A compound bacterium for relieving stress and promoting sleep and its application

Abstract

The present invention relates to the field of microbial technology, and particularly to a composite bacterium and its application. The present invention provides a composite bacterium and its application, wherein the composite bacterium includes Saccharomyces cerevisiae and Lactobacillus acidophilus. This composite bacterium can effectively relieve stress levels, improve fatigue, enhance stress resistance, extend sleep duration, and improve sleep quality. Moreover, when the amount of bacteria used remains the same, compared with single Saccharomyces cerevisiae or single Lactobacillus acidophilus, the combined effect of the two strains is better, that is, Saccharomyces cerevisiae and Lactobacillus acidophilus can cooperate with each other and have a significant synergistic effect in relieving stress and promoting sleep.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and in particular to a compound bacterium for relieving stress and promoting sleep and its application. Background Art

[0002] Sleep disorder refers to abnormal sleep quantity and abnormal behaviors during sleep, including sleep disorders (excessive / insufficient sleep quantity) and parasomnias (abnormal behaviors or physiology, such as sleepwalking and talking in sleep). With the continuous acceleration of the life rhythm, the number of people with sleep disorders is increasing day by day.

[0003] Currently, drugs are often used to relieve anxiety, depression or insomnia symptoms. Commonly used anti-anxiety and anti-depressant drugs in clinical practice include amitriptyline, sertraline, maprotiline, etc. These drugs mainly play a role by regulating the reabsorption of 5-hydroxytryptamine. Although they can significantly relieve symptoms, long-term use will cause side effects such as nausea and vomiting, and they are not suitable for mild symptoms and sensitive patients. Insomnia patients mainly rely on sleeping pills to relieve symptoms, but long-term use of sleeping pills will cause drug addiction and even affect physical health. Therefore, there is an urgent need to find a safe and effective functional supplement that can be taken for a long time and has the effects of relieving stress and promoting sleep.

[0004] The intestine contains hundreds of millions of microorganisms, including beneficial bacteria and harmful bacteria, which affect the function of the brain, our mood and behavior. These intricate and interconnected communication channels constitute what we know as the "gut-brain axis". In recent years, the "gut-brain axis" has become a research hotspot. More and more scientific studies have found that gut microbiota communicate with the central nervous system through neural, immune and endocrine pathways, thus affecting brain function and behavior. Probiotics, as an effective means to regulate the balance of gut microbiota, show great potential in the prevention and treatment of intricate central nervous system diseases.

[0005] It can be seen that the importance of probiotics in regulating the nervous system, improving sleep or improving emotional health has been gradually recognized. However, in recent years, scientists have also begun to explore the potential role of probiotics in improving sleep, and this discovery has attracted extensive attention. Although a large number of studies have pointed to the potential link between probiotics and mood regulation, the reality is that there are still very few probiotic products on the market that can truly provide effective regulation for sleep quality. This market gap not only reflects the depth of scientific research and the lag in product development, but also reveals an important development direction for the future probiotic industry. Therefore, developing more efficient probiotic products targeting sleep quality will become an important topic in the health field. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a compound bacterium for relieving stress and promoting sleep and its application.

[0007] The compound bacterium provided by the present invention is composed of Saccharomyces cerevisiae and Lactobacillus acidophilus.

[0008] In the present invention, the quantity ratio of Saccharomyces cerevisiae to Lactobacillus acidophilus is 1:5 to 5:1.

[0009] In some embodiments, the quantity ratio of Saccharomyces cerevisiae to Lactobacillus acidophilus is 1:(0.2 - 5). In specific embodiments, the quantity ratio of Saccharomyces cerevisiae to Lactobacillus acidophilus is 1:1, 1:5 or 5:1.

[0010] The yeast probiotic agent provided by the present invention can effectively relieve stress levels, improve fatigue, enhance stress resistance, extend sleep time, and improve sleep quality. And when the amount of bacteria used is kept the same, compared with single Saccharomyces cerevisiae or single Lactobacillus acidophilus, the combined effect of the two bacteria is better, that is, Saccharomyces cerevisiae and Lactobacillus acidophilus can cooperate with each other. Especially when the quantity ratio of the two is 1:5 to 5:1, they have a significant synergistic effect in relieving stress and promoting sleep.

[0011] In some specific embodiments, the Saccharomyces cerevisiae is Saccharomyces cerevisiae with the preservation number of CCTCC NO:M2019643.

[0012] In some specific embodiments, the Lactobacillus acidophilus is Lactobacillus acidophilus with the preservation number of CCTCC NO:M2023768.

[0013] The present invention also provides a probiotic agent, which includes the compound bacterium as described above and a protective agent.

[0014] The probiotic agent described in the present invention is prepared from the compound bacterium as described above and a protective agent, and it is in the form of powder, emulsion, tablet or capsule.

[0015] In the probiotic agent described in the present invention, the total viable count is not less than 1×10 10 CFU / ml or not less than 1×10 10 CFU / g.

[0016] In the probiotic agent described in the present invention, the protective agent includes at least one of milk powder, starch, gelatin, dextrin, vegetable oil, sorbitol, sorbitan monostearate, sucrose, lactose, polyvinylpyrrolidone. In some specific embodiments, the protective agent is dry starch, or milk powder, or dextrin.

[0017] The present invention also provides a preparation method of the probiotic agent, including:

[0018] Saccharomyces cerevisiae and Lactobacillus acidophilus were separately inoculated into a culture medium for cultivation, and then the cells were separately centrifuged to obtain cell bodies; the obtained cell bodies were granulated or freeze-dried after adding a cryoprotectant.

[0019] In the preparation method of the present invention, the culture medium composition of Saccharomyces cerevisiae An-15 is: sucrose 100 g / L, yeast extract 20 g / L, KH2PO4 1 g / L, MgSO4 0.5 g / L. The culture conditions include activation by culturing at 30 °C for 26 h to obtain an activation solution, and the activated fermentation broth is inoculated into a liquid medium for shake flask culture.

[0020] In the preparation method of the present invention, the culture medium composition of Lactobacillus acidophilus is: peptone 10 g, beef extract 10 g, glucose 20 g, sodium acetate 2 g, yeast powder 5 g, diammonium hydrogen citrate 2 g, K2PO4·3H2O 2.6 g, MgSO4·7H2O 0.1 g, MnSO4 0.05 g, and cysteine hydrochloride 0.5 g. The culture conditions include activation by culturing at 37 °C for 24 h, and continuous activation for 2 times to obtain an activation solution; the activation solution is inoculated into MRS liquid medium at an inoculation amount of 2% (v / v) and cultured at 37 °C for 24 h to obtain a bacterial solution; the bacterial solution is centrifuged to obtain Lactobacillus acidophilus cell bodies.

[0021] In the described preparation method, the centrifugation conditions include 10000 r / min for 10 min, and the mass ratio of the cell bodies to the cryoprotectant is 1:6. The granulation conditions include adding an emulsifier and a cryoprotectant, and the freeze-drying conditions include a temperature of -80 °C for 24 h.

[0022] Furthermore, the present invention also provides the application of the composite bacteria as described above, the probiotic agent as described above, or the probiotic agent prepared by the preparation method as described above, in the preparation of a product for improving stress resistance and / or improving sleep.

[0023] In the present invention, the improvement of stress resistance includes improving stress resistance, reducing the stress index and / or reducing the fatigue index.

[0024] In the present invention, the improvement of sleep includes prolonging the deep sleep time and / or increasing the sleep quality score.

[0025] Even further, the present invention also provides a product for improving stress resistance and / or improving sleep, which includes: the composite bacteria as described above, the probiotic agent as described above, or the probiotic agent prepared by the preparation method as described above.

[0026] The product for improving stress resistance and / or improving sleep according to the present invention further includes prebiotics and / or excipients.

[0027] In the present invention, the prebiotic includes at least one of fructooligosaccharide, galactooligosaccharide, isomaltooligosaccharide, xylooligosaccharide, stachyose, inulin, polydextrose, resistant dextrin, soybean oligosaccharide, and yeast β-glucan.

[0028] In the present invention, the excipient includes at least one of a filler, a binder, a wetting agent, a disintegrant, an emulsifier, a cosolvent, a solubilizer, a colorant, an antioxidant, and a flavoring agent.

[0029] The filler plays a role in increasing the volume of the drug, improving the physical properties of the drug, facilitating the shaping and stability of the drug in pharmaceutical preparations. In some embodiments, the filler is selected from at least one of starch, dextrin, pregelatinized starch (also known as compressible starch), powdered sugar, lactose, mannitol, microcrystalline cellulose (MCC), inorganic salts, sorbitol, or calcium hydrogen phosphate.

[0030] The binder is used to impart appropriate viscosity to materials with no or insufficient viscosity. In the present invention, the binder is selected from at least one of hydroxypropyl methylcellulose (HPMC), sodium carboxymethylcellulose (CMC-Na), corn starch, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), ethylcellulose (EC), polyvinyl alcohol (PVA), syrup, maltose syrup, or microcrystalline cellulose (MCC).

[0031] The wetting agent is a substance that reduces the surface tension or interfacial tension of water, enables water to spread on the surface of solid materials or penetrate into their surfaces, and thus wets the solid materials. In some embodiments, the wetting agent is selected from at least one of water, ethanol, tea seed cake, Nekal, soybean lecithin, sulfonated oil, methylcellulose, or glycerol.

[0032] The main function of the disintegrant is to accelerate the disintegration and dissolution of pharmaceutical preparations after administration, so as to ensure that the active ingredients of the drug can be rapidly released and absorbed by the human body, thereby improving the bioavailability and efficacy of the drug. In some embodiments, it is at least one of dry starch, sodium carboxymethyl starch (CMS-Na), low-substituted hydroxypropyl cellulose (L-HPC), cross-linked polyvinylpyrrolidone (also known as cross-linked PVP), cross-linked sodium carboxymethyl cellulose (CCNa), effervescent disintegrant (usually composed of sodium bicarbonate and acidic substances such as citric acid), microcrystalline cellulose (MCC), lactose, calcium sulfate (as a kind of inorganic salt disintegrant), or mannitol.

[0033] The emulsifiers include anionic emulsifiers, non-ionic emulsifiers or natural emulsifiers. Among them, the anionic emulsifiers are selected from sodium dodecyl sulfate (SDS), stearates or oleates; the non-ionic emulsifiers are selected from span types (sorbitan fatty acid esters), tween types (polysorbates), poloxamer (Pluronic F68), sucrose stearate, benzethonium or myrj. The natural emulsifiers are selected from gum arabic, tragacanth or gelatin.

[0034] The cosolvent is a complex, association compound or double salt formed by a poorly soluble drug and a third substance added in a solvent to increase the solubility of the drug in the solvent. In some embodiments, the cosolvent is selected from sodium benzoate, sodium salicylate, p-aminobenzoic acid, urea, nicotinamide, acetamide, potassium iodide, citric acid (also known as lemon acid), polyvinylpyrrolidone or ethylenediamine.

[0035] The solubilizer refers to a surfactant with solubilizing ability. In some embodiments, the solubilizer is selected from liquid solubilizers, such as petroleum ether, ethers (such as diethyl ether), alcohols (such as ethanol), acetone; or solid solubilizers, such as anilines, chloroform, vinyl chloride, trichloroethylene, tetrachloroethylene, trimethylbenzene; or surfactant-based solubilizers, such as polysorbates (such as Tween 80), polyoxyethylene fatty acid esters, PEG-40 hydrogenated castor oil (trade name: CO 40 or RH-40), PEG-50 hydrogenated castor oil or SP-115C.

[0036] The colorant is selected from beet red, turmeric, lac red, β-carotene, amaranth, carmine, lemon yellow, indigo or β-carotene.

[0037] The antioxidant is used to prevent the adverse effects of oxygen and is selected from butylated hydroxyanisole (BHA), dibutylhydroxytoluene (BHT), ascorbic acid (vitamin C), vitamin E, carotenoids, ascorbyl palmitate, tea polyphenols or phytic acid.

[0038] The flavoring agent is a pharmaceutical excipient that improves or masks the unpleasant odor and taste of drugs, including fruit powders, edible flavors, sweeteners or acidulants. Preferably, the fruit powder is one or a mixture of two or more of orange powder, orange powder, lemon powder, cherry powder, apple powder or coconut powder. The edible flavor is one or a mixture of two or more of orange essence, orange essence, lemon essence, cherry essence, menthol, apple essence or coconut essence. The sweetener is one or a mixture of two or more of sucralose, acesulfame potassium, aspartame or mogroside. The acidulant is one or a mixture of two or more of citric acid, malic acid, lactic acid or citric acid.

[0039] In some embodiments, the dosage forms of the products as described above are tablets, pills, oral liquid preparations, capsules, syrups, dripping pills or granules. For example, the capsules are hard capsules or soft capsules. The tablets are oral tablets or buccal tablets. In the present invention, oral tablets refer to tablets for oral administration. In most such tablets, the drugs are absorbed through the gastrointestinal tract to exert their effects, and in some tablets, the drugs exert their effects locally in the gastrointestinal tract. In some embodiments provided by the present invention, the oral tablets are ordinary compressed tablets, dispersible tablets, effervescent tablets, chewable tablets, coated tablets or sustained-release or controlled-release tablets.

[0040] Furthermore, the present invention also provides a method for improving stress resistance and / or improving sleep, which includes administering the composite bacteria, probiotic agent or product as described above.

[0041] The administration in the present invention can be implemented by any suitable method known in the art, including but not limited to oral, buccal, sublingual, ocular, topical, parenteral, rectal, intrathecal, intracisternal, inguinal, intravesical, topical (such as powders, ointments or drops), or nasal routes, preferably oral.

[0042] The present invention provides a composite bacteria and its application. The composite bacteria include Saccharomyces cerevisiae and Lactobacillus acidophilus. The composite bacteria can effectively relieve stress levels, improve fatigue, enhance stress resistance, prolong sleep time, and improve sleep quality. And when the amount of bacteria used is kept the same, compared with single Saccharomyces cerevisiae or single Lactobacillus acidophilus, the combined effect of the two strains is better, that is, Saccharomyces cerevisiae and Lactobacillus acidophilus can cooperate with each other and have a significant synergistic effect in relieving stress and promoting sleep. Description of the Drawings

[0043] Figure 1 Comparison of stress resistance before and after the trial diet;

[0044] Figure 2 Comparison of stress index before and after the trial diet;

[0045] Figure 3 Comparison of fatigue index before and after the trial diet;

[0046] Figure 4 Change in deep sleep time (h) before and after the trial diet;

[0047] Figure 5 Change in sleep quality score before and after the trial diet. Detailed Embodiments

[0048] The present invention provides a compound bacterium for relieving stress and promoting sleep and its application. Those skilled in the art can draw on the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments. It is obvious that relevant personnel can make changes or appropriate alterations and combinations to the methods and applications in this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0049] The numerical ranges and parameters involved in the present invention have been presented as precisely as possible in the specific embodiments. However, any numerical value inevitably contains standard deviations caused by individual testing methods. Therefore, unless otherwise clearly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have certain reasonable deviations within a certain range, such as within ±10%, ±5%, ±1% or ±0.5%.

[0050] Some cases are recorded in the examples and comparative examples of the present invention. Among them, the examples show certain implementation manners of the present invention. However, this does not mean that the effects of the present invention can only be achieved in these cases. In fact, any concentration between the two endpoint values shown in the examples for each component can achieve a good effect of improving sleep quality, and the case with the best effect is Group 6. Only some cases with poor effects in the tests are listed in each comparative example. In addition, many attempts have been made during the research and development process, such as using different components, different ratios, different culture media or different addition times, but the effects of these attempts are not as good as those of Group 6, and will not be elaborated here.

[0051] The test materials used in the present invention are all ordinary commercially available products and can be purchased in the market.

[0052] Reagents and instruments used in the examples

[0053]

[0054]

[0055] Among them, the Saccharomyces cerevisiae in the probiotic agent is Saccharomyces cerevisiae An-15 with the preservation number of CCTCC NO: M 2019643, and the Lactobacillus acidophilus in the probiotic agent is Lactobacillus acidophilus LA15 with the preservation number of CCTCC NO: M 2023768. The present invention will be further elaborated below in combination with the examples:

[0056] Example 1 Preparation of probiotic agent

[0057] Inoculate Saccharomyces cerevisiae An-15 into a liquid medium and culture it at 37 °C for 26 h for activation. Then inoculate the activated solution into a liquid medium and culture it at 30 °C for 24 h to obtain a seed fermentation broth. Centrifuge the fermentation broth (8000 r / min, 10 min) to obtain yeast cells. After adding an emulsifier and vegetable oil to the cells, add starch for granulation to obtain yeast cells (Saccharomyces cerevisiae 20 billion CFU / g).

[0058] Inoculate Lactobacillus acidophilus LA15 strain into MRS liquid medium and culture it at 37 °C for 24 h for activation. Activate it continuously twice to obtain an activated solution; inoculate the activated solution into MRS liquid medium at an inoculation amount of 2% (v / v) and culture it at 37 °C for 24 h to obtain a bacterial solution; centrifuge the bacterial solution at 10000 r / min for 10 min, wash it with physiological saline 2 - 3 times, and collect Lactobacillus acidophilus cells; resuspend the cells with an aqueous solution of skim milk powder with a mass concentration of 13% to obtain a cell suspension; pre-culture the cell suspension at 37 °C for 1 h and then freeze-dry it. The freeze-drying time is 24 h to obtain Lactobacillus acidophilus powder (Lactobacillus acidophilus 200 billion CFU / g).

[0059] Compound the two bacterial powder raw materials in a certain proportion to obtain a probiotic freeze-dried powder.

[0060] Control Example 1

[0061] According to the method described above, prepare a probiotic freeze-dried powder from Saccharomyces cerevisiae An-15 and Lactobacillus casei Zhang.

[0062] Efficacy Verification

[0063] 1. Mental Stress Test

[0064] 1.1 Experimental Method

[0065] 1.1.1. Volunteer Recruitment and Screening

[0066] Source: Inside the company (including the Nutrition and Health Technology Center, Production Department, Technical Quality Department, Marketing Department)

[0067] First, use a mental stress analyzer to conduct a preliminary test on the company's internal employees, and screen volunteers according to the test results. Among them, the stress resistance range: 50 - 70 extremely poor; 70 - 90 poor; 90 - 110 normal; 110 - 130 good; 130 - 150 excellent. The stress index range: 130 - 150 extremely poor; 110 - 130 poor; 90 - 110 normal; 70 - 90 good; 50 - 70 excellent. The fatigue index range: 130 - 150 extremely poor; 110 - 130 poor; 90 - 110 normal; 70 - 90 good; 50 - 70 excellent.

[0068] The inclusion criteria are as follows: Any two of the following can be included in the group (compressive capacity ≤ 90, stress index ≥ 110, fatigue index ≥ 110).

[0069] 1.1.2 Method

[0070] Through screening, 180 eligible volunteers were selected to conduct a 14-day trial food experiment.

[0071] The volunteers were randomly divided into six groups (30 people in each group):

[0072] Control group (denoted as group 0),

[0073] An-15 group (denoted as group 1),

[0074] LA15 group (denoted as group 2),

[0075] Commercially available probiotic strains (denoted as group 3),

[0076] An-15 + LA15 (viable count ratio 1:1, denoted as group 4),

[0077] Commercially available strain Lactobacillus casei Zhang + An-15 (viable count ratio 1:1, denoted as group 6).

[0078] The above control group received placebo (maltodextrin) to exclude the interference of environmental factors. The other five groups received the corresponding probiotic powder containing 1*10 10 CFU / g.

[0079] After the administration, a mental stress analysis test was conducted on them using a mental stress analyzer. Data analysis was performed using Minitab software.

[0080] 1.2 Result analysis

[0081] The influence results of the probiotic agent on the mental stress of the subjects are shown respectively as Figures 1 to 3 and Tables 1 - 3.

[0082] Table 1 Compressive capacity

[0083]

[0084] Note: NS indicates no significant difference before and after the trial food;

[0085] * indicates a significant difference before and after the trial food, p < 0.05;

[0086] ** indicates a significant difference before and after the trial food, p < 0.01.

[0087] Table 2 Stress index

[0088]

[0089] Note: NS indicates no significant difference before and after the trial food intake;

[0090] * indicates a significant difference before and after the trial food intake, p < 0.05;

[0091] ** indicates a significant difference before and after the trial food intake, p < 0.01.

[0092] Table 3 Fatigue Index

[0093]

[0094]

[0095] Note: NS indicates no significant difference before and after the trial food intake;

[0096] * indicates a significant difference before and after the trial food intake, p < 0.05;

[0097] ** indicates a significant difference before and after the trial food intake, p < 0.01.

[0098] According to the results of the trial food intake, there was no obvious change in the stress state of the subjects in the control group before and after taking the placebo. In addition, for the test groups taking probiotics, the stress resistance and stress index of groups 2 and 5 were significantly improved before and after the trial food intake, and the fatigue index of all subjects taking probiotics was significantly improved (p < 0.05). Among these groups, especially the improvement effect of group 5 was particularly significant (p < 0.01), indicating that compared with single components, Saccharomyces cerevisiae An-15 and Lactobacillus acidophilus LA15 can cooperate with each other and synergistically enhance the effect in regulating stress.

[0099] 2. Sleep Effect Test

[0100] 2.1 Experimental Method

[0101] The enrolled volunteers in the previous mental stress test experiment were simultaneously monitored for sleep quality, and the analysis of the sleep influence results was carried out based on the data before and after the trial food intake.

[0102] 2.2 Result Analysis

[0103] The influence results of the probiotics on the sleep status of the subjects are shown respectively in Figures 4 to 5 and Tables 4 - 5.

[0104] Table 4 Deep Sleep Time

[0105]

[0106] Note: NS indicates no significant difference before and after the trial food intake;

[0107] *There was a significant difference before and after the trial diet, p < 0.05;

[0108] **There was a significant difference before and after the trial diet, p < 0.01.

[0109] Table 5 Sleep Quality

[0110]

[0111] Note: NS indicates that there is no significant difference before and after the trial diet;

[0112] *There was a significant difference before and after the trial diet, p < 0.05;

[0113] **There was a significant difference before and after the trial diet, p < 0.01.

[0114] According to the results of the trial diet, there was no obvious change in the sleep situation of the subjects in the control group before and after taking the placebo. In addition, in the test group taking the probiotic agent, the deep sleep time in groups 2, 5, and 6 was significantly prolonged before and after the trial diet (p < 0.05). However, only the sleep scores of groups 2 and 5 increased significantly. Among these groups, especially in group 5, the effect of improving the sleep condition was particularly obvious (p < 0.01), indicating that compared with single components, Saccharomyces cerevisiae An-15 and Lactobacillus acidophilus LA15 can cooperate with each other and synergistically enhance the effect in improving the sleep condition.

[0115] Example 2 Preparation of a compound probiotic solid beverage

[0116] This example discloses a preparation method of a solid beverage containing Saccharomyces cerevisiae An-15 and Lactobacillus acidophilus LA15: The formula is as follows (weight components per 100 g): 60 g of whole milk powder, 10 g of fructooligosaccharide, 16.25 g of isomaltooligosaccharide, 12.5 g of Saccharomyces cerevisiae An-15, and 1.25 g of Lactobacillus acidophilus LA15. After accurately weighing the above materials, the other excipients except the bacterial powder are dried to a water activity < 0.2 at 70 °C by a fluidized bed granulator, then the excipients are cooled to room temperature, and mixed with the bacterial powder in a hopper mixer for 30 min and then filled by a filling machine to obtain the compound probiotic solid beverage.

[0117] Example 3 Preparation of a compound probiotic chewable candy

[0118] This embodiment discloses a preparation method of a chewable candy containing Saccharomyces cerevisiae An-15 and Lactobacillus acidophilus LA15: The formula is as follows (weight components per 100 g): sorbitol 49.25 g, isomaltulose 20 g, resistant dextrin 10 g, strawberry powder 5 g, Saccharomyces cerevisiae 12.5 g, Lactobacillus acidophilus LA15 1.25 g, citric acid 1 g, magnesium stearate 1 g. After accurately weighing the above materials, put them into an air flow mixer and mix evenly. After tabletting with a rotary tableting machine, a compound probiotic chewable candy is obtained.

[0119] Example 4 Preparation of a compound probiotic drops

[0120] This embodiment discloses a preparation method of a probiotic drops containing Saccharomyces cerevisiae An-15 and Lactobacillus acidophilus LA15: medium-chain triglyceride 85 g, mono- and diglycerol fatty acid esters 1 g, Saccharomyces cerevisiae An-15 12.5 g, Lactobacillus acidophilus LA15 1.25 g. Weigh the materials accurately according to the formula amount, and sieve all the bacterial powders for later use.

[0121] Stir and mix the mono- and diglycerol fatty acid esters in the formula amount with the medium-chain triglyceride, and then heat to completely melt the mono- and diglycerol fatty acid esters to obtain a mixed oil solution; then cool the mixed oil solution to 45°C ± 5°C, and then use a homogenizer for homogenization, homogenize 2-4 times to obtain a probiotic carrier; pour the probiotic carrier and the sieved bacterial powder into a stirring tank and stir evenly, and finally fill and package to obtain the compound probiotic drops.

[0122] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Composite bacteria, composed of Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) and Lactobacillus acidophilus ( Lactobacillus acidophilus )composition; The cerevisiae yeast is the cerevisiae yeast with a deposit number of CCTCC NO: M 2019643; The Lactobacillus acidophilus is a Lactobacillus acidophilus with a preservation number of CCTCC NO: M 2023768.

2. The composite bacteria according to claim 1, characterized in that The quantity ratio of the saccharomyces cerevisiae to Lactobacillus acidophilus is 1:5 to 5:

1.

3. A probiotic comprising the composite bacteria according to claim 1 or 2 and a protective agent.

4. The probiotic according to claim 3, characterized in that The total number of viable bacteria is not less than 1×10 10 CFU / ml or not less than 1×10 10 CFU / g.

5. The probiotic according to claim 3, characterized in that The protective agent includes at least one of milk powder, starch, gelatin, dextrin, vegetable oil, sorbitol, sorbitol monostearate, sucrose, lactose and polyvinyl pyrrolidone.

6. The method for preparing the probiotic according to any one of claims 3 to 5, comprising: Saccharomyces cerevisiae and Lactobacillus acidophilus are inoculated into culture fluids for cultivation, and then centrifuged to obtain bacterial cells; The obtained bacterial cells are added with a protective agent and then granulated or freeze-dried.

7. Use of the composite bacteria according to claim 1 or 2, the probiotic according to any one of claims 3 to 5, or the probiotic prepared by the preparation method according to claim 6 in the preparation of a pharmaceutical preparation for improving stress resistance and / or improving sleep.

8. A pharmaceutical preparation for improving stress resistance and / or improving sleep, comprising: The composite bacteria according to claim 1 or 2, the probiotic according to any one of claims 3 to 5, or the probiotic prepared by the preparation method according to claim 6.

9. The pharmaceutical preparation according to claim 8, characterized in that It also includes prebiotics and / or excipients; The prebiotics include at least one of fructooligosaccharides, galacto-oligosaccharides, isomaltooligosaccharides, xylo-oligosaccharides, stachyose, inulin, polydextrose, resistant dextrin, soybean oligosaccharides, and yeast β-glucan; The auxiliary materials include at least one of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, colorants, antioxidants, and flavoring agents.

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