CN108823099A - A kind of preparation method and its preparation of solid-state probiotic preparation - Google Patents
A kind of preparation method and its preparation of solid-state probiotic preparation Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods of solid-state probiotic preparation, belong to dairy products manufacturing field.In order to overcome cell mortality in freeze drying process in the prior art higher, there are the deficiencies of security risk for finished product, the present invention provides the preparation method and its preparation of a kind of solid-state probiotic preparation, it includes the freezing tolerance that glycine betaine and the auxiliary propylhomoserin of hydroxyl are added into prebiotic bacteria suspension and improves probiotics by cooling step by step, then glucose and sorbierite freeze-drying are added thereto and obtains, the survival rate for the probiotics in preparation that the probiotics solid pharmaceutical preparation is prepared is 85% or more, it is significantly better than the probiotics survival rate that probiotics solid pharmaceutical preparation is obtained according to the prior art, its step is simple, short processing time, it is suitable for being promoted and applied in prior art.
Description
Technical field
The present invention relates to a kind of preparation methods of solid-state probiotic preparation, belong to dairy products manufacturing field.
Background technique
Probiotics is a kind of active microorganism beneficial to host, is to be colonized in human body intestinal canal, in reproductive system, can generate
Definite health efficacy so as to improve host's microecological balance, play the active beneficial microorganism general name of beneficial effect.Human body moves
Beneficial bacterium or fungi mainly have in object：Clostridium butyricum, lactobacillus, Bifidobacterium, actinomyces, saccharomycete etc..Generation at present
The most powerful product of the function of being studied in boundary be mainly the above all kinds of microorganism groups at composite reactive probiotics, be widely applied
In bioengineering, industrial or agricultural, food safety and life and health field.
Rationally effective probiotics preparation can effectively repair intestinal flora ecosystem balance, be to promote humans and animals machine
Body is kept fit the keys of growth conditions.Solid-state version probiotics preparation has the characteristics that be easy to save, transport, apply, and closes at present
In the animal probiotics preparation of commercialization be often liquid, in traditional solid-state probiotic preparation probiotics due to drying it is easily dead
It dies, and is unable to long-term preservation, to lose utility value.
CN103690568A discloses a kind of method using hypertonic oligosaccharide solution preparation solid-state probiotic preparation, including
Following steps：1) probiotics bacterial liquid that volume is Vml is measured, protective agent is added after mixing into probiotics bacterial liquid, then plus
Enter oligosaccharide to be sufficiently stirred, obtain probiotics mixed liquor, it is 1~8%Vg, the oligosaccharide being added that protectant quality, which is added,
Quality is 30~70%Vg；2) by probiotics mixed liquor under conditions of 20~42 DEG C 30~240min of isothermal holding；3) to warp
Foaming leavening agent is added in the probiotics mixed liquor of isothermal holding after mixing, adds auxiliary material and is sufficiently stirred, obtain prebiotic
Bacterium mixed material, the foaming leavening agent quality being added are 0.1~0.8%Vg, and the quality of auxiliary material be added is 40~80%Vg；
4) probiotic composition material is dried in vacuo at 30~37 DEG C, is then crushed to get solid-state probiotic preparation is arrived.The work
Make cell dehydration using the osmotic pressure effect of oligosaccharide in skill, is easy to cause cell death, and its protective agent is in freeze-drying work
Protecting effect in skill is poor, therefore its practical application effect is not good enough.
CN104250630A discloses the preparation method of probiotics solid formulation, includes the following steps：It prepares respectively and contains fine jade
Rouge LB culture medium or physiological saline containing agar and LB culture medium or physiological saline, and high pressure sterilization；Take probiotics high speed centrifugation
Precipitating, discards upper layer bacterium solution, is resuspended with the LB culture medium or physiological saline of 35~37 DEG C of pre-temperatures；The probiotics bacterial liquid of resuspension is connect
Enter in liquid LB containing agar culture medium or physiological saline containing agar, mixes；It include the LB culture containing agar of probiotics by liquid
Base or liquid include that the physiological saline containing agar of probiotics is dispensed into mold, and room temperature is cooling, that is, is frozen into mold shape
Probiotics solid formulation.The matrix of the product is LB culture medium, and there are biggish security risks for component long-term use therein.
Summary of the invention
In order to overcome in freeze drying process in the prior art, cell mortality is higher, there are security risks for finished product not
Foot, the present invention provide the preparation method and its preparation of a kind of solid-state probiotic preparation comprising sweet tea is added into prebiotic bacteria suspension
Dish alkali and the auxiliary propylhomoserin of hydroxyl and the freezing tolerance that probiotics is improved by cooling step by step, are then added glucose and sorbierite thereto
Freeze-drying and obtain, the survival rate for the probiotics in preparation which is prepared 85% or more,
It is significantly better than the probiotics survival rate that probiotics solid pharmaceutical preparation is obtained according to the prior art, step is simple, short processing time, fits
Preferably promoted and applied in prior art.
The present invention is achieved through the following technical solutions above-mentioned technical effect：
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
It 1) is 8000-10000rpm in centrifugal rotational speed by probiotics fermention liquid, centrifugation time is to be centrifuged under the conditions of 3-5 minutes
Probiotic cell is obtained, probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
For 0.1%-0.2%, the final concentration of 0.06%-0.12% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is 0 DEG C -- it 5 DEG C, holds
It is 2-4 hours continuous, adjust the temperature to -15 DEG C -- 20 DEG C, continue 40min-60min；Adjust the temperature to -30 DEG C -- 40 DEG C, continue 4-6
Hour；Recovery temperature is to room temperature；
3) sorbierite and 4%-6% of probiotic cell 4%-6% weight percentage are added to probiotic cell suspension
The glucose of weight percentage, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -25 DEG C -- and 35
DEG C, sublimation drying is 18-24 hours.
The preparation method of solid-state probiotic preparation described above, glycine betaine in probiotic cell suspension in the step 2)
Weight ratio with the auxiliary propylhomoserin of hydroxyl is 5:3.Preferably, in the step 2) in probiotic cell suspension glycine betaine it is final concentration of
0.15%, the final concentration 0.09% of the auxiliary propylhomoserin of hydroxyl.
The preparation method of solid-state probiotic preparation described above, sorbierite is in probiotic cell suspension in the step 3)
Percentage composition be 5%, glucose probiotic cell suspension percentage composition be 5%.
The preparation method of solid-state probiotic preparation described above can also be added in the step 3) before freeze-drying
Auxiliary material, it is preferable that the auxiliary material is one kind of soluble starch, starch, skimmed milk power, konjaku powder, konjac glucomannan, corn flour
Or it is a variety of.
The preparation method of solid-state probiotic preparation described above, the middle freeze-drying temperature of the step 3) is -30 DEG C, cold
Freezing drying time is 21 hours.
The present invention also provides a kind of probiotics solid pharmaceutical preparations that above-mentioned preparation method is prepared.
It is experimentally confirmed that the survival of the probiotics in the preparation being prepared according to probiotics solid pharmaceutical preparation of the present invention
Rate is 85% or more, and wherein the effect of embodiment 3 is best, is significantly better than and obtains probiotics solid pharmaceutical preparation according to the prior art
Probiotics survival rate.From comparative experiments as can be seen that step 2 is committed step of the invention, the survival for probiotics
Rate plays a significant role, wherein glycine betaine is not added and the auxiliary propylhomoserin of hydroxyl will lead to being remarkably decreased for probiotics survival rate.
The present invention has the advantage that compared with prior art：
The survival rate for the probiotics in preparation that probiotics solid pharmaceutical preparation of the present invention is prepared 85% or more, wherein
The effect of embodiment 3 is best, is significantly better than the probiotics survival rate that probiotics solid pharmaceutical preparation is obtained according to the prior art.It is walked
Rapid simple, short processing time is suitable for promoting and applying in prior art.
Specific embodiment
The present invention is further described below by way of specific embodiment, but the embodiment does not limit this hair in any way
The range of bright patent protection.
Embodiment 1
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
1) by probiotics fermention liquid centrifugal rotational speed be 8000rpm, centrifugation time be 3 minutes under the conditions of centrifugation obtain it is prebiotic
Probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension by bacterium cell；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
It is 0.1%, final concentration of the 0.06% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is 0 DEG C, continues 2 hours, adjusts temperature
To -15 DEG C, continue 40min；- 30 DEG C are adjusted the temperature to, continues 4 hours；Recovery temperature is to room temperature；
3) sorbierite and 4% weight percent of 4% weight percentage of probiotic cell are added to probiotic cell suspension
The glucose of content, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -25 DEG C -- and 35 DEG C, freezing
Drying time is 18-24 hours.
Auxiliary material can also be added in the step 3) before freeze-drying, the auxiliary material is soluble starch, starch, takes off
Rouge milk powder, konjaku powder, konjac glucomannan, corn flour it is one or more.
Embodiment 2
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
It 1) is 10000rpm in centrifugal rotational speed by probiotics fermention liquid, centrifugation time is that centrifugation obtains benefit under the conditions of 5 minutes
Raw bacterium cell, probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
It is 0.2%, final concentration of the 0.12% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is -5 DEG C, continues 4 hours, adjusts temperature
To -20 DEG C, continue 60min；- 40 DEG C are adjusted the temperature to, continues 6 hours；Recovery temperature is to room temperature；
3) sorbierite and 6% weight percent of 6% weight percentage of probiotic cell are added to probiotic cell suspension
The glucose of content, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -35 DEG C, when freeze-drying
Between be 24 hours.
Auxiliary material can also be added in the step 3) before freeze-drying, the auxiliary material is soluble starch, starch, takes off
Rouge milk powder, konjaku powder, konjac glucomannan, corn flour it is one or more.
Embodiment 3
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
1) by probiotics fermention liquid centrifugal rotational speed be 9000rpm, centrifugation time be 4 minutes under the conditions of centrifugation obtain it is prebiotic
Probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension by bacterium cell；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
It is 0.15%, final concentration of the 0.09% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is -5 DEG C, continues 3 hours, adjusts temperature
To -18 DEG C, continue 50min；- 35 DEG C are adjusted the temperature to, continues 5 hours；Recovery temperature is to room temperature；
3) sorbierite and 5% weight percent of 5% weight percentage of probiotic cell are added to probiotic cell suspension
The glucose of content, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -30 DEG C, when freeze-drying
Between be 22 hours.
Auxiliary material can also be added in the step 3) before freeze-drying, the auxiliary material is soluble starch, starch, takes off
Rouge milk powder, konjaku powder, konjac glucomannan, corn flour it is one or more.
Embodiment 4
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
1) by probiotics fermention liquid centrifugal rotational speed be 8000rpm, centrifugation time be 5 minutes under the conditions of centrifugation obtain it is prebiotic
Probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension by bacterium cell；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
It is 0.1%, final concentration of the 0.12% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is 0 DEG C, continues 4 hours, adjusts temperature
To -15 DEG C, continue 60min；- 30 DEG C are adjusted the temperature to, continues 6 hours；Recovery temperature is to room temperature；
3) sorbierite and 6% weight percent of 4% weight percentage of probiotic cell are added to probiotic cell suspension
The glucose of content, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -25 DEG C, when freeze-drying
Between be 24 hours.
Auxiliary material can also be added in the step 3) before freeze-drying, the auxiliary material is soluble starch, starch, takes off
Rouge milk powder, konjaku powder, konjac glucomannan, corn flour it is one or more.
Embodiment 5
A kind of preparation method of solid-state probiotic preparation, specifically includes following step：
It 1) is 10000rpm in centrifugal rotational speed by probiotics fermention liquid, centrifugation time is that centrifugation obtains benefit under the conditions of 3 minutes
Raw bacterium cell, probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension；
2) final concentration for the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
It is 0.2%, final concentration of the 0.06% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is -5 DEG C, continues 2 hours, adjusts temperature
To -20 DEG C, continue 40min；- 40 DEG C are adjusted the temperature to, continues 4 hours；Recovery temperature is to room temperature；
3) sorbierite and 4% weight percent of 6% weight percentage of probiotic cell are added to probiotic cell suspension
The glucose of content, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -35 DEG C, when freeze-drying
Between be 18 hours.
Auxiliary material can also be added in the step 3) before freeze-drying, the auxiliary material is soluble starch, starch, takes off
Rouge milk powder, konjaku powder, konjac glucomannan, corn flour it is one or more.
Comparative example 1
The probiotics solid pharmaceutical preparation being prepared according to technique described in CN104250630A embodiment 1.
Comparative example 2
The probiotics solid pharmaceutical preparation being prepared according to CN103690568A embodiment 1.
Comparative example 3
Preparation method is with embodiment 3, but default step 2.
Comparative example 4
Glycine betaine and the auxiliary propylhomoserin of hydroxyl is not added with embodiment 3, but in step 2) in preparation method.
The preparation method effect assessment of the solid-state probiotic preparation of the present invention of embodiment 6
Handle the cell suspension of bifidobacterium longum and Lactobacillus casei respectively according to technique of the present invention.Bacterium colony counts
It is carried out according to method described in GB 4789.35-2010 national food safety standard food microbiological examination, calculates its survival
Rate, survival rate are the ratio of the total number of bacteria before total number of bacteria and freeze-drying after freeze-drying.The following institute of its measurement result
Show.
Protective effect of the composite protectant of the present invention of table 1 to probiotics before and after freeze drying process
| Probiotics survival rate (%) | |
| Embodiment 1 | 88.6 |
| Embodiment 2 | 86.9 |
| Embodiment 3 | 92.3 |
| Embodiment 4 | 85.7 |
| Embodiment 5 | 89.6 |
| Comparative example 1 | 55.2 |
| Comparative example 2 | 74.6 |
| Comparative example 3 | 25.3 |
| Comparative example 4 | 45.8 |
It can be seen that the survival of the probiotics in the preparation being prepared according to probiotics solid pharmaceutical preparation of the present invention
Rate is 85% or more, and wherein the effect of embodiment 3 is best, is significantly better than and obtains probiotics solid pharmaceutical preparation according to the prior art
Probiotics survival rate.From comparative experiments as can be seen that step 2 is committed step of the invention, the survival for probiotics
Rate plays a significant role, wherein glycine betaine is not added and the auxiliary propylhomoserin of hydroxyl will lead to being remarkably decreased for probiotics survival rate.
Claims (7)
1. a kind of preparation method of solid-state probiotic preparation, specifically includes following step：
It 1) is 8000-10000rpm in centrifugal rotational speed by probiotics fermention liquid, centrifugation time is to be centrifuged to obtain under the conditions of 3-5 minutes
Probiotic cell is suspended in 0.9% physiological saline and obtains probiotic cell suspension by probiotic cell；
2) the final concentration of of the glycine betaine that glycine betaine and the auxiliary propylhomoserin of hydroxyl make is added into probiotic cell suspension under room temperature
0.1%-0.2%, the final concentration of 0.06%-0.12% of the auxiliary propylhomoserin of hydroxyl, the temperature for controlling cell suspension is 0 DEG C -- 5 DEG C, continue
2-4 hours, adjust the temperature to -15 DEG C -- 20 DEG C, continue 40min-60min；Adjust the temperature to -30 DEG C -- 40 DEG C, it is small to continue 4-6
When；Recovery temperature is to room temperature；
3) sorbierite and 4%-6% weight of probiotic cell 4%-6% weight percentage are added to probiotic cell suspension
The glucose of percentage composition, is put into vacuum freeze drier and is freeze-dried, and control freeze-drying temperature is -25 DEG C -- and 35 DEG C,
Sublimation drying is 18-24 hours.
2. the preparation method of solid-state probiotic preparation according to claim 1, which is characterized in that prebiotic in the step 2)
The weight ratio of glycine betaine and the auxiliary propylhomoserin of hydroxyl is 5 in bacterium cell suspension:3.
3. the preparation method of solid-state probiotic preparation according to claim 1, which is characterized in that prebiotic in the step 2)
Final concentration of the 0.15% of glycine betaine in bacterium cell suspension, the final concentration 0.09% of the auxiliary propylhomoserin of hydroxyl.
4. the preparation method of solid-state probiotic preparation according to claim 1, which is characterized in that sorb in the step 3)
Alcohol is 5% in the percentage composition of probiotic cell suspension, and glucose is 5% in the percentage composition of probiotic cell suspension.
5. the preparation method of solid-state probiotic preparation according to claim 1, which is characterized in that cold in the step 3)
Be lyophilized it is dry before be additionally added auxiliary material.
6. the preparation method of solid-state probiotic preparation according to claim 1, which is characterized in that the auxiliary material is solvable
One kind of property starch, starch, skimmed milk power, konjaku powder, konjac glucomannan, corn flour.
7. a kind of probiotics solid pharmaceutical preparation that preparation method described in -6 is prepared according to claim 1.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151109A (en) * | 2021-05-19 | 2021-07-23 | 天津大学 | Probiotic low-temperature freezing composite protective agent and application thereof |
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