CN109943506A - A kind of culture medium and its fermentation process of suitable bacillus coagulans BC01 - Google Patents
A kind of culture medium and its fermentation process of suitable bacillus coagulans BC01 Download PDFInfo
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- CN109943506A CN109943506A CN201910227895.5A CN201910227895A CN109943506A CN 109943506 A CN109943506 A CN 109943506A CN 201910227895 A CN201910227895 A CN 201910227895A CN 109943506 A CN109943506 A CN 109943506A
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- fermentation
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Abstract
The invention discloses the culture medium of suitable bacillus coagulans BC01 a kind of, each component by weight percentage are as follows: lactose: 1%-5%, glucose: 2%-5%, peptone: 1%-3%, yeast powder: 1%-3%, magnesium sulfate: 0.1%-1%, manganese sulfate: 0.01%-1%, Tween 80: 0.1%-1%, biotin: 0.1%-1%, calcium chloride: 0.2%-1%.For the present invention under specific fermentation condition, the cell density in fermentation liquid can achieve the CFU/mL of 2,000,000,000 CFU/mL~7,000,000,000.And when cell density reaches maximum, it is gemma body cell that there are also enough nutrition by nutrition somatic cell transformation.In fermentation latter stage, the brood-gemma rate of fermentation liquid is greater than 85%.
Description
Technical field
The invention belongs to fermentation engineering fields, more specifically to a kind of suitable bacillus coagulans BC01's
The culture medium of high density fermentation and high yield gemma rate.
Background technique
Bacillus coagulans BC01 is that our company screen from Pickles, Sichuan Style one plant has very big prebiotic potentiality
Bacterial strain.
Bacillus coagulans BC01 belongs to bacillus on taxology, and there are two kinds of life shapes in its life cycle
State:
(1) bacillus coagulans can be with growth and breeding and the various metabolites of generation under the form for nutrition volume morphing.
But the bacillus coagulans anti-adversity of nutrition volume morphing is poor, and the ability of non-refractory, acid and alkali-resistance and chemical reagent also compares
It is weaker.
(2) the gemma volume morphing form is the suspend mode form of bacillus coagulans, and thallus cannot be grown when in the form
Breeding, but resistance with super strength, high temperature resistant, acid and alkali-resistance, and nutrition can be re-converted into when nutritional condition is suitable
Volume morphing plays beneficial function.
Therefore, when bacillus coagulans are applied to health industry, we wishes that its product form is brood-gemma, in this way
Convenient for the storage of product, transport and the application scenarios for expanding product.When gemma posture bacillus coagulans by people take in into
It after in vivo, can be sprouted again in small intestine and large intestine, be transformed into nutrition volume morphing and play beneficial function.
Therefore, when preparing coagulated bacillus living powder, the gemma rate for making great efforts to improve fermentation liquid thallus just seems
It is particularly important.But bacillus coagulans produce gemma and need specific fermentation condition and nutritional condition, under specific fermentation condition,
Nutritional condition determines cell density and spore forming rate in fermentation liquid.Nutritional condition is too abundant to be difficult to form gemma, battalion
The cell density of the too poor then fermentation liquid of feeding condition is low, high production cost and also is difficult to form gemma.
Summary of the invention
In view of the deficiencies of the prior art, the present invention intends to provide a kind of suitable bacillus coagulans BC01
Culture medium, under specific fermentation condition, the cell density in fermentation liquid can achieve the CFU/ of 2,000,000,000 CFU/mL~7,000,000,000
mL.And when cell density reaches maximum, it is gemma body cell that there are also enough nutrition by nutrition somatic cell transformation.It is fermenting
The brood-gemma rate in latter stage, fermentation liquid is greater than 85%.
To achieve the above object, the present invention provides the following technical scheme that
A kind of culture medium of suitable bacillus coagulans BC01, it is characterised in that: its each component by weight percentage are as follows:
Lactose: 1%-5%, glucose: 2%-5%, peptone: 1%-3%, yeast powder: 1%-3%, magnesium sulfate: 0.1%-1%, sulphur
Sour manganese: 0.01%-1%, Tween 80: 0.1%-1%, biotin: 0.1%-1%, calcium chloride: 0.2%-1%.
As a kind of technical solution of optimization, each component by weight percentage are as follows:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%；
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, sulphur
Sour manganese 0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%.
As a kind of technical solution of optimization, each component by weight percentage are as follows:
Seed culture medium: lactose 3%, glucose 3%, peptone 2%, yeast powder 2%, magnesium sulfate 0.3%, manganese sulfate
0.11%, Tween 80: 0.2%, biotin 0.2%；
Fermentation medium: lactose 5%, glucose 5%, peptone 3%, yeast powder 2%, magnesium sulfate 0.8%, manganese sulfate
0.45%, Tween 80: 0.6%, biotin 0.3%, calcium chloride 0.5%.
The present invention also provides a kind of fermentation process of the culture medium of suitable bacillus coagulans BC01, it is characterised in that:
Steps are as follows:
(1) seed culture medium and fermentation medium are configured:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%；
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, sulphur
Sour manganese 0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%；
After configuration, culture medium initial pH to 6.5 is adjusted, 121 DEG C sterilize 20 minutes.
(2) seed culture medium inoculation and culture
When seed culture medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into from glycerol tube a certain amount of
Seed culture medium, 0.5%, 40 DEG C of inoculum concentration, shaking speed: 220rpm, culture is to seed OD562 > 4.0.
(3) inoculation and culture of fermentation medium
When fermentation medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into hair from seed culture medium
Ferment culture medium；Inoculum concentration 0.5%, fermentation temperature: 40 DEG C, revolving speed 300rpm, ventilatory capacity: 15L/min, fermentation pH:5.8, fermentation
Process DO > 85%.
(4) under this fermentation condition and fermentation medium, the final gemma bulk concentration of the fermentation liquid of bacillus coagulans BC01
It can be with 3,000,000,000 CFU/mL of >, gemma rate > 85%.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
In the present invention, lactose is the crystallinity particle or powder of white；It is odorless, it is slightly sweet.Sugariness is the 15% of sucrose, and
Beta lactose is bigger than the sugariness of alpha-lactose.In the market there are many lactose supply: anhydrous alpha-lactose, alpha-lactose monohydrate and cry on a small quantity
Anhydrous beta lactose.
Glucose has critical role in field of biology, be living cells energy source and metabolic intermediate product,
That is biological main Energy supply material, in candy making industry and field of medicaments extensive application.
Peptone be will it is dry after meat, casein or gelatin acid or protease hydrolytic made of appearance in flaxen powder
Agent, the special breath with meat perfume.Protein can also form peptone after acid, alkali or proteases for decomposing.In stomach internal protein
One of protopepsia product be exactly peptone.Peptone is rich in organonitrogen compound, also contains some vitamins and carbohydrate.It
The primary raw material that can be used as microbiological culture media, in antibiotic, medical industry, fermentation industry, biochemical product and microbiology
Dosage in the fields such as scientific research is very big.
Tween 80, i.e., are as follows: polyoxyethylene sorbitan monooleate, abbreviation Tween-80.Tween 80 is soluble in
Water is dissolved in ethyl alcohol, vegetable oil, ethyl acetate, methanol, toluene, does not dissolve in mineral oil.Agglutination when low temperature is restored after heated.Have
Special smell, mildly bitter flavor are dispersed with certain help to phthalocyanine green.
Biotin, i.e., well known biotin, it can make body by food conversion self-energy.Biology is known as
Conducive to the health and regeneration of cell.
Calcium chloride can increase the permeability of bacteria cell wall.
By adopting the above-described technical solution, compared with prior art, the present invention is under specific fermentation condition, fermentation liquid
In cell density can achieve the CFU/mL of 2,000,000,000 CFU/mL~7,000,000,000.And when cell density reaches maximum, there are also enough
Nutrition by nutrition somatic cell transformation be gemma body cell.In fermentation latter stage, the brood-gemma rate of fermentation liquid is greater than 85%.
Referring to drawings and examples, the present invention will be further described.
Detailed description of the invention
Fig. 1 is that the bacillus coagulans trophosome and brood-gemma viable count of fermentation process in an embodiment of the present invention change
Tendency chart；
Fig. 2 is the gemma rate sediments microscope inspection picture of bacillus coagulans BC01 in an embodiment of the present invention.
Specific embodiment
Embodiment
A kind of culture medium of suitable bacillus coagulans BC01, by weight percentage each component are as follows:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%.
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, sulphur
Sour manganese 0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%.
Above-mentioned weight percent is the optimal proportion in normal temperature and pressure.If it is in highlands, air pressure compared with
Low, oxygen content is told somebody what one's real intentions are, and optimal mass percent is:
Seed culture medium: lactose 3%, glucose 3%, peptone 2%, yeast powder 2%, magnesium sulfate 0.3%, manganese sulfate
0.11%, Tween 80: 0.2%, biotin 0.2%；
Fermentation medium: lactose 5%, glucose 5%, peptone 3%, yeast powder 2%, magnesium sulfate 0.8%, manganese sulfate
0.45%, Tween 80: 0.6%, biotin 0.3%, calcium chloride 0.5%.
Highlands is only specific use.
A kind of fermentation process of the culture medium of suitable bacillus coagulans BC01, it is characterised in that:
Steps are as follows:
(1) seed culture medium and fermentation medium are configured:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%；
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, sulphur
Sour manganese 0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%；
After configuration, culture medium initial pH to 6.5 is adjusted, 121 DEG C sterilize 20 minutes.
(2) seed culture medium inoculation and culture
When seed culture medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into from glycerol tube a certain amount of
Seed culture medium, 0.5%, 40 DEG C of inoculum concentration, shaking speed: 220rpm, culture is to seed OD562 > 4.0.
(3) inoculation and culture of fermentation medium
When fermentation medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into hair from seed culture medium
Ferment culture medium；Inoculum concentration 0.5%, fermentation temperature: 40 DEG C, revolving speed 300rpm, ventilatory capacity: 15L/min, fermentation pH:5.8, fermentation
Process DO > 85%.
(4) under this fermentation condition and fermentation medium, the final gemma bulk concentration of the fermentation liquid of bacillus coagulans BC01
It can be with 3,000,000,000 CFU/mL of >, gemma rate > 85%.
Attached drawing 1 is the bacillus coagulans trophosome and brood-gemma viable count trend chart of fermentation process in step (3).
When attached drawing 2 is that fermentation was to 20 hours in step (4), the gemma rate sediments microscope inspection figure of bacillus coagulans BC01
Piece.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
For the present invention under specific fermentation condition, the cell density in fermentation liquid can achieve 2,000,000,000 CFU/mL~7,000,000,000
CFU/mL.And when cell density reaches maximum, it is gemma body cell that there are also enough nutrition by nutrition somatic cell transformation.?
It ferments latter stage, the brood-gemma rate of fermentation liquid is greater than 85%.
Protection scope of the present invention is not limited merely to above-described embodiment, and all technical solutions belonged under thinking of the present invention are equal
It belongs to the scope of protection of the present invention.It should be pointed out that for those skilled in the art, not departing from the present invention
Several improvements and modifications under the premise of principle, these modifications and embellishments should also be considered as the scope of protection of the present invention.
Claims (4)
1. a kind of culture medium of suitable bacillus coagulans BC01, it is characterised in that: its each component by weight percentage are as follows:
Lactose: 1%-5%, glucose: 2%-5%, peptone: 1%-3%, yeast powder: 1%-3%, magnesium sulfate: 0.1%-
1%, manganese sulfate: 0.01%-1%, Tween 80: 0.1%-1%, biotin: 0.1%-1%, calcium chloride: 0.2%-1%.
2. the culture medium of suitable bacillus coagulans BC01 according to claim 1 a kind of, it is characterised in that:
Its each component by weight percentage are as follows:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%；
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, manganese sulfate
0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%.
3. the culture medium of suitable bacillus coagulans BC01 according to claim 1 a kind of, it is characterised in that:
Its each component by weight percentage are as follows:
Seed culture medium: lactose 3%, glucose 3%, peptone 2%, yeast powder 2%, magnesium sulfate 0.3%, manganese sulfate
0.11%, Tween 80: 0.2%, biotin 0.2%；
Fermentation medium: lactose 5%, glucose 5%, peptone 3%, yeast powder 2%, magnesium sulfate 0.8%, manganese sulfate
0.45%, Tween 80: 0.6%, biotin 0.3%, calcium chloride 0.5%.
4. a kind of fermentation process of the culture medium of suitable bacillus coagulans BC01 according to claim 1, feature exist
In:
Steps are as follows:
(1) seed culture medium and fermentation medium are configured:
Seed culture medium: lactose 1%, glucose 2%, peptone 1%, yeast powder 1%, magnesium sulfate 0.1%, manganese sulfate
0.01%, Tween 80: 0.1%, biotin 0.1%；
Fermentation medium: lactose 2%, glucose 2%, peptone 2.5%, yeast powder 2.5%, magnesium sulfate 0.5%, manganese sulfate
0.05%, Tween 80: 0.5%, biotin 0.2%, calcium chloride 0.5%；
After configuration, culture medium initial pH to 6.5 is adjusted, 121 DEG C sterilize 20 minutes；
(2) seed culture medium inoculation and culture
When seed culture medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into a certain amount of kind from glycerol tube
Sub- culture medium, 0.5%, 40 DEG C of inoculum concentration, shaking speed: 220rpm, culture to seed OD562 > 4.0；
(3) inoculation and culture of fermentation medium
When fermentation medium temperature is down to 45 DEG C or less, bacillus coagulans BC01 is inoculated into fermentation training from seed culture medium
Support base；Inoculum concentration 0.5%, fermentation temperature: 40 DEG C, revolving speed 300rpm, ventilatory capacity: 15L/min, fermentation pH:5.8, fermentation process
DO > 85%；
(4) under this fermentation condition and fermentation medium, the final gemma bulk concentration of the fermentation liquid of bacillus coagulans BC01 can be with
3,000,000,000 CFU/mL of >, gemma rate > 85%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114836349A (en) * | 2022-05-13 | 2022-08-02 | 江苏新申奥生物科技有限公司 | Lactobacillus acidophilus LA16 for antagonizing helicobacter pylori and application thereof |
| CN115125171A (en) * | 2022-07-06 | 2022-09-30 | 呼伦贝尔东北阜丰生物科技有限公司 | High-temperature culture process of glutamic acid zymocyte |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114836349A (en) * | 2022-05-13 | 2022-08-02 | 江苏新申奥生物科技有限公司 | Lactobacillus acidophilus LA16 for antagonizing helicobacter pylori and application thereof |
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| CN115125171A (en) * | 2022-07-06 | 2022-09-30 | 呼伦贝尔东北阜丰生物科技有限公司 | High-temperature culture process of glutamic acid zymocyte |
| CN115125171B (en) * | 2022-07-06 | 2023-06-27 | 呼伦贝尔东北阜丰生物科技有限公司 | High-temperature culture process of glutamic acid fermentation bacteria |
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