CN108913747A - A kind of high density fermentation vitamin B2Method - Google Patents
A kind of high density fermentation vitamin B2Method Download PDFInfo
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Abstract
The invention belongs to technical field of biological fermentation, disclose a kind of high density fermentation vitamin B2Method, using bacillus subtilis as fermentation strain, pass through high cell density fermentation, improve vitamin B2Fermentation level, include the following steps：1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized, and at 37~42 DEG C, 120~140rpm, 12~18h of aerobic culture obtains seed culture fluid；2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, the concentration of initial glucose is 15-25g/L；3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.High density fermentation vitamin B can be realized using the above method2, improve vitamin B2Yield.
Description
Technical field
The invention belongs to technical field of biological fermentation, and in particular to a kind of high density fermentation vitamin B2Method.
Background technique
Vitamin B2Also known as riboflavin, it is the component part of internal yellow enzymes prothetic group（Yellow enzyme is sent out in biological oxidation
It waves and passs hydrogen effect）, when lacking, the biological oxidation of body is just influenced, makes metabolism that obstacle occur.It is mainly used for prevention and treatment riboflavin to lack
Weary disease, such as angular stomatitis, cheilitis, glossitis and eye conjunctivitis.In recent years, also some researches show that riboflavin also has diuresis, drop blood
The effects of rouge and improvement cardiac function.The synthesis of riboflavin mainly has chemical synthesis and microbe fermentation method, since chemistry closes
At method, complex and production cost is higher, and riboflavin mainly uses microbe fermentation method to be produced at present.
But current fermentation method, there are strain growth situation is poor, fermentation period is longer, the lower feelings of riboflavin production
Condition, main cause include：Strain does not screen, and spawn degeneration is serious；Fermentation medium is ineffective；Fermentation condition, it is such as molten
The control that oxygen concentration, cultivation temperature, the pH value of fermentation liquid, feed profile and fermentation liquid rheological properties etc. are not got well.
Summary of the invention
For above situation, the object of the present invention is to provide a kind of high density fermentation vitamin Bs2Method, by hair
The restriction of yeast-like fungi strain, seed culture medium, fermentation medium and fermentation process and parameter, high density fermentation produce vitamin B2, real
Existing vitamin B2The raising of yield.
The present invention provides a kind of high density fermentation vitamin B2Method pass through using bacillus subtilis as fermentation strain
High cell density fermentation improves vitamin B2Fermentation level, include the following steps：
1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized,
At 37~42 DEG C, 120~140rpm, 12~18h of aerobic culture obtains seed culture fluid；The group of seed culture medium becomes：Sugarcane
Sugared 18-24g/L, sodium nitrate 8-12g/L, ammonium nitrate 5-8g/L, epsom salt 1-2g/L, potassium dihydrogen phosphate 0.8-1.2g/L,
Dipotassium hydrogen phosphate 1.5-2.5g/L, yeast extract 3-5g/L, frerrous chloride 1-3g/L, zinc sulfate 1-3g/L, manganese chloride 2.5-
5g/L, pH are 6.7~6.9；
2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, is fermented
The group of culture medium becomes：Sucrose 90-120g/L, yeast extract 10-14g/L, sodium nitrate 8-12g/L, ammonium nitrate 6-8g/L, phosphorus
Acid dihydride potassium 7-9g/L, dipotassium hydrogen phosphate 3-5g/L, magnesium sulfate 2-4g/L, frerrous chloride 1.5-3g/L, zinc sulfate 2-4g/L, chlorine
Change manganese 3-5g/L, glycine betaine 0.2-1.5g/L, pH are 6.8~7.2, and the concentration of initial glucose is 15-25g/L；
Fermentation is divided into two stages progress, and in the first stage, keeping fermentation temperature is 37~40 DEG C, with liquefied ammonia adjust pH be 6.8~
7.2, control ventilation ratio is 1 ︰ 0.6~0.9, and dissolved oxygen amount is made to reach 15~30%, and fermentation keeps glucose consumption complete, continues the 2-3 that ferments
Hour；In second stage, keeping fermentation temperature is 37~39 DEG C, and adjusting pH is 7.0~7.2, and control ventilation ratio is 1：1.0~
1.5, control dissolved oxygen amount adds sterile dextrose liquid 20~30%, concentration of glucose is controlled between 5-15g/L, until bacterial strain
Aging, the glucose consumption added is complete, terminates fermentation；
3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.
In the present invention, the bacillus subtilis slant strains of use are made by following methods：Bacillus subtilis is original
The bacterium solution of bacterial strain aseptically carries out ultraviolet mutagenesis, obtains ultraviolet mutagenesis bacterial strain, then carry out chemistry with ethylmethane sulfonate
Mutagenesis obtains complex mutation bacterial strain；Then complex mutation bacterial strain is successively inoculated into metabolism substrate analog 8-anaguanine, 8-
Azaadenine, 6- azathymine, 6- thioguanine, DL- METHIONINE sulfoxide, rose flavine minimal medium in into
Row culture domestication, is made resistant mutant strain；Processed strain inoculated is cultivated into nutrient medium, obtained nutrition
Culture inoculates to be cultivated in strain fermentation culture medium, filters out production vitamin B2Highest bacterial strain.Pass through the party
Strain made from method obtains higher vitamin B by special induction mode, and by screening2Produce bacterial strain.
Preferably, the group of seed culture medium becomes：Sucrose 18-20g/L, sodium nitrate 9-11g/L, ammonium nitrate 6-8g/L, seven
Water magnesium sulfate 1-1.5g/L, potassium dihydrogen phosphate 0.9-1g/L, dipotassium hydrogen phosphate 1.5-2g/L, yeast extract 4-5g/L, chlorination
Ferrous 1-2g/L, zinc sulfate 2-3g/L, manganese chloride 3-5g/L, pH are 6.7~6.9.
It is further preferred that the group of seed culture medium becomes：Sucrose 20g/L, sodium nitrate 10g/L, ammonium nitrate 7g/L, seven water
Magnesium sulfate 1.5g/L, potassium dihydrogen phosphate 1g/L, dipotassium hydrogen phosphate 2g/L, yeast extract 4g/L, frerrous chloride 2g/L, zinc sulfate
2g/L, manganese chloride 4g/L.
Preferably, the group of fermentation medium becomes：Sucrose 110-120g/L, yeast extract 12-14g/L, sodium nitrate 9-
10g/L, ammonium nitrate 7-8g/L, potassium dihydrogen phosphate 8-9g/L, dipotassium hydrogen phosphate 3-4g/L, magnesium sulfate 2-3g/L, frerrous chloride 2-
3g/L, zinc sulfate 2-3g/L, manganese chloride 3-4g/L, glycine betaine 0.5-1.2g/L, pH are 6.8~7.2.
It is further preferred that the group of fermentation medium becomes：Sucrose 110g/L, yeast extract 12g/L, sodium nitrate 10g/
L, ammonium nitrate 7g/L, potassium dihydrogen phosphate 8g/L, dipotassium hydrogen phosphate 4g/L, magnesium sulfate 3g/L, frerrous chloride 2g/L, zinc sulfate 3g/
L, manganese chloride 4g/L, glycine betaine 1g/L.
Preferably, the dosage of defoaming agent is 0.01~0.03wt% of total material in fermentor.
Preferably, the concentration of initial glucose is 18-22g/L.
Preferably, in second stage, the concentration of glucose is divided into two-stage control, specifically, the 0-20 hour of fermentation,
The concentration for controlling glucose is 8-10g/L, and the 21-40 hour of fermentation, the concentration for controlling glucose is 3-6g/L.It is controlled using this kind
System can more effectively realize high density fermentation, suit the practical upgrowth situation of strain.
According to the present invention, in fermentation process, with the water of sterile water supplement evaporation, and keep fermentor tank pressure for 0.03~
0.10MPa。
High density fermentation in the present invention is a relative concept, refers generally in culture solution engineering bacteria stand density in 50g/
L（DWC）More than.
In the present invention, the parameter limited and processing step is not added to select ordinary skill in the art hand in fermentation process
Section, for example, material additive amount, strain inoculum concentration etc. in fermentor.
In the present invention, conventional use of separating-purifying side in the prior art is can be used in separation and Extraction riboflavin from fermentation liquid
Method.
Compared with prior art, the present invention has following beneficial to point：
The present invention by screening strain, limit seed culture medium and fermentation medium, and processing step to fermentation and parameter into
It has gone restriction, has realized high density fermentation vitamin B2, improve vitamin B2Yield.
Specific embodiment
Technical solution of the present invention is clearly and completely described below in conjunction with the embodiment of the present invention, it is clear that retouched
The embodiment stated is only a part of the embodiment of the present invention, instead of all the embodiments.
Embodiment 1-3 is for illustrating high density fermentation vitamin B of the invention2Method.
Embodiment 1
A kind of high density fermentation vitamin B2Method, using bacillus subtilis as fermentation strain, by high cell density fermentation,
Improve vitamin B2Fermentation level, include the following steps：
1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized,
At 37~40 DEG C, 120~140rpm, 15~18h of aerobic culture obtains seed culture fluid；The group of seed culture medium becomes：Sugarcane
Sugared 18g/L, sodium nitrate 8g/L, ammonium nitrate 5g/L, epsom salt 1g/L, potassium dihydrogen phosphate 0.8g/L, dipotassium hydrogen phosphate 1.5g/
L, yeast extract 3g/L, frerrous chloride 1g/L, zinc sulfate 1g/L, manganese chloride 2.5g/L, pH are 6.7~6.9；
2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, is defoamed
The dosage of agent is the 0.01wt% of total material in fermentor, and the group of fermentation medium becomes：Sucrose 100g/L, yeast extract
10g/L, sodium nitrate 8g/L, ammonium nitrate 6g/L, potassium dihydrogen phosphate 7g/L, dipotassium hydrogen phosphate 3g/L, magnesium sulfate 2g/L, frerrous chloride
1.5g/L, zinc sulfate 2g/L, manganese chloride 3g/L, glycine betaine 0.5g/L, pH are 6.8~7.0, and the concentration of initial glucose is 15g/
L；
Fermentation is divided into two stages progress, and in the first stage, keeping fermentation temperature is 38~40 DEG C, with liquefied ammonia adjust pH be 6.9~
7.1, control ventilation ratio is 1 ︰ 0.8, and dissolved oxygen amount is made to reach 20~30%, and fermentation keeps glucose consumption complete in 6 hours, and it is small to continue fermentation 2
When；In second stage, keeping fermentation temperature is 37~39 DEG C, and adjusting pH is 7.0~7.2, and control ventilation ratio is 1：1.2, control
Dissolved oxygen amount adds sterile dextrose liquid, the 0-20 hour of fermentation, the concentration for controlling glucose is 8-10g/L, hair 25~30%
The 21-40 hour of ferment, the concentration for controlling glucose is 3-6g/L, until bacterial strain aging, the glucose consumption added is complete, terminates hair
Ferment；In fermentation process, with the water of sterile water supplement evaporation, and keeping fermentor tank pressure is 0.05MPa；
3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.
Wherein, the bacillus subtilis slant strains of use are made by following methods：By bacillus subtilis original strain
Bacterium solution aseptically carry out ultraviolet mutagenesis, obtain ultraviolet mutagenesis bacterial strain, then carry out chemical mutagenesis with ethylmethane sulfonate,
Obtain complex mutation bacterial strain；Then complex mutation bacterial strain is successively inoculated into metabolism substrate analog 8-anaguanine, 8- azepine
Adenine, 6- azathymine, 6- thioguanine, DL- METHIONINE sulfoxide, rose flavine minimal medium in trained
Resistant mutant strain is made in domesticating；Processed strain inoculated is cultivated into nutrient medium, obtained nutrition culture
Object inoculates to be cultivated in strain fermentation culture medium, filters out production vitamin B2Highest bacterial strain.
Using vitamin B obtained by the above method2Fermentation titer be 28.05g/L.
Embodiment 2
A kind of high density fermentation vitamin B2Method, using bacillus subtilis as fermentation strain, by high cell density fermentation,
Improve vitamin B2Fermentation level, include the following steps：
1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized,
At 37~40 DEG C, 120~140rpm, 15~18h of aerobic culture obtains seed culture fluid；The group of seed culture medium becomes：Sugarcane
Sugared 20g/L, sodium nitrate 10g/L, ammonium nitrate 7g/L, epsom salt 1.5g/L, potassium dihydrogen phosphate 1g/L, dipotassium hydrogen phosphate 2g/
L, yeast extract 4g/L, frerrous chloride 2g/L, zinc sulfate 2g/L, manganese chloride 4g/L, pH are 6.7~6.9；
2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, is defoamed
The dosage of agent is the 0.03wt% of total material in fermentor, and the group of fermentation medium becomes：Sucrose 110g/L, yeast extract
12g/L, sodium nitrate 10g/L, ammonium nitrate 7g/L, potassium dihydrogen phosphate 8g/L, dipotassium hydrogen phosphate 4g/L, magnesium sulfate 3g/L, protochloride
Iron 2g/L, zinc sulfate 3g/L, manganese chloride 4g/L, glycine betaine 1g/L, pH are 6.8~7.0, and the concentration of initial glucose is 20g/L；
Fermentation is divided into two stages progress, and in the first stage, keeping fermentation temperature is 38~40 DEG C, with liquefied ammonia adjust pH be 6.9~
7.1, control ventilation ratio is 1 ︰ 0.8, and dissolved oxygen amount is made to reach 20~30%, and fermentation keeps glucose consumption complete in 8 hours, and it is small to continue fermentation 2
When；In second stage, keeping fermentation temperature is 37~39 DEG C, and adjusting pH is 7.0~7.2, and control ventilation ratio is 1：1.5, control
Dissolved oxygen amount adds sterile dextrose liquid, the 0-20 hour of fermentation, the concentration for controlling glucose is 8-10g/L, hair 25~30%
The 21-40 hour of ferment, the concentration for controlling glucose is 3-6g/L, until bacterial strain aging, the glucose consumption added is complete, terminates hair
Ferment；In fermentation process, with the water of sterile water supplement evaporation, and keeping fermentor tank pressure is 0.06MPa；
3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.
Wherein, the bacillus subtilis slant strains of use are made by following methods：By bacillus subtilis original strain
Bacterium solution aseptically carry out ultraviolet mutagenesis, obtain ultraviolet mutagenesis bacterial strain, then carry out chemical mutagenesis with ethylmethane sulfonate,
Obtain complex mutation bacterial strain；Then complex mutation bacterial strain is successively inoculated into metabolism substrate analog 8-anaguanine, 8- azepine
Adenine, 6- azathymine, 6- thioguanine, DL- METHIONINE sulfoxide, rose flavine minimal medium in trained
Resistant mutant strain is made in domesticating；Processed strain inoculated is cultivated into nutrient medium, obtained nutrition culture
Object inoculates to be cultivated in strain fermentation culture medium, filters out production vitamin B2Highest bacterial strain.
Using vitamin B obtained by the above method2Fermentation titer be 29.23g/L.
Embodiment 3
A kind of high density fermentation vitamin B2Method, using bacillus subtilis as fermentation strain, by high cell density fermentation,
Improve vitamin B2Fermentation level, include the following steps：
1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized,
At 37~40 DEG C, 120~140rpm, 15~18h of aerobic culture obtains seed culture fluid；The group of seed culture medium becomes：Sugarcane
Sugared 22g/L, sodium nitrate 10g/L, ammonium nitrate 7g/L, epsom salt 1g/L, potassium dihydrogen phosphate 1.2g/L, dipotassium hydrogen phosphate
2.5g/L, yeast extract 5g/L, frerrous chloride 3g/L, zinc sulfate 2g/L, manganese chloride 3g/L, pH are 6.7~6.9；
2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, is defoamed
The dosage of agent is the 0.02wt% of total material in fermentor, and the group of fermentation medium becomes：Sucrose 120g/L, yeast extract
14g/L, sodium nitrate 10g/L, ammonium nitrate 7g/L, potassium dihydrogen phosphate 7g/L, dipotassium hydrogen phosphate 3g/L, magnesium sulfate 4g/L, protochloride
Iron 2g/L, zinc sulfate 3g/L, manganese chloride 4g/L, glycine betaine 1g/L, pH are 6.8~7.0, and the concentration of initial glucose is 25g/L；
Fermentation is divided into two stages progress, and in the first stage, keeping fermentation temperature is 38~40 DEG C, with liquefied ammonia adjust pH be 6.9~
7.1, control ventilation ratio is 1 ︰ 0.8, and dissolved oxygen amount is made to reach 20~30%, and fermentation keeps glucose consumption complete in 10 hours, continues fermentation 2
Hour；In second stage, keeping fermentation temperature is 37~39 DEG C, and adjusting pH is 7.0~7.2, and control ventilation ratio is 1：1.2, control
Dissolved oxygen amount processed adds sterile dextrose liquid 25~30%, and the 0-20 hour of fermentation, the concentration for controlling glucose is 8-10g/L,
The 21-40 hour of fermentation, the concentration for controlling glucose is 3-6g/L, until bacterial strain aging, the glucose consumption added is complete, terminates
Fermentation；In fermentation process, with the water of sterile water supplement evaporation, and keeping fermentor tank pressure is 0.03MPa；
3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.
Wherein, the bacillus subtilis slant strains of use are made by following methods：By bacillus subtilis original strain
Bacterium solution aseptically carry out ultraviolet mutagenesis, obtain ultraviolet mutagenesis bacterial strain, then carry out chemical mutagenesis with ethylmethane sulfonate,
Obtain complex mutation bacterial strain；Then complex mutation bacterial strain is successively inoculated into metabolism substrate analog 8-anaguanine, 8- azepine
Adenine, 6- azathymine, 6- thioguanine, DL- METHIONINE sulfoxide, rose flavine minimal medium in trained
Resistant mutant strain is made in domesticating；Processed strain inoculated is cultivated into nutrient medium, obtained nutrition culture
Object inoculates to be cultivated in strain fermentation culture medium, filters out production vitamin B2Highest bacterial strain.
Using vitamin B obtained by the above method2Fermentation titer be 27.35g/L.
The embodiment of the present invention is described above, above description is exemplary, and non-exclusive, and also not
It is limited to disclosed embodiment.Without departing from the scope and spirit of illustrated embodiment, for the art
Many modifications and changes are obvious for those of ordinary skill.
Claims (8)
1. a kind of high density fermentation vitamin B2Method pass through high density fermentation work using bacillus subtilis as fermentation strain
Skill improves vitamin B2Fermentation level, which is characterized in that include the following steps：
1）The bacillus subtilis slant strains for taking fresh activation are inoculated in the seeding tank equipped with seed culture medium that sterilized,
At 37~42 DEG C, 120~140rpm, 12~18h of aerobic culture obtains seed culture fluid；The group of seed culture medium becomes：Sugarcane
Sugared 18-24g/L, sodium nitrate 8-12g/L, ammonium nitrate 5-8g/L, epsom salt 1-2g/L, potassium dihydrogen phosphate 0.8-1.2g/L,
Dipotassium hydrogen phosphate 1.5-2.5g/L, yeast extract 3-5g/L, frerrous chloride 1-3g/L, zinc sulfate 1-3g/L, manganese chloride 2.5-
5g/L, pH are 6.7~6.9；
2）Seed culture fluid is moved into the fermentor equipped with fermentation medium, defoaming agent and Glucose Liquid and starts to ferment, is fermented
The group of culture medium becomes：Sucrose 90-120g/L, yeast extract 10-14g/L, sodium nitrate 8-12g/L, ammonium nitrate 6-8g/L, phosphorus
Acid dihydride potassium 7-9g/L, dipotassium hydrogen phosphate 3-5g/L, magnesium sulfate 2-4g/L, frerrous chloride 1.5-3g/L, zinc sulfate 2-4g/L, chlorine
Change manganese 3-5g/L, glycine betaine 0.2-1.5g/L, pH are 6.8~7.2, and the concentration of initial glucose is 15-25g/L；
Fermentation is divided into two stages progress, and in the first stage, keeping fermentation temperature is 37~40 DEG C, with liquefied ammonia adjust pH be 6.8~
7.2, control ventilation ratio is 1 ︰ 0.6~0.9, and dissolved oxygen amount is made to reach 15~30%, and fermentation keeps glucose consumption complete, continues the 2-3 that ferments
Hour；In second stage, keeping fermentation temperature is 37~39 DEG C, and adjusting pH is 7.0~7.2, and control ventilation ratio is 1：1.0~
1.5, control dissolved oxygen amount adds sterile dextrose liquid 20~30%, concentration of glucose is controlled between 5-15g/L, until bacterial strain
Aging, the glucose consumption added is complete, terminates fermentation；
3）Step 2）Obtained fermentation liquid enters abstraction process and extracts riboflavin.
2. high density fermentation vitamin B according to claim 12Method, which is characterized in that the bacillus subtilis of use
Bacterium slant strains are made by following methods：The bacterium solution of bacillus subtilis original strain is aseptically subjected to ultraviolet lure
Become, obtains ultraviolet mutagenesis bacterial strain, then carry out chemical mutagenesis with ethylmethane sulfonate, obtain complex mutation bacterial strain；It then will be compound
Mutagenic strain is successively inoculated into metabolism substrate analog 8-anaguanine, 8- azaadenine, 6- azathymine, 6- sulfydryl
Guanine, DL- METHIONINE sulfoxide, rose flavine minimal medium in carry out culture domestication, be made resistant mutant strain；It will processing
The strain inoculated crossed is cultivated into nutrient medium, obtained vegetative cultures inoculate in strain fermentation culture medium into
Row culture, filters out production vitamin B2Highest bacterial strain.
3. high density fermentation vitamin B according to claim 12Method, which is characterized in that the composition of seed culture medium
For：Sucrose 18-20g/L, sodium nitrate 9-11g/L, ammonium nitrate 6-8g/L, epsom salt 1-1.5g/L, potassium dihydrogen phosphate 0.9-
1g/L, dipotassium hydrogen phosphate 1.5-2g/L, yeast extract 4-5g/L, frerrous chloride 1-2g/L, zinc sulfate 2-3g/L, manganese chloride 3-
5g/L, pH are 6.7~6.9.
4. high density fermentation vitamin B according to claim 12Method, which is characterized in that the composition of fermentation medium
For：Sucrose 110-120g/L, yeast extract 12-14g/L, sodium nitrate 9-10g/L, ammonium nitrate 7-8g/L, potassium dihydrogen phosphate 8-
9g/L, dipotassium hydrogen phosphate 3-4g/L, magnesium sulfate 2-3g/L, frerrous chloride 2-3g/L, zinc sulfate 2-3g/L, manganese chloride 3-4g/L,
Glycine betaine 0.5-1.2g/L, pH are 6.8~7.2.
5. high density fermentation vitamin B according to claim 12Method, which is characterized in that the dosage of defoaming agent be hair
0.01~0.03wt% of total material in fermentation tank.
6. high density fermentation vitamin B according to claim 12Method, which is characterized in that the concentration of initial glucose
For 18-22g/L.
7. high density fermentation vitamin B according to claim 12Method, which is characterized in that in second stage, fermentation
0-20 hours, the concentration for controlling glucose was 8-10g/L, and the 21-40 hour of fermentation, the concentration for controlling glucose is 3-6g/L.
8. high density fermentation vitamin B according to claim 12Method, which is characterized in that in fermentation process, with sterile
The water of water supplement evaporation, and keeping fermentor tank pressure is 0.03~0.10MPa.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402210A (en) * | 2018-12-12 | 2019-03-01 | 宁夏启元药业有限公司 | Culture medium and method used in a kind of riboflavin clean manufacturing |
| CN109609580A (en) * | 2018-12-26 | 2019-04-12 | 河南巨龙生物工程股份有限公司 | A kind of fermentation medium and its fermentation process of riboflavin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402210A (en) * | 2018-12-12 | 2019-03-01 | 宁夏启元药业有限公司 | Culture medium and method used in a kind of riboflavin clean manufacturing |
| CN109609580A (en) * | 2018-12-26 | 2019-04-12 | 河南巨龙生物工程股份有限公司 | A kind of fermentation medium and its fermentation process of riboflavin |
| CN109609580B (en) * | 2018-12-26 | 2022-04-01 | 河南巨龙生物工程股份有限公司 | Fermentation medium and fermentation method of riboflavin |
| CN109837322A (en) * | 2019-03-27 | 2019-06-04 | 山东泓达生物科技有限公司 | A kind of simple fermentation medium using bacillus subtilis production vitamin B2 |
| CN110564804A (en) * | 2019-09-12 | 2019-12-13 | 河南巨龙生物工程股份有限公司 | Clear liquid fermentation medium for producing riboflavin and fermentation method |
| CN112280679A (en) * | 2020-11-04 | 2021-01-29 | 赤峰制药股份有限公司 | Method for producing vitamin B2 by fermentation method |
| CN113249261A (en) * | 2021-05-26 | 2021-08-13 | 浙江新和成股份有限公司 | Bacillus subtilis and application thereof in production of riboflavin |
| CN113801910A (en) * | 2021-08-30 | 2021-12-17 | 湖北广济药业股份有限公司 | Preparation method of pure natural riboflavin |
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