CN108192835A - A kind of probiotics strain preparation and its cultural method - Google Patents
A kind of probiotics strain preparation and its cultural method Download PDFInfo
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- CN108192835A CN108192835A CN201711354283.XA CN201711354283A CN108192835A CN 108192835 A CN108192835 A CN 108192835A CN 201711354283 A CN201711354283 A CN 201711354283A CN 108192835 A CN108192835 A CN 108192835A
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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
- 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
- C12N1/205—Bacterial isolates
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- C12R2001/225—Lactobacillus
- C12R2001/245—Lactobacillus casei
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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
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Abstract
The invention discloses a kind of probiotics strain preparations, and the probiotics strain is obtained by the fermenting raw materials as follows in terms of mass percentage：Skimmed milk power 11.5 12.5%, white sugar 4.5 5.5%, glucose 6.5 7.2%, Lactobacillus casei 0.50 0.55 ‰ and surplus purified water；The sum of above-mentioned raw materials mass percentage is 100%.The present invention also provides a kind of cultural methods of probiotics strain preparation.Increase skimmed milk power, white sugar, glucose in the probiotics strain preparation of the present invention and provide suitable nutrient matrix for Lactobacillus casei so that Lactobacillus casei can rapidly enter logarithmic phase and ferment, and put into production, and can effectively improve production efficiency, save cost；Improve the active lactic acid bacterium number of product.
Description
Technical field
The present invention relates to a kind of probiotics more particularly to a kind of probiotics strain preparations and its cultural method.
Background technology
Strain is the microorganism as living-cell catalyst for fermentation process, including bacterium, actinomyces, saccharomycete and mould
Four major class of bacterium from a large amount of microorganism of nature, therefrom through detaching and filtering out useful strain, then improved, is stored
It is ready to use in production.Probiotics is a kind of active microorganism beneficial to host, is to be colonized in human body intestinal canal, in reproductive system, energy
Generate active beneficial microorganism general name of the definite health efficacy so as to improve host's microecological balance, performance beneficial effect.People
Beneficial bacterium or fungi mainly have in body, animal body：Clostridium butyricum, lactobacillus, Bifidobacterium, actinomyces, saccharomycete etc..Mesh
The most powerful product of the function of being studied on former world be mainly above all kinds of microorganism groups into composite reactive probiotics, extensively
Applied to bioengineering, industrial or agricultural, food security and life and health field.
It is powdered in direct putting type using strain in food industry at present, it is made using dry dormant technology, strain access base
Experience activation is needed in material, activationary time is long, causes the production cycle long, low production efficiency.
Invention content
For overcome the deficiencies in the prior art, one of the objects of the present invention is to provide a kind of probiotics strain preparations, should
Increase skimmed milk power, white sugar, glucose in probiotics strain preparation and provide suitable nutrient matrix for Lactobacillus casei so that is dry
Lactobacillus paracasei can rapidly enter logarithmic phase and ferment, and put into production, and can effectively improve production efficiency, save cost；It improves
The active lactic acid bacterium number of product.
Second object of the present invention is to provide for a kind of cultural method of probiotics strain preparation, this method pass through by
Tradition accesses and proliferation is cultivated in special nutrient matrix, plant active lactic acid bacterium number in liquid and be rapidly achieved logarithm in suspend mode shape strain
Phase is produced.
An object of the present invention adopts the following technical scheme that realization：
A kind of probiotics strain preparation, the probiotics strain is obtained by the fermenting raw materials as follows in terms of mass percentage：
The sum of above-mentioned raw materials mass percentage is 100%.
Further, the total solids content in the raw material is 22-24%.
Realize that second object of the present invention can reach by adopting the following technical scheme that：
A kind of cultural method of probiotics strain preparation, including,
Water-quality control step：By Drinking Water by triple filter and reverse osmosis, it is made as purified water；
Material addition step：Purified water is warming up to 50-60 DEG C, skimmed milk power, white granulated sugar and glucose, unlatching is then put into and follows
Ring pump circulation mixes material；
Dissolving step：Stop circulating pump after feeding intake, stirring makes material obtain milk after fully dissolving recovery, in 50-
55 DEG C of heat preservations；
Homogenizing step：Milk after heat preservation is warming up to 60-65 DEG C, then carries out homogeneous；
Sterilization steps：Milk after homogeneous is warming up to 95-97 DEG C, milk is all pumped into kind of a tank, is kept the temperature in kind of tank,
Then it stirs, 35-37 DEG C is cooled to after stirring；
Inoculation step：Lactobacillus casei is poured into the milk of sterilizing, be slowly stirred uniformly, obtain fermenation raw liquid；
Fermentation step：By fermenation raw liquid in 35-37 DEG C of culture, incubation time 26-30h terminates fermentation, obtains zymotic fluid；
Cryopreservation step：Zymotic fluid is moved into 2-6 DEG C of cold-storage jar and is refrigerated, cold preservation time is less than or equal to 7 days, after detection is up to standard
Obtain probiotics strain preparation.
Further, in Water-quality control step, the pH value of purified water is 6.60-6.90.
Further, in dissolving step, start stirring 1 time per 20min, stir 3 times altogether, each mixing time 30s.
Further, in dissolving step, soaking time 60min.
Further, in homogenizing step, homogenization pressure：Level-one is 4MPa, two level 20MPa.
Further, in sterilization steps, soaking time 100min.
Further, in sterilization steps, stirring is opened per 20min once, is stirred 5 times altogether, each mixing time 30s.
Further, in fermentation step, when acidity is 130-150 ° of T, pH value is 3.80-4.00, fermentation is terminated.
The beneficial effects of the present invention are：
1st, probiotics strain preparation of the invention, increasing skimmed milk power, white sugar, glucose in the probiotics strain preparation is
Lactobacillus casei provides suitable nutrient matrix so that Lactobacillus casei can rapidly enter logarithmic phase and ferment, input life
Production can effectively improve production efficiency, save cost；Improve the active lactic acid bacterium number of product；
2nd, the cultural method of probiotics strain preparation of the present invention, this method is by the way that tradition in suspend mode shape strain, is accessed special
It cultivates and is proliferated in the nutrient matrix of system, active lactic acid bacterium number is rapidly achieved logarithmic phase and is produced in kind liquid.
Specific embodiment
In the following, with reference to specific embodiment, the present invention is described further, it should be noted that is do not collided
Under the premise of, new embodiment can be formed between various embodiments described below or between each technical characteristic in any combination.
A kind of probiotics strain preparation, the probiotics strain is obtained by the fermenting raw materials as follows in terms of mass percentage：
The sum of above-mentioned raw materials mass percentage is 100%.
As the mode that further carries out, the total solids content in raw material is 22-24%.
The cultural method of above-mentioned probiotics strain preparation, including,
Water-quality control step：By Drinking Water by triple filter and counter-infiltration system, it is made as purified water；
Material addition step：Purified water is warming up to 50-60 DEG C, skimmed milk power, white granulated sugar and glucose, unlatching is then put into and follows
Ring pump circulation mixes material；
Dissolving step：Stop circulating pump after feeding intake, stirring makes material obtain milk after fully dissolving recovery, in 50-
55 DEG C of heat preservations；
Homogenizing step：Milk after heat preservation is warming up to 60-65 DEG C, then carries out homogeneous；
Sterilization steps：Milk after homogeneous is warming up to 95-97 DEG C, milk is all pumped into kind of a tank, is kept the temperature in kind of tank,
Then it stirs, 35-37 DEG C is cooled to after stirring；
Inoculation step：Lactobacillus casei is poured into the milk of sterilizing, be slowly stirred uniformly, obtain fermenation raw liquid；
Fermentation step：By fermenation raw liquid in 35-37 DEG C of culture, incubation time 26-30h terminates fermentation, obtains zymotic fluid；
Cryopreservation step：Zymotic fluid is moved into 2-6 DEG C of cold-storage jar and is refrigerated, cold preservation time is less than or equal to 7 days, after detection is up to standard
Obtain probiotics strain preparation.
As the mode that further carries out, in Water-quality control step, the pH value of purified water is 6.60-6.90.
As the mode that further carries out, in dissolving step, start stirring 1 time per 20min, stir 3 times altogether, stir every time
Time 30s.
As the mode that further carries out, in dissolving step, soaking time 60min.
As the mode that further carries out, in homogenizing step, homogenization pressure：Level-one is 4MPa, two level 20MPa.
As the mode that further carries out, in sterilization steps, soaking time 100min.
As the mode that further carries out, in sterilization steps, it is primary to open stirring per 20min, stirs 5 times, stirs every time altogether
Mix time 30s.
As the mode that further carries out, in fermentation step, when acidity is 130-150 ° of T, pH value is 3.80-4.00,
Terminate fermentation.
Gill Li Erdu (° T) represents the acidity of milk：° T refers to titration 100mL milk samples, consumes the NaOH of 0.1mol/L
Amount.
Wherein, in inoculation step, Lactobacillus casei uses freeze-drying lyophilization granular bacteria strain, in use, by freeze-drying lyophilization particle
Strain is introduced directly into the milk of sterilizing.Through it was discovered by researchers that can be easy to operation using freeze-drying lyophilization granular bacteria strain, reducing
The probability of cross contamination shortens the time into adjustment period, rapidly enters logarithmic phase.
It is specific embodiment of the present invention below, used raw material, equipment etc. remove special limit in the following embodiments
It can be obtained outside fixed by buying pattern.
Embodiment 1-3：
The probiotics strain preparation of embodiment 1-3, the probiotics strain are sent out by the raw material as follows in terms of mass percentage
Obtained by ferment：
1 embodiment 1-3 probiotics strains preparation of table matches
The probiotics strain preparation of embodiment 1-3 is carried out using following cultural method, and method includes,
Water-quality control step：Drinking Water is passed through into triple filter and counter-infiltration system, is made as purified water, purified water
PH value is 6.60-6.90；
Material addition step：Purified water is warming up to 50-60 DEG C, skimmed milk power, white granulated sugar and glucose, unlatching is then put into and follows
Ring pump circulation mixes material；
Dissolving step：Stop circulating pump after feeding intake, stirring makes material obtain milk, dissolving step after fully dissolving recovery
In rapid, start stirring 1 time per 20min, stir 3 times, each mixing time 30s altogether, then at 50-55 DEG C of heat preservation 60min；
Homogenizing step：Milk after heat preservation is warming up to 60-65 DEG C, then carries out homogeneous, first progress level-one homogeneous, then into
Row double-stage homogenization, homogenization pressure：Level-one is 4MPa, two level 20MPa；
Sterilization steps：Milk after homogeneous is warming up to 95-97 DEG C, milk is all pumped into kind of a tank, is kept the temperature in kind of tank
Then 100min is stirred, stirring is opened per 20min once, is stirred 5 times altogether, each mixing time 30s is cooled to 35- after stirring
37℃；
Inoculation step：Lactobacillus casei is poured into the milk of sterilizing, be slowly stirred uniformly, obtain fermenation raw liquid；
Fermentation step：By fermenation raw liquid in 35-37 DEG C of culture, incubation time 26-30h, when acidity is 130-150 ° of T, pH
When being worth for 3.80-4.00, fermentation is terminated, obtains zymotic fluid；
Cryopreservation step：Zymotic fluid is moved into 2-6 DEG C of cold-storage jar and is refrigerated, cold preservation time is less than or equal to 7 days, after detection is up to standard
Obtain probiotics strain preparation.
Effect assessment and performance detection
First, embodiment 1-3 microorganisms detection situation
2 embodiment 1-3 microorganisms of table detect situation
| Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Total plate count (cfu/ml) | <1 | <1 | <1 |
| Saccharomycete, mould (cfu/ml) | <1 | <1 | <1 |
| Lactic acid bacteria number Cfu/ml | 7.4×109 | 6.9×109 | 7.3×109 |
2nd, the effect after implementing compares
With the original species without activation process in embodiment 1, added in physiological saline after outer packing disinfection and suspension is made
Production as a comparison case 1 is directly used in input milk.This strain market price：25g/720 members, comparative example 1 and comparative example 1
Effect.
3 implementation result of table compares
As can be seen from Table 3, the cost of probiotics strain preparation of the invention is much smaller than the cost of the strain of comparative example 1.
3rd, fermentation time and terminal acidity change
By the original species without activation process respectively of the Lactobacillus casei in the strain preparation of embodiment 1-3, through outer packing
It adds in physiological saline to be made in suspension input milk after disinfection and comparative example 1-3, comparing embodiment 1-3 and comparative example 1- is made
3 fermentation time changes with terminal acidity.
4 fermentation time of table and terminal acidity result of variations
By table 4 as it can be seen that the strain fermentation time of embodiment 1-3 is short, and fermentation stability, terminal acidity fluctuates small, lactic acid bacteria
Number is 1.0 × 109More than cfu/ml, the quality of product are secure；And the strain fermentation time of comparative example 1-3 is long, terminal acid
Spend relatively low, the lactic acid bacteria number of product is on the low side, and production process quality fluctuation is big.
To sum up, increase skimmed milk power, white sugar, glucose in probiotics strain preparation of the invention to provide for Lactobacillus casei
Suitable nutrient matrix so that Lactobacillus casei can rapidly enter logarithmic phase and ferment, and cultural method by being in by tradition
Suspend mode shape strain accesses and proliferation is cultivated in special nutrient matrix, puts into production, and can effectively improve production efficiency, saves cost；
Improve the active lactic acid bacterium number of product.
The above embodiment is only the preferred embodiment of the present invention, it is impossible to the scope of protection of the invention is limited with this,
The variation and replacement for any unsubstantiality that those skilled in the art is done on the basis of the present invention belong to institute of the present invention
Claimed range.
Claims (10)
1. a kind of probiotics strain preparation, which is characterized in that the probiotics strain is by the raw material as follows in terms of mass percentage
Fermentation gained：
The sum of above-mentioned raw materials mass percentage is 100%.
2. probiotics strain preparation as described in claim 1, which is characterized in that the total solids content in the raw material is
22-24%.
3. a kind of cultural method of probiotics strain preparation as claimed in claim 1 or 2, it is characterised in that including,
Water-quality control step：By Drinking Water by triple filter and counter-infiltration system, it is made as purified water；
Material addition step：Purified water is warming up to 50-60 DEG C, then puts into skimmed milk power, white granulated sugar and glucose, open the circulation pump
Cycle mixing material；
Dissolving step：Stop circulating pump after feeding intake, stirring makes material obtain milk after fully dissolving recovery, in 50-55 DEG C
Heat preservation；
Homogenizing step：Milk after heat preservation is warming up to 60-65 DEG C, then carries out homogeneous；
Sterilization steps：Milk after homogeneous is warming up to 95-97 DEG C, milk is all pumped into kind of a tank, is kept the temperature in kind of tank, then
Stirring, 35-37 DEG C is cooled to after stirring；
Inoculation step：Lactobacillus casei is poured into the milk of sterilizing, be slowly stirred uniformly, obtain fermenation raw liquid；
Fermentation step：By fermenation raw liquid in 35-37 DEG C of culture, incubation time 26-30h terminates fermentation, obtains zymotic fluid；
Cryopreservation step：Zymotic fluid is moved into 2-6 DEG C of cold-storage jar and is refrigerated, cold preservation time is less than or equal to 7 days, is obtained after detection is up to standard
Probiotics strain preparation.
4. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that pure in Water-quality control step
The pH value for changing water is 6.60-6.90.
5. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in dissolving step, per 20min
Start stirring 1 time, stir 3 times altogether, each mixing time 30s.
6. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in dissolving step, during heat preservation
Between be 60min.
7. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in homogenizing step, homogeneous pressure
Power：Level-one is 4MPa, two level 20MPa.
8. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in sterilization steps, during heat preservation
Between be 100min.
9. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in sterilization steps, per 20min
It is primary to open stirring, stirs 5 times altogether, each mixing time 30s.
10. the cultural method of probiotics strain preparation as claimed in claim 3, which is characterized in that in fermentation step, work as acidity
When for 130-150 ° of T, pH value being 3.80-4.00, fermentation is terminated.
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2017
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| CN102361969A (en) * | 2009-03-31 | 2012-02-22 | 株式会社益力多本社 | Method for culturing lactic acid bacteria, and a food and drink product |
| CN104152354A (en) * | 2013-05-14 | 2014-11-19 | 仙农生物科技(上海)有限公司 | Compound probiotics as well as production process and application thereof to leavening agent |
| CN103734309A (en) * | 2013-12-19 | 2014-04-23 | 山东得益乳业股份有限公司 | Brown lactobacillus beverage and preparation method thereof |
| CN104938769A (en) * | 2015-07-27 | 2015-09-30 | 江西省科学院微生物研究所 | Method for preparing rice bran strain preparation and applying rice bran strain preparation to feed fermentation |
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Application publication date: 20180622 |