CN107629968A - A kind of purification of fermented bean curd strain and its application process - Google Patents
A kind of purification of fermented bean curd strain and its application process Download PDFInfo
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- CN107629968A CN107629968A CN201711043872.6A CN201711043872A CN107629968A CN 107629968 A CN107629968 A CN 107629968A CN 201711043872 A CN201711043872 A CN 201711043872A CN 107629968 A CN107629968 A CN 107629968A
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Abstract
The present invention relates to strain purification technique field, and in particular to a kind of purification of fermented bean curd strain and its application process, comprises the following steps：S1, sample preparation：Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, then the fluid sample in physiological saline is inoculated in sterile PDA plate culture medium, is cultivated 34 days；S2, sample dilution：1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate culture medium, cultivate 34 days；S3, sample are cultivated again：The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 34 days；And in the form of geometry multiple, purifying culture respectively, until obtaining sufu fermentation probioticses；S4, strain idenfication：The strain that S3 is obtained carries out DNA molecular spiral measure, is defined as sufu fermentation probioticses M60；The strain on original fermented bean curd can be utilized, is isolated and purified, in the case of ensureing sanitary condition, can be produced throughout the year.
Description
Technical field
The present invention relates to strain purification technique field, and in particular to a kind of purification of fermented bean curd strain and its application process.
Background technology
Fermented bean curd is also known as fermented bean curd, is the Traditional Folk cuisines that thousands of years are spread in China, because its is in good taste, nutrition is high, it is deep by
China common people and the south east asia people's likes, is unfailing delicious food together.Fermented bean curd is the tune of China's original creation
Taste product, there are the kinds such as preserved red beancurd, green-rot breast, white rot breast, sauce fermented bean curd, pattern fermented bean curd, it both can individually eat, it is also possible to cook
The dish of unique flavor.Fermented bean curd is the healthy food that nutritionist praises highly energetically.Its raw material --- dried bean curd is originally
The very high bean product of nutritive value, protein content is suitable with meat up to 15%~20%, simultaneously containing abundant calcareous.It is rotten
The fermentation of mould is have passed through in the manufacturing process of breast, makes that the digestibility of protein is higher, and vitamin content is more rich.Because
Phytic acid in microorganism decomposition beans so that the mineral matter such as the very low iron of script absorptivity, zinc is easier by human body in soybean
Absorb.Simultaneously as the Microbe synthesis unexistent vitamin B12 of general vegetable food, the people of vegetarian diet often eat a little corruption
Breast, pernicious anaemia can be prevented.
But existing sufu fermentation is all to carry out fresh bean curd fermentation, still, Ci Zhongfa using the natural strain on straw
Ferment mode, climate have a great influence, it is impossible to meet the needs of fermented bean curd produces throughout the year, in addition, such a fermentation method sanitary condition compared with
Difference, easily polluted by other bacteriums.
The content of the invention
It is an object of the invention to provide a kind of purification of fermented bean curd strain and its application process, can utilize on original fermented bean curd
Strain, isolated and purified, ensure sanitary condition in the case of, can produce throughout the year.
To solve above-mentioned technical problem, the present invention uses following technical scheme：
A kind of method of purification of fermented bean curd strain, comprises the following steps：
S1, sample preparation：
Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, the fluid sample in physiological saline is then inoculated in nothing
In bacterium PDA plate culture medium, cultivate 3-4 days；
S2, sample dilution：
1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate
In culture medium, cultivate 3-4 days；
S3, sample are cultivated again：
The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 3-4 days；And with geometry times
Number form formula, purifying culture respectively, until obtaining sufu fermentation probioticses；
Further, in S1 and S2 steps, the mass concentration of physiological saline is 0.85%.
Further, in S1, S2 and S3 step, the condition of culture is 16-32 DEG C of temperature, humidity 60%.
Further, the condition of strain M60 daily preservation is：16-28 DEG C of temperature, humidity 60%.
Further, the condition of strain M60 long-term preservation is：6-12 DEG C of temperature, humidity 60%.
A kind of fermented bean curd strain application process, in gnotobasis, the strain M60 of culture is inoculated in 0.85% physiology salt
In water, 20min is stirred, spore suspension is prepared into, the spore suspension of preparation is inoculated in soya-bean milk, glucose and sterilized water
In fluid nutrient medium, cultivate 2 days, 16-32 DEG C of temperature, obtain liquid spawn, the liquid spawn of preparation is placed under microscope and surveyed
Spore concentration is determined, until spore concentration is 107-108, choose concentration strain up to standard and be sprayed on fresh bean curd, after sprinkling strain
Fresh bean curd fermented under the conditions of 16-30 DEG C 48-60h, complete fermentation.
Further, strain M60 accesses amount is the 0.5% of sterile water volume.
Further, the volume ratio of soya-bean milk and sterilized water is 1:30.
Further, glucose addition accounts for the 4% of sterile water volume.
Compared with prior art, the present invention one of at least has the advantages that：
1. the single culture skill beneficial to fermented bean curd later stage fermentation is obtained after being isolated and purified using the strain adhered on original fermented bean curd
Art, the limitation of season and weather etc. is broken through, long-term continuous production can be realized.
2. cultivated using the strain on original fermented bean curd, it is possible to increase the mouthfeel and quality of fermented bean curd.
3. the entrance of foreign matter can be reduced, microorganism pollution is avoided, increases cleanliness factor.
4. shortening fermenting step and production cycle, increase benefit.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment 1
A kind of method of purification of fermented bean curd strain, comprises the following steps：
S1, sample preparation：
Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, the fluid sample in physiological saline is then inoculated in nothing
In bacterium PDA plate culture medium, cultivate 3 days；
S2, sample dilution：
1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate
In culture medium, cultivate 3 days；
S3, sample are cultivated again：
The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 3 days；And with geometry multiple
Form, purifying culture respectively, until obtaining sufu fermentation probioticses；
S4, strain idenfication：
The strain that S3 is obtained carries out DNA molecular spiral measure, is defined as sufu fermentation probioticses M60.
The single culture beneficial to fermented bean curd later stage fermentation is obtained after being isolated and purified using the strain adhered on original fermented bean curd
Technology, the limitation of season and weather etc. is broken through, long-term continuous production can be realized.
Embodiment 2
A kind of method of purification of fermented bean curd strain, comprises the following steps：
S1, sample preparation：
Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, the fluid sample in physiological saline is then inoculated in nothing
In bacterium PDA plate culture medium, cultivate 3.5 days；
S2, sample dilution：
1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate
In culture medium, cultivate 3.5 days；
S3, sample are cultivated again：
The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 3.5 days；And with geometry times
Number form formula, purifying culture respectively, until obtaining sufu fermentation probioticses；
S4, strain idenfication：
The strain that S3 is obtained carries out DNA molecular spiral measure, is defined as sufu fermentation probioticses M60.
The single culture beneficial to fermented bean curd later stage fermentation is obtained after being isolated and purified using the strain adhered on original fermented bean curd
Technology, the limitation of season and weather etc. is broken through, long-term continuous production can be realized.
Embodiment 3
A kind of method of purification of fermented bean curd strain, comprises the following steps：
S1, sample preparation：
Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, the fluid sample in physiological saline is then inoculated in nothing
In bacterium PDA plate culture medium, cultivate 4 days；
S2, sample dilution：
1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate
In culture medium, cultivate 4 days；
S3, sample are cultivated again：
The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 4 days；And with geometry multiple
Form, purifying culture respectively, until obtaining sufu fermentation probioticses；
S4, strain idenfication：
The strain that S3 is obtained carries out DNA molecular spiral measure, is defined as sufu fermentation probioticses M60.
The single culture beneficial to fermented bean curd later stage fermentation is obtained after being isolated and purified using the strain adhered on original fermented bean curd
Technology, the limitation of season and weather etc. is broken through, long-term continuous production can be realized.
Embodiment 4
On the basis of embodiment 1-3, in S1 and S2 steps, the mass concentration of physiological saline is 0.85%, the suitable physiology of concentration
Salt solution can increase the survival rate of strain.
Embodiment 5
On the basis of embodiment 1-4, in S1, S2 and S3 step, the condition of culture is 16 DEG C of temperature, humidity 60%, then this training
Under the conditions of supporting, the rate of vaccination of strain can be increased.
Embodiment 6
On the basis of embodiment 1-4, in S1, S2 and S3 step, the condition of culture is 25 DEG C of temperature, humidity 60%, then this training
Under the conditions of supporting, the rate of vaccination of strain can be increased.
Embodiment 7
On the basis of embodiment 1-4, in S1, S2 and S3 step, the condition of culture is 32 DEG C of temperature, humidity 60%, then this training
Under the conditions of supporting, the rate of vaccination of strain can be increased.
Embodiment 8
On the basis of embodiment 1-7, the condition of strain M60 daily preservation is：16 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 9
On the basis of embodiment 1-7, the condition of strain M60 daily preservation is：22 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 10
On the basis of embodiment 1-7, the condition of strain M60 daily preservation is：28 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 11
On the basis of embodiment 1-7, the condition of strain M60 long-term preservation is：6 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 12
On the basis of embodiment 1-7, the condition of strain M60 long-term preservation is：9 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 13
On the basis of embodiment 1-7, the condition of strain M60 long-term preservation is：12 DEG C of temperature, humidity 60%, can extend
The holding time of strain.
Embodiment 14
A kind of fermented bean curd strain application process, in gnotobasis, the strain M60 of culture is inoculated in 0.85% physiological saline,
20min is stirred, spore suspension is prepared into, the spore suspension of preparation is inoculated in the liquid of soya-bean milk, glucose and sterilized water
In culture medium, cultivate 2 days, 16 DEG C of temperature, obtain liquid spawn, it is dense that the liquid spawn of preparation is placed in into measure spore under microscope
Degree, until spore concentration is 107-108, choose concentration strain up to standard and be sprayed on fresh bean curd, by the fresh bean curd after sprinkling strain
Ferment 48h under the conditions of 16 DEG C, completes fermentation, is cultivated using the strain on original fermented bean curd, it is possible to increase the mouthfeel of fermented bean curd
And quality.
Embodiment 15
A kind of fermented bean curd strain application process, in gnotobasis, the strain M60 of culture is inoculated in 0.85% physiological saline,
20min is stirred, spore suspension is prepared into, the spore suspension of preparation is inoculated in the liquid of soya-bean milk, glucose and sterilized water
In culture medium, cultivate 2 days, 26 DEG C of temperature, obtain liquid spawn, it is dense that the liquid spawn of preparation is placed in into measure spore under microscope
Degree, until spore concentration is 107-108, choose concentration strain up to standard and be sprayed on fresh bean curd, by the fresh bean curd after sprinkling strain
Ferment 54h under the conditions of 24 DEG C, completes fermentation, is cultivated using the strain on original fermented bean curd, it is possible to increase the mouthfeel of fermented bean curd
And quality.
Embodiment 16
A kind of fermented bean curd strain application process, in gnotobasis, the strain M60 of culture is inoculated in 0.85% physiological saline,
20min is stirred, spore suspension is prepared into, the spore suspension of preparation is inoculated in the liquid of soya-bean milk, glucose and sterilized water
In culture medium, cultivate 2 days, 32 DEG C of temperature, obtain liquid spawn, it is dense that the liquid spawn of preparation is placed in into measure spore under microscope
Degree, until spore concentration is 107-108, choose concentration strain up to standard and be sprayed on fresh bean curd, by the fresh bean curd after sprinkling strain
Ferment 60h under the conditions of 30 DEG C, completes fermentation, is cultivated using the strain on original fermented bean curd, it is possible to increase the mouthfeel of fermented bean curd
And quality.
Embodiment 17
On the basis of embodiment 1-16, strain M60 accesses amount is the 0.5% of sterile water volume, and strain access amount is limited
It is fixed, enable to ferment effect preferable.
Embodiment 18
On the basis of embodiment 1-17, the volume ratio of soya-bean milk and sterilized water is 1:30, enable to ferment effect preferable.
Embodiment 19
On the basis of embodiment 1-18, glucose addition accounts for the 4% of sterile water volume, can be survived for strain and provide nutrition
Composition.
Although reference be made herein to invention has been described for multiple explanatory embodiments of the invention, however, it is to be understood that
Those skilled in the art can be designed that a lot of other modifications and embodiment, and these modifications and embodiment will fall in this Shen
Please be within disclosed spirit and spirit.More specifically, can be to master in the range of disclosure and claim
The building block and/or layout for inscribing composite configuration carry out a variety of variations and modifications.Except what is carried out to building block and/or layout
Outside modification and improvement, to those skilled in the art, other purposes also will be apparent.
Claims (9)
- A kind of 1. method of purification of fermented bean curd strain, it is characterised in that：Comprise the following steps：S1, sample preparation：Bean curd of fermented 100g after common fermentation is dissolved in physiological saline, the fluid sample in physiological saline is then inoculated in nothing In bacterium PDA plate culture medium, cultivate 3-4 days；S2, sample dilution：1000 times of normal saline dilution of sample after the completion of S1 is cultivated, by the sample inoculation after dilution in sterile PDA plate In culture medium, cultivate 3-4 days；S3, sample are cultivated again：The mucor strain that S2 cultures are completed is seeded in sterile PDA plate culture medium again, cultivated 3-4 days；And with geometry times Number form formula, purifying culture respectively, until obtaining sufu fermentation probioticses.
- A kind of 2. method of purification of fermented bean curd strain according to claim 1, it is characterised in that：In S1 and S2 steps, physiology The mass concentration of salt solution is 0.85%.
- A kind of 3. method of purification of fermented bean curd strain according to claim 1, it is characterised in that：In S1, S2 and S3 step, training Foster condition is 16-32 DEG C of temperature, humidity 60%.
- A kind of 4. method of purification of fermented bean curd strain according to claim 1, it is characterised in that：Strain M60 daily preservation Condition be：16-28 DEG C of temperature, humidity 60%.
- A kind of 5. method of purification of fermented bean curd strain according to claim 1, it is characterised in that：Strain M60 long-term preservation Condition be：6-12 DEG C of temperature, humidity 60%.
- A kind of 6. fermented bean curd strain application process according to claim 1, it is characterised in that：In gnotobasis, it will cultivate Strain M60 be inoculated in 0.85% physiological saline, stir 20min, spore suspension is prepared into, by the spore suspension of preparation Liquid is inoculated in the fluid nutrient medium of soya-bean milk, glucose and sterilized water, is cultivated 2 days, 16-32 DEG C of temperature, is obtained liquid spawn, will The liquid spawn of preparation is placed under microscope and determines spore concentration, until spore concentration is 107-108, choose concentration bacterium up to standard Kind it is sprayed on fresh bean curd, the fresh bean curd after strain will be sprayed and fermented under the conditions of 16-30 DEG C 48-60h, complete fermentation.
- A kind of 7. fermented bean curd strain application process according to claim 6, it is characterised in that：Strain M60 accesses amount is sterile The 0.5% of water volume.
- A kind of 8. fermented bean curd strain application process according to claim 6, it is characterised in that：The volume ratio of soya-bean milk and sterilized water For 1:30.
- A kind of 9. fermented bean curd strain application process according to claim 6, it is characterised in that：Glucose addition accounts for sterilized water The 4% of volume.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108740062A (en) * | 2018-08-20 | 2018-11-06 | 江南大学 | A kind of preparation method of the oil preserved bean curd of the high yield fragrance of a flower and fruity |
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| CN102488019A (en) * | 2011-12-02 | 2012-06-13 | 黄山市万安食品有限公司 | Preparation method for villi bean curd |
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| CN106954680A (en) * | 2017-04-17 | 2017-07-18 | 广西大学 | A kind of manufacture craft of bean curd of fermented |
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2017
- 2017-10-31 CN CN201711043872.6A patent/CN107629968A/en active Pending
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| CN102488019A (en) * | 2011-12-02 | 2012-06-13 | 黄山市万安食品有限公司 | Preparation method for villi bean curd |
| CN104087511A (en) * | 2014-04-11 | 2014-10-08 | 湖南农业大学 | Mucor racemosus strain and its application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108740062A (en) * | 2018-08-20 | 2018-11-06 | 江南大学 | A kind of preparation method of the oil preserved bean curd of the high yield fragrance of a flower and fruity |
| CN108740062B (en) * | 2018-08-20 | 2021-10-22 | 江南大学 | Preparation method of oil fermented bean curd with high flowery flavor and fruit flavor yield |
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