CN108676759A - A kind of lactic acid bacteria freeze drying protective agent and preparation method thereof - Google Patents
A kind of lactic acid bacteria freeze drying protective agent and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of lactic acid bacteria freeze drying protective agents and preparation method thereof.The present invention includes the following steps：Freeze drying protectant is prepared first; mainly contain skimmed milk powder 10%; sodium glutamate 6%, trehalose 12%, lactose 4%; inulin 6%; then the lactic acid bacteria bacterium mud that fermentation centrifugation obtains is added to mixing in the freeze drying protectant after preparing and sterilizing cooling by glycine betaine 3%, Cordyceps sinensis polysaccharide 2%; wherein protectant ratio is prepared by 4 times of bacterium mud quality, and finally the bacteria suspension sabot after mixing is lyophilized.Wherein lactic acid bacteria includes mainly lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, lactobacillus bulgaricus etc..Using the protective agent, the survival rate of lactic acid bacteria freeze drying bacterium powder is all 85% or more, and bacterium powder viable count is 1011CFU/mL or more.
Description
Technical field
The present invention relates to field of microbial fermentation, and in particular to a kind of lactic acid bacteria freeze drying protective agent and preparation method thereof.
Background technology
Lactic acid bacteria is closely bound up with our life as a kind of important probiotics.It is numerous studies have shown that probiotic lactic acid
Bacterium tool adjusts intestinal flora balance, prevention and treatment diarrhea, alleviates intestinal inflammatory, treatment lactose intolerance, reducing blood lipid, drop blood
The different physiological roles such as pressure, activation body immune system.With the continuous development of biotechnology, stablizing for the market demand increases,
Probiotic lactobacillus is all widely used in food, health care, medicine, agricultural and animal husbandry etc..
By years of researches, the preparation of lactic acid bacteria bacterium powder is mainly cold by spray drying after enrichment of fermenting or vacuum
Dry acquisition lactic acid bacteria bacterium powder is lyophilized, it is larger to thalline damage since in spray-drying process, temperature is higher, obtain depositing for bacterium powder
Motility rate is relatively low, this method less use；The lactic acid bacteria bacterium powder of high-quality is presently mainly obtained by vacuum freeze drying.Vacuum is cold
Freezing dry technology was invented for the first time in 1813 by Wollaston, and Heckly and Hammer for the first time answers freeze drying technology within 1911
Preparation for microorganism preserves, and has the research history of last 100 years so far.Domestic and foreign scholars are studies have shown that dry compared to other
Dry technology, Vacuum Freezing ＆ Drying Technology have enzymatic activity in protection microbial body, reduce the death rate in microorganism drying process, is de-
Water is thorough, is easy to the advantages that preservation.But it with the further research to freeze drying technology, finds in freeze-drying process, the change of temperature
Change, the formation of ice crystal, intracellular protein combination water such as lose at the activity that will all influence cell, such as the shape of intracellular irregular crystal
Lead to protein denaturation at meeting damaging cells film, too low temperature, lose change protein folding structure, intracellular pH in conjunction with water
Variation etc. will all reduce the bioactivity after lactic acid bacteria bacterium powder rehydration and pot-life.
And interaction between the solution properties, protective agent and protein molecule by protective agent to change bacteria suspension,
The destructive power in Ice Formation Process is weakened, damage of the freezing dry process to somatic cells can be effectively reduced.Therefore, it works out
A kind of protective agent of suitable lactic acid bacteria vacuum freeze drying just seems most important to the lactic acid bacteria freeze drying bacterium powder for obtaining high-quality.
A kind of suitable freeze drying protectant simultaneously, can effectively improve survival rate, the bioactivity of lactic acid bacteria freeze drying bacterium powder, make it easier to store up
It deposits and transports.
Invention content
In view of this, a kind of lactic acid bacteria freeze drying protective agent of present invention offer and preparation method thereof.Freeze-drying provided by the invention
Protective agent effect is good, and can effectively improve the survival rate of lactic acid bacteria freeze drying bacterium powder, bioactivity.
First aspect present invention provides a kind of lactic acid bacteria freeze drying protective agent, includes mainly following component：Skimmed milk powder, paddy
Propylhomoserin sodium, trehalose, lactose, inulin, glycine betaine, Cordyceps sinensis polysaccharide.
The present invention an embodiment in, the lactic acid bacteria freeze drying protective agent include following weight percent at
Point：Skimmed milk powder 10~20%, sodium glutamate 2~10%, trehalose 8~16%, lactose 2~6%, inulin 2~10%, beet
Alkali 1~5%, Cordyceps sinensis polysaccharide 1~5%.
Preferably, the lactic acid bacteria freeze drying protective agent, including following component：Skimmed milk powder 10%, sodium glutamate 6%,
Trehalose 12%, lactose 4%, inulin 6%, glycine betaine 3%, Cordyceps sinensis polysaccharide 2%.
Second aspect of the present invention provides a kind of protectant preparation method of lactic acid bacteria freeze drying, includes the following steps：
Each group assignment system freeze drying protectant is weighed according to proportioning
(1) skimmed milk powder is prepared：In the purified water that the skimmed milk powder weighed up is completely dissolved in, then go out high temperature is damp and hot
5~15min of bacterium, is then rapidly cooled to room temperature, is placed in 4 DEG C of pre- cold standbies of refrigerator；
(2) inulin is prepared：In the purified water that the inulin weighed up is completely dissolved in, then high-temperature heat sterilization 10~
Then 30min is rapidly cooled to room temperature, be placed in 4 DEG C of pre- cold standbies of refrigerator；
(3) glycine betaine weighed up, Cordyceps sinensis polysaccharide, sodium glutamate, trehalose, lactose are dissolved separately in purified water,
10~30min of high-temperature heat sterilization, then cools to room temperature, and is placed in 4 DEG C of pre- cold standbies of refrigerator；
(4) it by each component obtained by above-mentioned steps, is uniformly mixed, obtains freeze drying protectant.
In the embodiment of the present invention, in the frozen-dried protective preparation method step (1), the temperature of purified water is 50
~60 DEG C.
In the embodiment of the present invention, in the frozen-dried protective preparation method step (1), the temperature of high-temperature heat sterilization
Degree is 115 DEG C, sterilization time 10min.
In the embodiment of the present invention, in the frozen-dried protective preparation method step (2), the temperature of purified water is 50
~60 DEG C.
In the embodiment of the present invention, in the frozen-dried protective preparation method step (2), the temperature of high-temperature heat sterilization
Degree is 115 DEG C, sterilization time 20min.
In the embodiment of the present invention, in the frozen-dried protective preparation method step (3), the temperature of high-temperature heat sterilization
Degree is 121 DEG C, sterilization time 20min.
Third aspect present invention provides lactic acid bacteria freeze drying protective agent as described in claim 1, as claimed in claim 3
Application of the protectant preparation method of lactic acid bacteria freeze drying in preparing lactic acid bacteria freeze drying bacterium powder.
In the embodiment of the present invention, the preparation of the lactic acid bacteria freeze drying bacterium powder includes the following steps：By lactic acid
Bacterium activation, culture, are enriched with to obtain bacterium mud, by bacterium mud and freeze drying protectant 1：It is equal that freeze drying protectant mixing is added in 1~10 weight ratio
Even, vacuum freeze drying obtains lactic acid bacteria freeze drying bacterium powder.
Preferably, the bacterium mud and the weight ratio of freeze drying protectant are 1：4.
In the embodiment of the present invention, the lactic acid bacteria includes Lactococcus species, Lactobacillus species, hammer
Pseudomonas bacterium is one or more respectively, in Pediococcus species, Bifidobacterium strain, Leuconostoc strain.
The freeze drying protectant of freeze-drying bacterium powder can be divided into small molecule protective agent (such as oligosaccharides, alcohols, buffering salt, amino
Acid and vitamins) and macromolecular protective agent (such as protein, polypeptide and polysaccharide).It is general to have as small molecule protective agent
There is very strong hydrophily, molecular structure contains 3 or more hydrogen bonds, can be with somatic cells membrane phospholipid in freezing or drying process
In phosphate group or bacterium protein polar group form hydrogen bond, protect the complete of cell membrane and protein structure and function
Property.And macromolecular protective agent protects thalline by " package " form, meanwhile, promote low molecule protective agent to play a role.The present invention
Added inulin, glycine betaine and Cordyceps sinensis polysaccharide on the basis of conventional formulation, and by experimental verification be added inulin, glycine betaine with
And the freeze drying protectant of Cordyceps sinensis polysaccharide has better protective effect.
The present invention, which has, is implemented as follows effect：
One, lactic acid bacteria of the invention includes lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, bulgarian milk bar
Bacterium etc., protection agent prescription of the invention can be used for preparation that is above-mentioned but being not limited to above-mentioned lactic acid bacteria freeze drying bacterium powder, use the protection
Can be produced after agent high viable count, high activity lactobacillus preparation, it is easy to operate, of low cost, applicability is wide, can scale metaplasia
Production.
Two, protective agent of the present invention can apply to lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, Bulgaria
The preparation of the lactic acid bacteria freeze dryings bacterium powder such as lactobacillus, the bacterium powder viable count of acquisition is 1011CFU/mL or more, freeze-drying survival rate is 85%
More than.
Specific implementation mode
As described below is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
The preferred embodiment of the present invention will be described in detail below.The experiment of actual conditions is not specified in embodiment
Method, usually according to normal condition.In the embodiment of the present invention unless otherwise noted, agents useful for same and consumptive material are commercial goods.
Lactic acid bacteria freeze drying protective agent provided by the invention includes mainly following components：
Protectant preparation, wherein skimmed milk powder 10~20%, inulin 2~10%, beet are carried out by 4 times of bacterium mud quality
Alkali 1~5%, sodium glutamate 2~10%, trehalose 8~16%, lactose 2~6%, Cordyceps sinensis polysaccharide 1~5%.
Lactic acid bacteria freeze drying protective agent provided by the invention, preparation method mainly include the following steps that：
(1) skimmed milk powder is prepared, and the skimmed milk powder weighed up is completely dissolved in suitable 50~60 DEG C of purified water, so
Afterwards at 115 DEG C, then moist heat sterilization 10min is rapidly cooled to room temperature, is placed in 4 DEG C of pre- cold standbies of refrigerator；
(2) inulin is prepared, and the inulin weighed up is completely dissolved in suitable 50~60 DEG C of purified water, then 115
At DEG C, then moist heat sterilization 20min is rapidly cooled to room temperature, is placed in 4 DEG C of pre- cold standbies of refrigerator；
(3) glycine betaine weighed up, sodium glutamate, Cordyceps sinensis polysaccharide, trehalose, lactose are dissolved separately in appropriate purified water
In, at 121 DEG C, moist heat sterilization 20min is then cooled to room temperature, and is placed in 4 DEG C of pre- cold standbies of refrigerator；
(4) centrifugation obtain bacterium mud after, by be pre-chilled finish various protective ingredients (skimmed milk powder, inulin, trehalose,
Sodium glutamate, lactose, Cordyceps sinensis polysaccharide) it is uniformly mixed, you can as lactic acid bacteria freeze drying protective agent.
The lactic acid bacteria freeze drying protective agent of the present invention can be used for vacuum freeze-drying method and prepare lactic acid bacteria preparation lactic acid bacteria bacterium powder,
Specific embodiment is as follows：
Embodiment 1：Vacuum freeze-drying method prepares lactobacillus plantarum bacterium powder
(1) High Density Cultivation
Lactobacillus plantarum strain after activation is inoculated in proliferated culture medium (this training of lactobacillus plantarum in 10% ratio
Foster base is the lactobacillus plantarum culture medium by optimization), control 37 DEG C, pH6.0 of temperature, monitor fermentation liquid OD600It is in steady to it
Fixed, the viable count that ferments at this time is 109~1010Between.
(2) protective agent prepares
Contain skimmed milk powder 10%, sodium glutamate 6%, trehalose 12%, lactose 4% in protective agent A；Each ingredient is individually molten
Solution and individually sterilizing, until being remixed before use, after wherein skimmed milk powder need to fully be dissolved with 50~60 DEG C of purified waters, in 115 DEG C
Under, then 8~10min of moist heat sterilization is quickly cooled down；Remaining ingredient is at 121 DEG C, moist heat sterilization 20min.
Contain in protective agent B containing skimmed milk powder 10%, sodium glutamate 6%, trehalose 12%, lactose 4%, inulin 6%,
Glycine betaine 3%, Cordyceps sinensis polysaccharide 2%；Individually dissolving and the individually sterilizing of each ingredient, until being remixed before use, wherein skimmed milk powder needs
After fully being dissolved with 50~60 DEG C of purified waters, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to use 50
After~60 DEG C of purified waters fully dissolve, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is at 121 DEG C
Under, moist heat sterilization 20min.
Contain in protective agent C containing skimmed milk powder 10%, sodium glutamate 6%, trehalose 12%, lactose 4%, inulin 6%,
Glycine betaine 3%,；Individually dissolving and the individually sterilizing of each ingredient, until remixed before use, wherein skimmed milk powder need to 50~60 DEG C it is pure
After change water fully dissolves, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to use 50~60 DEG C of purified waters
Fully after dissolving, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is at 121 DEG C, moist heat sterilization
20min。
Contain skimmed milk powder 20%, sodium glutamate 6%, trehalose 12%, lactose 4%, inulin 6% in protective agent D；
Individually dissolving and the individually sterilizing of each ingredient, until being remixed before use, wherein skimmed milk powder need to be fully molten with 50~60 DEG C of purified waters
Xie Hou, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to fully be dissolved with 50~60 DEG C of purified waters
Afterwards, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is at 121 DEG C, moist heat sterilization 20min.
(3) acquisition of lactobacillus plantarum bacterium mud and protectant addition
Zymotic fluid is centrifuged using the tube centrifuge that rotating speed is 10000rpm/min, obtains lactobacillus plantarum bacterium
Bacterium mud is uniformly divided into 4 parts by mud after obtaining bacterium mud, after each ingredient of protective agent is mixed, is separately added by 4 times of bacterium mud quality
Mixed freeze drying protectant A, B, C, D, homogeneous 10min makes bacterium mud be mixed well with protective agent at 8000rpm later, prepares
At freeze-drying bacteria suspension.
(4) vacuum freeze drying
It by the bacteria suspension after mixing, is laid in lyophilized plate by the thickness of 3~5mm, controls -35 DEG C of precooling temperature, pre-freeze speed
0.6 DEG C/min, drying chamber pressure 32Pa are spent, heat up 30 DEG C of maximum temperature, carries out vacuum freeze drying, obtains bacterium powder water content and exists
Within 5%.
(5) result is lyophilized
Bacterium powder is lyophilized in the lactobacillus plantarum that freeze-drying acquisition is carried out using protective agent A, and viable count is 2.1 × 1011Cfu/g freezes
Dry survival rate is 70%；
Bacterium powder is lyophilized in the lactobacillus plantarum that freeze-drying acquisition is carried out using protective agent B, and viable count is 3.1 × 1011Cfu/g freezes
Dry survival rate is 88%；
Bacterium powder is lyophilized in the lactobacillus plantarum that freeze-drying acquisition is carried out using protective agent C, and viable count is 2.6 × 1011Cfu/g freezes
Dry survival rate is 75%；
Bacterium powder is lyophilized in the lactobacillus plantarum that freeze-drying acquisition is carried out using protective agent D, and viable count is 1.8 × 1011Cfu/g freezes
Dry survival rate is 65%.
The result shows that the frozen-dried protective effect of protective agent B is best, i.e., in original formula (skimmed milk powder 10%, sodium glutamate
6%, trehalose 12%, lactose 4%) on the basis of add inulin 6%, glycine betaine 3%, Cordyceps sinensis polysaccharide 2% can improve freeze-drying
Protecting effect of the protective agent to lactobacillus plantarum.
Embodiment 2：Vacuum freeze-drying method prepares Lactobacillus rhamnosus bacterium powder
(1) High Density Cultivation
Lactobacillus rhamnosus strain after activation is inoculated in the proliferated culture medium of Lactobacillus rhamnosus in 10% ratio
(this culture medium is the Lactobacillus rhamnosus culture medium by optimization) controls 37 DEG C, pH5.7 of temperature, monitor fermentation liquid OD600Extremely
It, which is in, stablizes, and the viable count that ferments at this time is 109~1010Between.
(2) protective agent prepares
Contain skimmed milk powder 15%, sodium glutamate 8%, trehalose 14%, lactose 6% in protective agent A；Each ingredient is individually molten
Solution and individually sterilizing, until being remixed before use, after wherein skimmed milk powder need to fully be dissolved with 50~60 DEG C of purified waters, in 115 DEG C
Under, then 8~10min of moist heat sterilization is quickly cooled down；Remaining ingredient is at 121 DEG C, moist heat sterilization 20min.
Contain skimmed milk powder 10%, inulin 6%, glycine betaine 3%, sodium glutamate 6%, trehalose in protective agent B
12%, lactose 4%；Individually dissolving and the individually sterilizing of each ingredient, until being remixed before use, wherein skimmed milk powder need to use 50~60 DEG C
After purified water fully dissolves, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to be purified with 50~60 DEG C
After water fully dissolves, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is damp and hot to go out at 121 DEG C
Bacterium 20min.
Contain skimmed milk powder 10%, inulin 6%, glycine betaine 3%, sodium glutamate 6%, trehalose in protective agent C
12%, lactose 4%；Individually dissolving and the individually sterilizing of each ingredient, until being remixed before use, wherein skimmed milk powder need to use 50~60 DEG C
After purified water fully dissolves, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to be purified with 50~60 DEG C
After water fully dissolves, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is damp and hot to go out at 121 DEG C
Bacterium 20min.
Contain skimmed milk powder 20%, inulin 6%, sodium glutamate 6%, trehalose 12%, lactose 4% in protective agent D；
Individually dissolving and the individually sterilizing of each ingredient, until being remixed before use, wherein skimmed milk powder need to be fully molten with 50~60 DEG C of purified waters
Xie Hou, at 115 DEG C, then 8~10min of moist heat sterilization is quickly cooled down；Inulin need to fully be dissolved with 50~60 DEG C of purified waters
Afterwards, at 115 DEG C, then moist heat sterilization 20min is quickly cooled down；Remaining ingredient is at 121 DEG C, moist heat sterilization 20min.
(3) acquisition of Lactobacillus rhamnosus bacterium mud and protectant addition
Zymotic fluid is centrifuged using the tube centrifuge that rotating speed is 10000rpm/min, obtains Lactobacillus rhamnosus bacterium
Bacterium mud after obtaining bacterium mud, after each ingredient of protective agent is mixed, is divided into two parts by mud, presses 4 times of additions of bacterium mud quality respectively
Mixed freeze drying protectant A and freeze drying protectant B, homogeneous 10min keeps bacterium mud fully mixed with protective agent at 8000rpm later
It is even, it is prepared into freeze-drying bacteria suspension.
(4) vacuum freeze drying
It by the bacteria suspension after mixing, is laid in lyophilized plate by the thickness of 3~5mm, controls -35 DEG C of precooling temperature, pre-freeze speed
0.6 DEG C/min, drying chamber pressure 32Pa are spent, heat up 30 DEG C of maximum temperature, carries out vacuum freeze drying, obtains bacterium powder water content and exists
Within 5%.
(5) result is lyophilized
Bacterium powder is lyophilized in the Lactobacillus rhamnosus that freeze-drying acquisition is carried out using protective agent A, and viable count is 1.5 × 1011Cfu/g,
It is 70% that survival rate, which is lyophilized,；
Bacterium powder is lyophilized in the Lactobacillus rhamnosus that freeze-drying acquisition is carried out using protective agent B, and viable count is 2.5 × 1011Cfu/g,
It is 86% that survival rate, which is lyophilized,；
Bacterium powder is lyophilized in the Lactobacillus rhamnosus that freeze-drying acquisition is carried out using protective agent C, and viable count is 2.0 × 1011Cfu/g,
It is 77% that survival rate, which is lyophilized,；
Bacterium powder is lyophilized in the Lactobacillus rhamnosus that freeze-drying acquisition is carried out using protective agent D, and viable count is 1.1 × 1011Cfu/g,
It is 64% that survival rate, which is lyophilized,.
The result shows that the frozen-dried protective effect of protective agent B is best, i.e., in existing formula (skimmed milk powder 10%, sodium glutamate
6%, trehalose 12%, lactose 4%) on the basis of add inulin 6%, glycine betaine 3%, Cordyceps sinensis polysaccharide 2% can improve freeze-drying
Protecting effect of the protective agent to Lactobacillus rhamnosus.
To sum up, lactic acid bacteria freeze drying protective agent provided by the invention can be provided than existing formula more preferably preparing when that bacterium powder is lyophilized
Protecting effect.
The above embodiments are merely illustrative of the technical solutions of the present invention rather than limiting the scope of the invention, although ginseng
The present invention is explained in detail according to preferred embodiment, it will be understood by those of ordinary skill in the art that, it can be to the present invention's
Technical solution is modified or replaced equivalently, without departing from the spirit and scope of technical solution of the present invention.
Claims (9)
1. a kind of lactic acid bacteria freeze drying protective agent, which is characterized in that including following component：Skimmed milk powder, sodium glutamate, trehalose,
Lactose, inulin, glycine betaine, Cordyceps sinensis polysaccharide.
2. lactic acid bacteria freeze drying protective agent as described in claim 1, which is characterized in that include the ingredient of following weight percent：
Skimmed milk powder 10~20%, sodium glutamate 2~10%, trehalose 8~16%, lactose 2~6%, inulin 2~10%, glycine betaine 1
~5%, Cordyceps sinensis polysaccharide 1~5%.
3. a kind of protectant preparation method of lactic acid bacteria freeze drying, which is characterized in that include the following steps：
Each group assignment system freeze drying protectant is weighed according to proportioning
1) skimmed milk powder is prepared：In the purified water that the skimmed milk powder weighed up is completely dissolved in, then high-temperature heat sterilization 5~
Then 15min is rapidly cooled to room temperature, be placed in 4 DEG C of pre- cold standbies of refrigerator；
2) inulin is prepared：In the purified water that the inulin weighed up is completely dissolved in, then in 10~30min of high-temperature heat sterilization,
Then it is rapidly cooled to room temperature, is placed in 4 DEG C of pre- cold standbies of refrigerator；
3) glycine betaine weighed up, Cordyceps sinensis polysaccharide, sodium glutamate, trehalose, lactose are dissolved separately in purified water, it is high warm and humid
10~30min of heat sterilization, then cools to room temperature, and is placed in 4 DEG C of pre- cold standbies of refrigerator；
4) it by each component obtained by above-mentioned steps, is uniformly mixed, obtains freeze drying protectant.
4. preparation method as claimed in claim 3, which is characterized in that in the step 1), the temperature of high-temperature heat sterilization is
115 DEG C, sterilization time 10min.
5. preparation method as claimed in claim 3, which is characterized in that in the step 2), the temperature of high-temperature heat sterilization is
115 DEG C, sterilization time 20min.
6. preparation method as claimed in claim 3, which is characterized in that in the step 2), the temperature of high-temperature heat sterilization is
121 DEG C, sterilization time 30min.
7. lactic acid bacteria freeze drying protective agent as described in claim 1, the protectant system of lactic acid bacteria freeze drying as claimed in claim 3
Application of the Preparation Method in preparing lactic acid bacteria freeze drying bacterium powder.
8. the use as claimed in claim 7, which is characterized in that include the following steps：By lactic acid bacteria activation, culture, it is enriched with
Bacterium mud, by bacterium mud and freeze drying protectant 1：1~10 weight ratio is added freeze drying protectant and is uniformly mixed, and vacuum freeze drying obtains
Lactic acid bacteria freeze drying bacterium powder.
9. the use as claimed in claim 7, which is characterized in that the lactic acid bacteria includes Lactococcus species, lactobacillus
Strain, streptococcus bacterium are one or more respectively, in Pediococcus species, Bifidobacterium strain, Leuconostoc strain.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112457990A (en) * | 2020-12-18 | 2021-03-09 | 中国科学院合肥物质科学研究院 | Freeze-drying protective agent and application thereof |
| CN112501068A (en) * | 2020-12-11 | 2021-03-16 | 云南省农业科学院农产品加工研究所 | Preparation method of freeze-dried leaven for meat products |
| CN113201459A (en) * | 2021-06-17 | 2021-08-03 | 浙江海洋大学 | Method for quick freeze-drying preservation of microorganisms |
| CN114806972A (en) * | 2022-06-07 | 2022-07-29 | 中国海洋大学 | Method for preparing high-activity lactic acid bacteria starter by using natural eutectic solvent |
| CN117987270A (en) * | 2024-03-19 | 2024-05-07 | 华南农业大学 | Application of edible fungus polysaccharide in spray drying of pediococcus acidilactici |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660477A (en) * | 2012-05-10 | 2012-09-12 | 北京市农林科学院 | Lactobacillus brevis, freeze-dried powder of Lactobacillus brevis and application of freeze-dried powder |
| CN104212733A (en) * | 2013-05-29 | 2014-12-17 | 镇江拜因诺生物科技有限公司 | Photosynthetic bacterium freeze-drying protective agent |
| CN105219645A (en) * | 2015-11-06 | 2016-01-06 | 南京农业大学 | A kind of novel microorganism protective material and application thereof |
| CN106497847A (en) * | 2016-12-14 | 2017-03-15 | 曹书华 | A kind of lactobacillus rhamnosuss lyophilized powder and preparation method thereof |
-
2018
- 2018-07-09 CN CN201810747521.1A patent/CN108676759A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660477A (en) * | 2012-05-10 | 2012-09-12 | 北京市农林科学院 | Lactobacillus brevis, freeze-dried powder of Lactobacillus brevis and application of freeze-dried powder |
| CN104212733A (en) * | 2013-05-29 | 2014-12-17 | 镇江拜因诺生物科技有限公司 | Photosynthetic bacterium freeze-drying protective agent |
| CN105219645A (en) * | 2015-11-06 | 2016-01-06 | 南京农业大学 | A kind of novel microorganism protective material and application thereof |
| CN106497847A (en) * | 2016-12-14 | 2017-03-15 | 曹书华 | A kind of lactobacillus rhamnosuss lyophilized powder and preparation method thereof |
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| CN112501068A (en) * | 2020-12-11 | 2021-03-16 | 云南省农业科学院农产品加工研究所 | Preparation method of freeze-dried leaven for meat products |
| CN112457990A (en) * | 2020-12-18 | 2021-03-09 | 中国科学院合肥物质科学研究院 | Freeze-drying protective agent and application thereof |
| CN113201459A (en) * | 2021-06-17 | 2021-08-03 | 浙江海洋大学 | Method for quick freeze-drying preservation of microorganisms |
| CN113201459B (en) * | 2021-06-17 | 2023-06-16 | 浙江海洋大学 | Method for quick-freezing, drying and preserving microorganisms |
| CN114806972A (en) * | 2022-06-07 | 2022-07-29 | 中国海洋大学 | Method for preparing high-activity lactic acid bacteria starter by using natural eutectic solvent |
| CN114806972B (en) * | 2022-06-07 | 2023-12-08 | 中国海洋大学 | Method for preparing high-activity lactobacillus starter by using natural eutectic solvent |
| CN117987270A (en) * | 2024-03-19 | 2024-05-07 | 华南农业大学 | Application of edible fungus polysaccharide in spray drying of pediococcus acidilactici |
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