Probiotic composition with sleep aiding effect and application thereof

Abstract

The invention relates to a probiotic composition with a sleep aiding effect and application of the probiotic composition. A bacterial strain in the probiotic composition with the sleep aiding effect is composed of a lactobacillus fermentum GLF-217 bacterial strain and a lactobacillus plantarum FLP-215 bacterial strain. The probiotic composition with the sleep aiding effect has the advantages that the probiotic composition with the sleep aiding effect is prepared from the lactobacillus fermentum GLF-217 bacterial strain, the lactobacillus plantarum FLP-215 bacterial strain, the lactobacillus plantarum GLF-217 bacterial strain and the lactobacillus plantarum FLP-215 bacterial strain; potential interaction exists between the two strains, the two strains can be matched with each other, and synergistic interaction is achieved on the effects of improving the yield of gamma-aminobutyric acid and exopolysaccharide and improving the sleep quality. Meanwhile, when the probiotic composition is used for yoghourt fermentation, the acidity, the water retention rate, the viscosity, the flavor and the sensory comprehensive evaluation of the yoghourt can be remarkably improved. Compared with a single GLF-217 strain or a single FLP-215 strain, the probiotic composition provided by the invention has the advantages that under the condition that the use amount of bacteria is consistent, the effects are more excellent than those of the single GLF-217 strain or the single FLP-215 strain.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotics and relates to a probiotic composition with sleep-aiding effect and application thereof. Background Art

[0002] With the continuous development and progress of the current society, the phenomenon of insomnia caused by irregular life and various pressures in life, study, work, etc. is increasing. Insomnia affects a person's work, study efficiency and quality of life. In severe cases, it can also cause symptoms such as anxiety and depression. Therefore, high-quality sleep plays an extremely important role in the physical and mental health of the human body. At present, sedative hypnotics, melatonin and other drugs used to treat insomnia can easily affect the body if taken for a long time, and have a certain degree of dependence.

[0003] In recent years, scientists have also begun to explore the use of probiotics to improve insomnia. The main ways in which probiotics improve insomnia include: (1) directly changing the abundance and species diversity of intestinal microbial flora, restoring the disorder of intestinal microbial flora caused by insomnia, and then restoring the homeostasis of intestinal flora to play a biological role in regulating insomnia; (2) indirectly regulating the function of the gut-brain axis by regulating the functions of certain special flora in the intestine, such as secreting SCFAS and metabolizing bile salts, thereby improving the symptoms of insomnia patients.

[0004] One of the pathogenesis of insomnia is the downregulation of γ-aminobutyric acid (GABA) in the limbic system of the brain. γ-aminobutyric acid is a natural functional non-protein active amino acid widely found in animals, plants and microorganisms. γ-aminobutyric acid is an extremely important inhibitory neurotransmitter in the central nervous system of mammals such as the brain and spinal cord. It participates in brain circulation and various metabolic activities, and has many important physiological functions such as calming the nerves, resisting excitement, effectively relieving anxiety, improving memory, lowering blood pressure, treating epilepsy and alleviating Alzheimer's disease. Although GABA in the human body can be generated by glutamate in the body under the catalysis of glutamate decarboxylase with strong specificity, the content of GABA synthesized in the body continues to decrease with the continuous increase of age and the current fast-paced lifestyle and living conditions of people for a long time, so the body can only be maintained healthy through exogenous supplementation.

[0005] Traditional chemically synthesized γ-aminobutyric acid is less safe and poses a great safety hazard when used in food; the cost of collecting it through plant enrichment is relatively high; while the microbial fermentation method, which produces γ-aminobutyric acid during the growth and reproduction of microorganisms, is relatively safe and suitable for the application of food and pharmaceutical products. Therefore, it is very meaningful to develop more strains that can produce high γ-aminobutyric acid. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a probiotic composition with sleep-aiding effect and its application, specifically to a probiotic composition with sleep-aiding effect (high production of γ-aminobutyric acid and extracellular polysaccharides) and its application in the preparation of products for improving sleep quality, as well as a method for preparing yogurt by fermenting the probiotic composition.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a probiotic composition with a sleep-aiding effect, wherein the strains in the probiotic composition with a sleep-aiding effect are composed of a Lactobacillus fermentum GLF-217 strain with a preservation number of CGMCC No. 28336 and a Lactobacillus plantarum FLP-215 strain with a preservation number of CGMCC No. 28337.

[0009] The present invention has developed a new probiotic compounding method and a new probiotic strategy for improving sleep quality, that is, compounding the GLF-217 strain and the FLP-215 strain, and found that there is a potential interaction between the two strains, which can cooperate with each other to synergistically enhance the production of γ-aminobutyric acid and extracellular polysaccharides. Since the intake of a certain amount of γ-aminobutyric acid has the effect of improving the body's sleep quality, the probiotic combination also has excellent sleep-aiding and sleep quality improvement effects. At the same time, the probiotic combination is used for yogurt fermentation, which can significantly improve the acidity, water holding rate, viscosity, flavor and sensory comprehensive evaluation of yogurt. When the amount of bacteria is consistent, compared with a single GLF-217 strain or a single FLP-215 strain, the probiotic composition involved in the present invention is more excellent in the above-mentioned effects.

[0010] Preferably, the ratio of the bacterial count of the GLF-217 strain to that of the FLP-215 strain is 1:9-9:1, for example, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0011] Based on the potential cooperative relationship between the GLF-217 strain and the FLP-215 strain, when they meet the above-mentioned specific bacterial count ratio, the effects on increasing the production of γ-aminobutyric acid and extracellular polysaccharides, as well as improving the acidity, water holding capacity, viscosity, flavor and comprehensive sensory evaluation of yogurt are more prominent.

[0012] Preferably, the total bacterial count in the probiotic composition is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g, for example 1×10 6 CFU / mL (CFU / g), 5×10 6 CFU / mL (CFU / g), 1×10 7 CFU / mL (CFU / g), 5×10 7 CFU / mL (CFU / g), 1×10 8 CFU / mL (CFU / g), 5×10 8 CFU / mL (CFU / g), 1×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), etc.; other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0013] Preferably, the dosage form of the probiotic composition includes a solution or a powder.

[0014] When the probiotic composition is a powder, it is prepared by the following preparation method:

[0015] The relevant strains are inoculated into culture medium for activation and fermentation to obtain fermentation broth; the fermentation broth is centrifuged, mixed with a freeze-drying protective agent and freeze-dried to obtain bacterial powders of each strain; the bacterial powders of each strain are mixed according to the bacterial number ratio to obtain the product.

[0016] Preferably, the lyoprotectant comprises any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

[0017] In a second aspect, the present invention provides use of the probiotic composition with sleep-aiding effect according to the first aspect in preparing a product for improving sleep quality.

[0018] In a third aspect, the present invention provides use of the probiotic composition with sleep-aiding effect according to the first aspect in the preparation of γ-aminobutyric acid and / or extracellular polysaccharide.

[0019] In a fourth aspect, the present invention provides use of the probiotic composition with sleep-aiding effect according to the first aspect in preparing yogurt, wherein the preparation method of the yogurt comprises:

[0020] (1) Mixing raw milk and prebiotics, sterilizing and cooling, and using the mixture as a fermentation base;

[0021] (2) The probiotic composition and the basic starter described in the first aspect are inoculated into a fermentation base for fermentation to obtain yogurt.

[0022] Preferably, the prebiotics include any one of fructooligosaccharides, galacto-oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, soybean oligosaccharides, inulin, polydextrose, α-lactalbumin or lactoferrin, or a combination of at least two thereof.

[0023] The prebiotics account for 1-15% of the fermentation base, for example 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0024] Preferably, the inoculation amount of the probiotic composition is 1-4%, for example, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, etc.

[0025] The inoculation amount of the basic leavening agent is 0.02-0.04%, such as 0.02%, 0.025%, 0.03%, 0.035%, 0.04% and the like.

[0026] The bacterial strains in the basic fermentation agent are composed of thermophilic Streptococcus and Lactobacillus delbrueckii subspecies bulgaricus.

[0027] The composite bacterial agent composed of Streptococcus thermophilus and Lactobacillus delbrueckii subspecies bulgaricus is the most commonly used type of yogurt starter. The basic starter here is not limited to a specific type.

[0028] Preferably, the fermentation temperature is 35-40°C, such as 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, etc.; the fermentation time is 5-10h, such as 5h, 6h, 7h, 8h, 9h, 10h, etc. Other specific point values ​​within this numerical range can be selected, and they will not be repeated here.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The present invention also develops a new probiotic compounding method and a new probiotic strategy for improving sleep quality, that is, compounding the GLF-217 strain and the FLP-215 strain, and finds that there is a potential interaction between the two strains, which can cooperate with each other to synergistically enhance the effect of increasing the production of γ-aminobutyric acid and extracellular polysaccharides and improving sleep quality. At the same time, the probiotic combination is used for yogurt fermentation, which can significantly improve the acidity, water holding rate, viscosity, flavor and sensory comprehensive evaluation of yogurt. When the amount of bacteria is consistent, the probiotic composition involved in the present invention is more excellent in the above-mentioned effects compared with a single GLF-217 strain or a single FLP-215 strain.

[0031] The GLF-217 strain involved in the present invention is classified and named as Lactobacillus fermentum, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is September 4, 2023, the preservation number is CGMCC No.28336, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0032] The FLP-215 strain involved in the present invention is classified and named Lactobacillus plantarum, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is September 4, 2023, the preservation number is CGMCC No.28337, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only used to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0034] The culture medium formula involved in the following content is as follows:

[0035] MRS liquid or solid medium: peptone 10g / L, beef extract 10g / L, yeast extract 5g / L, K 2 HPO 4 2g / L, diammonium citrate 2g / L, sodium acetate 5g / L, glucose 20g / L, Tween 80 1mL, MgSO 4 7H 2 O 0.58g / L, MnSO 4 ·4H 2 O 0.25g / L, (agar 20g / L), calcium carbonate 5g / L, adjust pH to 6.4.

[0036] The GLF-217 strain involved below is classified and named as Lactobacillus fermentum, the preservation date is September 4, 2023, and the preservation number is CGMCC No.28336.

[0037] The FLP-215 strain involved below is classified and named Lactobacillus plantarum, the preservation date is September 4, 2023, and the preservation number is CGMCC No.28337.

[0038] Preparation Example 1

[0039] Preparation of single bacterial solution or mixed bacterial solution:

[0040] The seed liquid of GLF-217 strain and FLP-215 strain were inoculated into MRS medium at an inoculum amount of 3% of the total mass of the medium, and cultured at 37°C for 24 h to obtain bacterial suspensions. The bacterial suspensions were transferred to sterile centrifuge tubes and centrifuged (8000 rpm, 10 min, 4°C). The bacteria were collected and resuspended in sterile PBS buffer solution to obtain a live bacterial concentration of 5×10 6 CFU / mL of GLF-217 single bacterial liquid and FLP-215 single bacterial liquid.

[0041] Then, equal volumes of the GLF-217 single bacterial solution and the FLP-215 single bacterial solution were mixed to prepare a mixed bacterial solution 1;

[0042] Mix the GLF-217 single bacterial solution and the FLP-215 single bacterial solution in a volume ratio of 1:9 to obtain a mixed bacterial solution 2;

[0043] Mix the GLF-217 single bacterial solution and the FLP-215 single bacterial solution in a volume ratio of 9:1 to obtain a mixed bacterial solution 3;

[0044] Take an equal volume of commercially available Lactobacillus fermentum ATCC14931 single bacterial solution (5×10 6 CFU / mL and commercially available Lactobacillus plantarum ATCC14917 single bacterial solution (5×10 6 CFU / mL) to prepare mixed bacterial solution 4.

[0045] Example 1

[0046] Evaluation of γ-aminobutyric acid production:

[0047] The GLF-217 single bacterial solution, FLP-215 single bacterial solution, and mixed bacterial solutions 1-4 prepared in Preparation Example 1 were inoculated into sterilized raw milk at a rate of 5%, and fermented at 37°C for 24 hours, and then the whey was separated and the content of γ-aminobutyric acid was determined by Berthlot colorimetry. The results are shown in Table 1.

[0048] Table 1

[0049] Group GABA content (g / kg) GLF-217 single bacterial liquid 4.508±0.034 FLP-215 single bacterial liquid 4.519±0.072 Mixed bacterial solution 1 4.765±0.098 Mixed bacterial solution 2 4.584±0.010 Mixed bacterial solution 3 4.547±0.018 Mixed bacterial solution 4 4.449±0.018

[0050] It can be seen from the data results in Table 1 that the probiotic composition of the present invention has an excellent effect of high production of γ-aminobutyric acid, and the GLF-217 strain and the FLP-215 strain have synergistic effects in the above effect.

[0051] Example 2

[0052] Evaluation of extracellular polysaccharide production:

[0053] (1) The GLF-217 single bacterial solution, FLP-215 single bacterial solution, and mixed bacterial solutions 1-4 prepared in Preparation Example 1 were inoculated into sterilized raw milk at an inoculation rate of 5%, and fermented at a constant temperature of 37° C. for 24 hours to obtain fermented milk.

[0054] (2) Purification of extracellular polysaccharides: Take the fermented milk and heat it in a 95℃ water bath for 10 minutes to remove the enzymes that may degrade the polysaccharides, and cool it to 25℃. Then centrifuge it at 4℃ and 8000rpm for 30 minutes to remove cells and some protein precipitates. Add 80% TCA to the supernatant to a final concentration of 4% (w / v), stir for 5 minutes, let it stand at 4℃ for 18 hours, and centrifuge it at 12000×g at 4℃ for 40 minutes to remove proteins. Then concentrate it to 1 / 3, mix it with 3 times the amount of pre-cooled anhydrous ethanol, let it stand at 4℃ for 18 hours, and centrifuge it. Add pure water to dissolve it to obtain a polysaccharide solution. The polysaccharide solution is placed in a dialysis bag (molecular weight cutoff 14000Da), dialyzed with ultrapure water in a 4℃ refrigerator for 48 hours, and the water is changed every 8 hours. After freeze-drying for 24 hours, dry polysaccharides are obtained.

[0055] (3) Determination of extracellular polysaccharide content: The phenol-sulfuric acid method was used to determine the content of extracellular polysaccharide, and glucose was used as the standard to draw a standard curve y = 0.0095x-0.013 (R 2 =0.9984). The absorbance of the dry polysaccharide solution obtained in step (2) at 490 nm was measured in the same manner and the extracellular polysaccharide content was calculated by entering the regression equation. The results are shown in Table 2.

[0056] Table 2

[0057] Group Extracellular polysaccharide content (mg / L) GLF-217 single bacterial liquid 489.0±6.7 FLP-215 single bacterial liquid 503.6±7.4 Mixed bacterial solution 1 519.7±7.7 Mixed bacterial solution 2 514.0±4.0 Mixed bacterial solution 3 508.3±5.5 Mixed bacterial solution 4 476.2±5.1

[0058] It can be seen from the data results in Table 2 that the probiotic composition of the present invention has an excellent effect of high production of extracellular polysaccharides, and the GLF-217 strain and the FLP-215 strain have synergistic effects in the above effect.

[0059] Preparation Example 2

[0060] Preparation of single bacterial powder or mixed bacterial powder:

[0061] The seed liquid of GLF-217 strain and FLP-215 strain was inoculated into MRS medium at an inoculum amount of 3% of the total mass of the medium, and cultured at 37°C for 24 h to obtain bacterial suspension; the bacterial suspension was transferred to a sterile centrifuge tube, centrifuged (8000 rpm, 10 min, 4°C), the bacterial cells were collected, washed with sterile saline, centrifuged again to obtain the bacterial cells, and the washing was repeated once, and then a lyophilization protective agent was added to the bacterial mud, pre-frozen at -20°C for 5 h, and then placed in a freeze dryer for lyophilization for 24 h to obtain a live bacterial concentration of 2×10 6 CFU / g of GLF-217 single bacterial powder and FLP-215 single bacterial powder.

[0062] Then, equal masses of GLF-217 single bacterial powder and FLP-215 single bacterial powder were mixed to prepare mixed bacterial powder 1;

[0063] Mix the GLF-217 single bacterial powder and the FLP-215 single bacterial powder in a mass ratio of 1:9 to prepare mixed bacterial powder 2;

[0064] The GLF-217 single bacterial powder and the FLP-215 single bacterial powder were mixed in a mass ratio of 9:1 to prepare mixed bacterial powder 3.

[0065] Example 3

[0066] Effects of probiotics on the physical and chemical indicators and sensory properties of fermented milk:

[0067] (1) Preparation of fermented milk: Using direct-injection starter Yo-cul 904 (yogurt starter produced by Weikang Probiotics Co., Ltd.) as the starter, the starter was inoculated at 0.03% into a fermentation base (90% raw milk + 10% sucrose), and 2% of the GLF-217 single bacteria powder, FLP-215 single bacteria powder, and mixed bacteria powder 1-3 prepared in Preparation Example 2 were inoculated at the same time. The mixture was fermented at 37°C for 7 h, and the fermented yogurt was placed in a refrigerator at 4°C for 18 h.

[0068] (2) Determination of physical and chemical indicators of fermented milk:

[0069] (2.1) Determination of titrable acidity

[0070] Referring to the method in GB 5009.239-2016, weigh 10g of sample, place it in a 150mL conical flask, add 20mL of freshly boiled water cooled to 25°C, mix well, and titrate with 0.1mol / L sodium hydroxide standard solution until the reddish color does not disappear within 1min. During the titration, blow nitrogen into the conical flask to prevent the solution from absorbing carbon dioxide in the air. Record the number of milliliters of sodium hydroxide standard titration solution consumed V1, and substitute it into the following formula to calculate the acidity X1 of the sample. Perform three parallel experiments in each group and take the average value.

[0071] X1=[C1×(V1-V0)×100] / (m1×0.1);

[0072] Among them, X1 represents the acidity of the sample, in degrees;

[0073] C1 represents the molar concentration of the sodium hydroxide standard solution, in moles per liter (mol / L);

[0074] V1 represents the volume of sodium hydroxide standard solution consumed during titration, in milliliters (mL);

[0075] V0——The volume of sodium hydroxide standard solution consumed in the blank experiment, in milliliters (mL);

[0076] 100 means 100g sample;

[0077] m1 represents the mass of the sample, in grams (g);

[0078] 0.1 represents the molar concentration of sodium hydroxide, which is the theoretical definition of acidity, in moles per liter (mol / L).

[0079] (2.2) Determination of water holding capacity

[0080] Take 20g of fermented milk sample, centrifuge at 1000g for 3min at 10℃, pour off the supernatant, keep the centrifuge tube inverted for 10min, weigh immediately after the end, make three parallel tests for each sample, and take the arithmetic mean of the results. Water holding rate of fermented milk (%) = weight of centrifugal sediment (g) / sample weight (g) × 100%.

[0081] (2.3) Determination of viscosity

[0082] The viscosity was tested by rheometer at 15°C, and the test was performed three times at a speed of 30 rpm, and the data at 30 seconds was selected as the measured value. The three parallel experiments were performed and the average value was taken.

[0083] The results are shown in Table 3.

[0084] Table 3

[0085] Group Acidity(°T) Water holding rate (%) Viscosity (mPa·s) GLF-217 single bacteria powder 79.5±1.2 62.49±2.7 3619.63±20.35 FLP-215 single bacteria powder 80.3±0.4 60.30±2.53 3192.32±25.35 Mixed fungus powder 1 83.3±0.6 78.34±2.98 5379.41±12.28 Mixed bacteria powder 2 82.1±0.5 65.33±2.83 4869.36±15.36 Mixed bacteria powder 3 81.5±0.9 69.45±3.26 5076.24±16.34

[0086] It can be seen from the data results in Table 3 that the dairy products fermented by the probiotic composition involved in the present invention have excellent physical and chemical properties, including higher acidity, water holding capacity and viscosity value, and the GLF-217 strain and the FLP-215 strain synergize in the above effects.

[0087] (3) Sensory evaluation of fermented milk:

[0088] Weigh 20g of fermented milk into three tasting cups and evaluate its flavor. The flavor characteristics are described as sour aroma, milk aroma, cheese aroma, nut aroma, creamy flavor, and steamed flavor. After tasting one sample, the three assessors rinsed their mouths with mineral water, and each sample was repeated 3 times. Sensory scoring standard (0-9 points): 0 points (none); 1-3 points (weak); 4-6 points (medium intensity); 7-9 points (strong). The results are shown in Table 4.

[0089] Table 4

[0090] Group Sour aroma Milky flavor Cheese flavor Nutty aroma Creamy Steaming flavor GLF-217 single bacterial liquid 6.7±0.1 4.5±0.3 1.5±0.3 0.5±0.1 3.6±0.2 0.5±0.4 FLP-215 single bacterial liquid 6.2±0.3 3.9±0.1 1.2±0.2 1.3±0.4 2.7±0.4 0.6±0.2 Mixed bacterial solution 1 4.2±0.3 5.7±0.4 2.4±0.4 3.1±0.3 4.3±0.3 0.1±0.4 Mixed bacterial solution 2 4.4±0.2 5.3±0.2 2.3±0.3 2.9±0.2 3.5±0.1 0.2±0.1 Mixed bacterial solution 3 4.9±0.1 4.9±0.1 1.8±0.2 2.4±0.1 2.9±0.2 0.2±0.3

[0091] It can be seen from the data results in Table 4 that, compared with the single bacteria fermentation method, the dairy product fermented by the probiotic composition of the present invention has a high level of flavor evaluation score.

[0092] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0093] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0094] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A probiotic composition having a sleep-aiding effect, characterized in that: The strains in the probiotic composition with sleep-aiding effect are composed of Lactobacillus fermentum GLF-217 strain with a preservation number of CGMCC No.28336 and Lactobacillus plantarum FLP-215 strain with a preservation number of CGMCC No.28337.

2. The probiotic composition with sleep-aiding effect according to claim 1, characterized in that: The bacterial count ratio of the GLF-217 strain to the FLP-215 strain is 1:9-9:

1.

3. The probiotic composition with sleep-aiding effect according to claim 1, characterized in that: The total bacterial count in the probiotic composition is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

4. The probiotic composition with sleep-aiding effect according to claim 1, characterized in that: The dosage form of the probiotic composition includes solution or powder.

5. Use of the probiotic composition with sleep-aiding effect according to any one of claims 1 to 4 in the preparation of a product for improving sleep quality.

6. Use of the probiotic composition with sleep-aiding effect according to any one of claims 1 to 4 in the preparation of γ-aminobutyric acid and / or exopolysaccharide.

7. Use of the probiotic composition with sleep-aiding effect according to any one of claims 1 to 4 in preparing yogurt, the preparation method of the yogurt comprising: (1) Mixing raw milk and prebiotics, sterilizing and cooling, and using the mixture as a fermentation base; (2) The probiotic composition according to any one of claims 1 to 4 and a basic starter are inoculated into a fermentation base for fermentation to obtain yogurt.

8. The use according to claim 7, characterized in that: The prebiotics include any one of fructooligosaccharides, galacto-oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, soybean oligosaccharides, inulin, polydextrose, α-lactalbumin or lactoferrin, or a combination of at least two thereof; The prebiotics account for 1-15% of the fermentation base.

9. The use according to claim 7, characterized in that: The inoculation amount of the probiotic composition is 1-4%; The inoculation amount of the basic leavening agent is 0.02-0.04%; The bacterial strains in the basic fermentation agent are composed of thermophilic Streptococcus and Lactobacillus delbrueckii subspecies bulgaricus.

10. The use according to claim 7, characterized in that: The fermentation temperature is 35-40°C, and the fermentation time is 5-10 hours.