CN108048367A - White staphylococcus powder and its application - Google Patents
White staphylococcus powder and its application Download PDFInfo
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- CN108048367A CN108048367A CN201810072108.XA CN201810072108A CN108048367A CN 108048367 A CN108048367 A CN 108048367A CN 201810072108 A CN201810072108 A CN 201810072108A CN 108048367 A CN108048367 A CN 108048367A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N1/20—Bacteria; Culture media therefor
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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Abstract
The invention belongs to microbial technology fields, disclose a kind of White staphylococcus powder, and being inoculated into wheat bran medium fermented and cultured by white grape bacterium seed liquor is made.White staphylococcus powder of the present invention is prepared using special zymotechnique, of low cost, can be applied to prepare antitussive medicine.
Description
Technical field
The invention belongs to microbial technology fields, more particularly to White staphylococcus powder and its application.
Background technology
White staphylococcus powder is a kind of oral bacterial preparation of non-pathogenic, is to be trained by non-pathogenic White staphylococcus liquid
The dry thalline of inactivation obtained by supporting is made.Experiment shows that it has the antibechic intensity of approximate codeine, directly acts on medulla oblongata cough
It coughs maincenter, apparent to acute cough's antitussive effect, dosage is bigger, and antitussive effect is stronger, and has adjusting autonomic nervous system function,
Amount of expectoration can be significantly reduced.Fermentation method prepares unique way that White staphylococcus powder is pharmacy corporation production White staphylococcus powder
Footpath, but the cost of culture medium restricts the development of enterprise, at present compared with multiple enterprises using dextrose peptone medium, raw material into
This height, cell density, the profit of enterprise substantially reduces." with the optimised process item of solution fermentation production Vaccinum Bronehitidis piece
Part is studied, Shandong medical industry 1999 ", a kind of White staphylococcus culture medium is disclosed, including peptone 0.5%, urea 2%,
Corn pulp 1%, glucose 1%, sodium citrate 0.3%, magnesium sulfate 0.05% and sodium chloride 0.5%, the culture medium cost is higher, and
And the cell concentration obtained only reaches 109CFU/mL ranks.It is me to develop a kind of White staphylococcus powder preparation technique of low cost
The technical issues that need to address.
Wheat bran（Wheat bran）It is the principal by product of flour processing, in the past, wheat bran is primarily used to animal feed, economical
Value is not high, also has many producers arbitrarily to abandon wheat bran.Wheat bran contain abundant carbohydrate, protein,
Vitamin E, phenolic compound and inorganic mineral etc.（The functional component of wheat bran and its application study progress, Jia Tianguang, food
Research and development 2014）, be gradually applied to other field, including health products, food additives, extraction dietary fiber and
Dietary proteins etc., possess the prospect of being widely applied and Development volue.But due to wheat bran complicated component, treatment process is difficult to
It grasps.In Bacteria Culture field, existing research carries out simple process, the carbon source as bacteria culture media using wheat bran mostly
Part complements, for reducing the cost of culture medium；There are no occur using wheat bran as the white grape of primary raw material so far
Coccus culture medium.
The content of the invention
Present invention aim to address prior art White staphylococcus fermentation costs it is high the defects of, provide using new fermentation work
White staphylococcus powder and its application prepared by skill.
The present invention is achieved by the following technical solution：
A kind of White staphylococcus powder is inoculated into wheat bran medium fermented and cultured by white grape bacterium seed liquor and is made.
Further,
The wheat bran medium is prepared according to following technique：
（1）Wheat bran is ground, 100 mesh sieves are crossed, then with 1：15 material water quality ratio mixing, adds neutral starch enzyme,
Additive amount is 0.6U/g wheat bran, handles 15min under 70 DEG C of temperature conditionss, is then centrifuged for, and collects precipitation and supernatant liquid, will be upper
Layer liquid is placed under conditions of 90 DEG C, is kept the temperature 10min enzyme deactivations, is cooled to room temperature, obtains component A；
（2）Hydrochloric acid of the concentration for being added to 5 times of weight for 3M will be precipitated, when 80 DEG C of stirring hydrolysis 9 are small, mixing speed is
Filtrate is collected by filtration in 100rpm, then with, with remaining hydrochloric acid, adjustment pH value of solution is 7.5, obtains component B in ammonium hydroxide；
（3）Component A and component B are uniformly mixed so as to obtain wheat bran medium.
Further,
The White staphylococcus powder is prepared according to following technique：
1）White staphylococcus liquid is taken, is inoculated into according to the inoculum concentration of 2-3% in seed culture medium, 30-32 DEG C, the training of 100rpm shaking tables
18-24h is supported, obtains seed liquor；
2）Seed liquor is inoculated according to the inoculum concentration of 3-5% in the fermentation tank equipped with wheat bran medium and carries out fermented and cultured, is cultivated
30-32 DEG C of temperature, it is 20% that throughput, which keeps dissolved oxygen level, and passes through and add glucose control sugar content not less than 0.5wt%,
Fermented and cultured 48-60h obtains White staphylococcus zymotic fluid；
3）Into White staphylococcus zymotic fluid add formaldehyde inactivated, then by high-speed dish piece machine with the speed of 5000rpm from
Heart 6-8min, collect bacterial sediment, vacuum freeze drying to get.
Preferably,
The component of the seed culture medium is：Glucose 12g/L, agar 7g/L, peptone 3g/L, KH2PO40.5g/L,
K2HPO4 0.5g/L, MgSO40.1g/L, FeSO4 0.1g/L, pH7.5；
Preferably,
It is described inactivation parameter be：Control formaldehyde final concentration of 0.5wt%, 25 DEG C of inactivation 48h.
A kind of technique for preparing White staphylococcus powder is also claimed in the present invention.
Application of the White staphylococcus powder of above-mentioned preparation in antitussive medicine is prepared is also claimed in the present invention.
The advantageous effect that the present invention obtains mainly includes but is not limited to the following aspects：
White staphylococcus powder of the present invention is prepared using special zymotechnique, of low cost, and the added value of industry is high；Wheat bran
Skin contains abundant carbohydrate, protein, vitamin E and inorganic mineral, and the present invention has carried out special work to wheat bran
Skill processing, makes the nutriment of wheat bran release to greatest extent, and abundant carbon source, nitrogen source are contained in processed material, is dissociated
Amino acid, small peptide, vitamin and inorganic mineral comply fully with the growth demand of White staphylococcus, and culture effect is better than normal
The culture medium of rule, and it is of low cost, cost of material is saved to pharmacy corporation, improves the added value of industry；Wheat bran of the present invention
Enzymolysis processing is impregnated, obtains reduced sugar as carbon source using pulverizing and sieving first in treatment process, and dimension life will not be destroyed
Element and other active materials, while avoid the destruction of acid-base pair glucide；Contain protein in sediment, the present invention adopts
With using acid hydrolysis mode, proteolysis rate obtains more than 50%, then using alkali neutralization, while balance pH is spent, increases simultaneously
The supply of ammonium chloride nitrogen source is added, has killed two birds with one stone；Fermentation medium of the present invention is of low cost only with wheat bran, can take completely
Culture medium is often used for market, fermentation efficiency also accordingly improves.
Description of the drawings
Fig. 1：Influence of the temperature to concentration of reduced sugar；
Fig. 2：Influence of the enzyme additive amount to concentration of reduced sugar.
Specific embodiment
Those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.In particular, it should be pointed out that
All similar substitutions and modifications are apparent to those skilled in the art, they are considered as being included in this hair
It is bright.The product and method of the present invention is described by preferred embodiment, and related personnel can substantially not depart from this hair
Product as described herein and method are modified or suitably changed with combining in bright content, spirit and scope, to realize and answer
Use the technology of the present invention.For a further understanding of the present invention, with reference to embodiment, the present invention is described in detail.
Embodiment 1
A kind of White staphylococcus powder is inoculated into wheat bran medium fermented and cultured by white grape bacterium seed liquor and is made；
Specifically, the White staphylococcus powder is prepared according to following technique：
1）Take White staphylococcus liquid（5×107CFU/mL）, it is inoculated into according to 3% inoculum concentration in seed culture medium, 30 DEG C,
100rpm shaking table culture 18h obtain seed liquor；The component of the seed culture medium is：Glucose 12g/L, agar 7g/L, albumen
Peptone 3g/L, KH2PO4 0.5g/L, K2HPO4 0.5g/L, MgSO40.1g/L, FeSO4 0.1g/L, pH7.5；
2）Seed liquor is inoculated according to 5% inoculum concentration in the fermentation tank equipped with 50L wheat bran mediums and carries out fermented and cultured, is trained
30 DEG C of temperature is supported, it is 20% that throughput, which keeps dissolved oxygen level, and passes through and add glucose control sugar content not less than 0.2wt%, hair
Ferment culture 54h, obtains White staphylococcus zymotic fluid；
The wheat bran medium is prepared according to following technique：
（1）Wheat bran is ground, 100 mesh sieves are crossed, then with 1：15 material water quality ratio mixing, adds neutral starch enzyme,
Additive amount is 0.6U/g wheat bran, handles 15min under 70 DEG C of temperature conditionss, is then centrifuged for, and collects precipitation and supernatant liquid, will be upper
Layer liquid is placed under conditions of 90 DEG C, is kept the temperature 10min enzyme deactivations, is cooled to room temperature, obtains component A；
（2）Hydrochloric acid of the concentration for being added to 5 times of weight for 3M will be precipitated, when 80 DEG C of stirring hydrolysis 9 are small, mixing speed is
Filtrate is collected by filtration in 100rpm, then with, with remaining hydrochloric acid, adjustment pH value of solution is 7.5, obtains component B in ammonium hydroxide；
（3）Component A and component B are uniformly mixed so as to obtain wheat bran medium；
3）Formaldehyde is added into White staphylococcus zymotic fluid, final concentration of 0.5wt%, 25 DEG C of inactivation 48h is controlled, then passes through at a high speed
Disk machine centrifuges 8min with the speed of 5000rpm, collects bacterial sediment, vacuum freeze drying to get.
Embodiment 2
A kind of White staphylococcus powder is inoculated into wheat bran medium fermented and cultured by white grape bacterium seed liquor and is made；
Specifically, the White staphylococcus powder is prepared according to following technique：
1）Take White staphylococcus liquid（7×107CFU/mL）, it is inoculated into according to 2% inoculum concentration in seed culture medium, 32 DEG C,
100rpm shaking table cultures for 24 hours, obtain seed liquor；The component of the seed culture medium is：Glucose 12g/L, agar 7g/L, albumen
Peptone 3g/L, KH2PO4 0.5g/L, K2HPO4 0.5g/L, MgSO40.1g/L, FeSO4 0.1g/L, pH7.5；
2）Seed liquor is inoculated according to 3% inoculum concentration in the fermentation tank equipped with 50L wheat bran mediums and carries out fermented and cultured, is trained
32 DEG C of temperature is supported, it is 20% that throughput, which keeps dissolved oxygen level, and passes through and add glucose control sugar content not less than 0.2wt%, hair
Ferment culture 48h, obtains White staphylococcus zymotic fluid；
The wheat bran medium is prepared according to following technique：
（1）Wheat bran is ground, 100 mesh sieves are crossed, then with 1：15 material water quality ratio mixing, adds neutral starch enzyme,
Additive amount is 0.6U/g wheat bran, handles 15min under 70 DEG C of temperature conditionss, is then centrifuged for, and collects precipitation and supernatant liquid, will be upper
Layer liquid is placed under conditions of 90 DEG C, is kept the temperature 10min enzyme deactivations, is cooled to room temperature, obtains component A；
（2）Hydrochloric acid of the concentration for being added to 5 times of weight for 3M will be precipitated, when 80 DEG C of stirring hydrolysis 9 are small, mixing speed is
Filtrate is collected by filtration in 100rpm, then with, with remaining hydrochloric acid, adjustment pH value of solution is 7.5, obtains component B in ammonium hydroxide；
（3）Component A and component B are uniformly mixed so as to obtain wheat bran medium；
3）Formaldehyde is added into White staphylococcus zymotic fluid, final concentration of 0.5wt%, 25 DEG C of inactivation 48h is controlled, then passes through at a high speed
Disk machine centrifuges 6min with the speed of 5000rpm, collects bacterial sediment, vacuum freeze drying to get.
Embodiment 3
The content of reduced sugar in nitro-salicylic acid colorimetric determination component A：
By taking embodiment 1 as an example, the influence of temperature and enzyme concentration to content of reducing sugar is had detected, set temperature gradient is：40℃、
50℃、60℃、70℃、80℃；Set enzyme concentration gradient be：0.2U/g、0.4U/g、0.6U/g、0.8U/g、1.0U/g.Such as Fig. 1
Shown, when temperature is less than 70 DEG C, with the rise of temperature, the content of reduced sugar increases, and at 70 DEG C or so, hydrolyzes
The content highest of reduced sugar, the high level of comparison can be reached by illustrating the vigor of the medium temperature amylase at 70 DEG C；And reach 80 DEG C
Temperature reduction sugared content drastically reduce, this may be because high temperature makes the activity reduction of medium temperature amylase, while may also companion
With the consumption of substantial amounts of reduced sugar.As shown in Fig. 2, constantly increase with the additive amount of medium temperature amylase, starch in wheat bran
Hydrolysis degree constantly increases, and content of reducing sugar is continuously improved, when the additive amount of enzyme is 0.6U/g, content of reducing sugar highest；With
It enzyme additive amount to continue to increase, the incrementss of reduced sugar are not obvious, finally tend to be steady；In view of cost, final choice
The additive amount of 0.6U/g.
Embodiment 4
Influence of the different culture media used in zymotechnique of the present invention to bacteria concentration in zymotic fluid：
By taking embodiment 1 as an example, 1 fermentation medium of embodiment is wheat bran medium；Control group is set simultaneously：
Control group 1：Fermentation medium uses（Mass percent）：Peptone 0.5%, urea 2%, corn pulp 1%, glucose 1%, lemon
Lemon acid sodium 0.3%, magnesium sulfate 0.05% and sodium chloride 0.5%.Remaining technique is the same as embodiment 1.
Control group 2：Fermentation medium uses（Mass percent）：Ammonium chloride 1%, dusty yeast 1%, corn pulp 2%, glucose
2%, sodium chloride 0.05%, ferrous sulfate 0.01%, magnesium sulfate 0.01%.Remaining technique is the same as embodiment 1.
Control group 3：Fermentation medium uses（Mass percent）：Ammonium chloride 0.5%, dregs of beans 2%, wheat bran 1%, glucose 2%,
Sodium chloride 0.02%, ferrous sulfate 0.01%, magnesium sulfate 0.01%.Remaining technique is the same as embodiment 1.Have detected respectively embodiment 1 and
Bacteria concentration in zymotic fluid prepared by control group 1-3 sets three parallel groups per group, averages, concrete outcome is shown in Table 1：
Table 1
| Group | Embodiment 1 | Control group 1 | Control group 2 | Control group 3 |
| Bacteria concentration（CFU/mL） | 1.2×1011 | 9.7×109 | 4.1×1010 | 1.9×1010 |
Conclusion：The present invention, as fermentation medium, uses wheat bran as primary raw material, containing abundant using wheat bran medium
Carbon source, nitrogen source, amino acid, small peptide, vitamin and inorganic mineral comply fully with the growth demand of White staphylococcus, culture
Effect is better than conventional culture medium, and of low cost, saves cost of material.
Although above having used general explanation, specific embodiment and experiment, the present invention is described in detail,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (7)
1. a kind of White staphylococcus powder is inoculated into wheat bran medium fermented and cultured by white grape bacterium seed liquor and is made.
2. White staphylococcus powder according to claim 1, which is characterized in that the wheat bran medium is according to following technique system
It is standby and obtain：
（1）Wheat bran is ground, 100 mesh sieves are crossed, then with 1：15 material water quality ratio mixing, adds neutral starch enzyme,
Additive amount is 0.6U/g wheat bran, handles 15min under 70 DEG C of temperature conditionss, is then centrifuged for, and collects precipitation and supernatant liquid, will be upper
Layer liquid is placed under conditions of 90 DEG C, is kept the temperature 10min enzyme deactivations, is cooled to room temperature, obtains component A；
（2）Hydrochloric acid of the concentration for being added to 5 times of weight for 3M will be precipitated, when 80 DEG C of stirring hydrolysis 9 are small, mixing speed is
Filtrate is collected by filtration in 100rpm, then with, with remaining hydrochloric acid, adjustment pH value of solution is 7.5, obtains component B in ammonium hydroxide；
（3）Component A and component B are uniformly mixed so as to obtain wheat bran medium.
3. White staphylococcus powder according to claim 1 or 2, which is characterized in that the White staphylococcus powder is according to as follows
Technique is prepared：
1）White staphylococcus liquid is taken, is inoculated into according to the inoculum concentration of 2-3% in seed culture medium, 30-32 DEG C, the training of 100rpm shaking tables
18-24h is supported, obtains seed liquor；
2）Seed liquor is inoculated according to the inoculum concentration of 3-5% in the fermentation tank equipped with wheat bran medium and carries out fermented and cultured, is cultivated
30-32 DEG C of temperature, it is 20% that ventilation, which keeps dissolved oxygen level, and passes through and add glucose control sugar content not less than 0.5wt%, hair
Ferment culture 48-60h, obtains White staphylococcus zymotic fluid；
3）Into White staphylococcus zymotic fluid add formaldehyde inactivated, then by high-speed dish piece machine with the speed of 5000rpm from
Heart 6-8min, collect bacterial sediment, vacuum freeze drying to get.
4. White staphylococcus powder according to claim 3, which is characterized in that
The component of the seed culture medium is：Glucose 12g/L, agar 7g/L, peptone 3g/L, KH2PO40.5g/L,
K2HPO4 0.5g/L, MgSO40.1g/L, FeSO4 0.1g/L, pH7.5.
5. White staphylococcus powder according to claim 3, which is characterized in that it is described inactivation parameter be：Control formaldehyde dense eventually
It spends for 0.5wt%, 25 DEG C of inactivation 48h.
6. a kind of technique for preparing the White staphylococcus powder described in claim 1-5.
7. claim 1-5 is allowed to application of the White staphylococcus powder in antitussive medicine is prepared described in one.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468068A (en) * | 2019-08-19 | 2019-11-19 | 仁和堂药业有限公司 | A kind of preparation process of White staphylococcus powder |
| CN110499268A (en) * | 2019-08-19 | 2019-11-26 | 仁和堂药业有限公司 | White staphylococcus powder and its application |
| CN116531410A (en) * | 2023-07-06 | 2023-08-04 | 首都医科大学附属北京友谊医院 | Application of staphylococcus albus in preparation of composition |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468068A (en) * | 2019-08-19 | 2019-11-19 | 仁和堂药业有限公司 | A kind of preparation process of White staphylococcus powder |
| CN110499268A (en) * | 2019-08-19 | 2019-11-26 | 仁和堂药业有限公司 | White staphylococcus powder and its application |
| CN116531410A (en) * | 2023-07-06 | 2023-08-04 | 首都医科大学附属北京友谊医院 | Application of staphylococcus albus in preparation of composition |
| CN116531410B (en) * | 2023-07-06 | 2023-10-03 | 首都医科大学附属北京友谊医院 | Application of staphylococcus albus in preparation of composition |
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Application publication date: 20180518 |