CN112795558A - Three-layer embedding method for improving stability of lactic acid bacteria - Google Patents
Three-layer embedding method for improving stability of lactic acid bacteria Download PDFInfo
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Abstract
The invention relates to the technical field of microbial preparations, and provides a three-layer embedding method for improving the stability of lactic acid bacteria, which comprises the steps of S1, performing high-density culture on the lactic acid bacteria in a fermentation tank; s2, centrifugally separating the bacterial sludge from the fermentation liquor, wherein the water content of the bacterial sludge is 5-50%; s3, uniformly mixing the bacterial sludge and a first layer of embedding protective agent according to the mass ratio of 1 (1-3), wherein the first layer of embedding protective agent contains 5% -50% of skim milk powder; s4, burying the bacterial sludge and the second layer of embedding protective agent according to the mass ratio of 1: (1-3) uniformly mixing the mixture with S3; s5, mixing the bacterial sludge and the third layer of embedding protective agent according to the mass ratio of 1: (1-3) mixing the mixture with the mixture obtained in the step S4, and emulsifying uniformly to obtain an S5 emulsion; s6, carrying out cryogenic granulation on the S5 emulsion by using a liquid nitrogen freezing technology to enable the thalli to enter a dormant state instantly to obtain S6 cryogenic granules; s7, drying the S6 deep-frozen grains until the water content is 3% -8%. Through above-mentioned technical scheme, the problem of poor stability among the prior art has been solved.
Description
Technical Field
The invention relates to the field of microbial preparations, in particular to a three-layer embedding method for improving the stability of lactic acid bacteria.
Background
In the early 70 s of the last century, researchers in developed countries such as europe, america and japan and korea began to develop novel microecologics for partially or totally replacing antibiotics, and hope to improve human immunity, and after decades of development, certain results have been achieved in related fields. After the development, along with the economic development of China, people continuously deepen the understanding of the health system, and gradually realize that the lactobacillus microecologics have great advantages in the aspects of regulating the balance of intestinal flora, improving the immunity and the like.
However, because the stress resistance of the lactic acid bacteria is poor, even if the bacteria are placed under an undamaged precursor, the bacteria can easily die at room temperature, so that the shelf life of the lactic acid bacteria microecological preparation product is short, the number of viable bacteria in the product can be easily reduced to a lower level, and the product effect is influenced. The existing lactobacillus products are all obtained by vacuum freeze drying treatment. In the production process, because of the difference of the environment inside and outside the cell, the crystallization speed is different, the generation of ice crystals is different, the structures such as cell membranes and the like are greatly damaged, and the stability of the product in the shelf life is not facilitated. Therefore, there is an urgent need to develop a technology for preserving a microecological preparation of lactic acid bacteria at normal temperature for a long period of time.
The technology advocated at present is to carry out the embedding measure to the lactic acid bacteria when the product is produced to this reduces the external environment and to the stimulation of thallus, promotes product shelf life stability. The principle of the lactobacillus embedding method is that natural or synthetic macromolecular edible materials are utilized, and the lactobacillus is wrapped to form a micro-capsule structure by a certain technical means, so that the wrapping material has no adverse effect on the lactobacillus, and meanwhile, the hygroscopicity, the permeability and the like of the product are considered.
At present, the shelf life stability of the existing lactobacillus microecological preparation products on the market is poor, and the death rate of viable count is high after the lactobacillus microecological preparation products are digested by gastric juice and bile by consumers.
Disclosure of Invention
The invention provides a three-layer embedding method for improving the stability of lactic acid bacteria, and solves the problem of poor stability in the prior art.
The technical scheme of the invention is as follows:
a three-layer embedding method for improving the stability of lactic acid bacteria comprises the following steps:
s1, culturing lactobacillus in fermentation tank at high density to make total lactobacillus number in fermentation liquid reach 3.0 × 109～1.0×1010cfu/ml；
S2, centrifugally separating the bacterial sludge from the fermentation liquor, wherein the water content of the bacterial sludge is 5-50%;
s3, uniformly mixing the bacterial sludge and a first layer of embedding protective agent according to the mass ratio of 1 (1-3), wherein the first layer of embedding protective agent contains 5% -50% of skim milk powder;
s4, burying the bacterial sludge and the second layer of embedding protective agent according to the mass ratio of 1: (1-3) mixing the mixture obtained in the step S3 uniformly;
s5, mixing the bacterial sludge and the third layer of embedding protective agent according to the mass ratio of 1: (1-3) mixing the mixture with the mixture obtained in the step S4, and emulsifying uniformly to obtain an S5 emulsion;
s6, carrying out cryogenic granulation on the S5 emulsion by using a liquid nitrogen freezing technology to enable the thalli to enter a dormant state instantly to obtain S6 cryogenic granules;
s7, drying the S6 deep-frozen grains until the water content is 3% -8%.
As a further technical scheme, the second layer embedding protective agent comprises the following components in parts by weight: 2 to 20 percent of trehalose, 0.5 to 5 percent of mannitol, 0.5 to 5 percent of water-soluble vitamin E, 0.5 to 5 percent of sodium glutamate, 1 to 10 percent of beta-cyclodextrin, 1 to 10 percent of resistant dextrin, 0.1 to 1 percent of Tween 80 and the balance of water.
As a further technical scheme, the third layer of embedding protective agent comprises the following components in parts by weight: 0.4 to 4 percent of sodium alginate, 0.2 to 2 percent of Arabic gum and the balance of water.
As a further technical scheme, the deep cooling granulation is carried out in liquid nitrogen at the temperature of-200 to-190 ℃ until the radius is 0.2 cm-2 cm
As a further technical scheme, the drying is vacuum freeze drying, the primary sublimation temperature is 2-10 ℃, the vacuum degree is 0.01-0.05 mbar, the analytic drying temperature is 20-32 ℃, and the vacuum degree is 0.001-0.01 mbar.
The lactobacillus prepared by the three-layer embedding method for improving the stability of the lactobacillus is applied to the fields of food and health care products.
The working principle and the beneficial effects of the invention are as follows:
1. the invention provides a three-layer embedding method for improving the stability of lactic acid bacteria, the processed lactic acid bacteria have high fluidity, high dispersion property, low hygroscopicity, stable shelf life and strong acid resistance, the storage life activity of the lactic acid bacteria is prolonged, and the three-layer embedding method is suitable for development and use of various products. The lactic acid bacteria protected by multilayer embedding of the invention have better stability and survival rate than the lactic acid bacteria without embedding, the embedding steps of the invention need to carry out embedding and emulsification in layers in sequence, otherwise, the embedding rate of the bacteria can be reduced.
2. The skim milk powder in the first layer of embedding protective agent is mainly used as a freeze-drying filler during freeze-drying. The first layer of protective agent is used for uniformly dispersing thalli and facilitating the subsequent embedding of the embedding protective agent 2 and the embedding protective agent 3. Amino acid and micromolecular peptide in the skim milk powder can enter cells to play a role in protecting the interior of the cells, and the survival rate of thalli passing through gastric juice and bile salt is improved under the synergistic effect of the amino acid and the micromolecular peptide and the embedding protective agents 2 and 3.
3. According to the invention, trehalose and mannitol protect the cell membrane of the thallus, beta-cyclodextrin and resistant dextrin synergistically act to protect the cell wall of the thallus, the survival rate of the thallus during vacuum freeze drying is improved, water-soluble vitamin E and sodium glutamate provide antioxidation of the thallus during vacuum freeze drying and normal temperature storage, and the synergistic effect with other components is used for improving the shelf life stability of a lactobacillus microecological preparation product and the survival rate of the lactobacillus microecological preparation product when the lactobacillus microecological preparation product passes through gastric juice and bile salt.
4. Many free hydrophilic groups (hydroxyl and carboxyl) on the molecular chain of sodium alginate, Ca2+、Zn2+、Al3+The gel can occupy the hydrophilic space in the sodium alginate and chelate with oxygen atoms on carboxyl functional groups to form three-dimensional network structure gel, and the thalli are embedded in the gel. The reaction speed in the process is high, and the reaction condition is mild. Arabic gum is a natural high molecular product obtained by partially hydrolyzing collagen, and can form a gel structure under low temperature conditions. When the pH value of the environment is lower than the isoelectric point of gelatin, the positive charges and the negative charges on the surface can interact, so that the microcapsule structure is more compact. Meanwhile, the long-chain molecules of sodium alginate and Arabic gum are used as embedding protective agent wall materials, and the thalli and the embedding protective agents 1 and 2 are wrapped in the microcapsules, so that the stability of the thalli stored at normal temperature is improved.
5. The embedding method has the advantages of simple production process, low energy consumption, no three wastes, small investment and easy large-scale production.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any inventive step, are intended to be within the scope of the present invention.
The technique of high density culture of lactic acid bacteria in a fermenter is a conventional technique in the prior art and is not described herein. Survival was counted using a pour plate meter.
Three-layer embedding process of lactobacillus plantarum Lp45 strain:
example 1
S1, high-density culturing the strain in a fermentation tank according to the prior art to ensure that the viable count of the lactobacillus plantarum in the fermentation liquor reaches 8.5 multiplied by 109cfu/ml, centrifuging the fermentation liquor by using a butterfly centrifuge under the condition of ensuring that thalli are not damaged, and concentrating the fermentation liquor by 70 times to obtain bacterial sludge;
s2, adding an embedding protective agent 1 into the bacterial sludge according to the mass ratio of 1:3, wherein the embedding protective agent 1 is 5% of skimmed milk powder and 95% of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s3, preparing the bacterial sludge: adding an embedding protective agent 2 into the protective agent 2 according to the mass ratio of 1:3, wherein the embedding protective agent 2 comprises 2% of trehalose, 0.5% of mannitol, 0.5% of water-soluble vitamin E, 0.5% of sodium glutamate, 1% of beta cyclodextrin, 1% of resistant dextrin, 0.1% of Tween 80 and the balance of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s4, preparing the bacterial sludge: adding an embedding protective agent 3 into a protective agent 3 according to the mass ratio of 1:3, wherein the embedding protective agent 3 is 0.4% of sodium alginate, 0.2% of Arabic gum and 99.4% of water (mass ratio), fully and uniformly mixing an emulsion obtained by mixing the bacterial sludge and the protective agent by using a homogenizer, after the bacterial sludge and the three-layer protective agent are emulsified, dripping the emulsion into liquid nitrogen at minus 190 ℃ by using a liquid nitrogen dripping freezing technology for deep cooling granulation to enable the thalli to enter a dormant state instantly, and then carrying out vacuum freeze drying in a freeze dryer, wherein the primary sublimation temperature is 10 ℃, the vacuum degree is 0.015mbar, the analytic drying temperature is 32 ℃, the vacuum degree is 0.005mbar, and the freeze drying is finished until the water content of a product is 4.5%, thus obtaining the finished product.
Finished product fungus powder live bacteriaThe number is 6.0X 1011cfu/g。
Example 2
S1, high-density culturing the strain in a fermentation tank according to the prior art to ensure that the viable count of the lactobacillus plantarum in the fermentation liquor reaches 8.5 multiplied by 109cfu/ml, centrifuging the fermentation liquor by using a butterfly centrifuge under the condition of ensuring that thalli are not damaged, and concentrating the fermentation liquor by 70 times to obtain bacterial sludge;
s2, adding an embedding protective agent 1 into the bacterial sludge according to the mass ratio of 1:2, wherein the embedding protective agent 1 is 30% of skimmed milk powder and 70% of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s3, preparing the bacterial sludge: adding an embedding protective agent 2 into a protective agent 2 according to the mass ratio of 1:2, wherein the embedding protective agent 2 comprises 10% of trehalose, 3% of mannitol, 3% of water-soluble vitamin E, 3% of sodium glutamate, 6% of beta cyclodextrin, 6% of resistant dextrin, 0.5% of Tween 80 and the balance of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s4, preparing the bacterial sludge: adding an embedding protective agent 3 into a protective agent 3 according to the mass ratio of 1:2, wherein the embedding protective agent 3 is 2% of sodium alginate, 1% of Arabic gum and 97% of water (mass ratio), fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer, after the bacterial sludge and the three-layer protective agent are emulsified, dripping the emulsion into liquid nitrogen with the temperature of minus 196 ℃ by using a liquid nitrogen dripping freezing technology to carry out deep cooling granulation so that the bacterial cells enter a dormant state instantly, then carrying out vacuum freeze drying in a freeze dryer, setting the primary sublimation temperature to 10 ℃, the vacuum degree to 0.015mbar, the analytic drying temperature to 32 ℃, the vacuum degree to 0.005mbar, and finishing freeze drying until the product water content is 4.2%, thus obtaining the finished product.
The viable count of the finished product of the bacterial powder is 6.5 multiplied by 1011cfu/g。
Example 3
S1, high-density culturing the strain in a fermentation tank according to the prior art to ensure that the viable count of the lactobacillus plantarum in the fermentation liquor reaches 8.5 multiplied by 109cfu/ml, and is fermented by butterfly centrifuge under the condition of ensuring the undamaged thallusCentrifuging the fermentation liquor to concentrate the fermentation liquor by 70 times to obtain bacterial sludge;
s2, adding an embedding protective agent 1 into the bacterial sludge according to the mass ratio of 1:1, wherein the embedding protective agent 1 is 50% of skimmed milk powder and 50% of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s3, preparing the bacterial sludge: adding an embedding protective agent 2 into a protective agent 2 according to the mass ratio of 1:1, wherein the embedding protective agent 2 comprises 20% of trehalose, 5% of mannitol, 5% of water-soluble vitamin E, 5% of sodium glutamate, 10% of beta cyclodextrin, 10% of resistant dextrin, 1% of Tween 80 and the balance of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s4, preparing the bacterial sludge: adding an embedding protective agent 3 into a protective agent 3 according to the mass ratio of 1:1, wherein the embedding protective agent 3 is 4% of sodium alginate, 2% of Arabic gum and 94% of water (mass ratio), fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer, after the bacterial sludge and the three layers of protective agents are emulsified, dripping the emulsion into liquid nitrogen at 200 ℃ below zero by using a liquid nitrogen dripping freezing technology to carry out deep cooling granulation so that the bacterial cells enter a dormant state instantly, then carrying out vacuum freeze drying in a freeze dryer, setting the primary sublimation temperature to 10 ℃, the vacuum degree to 0.015mbar, the analytic drying temperature to 32 ℃, the vacuum degree to 0.005mbar, and finishing freeze drying until the product water content is 4%, thus obtaining the finished product.
The viable count of the finished product of the bacterial powder is 5.7 multiplied by 1011cfu/g。
Comparative example 1
The same procedure was followed except that the 3% water-soluble vitamin E in example 2 was replaced with water. The viable count of the obtained finished product bacteria powder is 2.9 multiplied by 1011cfu/g。
Comparative example 2
The 6% resistant dextrin of example 2 was replaced with water and the other operations were the same. The viable count of the obtained finished product bacteria powder is 2.6 multiplied by 1011cfu/g。
Comparative example 3
The third layer of the embedding protective agent in example 2 was removed and the other operations were the same. The viable count of the obtained finished product bacteria powder is 1.1 × 1011cfu/g。
Comparative example 4
The gum arabic was removed from example 2 and the procedure was otherwise the same. The viable count of the obtained finished product bacteria powder is 2.1 × 1011cfu/g。
A three-layer embedding process of lactobacillus rhamnosus LR863 strain comprises the following steps:
example 4
S1, high-density culturing the strain in a fermentation tank according to the prior art to ensure that the viable count of the lactobacillus plantarum in the fermentation liquor reaches 1.0 multiplied by 1010cfu/ml, centrifuging the fermentation liquor by using a butterfly centrifuge under the condition of ensuring that thalli are not damaged, and concentrating the fermentation liquor by 70 times to obtain bacterial sludge;
s2, adding an embedding protective agent 1 into the bacterial sludge according to the mass ratio of 1:3, wherein the embedding protective agent 1 is 5% of skimmed milk powder and 95% of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s3, preparing the bacterial sludge: adding an embedding protective agent 2 into the protective agent 2 according to the mass ratio of 1:3, wherein the embedding protective agent 2 comprises 2% of trehalose, 0.5% of mannitol, 0.5% of water-soluble vitamin E, 0.5% of sodium glutamate, 1% of beta cyclodextrin, 1% of resistant dextrin, 0.1% of Tween 80 and the balance of water (mass ratio), and fully and uniformly mixing the bacterial sludge and the protective agent mixed emulsion by using a homogenizer;
s4, preparing the bacterial sludge: adding an embedding protective agent 3 into a protective agent 3 according to the mass ratio of 1:3, wherein the embedding protective agent 3 is 0.4% of sodium alginate, 0.2% of Arabic gum and 99.4% of water (mass ratio), fully and uniformly mixing an emulsion obtained by mixing the bacterial sludge and the protective agent by using a homogenizer, after the bacterial sludge and the three layers of protective agents are emulsified, dripping the emulsion into liquid nitrogen with the temperature of minus 196 ℃ by using a liquid nitrogen dripping freezing technology for deep cooling granulation to enable the thalli to enter a dormant state instantly, and then carrying out vacuum freeze drying in a freeze dryer, wherein the primary sublimation temperature is 8 ℃, the vacuum degree is 0.01, the analytic drying temperature is 30 ℃, the vacuum degree is 0.008, and the freeze drying is finished until the water content of a product is 5.2%, thus obtaining the finished product.
The viable count of the finished product of the bacterial powder is 5.0 multiplied by 1011cfu/g。
Comparative example 5
The first layer of embedding medium of example 4 was removed and the procedure was otherwise the same. The viable count of the obtained finished product bacteria powder is 2.7 multiplied by 1011cfu/g。
Comparative example 6
The sodium alginate in example 4 was replaced with water and the other operations were the same. The viable count of the obtained finished product bacteria powder is 3.9 multiplied by 1011cfu/g。
Comparative example 7
The embedding protective agent 3 and the embedding protective agent 2 in example 4 were interchanged in sequence, and the other operations were the same. The viable count of the obtained finished product bacteria powder is 9.2 multiplied by 1010cfu/g。
The examples and comparative examples and the original strains were subjected to manual accelerated experiments. And (3) acceleration conditions: and (3) detecting the viable count of the bacterial powder at the temperature of 37 ℃ and the relative humidity of 75 percent for 56 days, and calculating the survival rate, wherein the survival rate refers to the ratio of the detected value of the viable count to the initial value after being stored for a period of time and is expressed by percentage. And the other group is stored for 56 days at normal temperature, the viable count of the bacterial powder is detected, and the survival rate is calculated.
TABLE 1 survival rate of examples and comparative examples at different temperatures of 56d
| Survival rate at different temperatures for 56d | Example 4 | Comparative example 5 | Comparative example 6 | Comparative example 7 | Lactobacillus rhamnosus LR863 |
| 25℃ | 46％ | 30％ | 31％ | 28％ | 25％ |
| 37℃ | 11％ | 7％ | 9％ | 6％ | 4％ |
The survival rates of the examples and comparative examples were counted after 1h testing in simulated gastric fluid and after 1h in medium containing bile salts.
TABLE 2 survival rates of different media of examples and comparative examples
The lactic acid bacteria of examples and comparative examples were subjected to a hygroscopicity test under test conditions: placing 5g of a test sample in a culture dish with the diameter of 9cm, covering the culture dish, placing the culture dish in a constant-temperature constant-humidity incubator at the temperature of 30 ℃, the relative humidity of 50% for 12h, calculating the moisture absorption rate, wherein the moisture absorption rate is the weight of the sample/the original mass of the sample and is expressed by percentage, and the specific results are shown in table 3.
TABLE 3 moisture absorption Rate of examples and comparative examples
| Sample (I) | Example 4 | Comparative example 5 | Comparative example 6 | Comparative example 7 | Lactobacillus rhamnosus LR863 |
| Moisture absorption rate% | 11％ | 16％ | 15％ | 22％ | 38％ |
Comparing example 2 with comparative examples 1 and 2, it is found that when vitamin E is absent or resistant dextrin is absent, the survival rate and stability of the obtained lactic acid bacteria are reduced, and the moisture absorption rate is increased. Compared with the embodiment of the invention, the survival rate of the prepared finished bacterial powder is greatly reduced, the stability is poor and the moisture absorption rate is high. The first layer of embedded protective agent is not arranged in the comparative example 5, the survival rate of the obtained lactobacillus is reduced, the stability is reduced, the moisture absorption rate is increased, amino acid and small molecular peptide in the skim milk powder can enter cells to play a role in protecting the inside of the cells, and the survival rate of the bacteria passing through gastric juice and bile salt is improved under the synergistic effect of the embedded protective agent 2 and the embedded protective agent 3. In the comparative example 6, sodium alginate is not added, the stability is reduced, long-chain molecules in the sodium alginate are used as embedding protective agent wall materials, and the thalli and the embedding protective agents 1 and 2 are wrapped in the microcapsules, so that the stability of the thalli stored at normal temperature is improved. Comparative example 7 the order of the two-layer and three-layer embedding protective agents was exchanged, the stability of the finished bacterial powder was also reduced, and the best protective effect on the bacterial powder could be achieved only when the three-layer embedding protective agents were sequentially embedded and emulsified. Therefore, the treatment method of the invention can only play the best stabilizing effect under the synergistic effect of the compounding of the components.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A three-layer embedding method for improving the stability of lactic acid bacteria is characterized by comprising the following steps:
s1, culturing lactobacillus in fermentation tank at high density to make total lactobacillus number in fermentation liquid reach 3.0 × 109～1.0×1010cfu/ml；
S2, centrifugally separating the bacterial sludge from the fermentation liquor, wherein the water content of the bacterial sludge is 5-50%;
s3, uniformly mixing the bacterial sludge and a first layer of embedding protective agent according to the mass ratio of 1 (1-3), wherein the first layer of embedding protective agent contains 5% -50% of skim milk powder;
s4, burying the bacterial sludge and the second layer of embedding protective agent according to the mass ratio of 1: (1-3) mixing the mixture obtained in the step S3 uniformly;
s5, mixing the bacterial sludge and the third layer of embedding protective agent according to the mass ratio of 1: (1-3) mixing the mixture with the mixture obtained in the step S4, and emulsifying uniformly to obtain an S5 emulsion;
s6, carrying out cryogenic granulation on the S5 emulsion by using a liquid nitrogen freezing technology to enable the thalli to enter a dormant state instantly to obtain S6 cryogenic granules;
s7, drying the S6 deep-frozen grains until the water content is 3% -8%.
2. The three-layer embedding method for improving the stability of lactic acid bacteria according to claim 1, wherein the second layer of embedding protective agent comprises the following components by weight: 2 to 20 percent of trehalose, 0.5 to 5 percent of mannitol, 0.5 to 5 percent of water-soluble vitamin E, 0.5 to 5 percent of sodium glutamate, 1 to 10 percent of beta-cyclodextrin, 1 to 10 percent of resistant dextrin, 0.1 to 1 percent of Tween 80 and the balance of water.
3. The three-layer embedding method for improving the stability of lactic acid bacteria according to claim 1, wherein the third layer of embedding protective agent comprises the following components by weight: 0.4 to 4 percent of sodium alginate, 0.2 to 2 percent of Arabic gum and the balance of water.
4. The three-layer embedding method for improving the stability of lactic acid bacteria according to claim 1, wherein the deep cooling granulation is performed in liquid nitrogen at-200 to-190 ℃ until the radius is 0.2cm to 2 cm.
5. The three-layer embedding method for improving the stability of lactic acid bacteria according to claim 1, wherein the drying is vacuum freeze drying, the primary sublimation temperature is 2 to 10 ℃, the vacuum degree is 0.01 to 0.05mbar, the analytic drying temperature is 20 to 32 ℃, and the vacuum degree is 0.001 to 0.01 mbar.
6. The application of the lactobacillus prepared by the three-layer embedding method for improving the stability of the lactobacillus according to any one of claims 1 to 5 in the fields of food and health care products.
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| CN115644448A (en) * | 2022-09-07 | 2023-01-31 | 西安诺众康健生物科技有限责任公司 | Embedding protection method for intestinal probiotics |
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| CN116121074A (en) * | 2023-03-16 | 2023-05-16 | 亩巴贝农业科技有限公司 | Three-layer embedding method for improving stability of microorganism bacteria and application thereof |
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