium culture, and bacterium solution is seeded to plating medium and grows bacterium colony, by bacterium colony, is placed in fermentor and expands
Fermentation obtains fermentation liquid；Fermentation liquid is centrifuged to obtain supernatant and be concentrated and dried supernatant and obtains glucose oxidase；It realizes
High-volume high efficiency extraction glucose oxidase.
In order to solve the above technical problems, the present invention is achieved by the following technical solutions:
The present invention is a kind of industrial fermentation extracting method of glucose oxidase, comprising: strain culturing and grape are glycoxidative
Enzyme extracts；
The strain culturing process includes the following:
Step 1: strain is put into constant temperature incubation generation spore in slant medium and spore is packed into Liquid Culture bottle
In, bacterium solution is made in culture after mixing well；
Step 2: bacterium solution is seeded in plating medium and is cultivated until growing bacterium colony；
Step 3: bacterium colony is placed in fermentor and expands fermented and cultured acquisition fermentation liquid；
The process that the glucose oxidase extracts includes the following:
Step 4: being that grape is glycoxidative by supernatant is obtained after the centrifugation of fermentation liquid 4000~8000r/min speed conditions
Enzyme solution；
Step 5: it is concentrated and dried the glucose oxidase liquid to obtain glucose oxidase.
Preferably, strain is aspergillus niger or Penicillium notatum in step 1；Slant medium cultivation temperature is 26-30 in step 1
DEG C, incubation time is 4-6 days；The concentration of bacterium solution is 1.5 × 10-3 × 10 spore/mL in step 1.
Preferably, cultivation temperature is 24-26 DEG C in plating medium in step 2, and incubation time is 3-5 days；In step 2
Plating medium has multiple, after the completion of culture, measures the glucose oxidase enzyme activity in each fermentation liquid, it is highest to filter out enzyme activity
Bacterial strain.
Preferably, in step 3 fermentor tank press 0.05-0.08Mpa, 28-30 DEG C of cultivation temperature, speed of agitator 300r/
Min, pH control 6.5-7.0；Ventilation quantity: 0-40h 30m3/ h, 40-58h 45m3It is 40m that/h, 58h-120h, which put tank,3/h；When
When pH rises to 7.0, start feed supplement, controls pH in 6.5-7.0.
Preferably, in 0~5 DEG C of 10~15min of centrifugation in step 4, obtained supernatant is glucose oxidase liquid.
Preferably, thickening temperature is 25-35 DEG C in step 5.
Preferably, the nutrition of the slant medium are as follows: glucose 20%, peptone 25%, (NH4)2SO45%,
K2HPO41%, MgSO4·7H2O 0.5%, pH 7.0；
The nutrition of the plating medium are as follows: glucose 40%, peptone 30%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO4·7H2O 0.5%, pH 7.2；
Nutrition in the fermentor are as follows: glucose 50%, sucrose 10%, corn pulp 10%, (NH4)2SO47.5%, K2HPO41.5%, MgSO47H-O 0.5%, calcium chloride 0.5%, pH 7.2.
The invention has the following advantages:
The present invention is put into slant medium culture by strain and bacterium solution is made, and bacterium solution is seeded to plating medium and is grown
Bacterium colony is placed in fermentor and expands fermentation acquisition fermentation liquid by bacterium colony；By fermentation liquid be centrifuged obtain supernatant and to supernatant it is dense
Contracting is dry to obtain glucose oxidase；Realize high-volume high efficiency extraction glucose oxidase.
Certainly, it implements any of the products of the present invention and does not necessarily require achieving all the advantages described above at the same time.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, will be described below to embodiment required
Attached drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for ability
For the those of ordinary skill of domain, without creative efforts, it can also be obtained according to these attached drawings other attached
Figure.
Fig. 1 is a kind of flow chart of the industrial fermentation extracting method of glucose oxidase of the invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts all other
Embodiment shall fall within the protection scope of the present invention.
Specific embodiment one:
Refering to Figure 1, a kind of industrial fermentation extracting method of glucose oxidase, comprising: strain culturing and grape
Carbohydrate oxidase extracts；
Strain culturing process includes the following:
Step 1: strain is put into constant temperature incubation generation spore in slant medium and spore is packed into Liquid Culture bottle
In, bacterium solution is made in culture after mixing well, wherein strain is aspergillus niger, and slant medium cultivation temperature is 30 DEG C, incubation time
It is 5 days；The concentration of bacterium solution is 1.5 × 10 spore/mL in step 1；
Step 2: bacterium solution is seeded in plating medium and is cultivated until growing bacterium colony；Wherein, it is cultivated in plating medium
Temperature is 26 DEG C, and incubation time is 5 days；Plating medium has multiple, after the completion of culture, measures the grape glycosyloxy in each fermentation liquid
Change enzyme enzyme activity, filters out the highest bacterial strain bacterium colony of enzyme activity；
Step 3: bacterium colony is placed in fermentor and expands fermented and cultured acquisition fermentation liquid, wherein the tank pressure of fermentor
0.08Mpa, 30 DEG C of cultivation temperature, speed of agitator 300r/min, pH control 7.0；Ventilation quantity: 0-40h 30m3/ h, 40-58h are
45m3It is 40m that/h, 58h-120h, which put tank,3/h；When pH rises to 7.0, start feed supplement, controls pH 7.0；
Glucose oxidase extract process include the following:
Step 4: being that grape is glycoxidative by supernatant is obtained after the centrifugation of fermentation liquid 4000~8000r/min speed conditions
Enzyme solution, in 0~5 DEG C of 10~15min of centrifugation, obtained supernatant is glucose oxidase liquid；
Step 5: it is concentrated and dried glucose oxidase liquid to obtain glucose oxidase.
Wherein, the nutrition of slant medium are as follows: glucose 20%, peptone 25%, (NH4)2SO45%,
K2HPO41%, MgSO4·7H2O 0.5%, pH 7.0；
The nutrition of plating medium are as follows: glucose 40%, peptone 30%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO4·7H2O 0.5%, pH 7.2；
Nutrition in fermentor are as follows: glucose 50%, sucrose 10%, corn pulp 10%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO47H-O 0.5%, calcium chloride 0.5%, pH 7.2.
Specific embodiment two:
Refering to Figure 1, a kind of industrial fermentation extracting method of glucose oxidase, comprising: strain culturing and grape
Carbohydrate oxidase extracts；
Strain culturing process includes the following:
Step 1: strain is put into constant temperature incubation generation spore in slant medium and spore is packed into Liquid Culture bottle
In, bacterium solution is made in culture after mixing well, wherein strain is Penicillium notatum, and slant medium cultivation temperature is 28 DEG C, incubation time
It is 6 days；The concentration of bacterium solution is 3 × 10 spore/mL in step 1；
Step 2: bacterium solution is seeded in plating medium and is cultivated until growing bacterium colony；Wherein, it is cultivated in plating medium
Temperature is 28 DEG C, and incubation time is 5 days；Plating medium has multiple, after the completion of culture, measures the grape glycosyloxy in each fermentation liquid
Change enzyme enzyme activity, filters out the highest bacterial strain bacterium colony of enzyme activity；
Step 3: bacterium colony is placed in fermentor and expands fermented and cultured acquisition fermentation liquid, wherein the tank pressure of fermentor
0.04Mpa, 28 DEG C of cultivation temperature, speed of agitator 300r/min, pH control 6.8；Ventilation quantity: 0-40h 30m3/ h, 40-58h are
45m3It is 40m that/h, 58h-120h, which put tank,3/h；When pH rises to 7.0, start feed supplement, controls pH 7.0；
Glucose oxidase extract process include the following:
Step 4: being that grape is glycoxidative by supernatant is obtained after the centrifugation of fermentation liquid 4000~8000r/min speed conditions
Enzyme solution, in 0~5 DEG C of 10~15min of centrifugation, obtained supernatant is glucose oxidase liquid；
Step 5: it is concentrated and dried glucose oxidase liquid to obtain glucose oxidase.
Wherein, the nutrition of slant medium are as follows: glucose 20%, peptone 25%, (NH4)2SO4 5%,
K2HPO41%, MgSO4·7H2O 0.5%, pH 7.0；
The nutrition of plating medium are as follows: glucose 40%, peptone 30%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO4·7H2O 0.5%, pH 7.2；
Nutrition in fermentor are as follows: glucose 50%, sucrose 10%, corn pulp 10%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO47H-O 0.5%, calcium chloride 0.5%, pH 7.2.
It is worth noting that, included each unit is only drawn according to function logic in the above system embodiment
Point, but be not limited to the above division, as long as corresponding functions can be realized；In addition, each functional unit is specific
Title is also only for convenience of distinguishing each other, the protection scope being not intended to restrict the invention.
In addition, those of ordinary skill in the art will appreciate that realizing all or part of the steps in the various embodiments described above method
It is that relevant hardware can be instructed to complete by program, corresponding program can store to be situated between in a computer-readable storage
In matter.
Present invention disclosed above preferred embodiment is only intended to help to illustrate the present invention.There is no detailed for preferred embodiment
All details are described, are not limited the invention to the specific embodiments described.Obviously, according to the content of this specification,
It can make many modifications and variations.These embodiments are chosen and specifically described to this specification, is in order to better explain the present invention
Principle and practical application, so that skilled artisan be enable to better understand and utilize the present invention.The present invention is only
It is limited by claims and its full scope and equivalent.
Claims (7)
1. a kind of industrial fermentation extracting method of glucose oxidase characterized by comprising strain culturing and grape are glycoxidative
Enzyme extracts；
The strain culturing process includes the following:
Step 1: strain is put into constant temperature incubation generation spore in slant medium and spore is fitted into Liquid Culture bottle, is filled
Divide culture after mixing that bacterium solution is made；
Step 2: bacterium solution is seeded in plating medium and is cultivated until growing bacterium colony；
Step 3: bacterium colony is placed in fermentor and expands fermented and cultured acquisition fermentation liquid；
The process that the glucose oxidase extracts includes the following:
Step 4: being glucose oxidase liquid by supernatant is obtained after the centrifugation of fermentation liquid 4000~8000r/min speed conditions；
Step 5: it is concentrated and dried the glucose oxidase liquid to obtain glucose oxidase.
2. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that step 1
Middle strain is aspergillus niger or Penicillium notatum；Slant medium cultivation temperature is 26-30 DEG C in step 1, and incubation time is 4-6 days；Step
The concentration of bacterium solution is 1.5 × 10-3 × 10 spore/mL in rapid one.
3. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that step 2
Cultivation temperature is 24-26 DEG C in middle plating medium, and incubation time is 3-5 days；There is multiple plating medium in step 2, culture
After the completion, the glucose oxidase enzyme activity in each fermentation liquid is measured, the highest bacterial strain of enzyme activity is filtered out.
4. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that step 3
The tank of middle fermentor presses 0.05-0.08Mpa, and 28-30 DEG C of cultivation temperature, speed of agitator 300r/min, pH control 6.5-7.0；It is logical
Air quantity: 0-40h 30m3/ h, 40-58h 45m3It is 40m that/h, 58h-120h120h, which put tank,3/h；When pH rises to 7.0, start
Feed supplement controls pH in 6.5-7.0.
5. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that step 4
In in 0~5 DEG C of 10~15min of centrifugation, obtained supernatant is glucose oxidase liquid.
6. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that step 5
Middle thickening temperature is 25-35 DEG C.
7. a kind of industrial fermentation extracting method of glucose oxidase according to claim 1, which is characterized in that
The nutrition of the slant medium are as follows: glucose 20%, peptone 25%, (NH4)2SO4 5%, K2HPO41%,
MgSO4·7H2O 0.5%, pH 7.0；
The nutrition of the plating medium are as follows: glucose 40%, peptone 30%, (NH4)2SO47.5%, K2HPO4
1.5%, MgSO4·7H2O 0.5%, pH 7.2；
Nutrition in the fermentor are as follows: glucose 50%, sucrose 10%, corn pulp 10%, (NH4)2SO47.5%,
K2HPO41.5%, MgSO47H-O 0.5%, calcium chloride 0.5%, pH 7.2.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334538A (en) * | 2020-03-20 | 2020-06-26 | 鄂州职业大学 | Method for producing gluconic acid by strengthening penicillium funiculosum fermentation glucose |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
-
2019
- 2019-06-03 CN CN201910476418.2A patent/CN110129293A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (1)
| Title |
|---|
| 郭云瑕等: "葡萄糖氧化酶产生菌的紫外诱变及发酵条件优化", 《食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334538A (en) * | 2020-03-20 | 2020-06-26 | 鄂州职业大学 | Method for producing gluconic acid by strengthening penicillium funiculosum fermentation glucose |
| CN111334538B (en) * | 2020-03-20 | 2023-04-11 | 鄂州职业大学 | Method for producing gluconic acid by strengthening penicillium funiculosum fermentation glucose |
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