Microbial composition and application thereof in preparation of selenium-enriched sleep-aiding goat milk powder

Abstract

The invention provides a microbial composition and application thereof in preparation of selenium-rich sleep-aiding goat milk powder, and belongs to the technical field of microbial fermentation. The microbial composition disclosed by the invention comprises bifidobacterium animalis, enterococcus durans and candida tropicalis. A food additive obtained by fermenting Chinese yam, selenium-rich black beans, Chinese wolfberry fruits, licorice roots, wheat and tremella fuciformis through the microbial composition has the sleep aiding effect, milk powder prepared by combining the microbial composition with other raw materials has the remarkable sleep improving effect, and the milk powder can be taken for a long time and is safe, nutritional and healthy.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and particularly relates to a microbial composition and application thereof in the preparation of selenium-enriched sleep-promoting goat milk powder. Background Art

[0002] The current fast-paced lifestyle and blue light interference from electronic devices disrupt the body's original biological clock rhythm and cause sleep disorders. In order to solve the above-mentioned sleep disorders, many fields such as medicine and nutrition have launched explorations. Although traditional drug treatments can relieve insomnia symptoms in the short term, they are often accompanied by side effects, such as dizziness, fatigue, and increased dependence, and are not suitable for long-term conditioning.

[0003] Microbial fermentation technology is widely used in the food industry and can improve the nutritional content, flavor and texture of food. By using microbial fermentation raw materials, macromolecules can be converted into small molecules, while improving the utilization rate and biological activity of the nutrients in the raw materials. However, the active functions of products obtained from different microbial fermentation raw materials are different, resulting in uneven active functions of products obtained from microbial fermentation. At present, there are few studies on combining microbial fermentation technology with the development of sleep-aid products. Using microbial fermentation technology to develop a safe, nutritious, healthy and long-term sleep-aid product is a technical problem that needs to be solved at this stage. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a microbial composition and its application in the preparation of selenium-enriched sleep-aiding goat milk powder. The microbial composition of the present invention can degrade the macromolecules in the raw materials into small molecules that aid sleep. The goat milk powder prepared using the microbial fermentation composition combined with special raw materials has a significant sleep-aiding effect.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: The invention provides a microbial composition, which comprises animal bifidobacterium, duractic enterococcus and tropical candida; the strain number of animal bifidobacterium is CICC 6165, the strain number of duractic enterococcus is CICC 20379, and the strain number of tropical candida is CICC 1779.

[0006] The invention provides application of the microbial composition in preparing food additives.

[0007] The invention provides a method for preparing a food additive using the microbial composition, comprising the following steps: mixing 15-25 parts by weight of yam with water, heating until the yam is cooked, and beating the mixture after cooling to obtain yam slurry; mixing 10-20 parts by weight of selenium-enriched black beans, 8-12 parts by weight of wolfberries, 1-4 parts by weight of liquorice, 5-11 parts by weight of wheat and 4-10 parts by weight of tremella with water, adding a mixed bacterial liquid of animal bifidobacterium, durable enterococci and tropical candida, fermenting, filtering and collecting the filtrate to obtain a fermentation liquid; mixing the yam slurry with the fermentation liquid, standing for 8-12 hours, and drying to obtain a food additive.

[0008] Preferably, the preparation of the mixed bacterial solution comprises the following steps: activating Bifidobacterium animalis, Enterococcus durvaluma and Candida tropicalis respectively, and culturing at 32-37°C until the bacterial concentration reaches 1×10 8 -6×10 8 cfu / mL of bacterial solution, and then mixed with animal Bifidobacterium bacterial solution, durable Enterococcus bacterial solution and tropical Candida solution in a volume ratio of 1: (0.1-3.5): (0.1-3.5).

[0009] Preferably, the fermentation temperature is 28-37° C., and the fermentation time is 12-84 hours.

[0010] The invention provides a food additive prepared by the method.

[0011] The present invention provides a food additive prepared by the method or application of the food additive in preparing a sleep-aiding product.

[0012] The invention provides a selenium-enriched sleep-aiding goat milk powder, which comprises the following raw materials in parts by weight: 70-110 parts of goat milk, 3-15 parts of the food additive, 2-20 parts of oligofructose, 8-30 parts of lactose, 0.005-0.20 parts of linseed oil, 0.005-0.20 parts of walnut oil and 0.005-0.20 parts of a composite bacterial agent.

[0013] Preferably, the composite bacterial agent is the mixed bacterial solution.

[0014] The invention provides a preparation method of the selenium-enriched sleep-promoting goat milk powder, comprising the following steps: mixing linseed oil, walnut oil and a composite bacterial agent, homogenizing to obtain a composite bacterial emulsion; mixing and dissolving goat milk, food additives, oligofructose and lactose, homogenizing to obtain a composite emulsion; mixing the composite bacterial emulsion and the composite emulsion, standing, homogenizing, and drying to obtain the selenium-enriched sleep-promoting goat milk powder.

[0015] The present invention provides a food additive prepared by the method, and uses of the food additive, the selenium-enriched sleep-aiding goat milk powder, or the selenium-enriched sleep-aiding goat milk powder obtained by the preparation method in preparing sleep-aiding products.

[0016] Compared with the prior art, the present invention has the following beneficial effects: The present invention proposes for the first time to use a combination of animal Bifidobacterium (CICC 6165), Enterococcus durvalume (CICC 20379) and Candida tropicalis (CICC 1779) to ferment yam, selenium-rich black beans, wolfberry, licorice, wheat and white fungus, degrade the macromolecules in the above raw materials into small molecules, and convert the selenium element in the selenium-rich black beans into organic selenium which is easier for the body to absorb. The additive obtained by fermentation has the function of nourishing yin and moistening the lungs, regulating the balance of yin and yang and qi and blood, and has a certain effect of improving sleep.

[0017] Furthermore, the present invention compounded the additive prepared above with oligofructose, lactose, linseed oil, walnut oil and composite bacterial agent, and continued to use animal bifidobacterium, durable enterococci and tropical candida in the preparation of milk powder to adjust the balance of intestinal flora, improve overall health, and further micro-ferment the macromolecules in other raw materials. Animal experiments have verified that the milk powder has a significant effect of improving and helping sleep. DETAILED DESCRIPTION

[0018] The present invention provides a microbial composition, which comprises Bifidobacterium animalis, Enterococcus durvastatus and Candida tropicalis; the strain number of Bifidobacterium animalis is CICC 6165, the strain number of Enterococcus durvastatus is CICC 20379, and the strain number of Candida tropicalis is CICC 1779. The Bifidobacterium (CICC6165), Enterococcus durvastatus (CICC 20379) and Candida tropicalis (CICC 1779) of the present invention are all purchased from the China Industrial Microbiological Culture Collection Administration Center through commercial channels.

[0019] The present invention also provides the use of the microbial composition in preparing food additives. The food of the present invention includes but is not limited to dairy products, cakes, and beverages.

[0020] The present invention also provides a method for preparing a food additive from the microbial composition, comprising the steps of: mixing 15-25 parts by weight of yam with water, heating until the yam is cooked, and beating after cooling to obtain yam slurry; mixing 10-20 parts by weight of selenium-enriched black beans, 8-12 parts by weight of wolfberry, 1-4 parts by weight of licorice, 5-11 parts by weight of wheat and 4-10 parts by weight of white fungus with water, adding the mixed bacterial liquid of animal bifidobacterium, durable enterococcus and tropical candida, fermenting, filtering and taking the filtrate to obtain a fermentation liquid; mixing the yam slurry with the fermentation liquid, standing for 8-12h, and drying to obtain a food additive. The fermentation temperature of the present invention is 28-37°C, preferably 30-35°C; the fermentation time is 12-84h, preferably 24-72h.

[0021] The present invention uses a mixed bacterial liquid prepared by animal bifidobacterium, enterococcus durvaluma and tropical candida to simultaneously ferment selenium-rich black beans, wolfberry, liquorice, wheat and white fungus, which can degrade the macromolecules in the above raw materials into small molecules, obtain small molecule effective components that help sleep, and at the same time, convert the selenium element in the selenium-rich black beans into organic selenium, which is more conducive to the body's absorption and utilization. The present invention is to mix the yam with the fermentation liquid after pulping and let it stand, in order to further react the components in the fermentation liquid with the components in the yam pulp, which is conducive to the escape of the effective components in the yam.

[0022] In the present invention, the Chinese yam has the effects of nourishing the spleen and stomach, promoting fluid production and benefiting the lungs, and nourishing the kidneys and astringing the essence; it can improve the intestines, regulate blood sugar and blood pressure, improve hormone levels, enhance immunity, and nourish yin and nourish the lungs. The selenium-rich black beans have the effects of benefiting the essence and improving eyesight, nourishing blood and dispelling wind, promoting diuresis and detoxification; it can resist oxidation, enhance immunity and protect the eyes, especially selenium can improve sleep. The wolfberry has the effects of nourishing the liver and kidneys and benefiting the essence and improving eyesight; it can resist fatigue, improve sleep, resist oxidation and delay aging. The licorice has the effects of nourishing the spleen and replenishing qi, relieving cough and removing phlegm, and relieving pain, and is a food with medicinal and edible properties that replenishes qi. The wheat has the effects of nourishing the heart and calming the mind, strengthening the spleen and stomach, moistening the intestines and relieving constipation, benefiting qi and stopping sweating, and consolidating the exterior and stopping sweating. The white fungus has the effects of moistening the lungs and promoting fluid production and nourishing yin and nourishing the stomach; it can improve blood circulation, enhance body vitality and improve respiratory immunity. The present invention adopts the above-mentioned combination of raw materials of medicinal and edible origin to adjust the balance of yin and yang of the body, harmonize qi and blood, enhance the vitality of the body, regulate the internal organs, make the body in a healthy state, and further help the body sleep in a resting state.

[0023] In the present invention, the preparation of the mixed bacterial solution comprises the following steps: activating Bifidobacterium animalis, Enterococcus durophilus and Candida tropicalis respectively, and culturing at 32-37°C until the bacterial concentration reaches 1×10 8 -6×10 8 cfu / mL of bacterial liquid, animal Bifidobacterium bacterial liquid, durable Enterococcus bacterial liquid and tropical Candida liquid are mixed in a ratio of 1:0.1-3.5:0.1-3.5. The animal Bifidobacterium, durable Enterococcus and tropical Candida activation of the present invention are activated twice using MRS solid culture medium, and the activation culture conditions are 32-38°C for 12-48h, preferably 37°C for 24h. After activation of the present invention, the active strains are inoculated into MRS liquid culture medium for shaking culture, and the culture conditions are preferably 36°C for culture until the bacterial concentration reaches 4×10 8 cfu / mL. The mixing ratio of the mixed bacterial solution of the present invention is preferably 1:0.5-2.0:0.5-2.0 in volume ratio of animal Bifidobacterium solution, hard-core Enterococcus solution and tropical Candida solution. The MRS solid culture medium and MRS liquid culture medium of the present invention are culture media well known in the art and can be obtained through commercial channels unless otherwise specified.

[0024] The present invention also provides a food additive prepared by the above preparation method. The food additive of the present invention has the effects of regulating sub-health of the body, enhancing immunity and helping sleep.

[0025] The present invention also provides a selenium-enriched sleep-aiding goat milk powder, comprising the following raw materials in parts by weight: 70-110 parts of goat milk, 3-15 parts of the food additive, 2-20 parts of oligofructose, 8-30 parts of lactose, 0.005-0.20 parts of linseed oil, 0.005-0.20 parts of walnut oil and 0.005-0.20 parts of compound bacterial agent; further preferably comprising the following raw materials in parts by weight: 80-100 parts of goat milk, 5-11 parts of the food additive, 3-13 parts of oligofructose, 10-20 parts of lactose, 0.005-0.15 parts of linseed oil, 0.005-0.15 parts of walnut oil and 0.01-0.1 parts of compound bacterial agent. The composite bacterial agent of the present invention is a mixed bacterial liquid prepared from the Bifidobacterium (CICC6165), Enterococcus durvaluma (CICC 20379) and Candida tropicalis (CICC 1779), and the preparation method of the mixed bacterial liquid is as described above.

[0026] The present invention also provides a preparation method of the selenium-enriched sleep-promoting goat milk powder, comprising the following steps: mixing linseed oil, walnut oil and a composite bacterial agent, homogenizing to obtain a composite bacterial emulsion; mixing and dissolving goat milk, food additives, oligofructose and lactose, homogenizing to obtain a composite emulsion; mixing the composite bacterial emulsion and the composite emulsion, standing, homogenizing, and drying to obtain selenium-enriched sleep-promoting goat milk powder. The present invention comprises the following conditions for homogenizing after mixing linseed oil, walnut oil and a composite bacterial agent: primary homogenization for 5-20min under 28-38°C and 8-35Mpa, and then secondary homogenization for 2-10min at 3-20Mpa at the same temperature; further preferably, primary homogenization for 8-15min under 30-36°C and 10-30Mpa, and then secondary homogenization for 3-8min at 5-15Mpa at the same temperature. The present invention comprises the following conditions for homogenizing after mixing the composite bacterial emulsion and the composite emulsion evenly and standing, which are consistent with the conditions for homogenizing after mixing linseed oil, walnut oil and a composite bacterial agent. The present invention comprises the following conditions for homogenizing after mixing and dissolving goat milk, food additives, oligofructose and lactose: homogenizing for 8-30 minutes at 50-80°C and 15-30Mpa; and further preferably: homogenizing for 10-25 minutes at 55-65°C and 20-25Mpa. The suitable homogenizing conditions in the preparation of the selenium-enriched sleep-aiding milk powder of the present invention make the milk powder more uniform and delicate, and the activity of the effective ingredients is not easily lost, which is more conducive to the performance of the active functions of the effective ingredients.

[0027] The present invention provides a food additive prepared by the method or the use of the food additive in preparing a sleep-aiding product. The product of the present invention includes but is not limited to food or health food.

[0028] In the present invention, unless otherwise specified, all components, reagents or culture media are commercially available products well known to those skilled in the art.

[0029] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] The strain number of Bifidobacterium animalis used in the following examples is CICC 6165, the strain number of Enterococcus durophilus is CICC 20379, the strain number of Candida tropicalis is CICC 1779, Saccharomyces cerevisiae (CICC 1251), Bifidobacterium longum (CICC 6193) and Bacillus licheniformis (CICC 10037) are all purchased from the China Industrial Microbiological Culture Collection Administration Center. Unless otherwise specified, other raw materials or culture media used can be purchased through commercial channels.

[0031] Example 1 1. Preparation of mixed bacterial solution: Bifidobacterium animalis (CICC 6165), Enterococcus durconium (CICC 20379) and Candida tropicalis (CICC 1779) were inoculated into MRS solid medium respectively and cultured at 37°C for 24 h. The strains were activated for two generations. The activated strains were inoculated into MRS liquid medium and cultured at 36°C with shaking until the bacterial concentration reached 4×10 8 cfu / mL of bacterial solution, respectively obtain the bacterial solutions of the above three strains; sequentially mix the bacterial solution of Bifidobacterium animalis, the bacterial solution of Enterococcus duractic and the bacterial solution of Candida tropicalis in a volume ratio of 1:0.8:1.5 to obtain a mixed bacterial solution of a microbial composition.

[0032] 2. The preparation of food additives includes the following steps: (1) Peeling the yam, mixing 20 parts by weight of the peeled yam with 40 parts by weight of water, heating and boiling until the yam is cooked, taking out the yam and the water, cooling them, and beating them to obtain a yam slurry; (2) Selenium-rich black beans, wolfberry, licorice, wheat and white fungus are dried and crushed separately until they pass through a 4-mesh sieve. Then, 15 parts by weight of selenium-rich black bean powder, 10 parts by weight of wolfberry powder, 2.5 parts by weight of licorice powder, 8 parts by weight of wheat flour and 7 parts by weight of white fungus powder are weighed and mixed evenly. Water (20 times the weight of the above materials) is added, mixed evenly, and soaked for 12 hours to obtain a material mixture.

[0033] (3) Add the mixed bacterial solution prepared in step 1 above to the material mixture at an inoculation rate of 1% (volume percentage), ferment at 33°C for 48 hours, and filter the filtrate through an 80-mesh sieve to obtain a fermentation liquid; (4) Evenly mix the yam slurry and the fermentation liquid, let stand for 8 hours, and spray dry at low temperature to obtain a food additive.

[0034] 3. Preparation of selenium-enriched sleep-aiding goat milk powder Selenium-enriched sleep-aiding goat milk powder is prepared from the following raw materials in parts by weight: 90 parts of goat milk, 8 parts of the above-prepared food additives, 8 parts of oligofructose, 15 parts of lactose, 0.01 parts of linseed oil, 0.01 parts of walnut oil and 0.05 parts of the above-prepared mixed bacterial liquid.

[0035] The preparation method of selenium-enriched sleep-aiding goat milk powder is as follows: (1) Mix the above-mentioned linseed oil, walnut oil and mixed bacterial solution in parts by weight, homogenize them at 32°C and 20 MPa for 10 min, and then homogenize them at the same temperature and 10 MPa for 5 min to obtain a composite bacterial emulsion; (2) Mix and dissolve the above-mentioned goat milk, food additives, oligofructose and lactose in parts by weight, homogenize at 55° C. and 20 MPa for 15 min, and sterilize to obtain a composite emulsion; (3) The composite bacterial emulsion and the composite emulsion were mixed evenly, allowed to stand for 12 h, homogenized at 32 °C and 20 MPa for 10 min, and then homogenized at 10 MPa for 5 min at the same temperature. The mixture was spray-dried at low temperature to obtain selenium-enriched sleep-promoting goat milk powder.

[0036] Example 2 1. The preparation of the mixed bacterial solution is the same as in Example 1.

[0037] 2. The preparation of food additives includes the following steps: (1) Peel the yam, mix 15 parts by weight of the yam with 30 parts by weight of water, heat and boil until the yam is cooked, and then pulp it after cooling to obtain a yam pulp; (2) Selenium-rich black beans, wolfberry, licorice, wheat and white fungus are dried and crushed separately until they pass through a 6-mesh sieve, 10 parts by weight of selenium-rich black bean powder, 8 parts by weight of wolfberry powder, 1 part by weight of licorice powder, 5 parts by weight of wheat flour and 4 parts by weight of white fungus powder are weighed and mixed evenly, and then 15 times the weight of water of the above materials is added, mixed evenly, and soaked for 8 hours to obtain a material mixture.

[0038] (3) Add the mixed bacterial solution prepared in step 1 above to the material mixture at an inoculation rate of 0.5% (volume percentage), ferment at 30° C. for 24 h, and filter the filtrate through a 90-mesh sieve to obtain a fermentation liquid; (4) Evenly mix the yam slurry and the fermentation liquid, let stand for 6 hours, and spray dry to obtain a food additive.

[0039] 3. Preparation of selenium-enriched sleep-aiding goat milk powder Selenium-enriched sleep-aiding goat milk powder is prepared from the following raw materials in parts by weight: 80 parts of goat milk, 5 parts of the food additives prepared above, 3 parts of oligofructose, 10 parts of lactose, 0.008 parts of linseed oil, 0.008 parts of walnut oil and 0.01 parts of the mixed bacterial solution prepared above.

[0040] The preparation method of selenium-enriched sleep-aiding goat milk powder is as follows: (1) Mix the above-mentioned linseed oil, walnut oil and mixed bacterial solution in parts by weight, homogenize for 8 minutes at 30°C and 10 MPa, and then homogenize for 3 minutes at 5 MPa at the same temperature to obtain a composite bacterial emulsion; (2) Mix and dissolve the above-mentioned goat milk, food additives, oligofructose and lactose in parts by weight, homogenize at 65° C. and 20 MPa for 10 min, and sterilize to obtain a composite emulsion; (3) The composite bacterial emulsion and the composite emulsion were mixed evenly, allowed to stand for 8 h, homogenized for 8 min at 30 °C and 10 MPa, and then homogenized for 3 min at 5 MPa at the same temperature. The mixture was spray-dried at low temperature to obtain selenium-enriched sleep-promoting goat milk powder.

[0041] Example 3 1. The preparation of the mixed bacterial solution is the same as in Example 1.

[0042] 2. The preparation of food additives includes the following steps: (1) Peel the yam, mix 25 parts by weight of the yam with 50 parts by weight of water, heat and boil until the yam is cooked, take out the yam and water, cool them, and pulp them to obtain yam pulp; (2) Selenium-rich black beans, wolfberry, licorice, wheat and white fungus are dried and crushed separately until they pass through an 8-mesh sieve, 20 parts by weight of selenium-rich black bean powder, 12 parts by weight of wolfberry powder, 4 parts by weight of licorice powder, 11 parts by weight of wheat flour and 10 parts by weight of white fungus powder are weighed and mixed evenly, and then 30 times the weight of water of the above materials is added, mixed evenly, and soaked for 18 hours to obtain a material mixture.

[0043] (3) Add the mixed bacterial solution prepared in step 1 above to the material mixture at an inoculation rate of 1.5% (volume percentage), ferment at 35°C for 72 hours, and filter the filtrate through a 100-mesh sieve to obtain a fermentation liquid; (4) Evenly mix the yam slurry and the fermentation liquid, let stand for 12 hours, and spray dry to obtain a food additive.

[0044] 3. Preparation of selenium-enriched sleep-aiding goat milk powder Selenium-enriched sleep-aiding goat milk powder is prepared from the following raw materials in parts by weight: 100 parts of goat milk, 11 parts of the food additives prepared above, 13 parts of oligofructose, 20 parts of lactose, 0.15 parts of linseed oil, 0.15 parts of walnut oil and 0.1 parts of the mixed bacterial liquid prepared above.

[0045] The preparation method of selenium-enriched sleep-aiding goat milk powder is as follows: (1) Mix the above-mentioned linseed oil, walnut oil and mixed bacterial solution in parts by weight, homogenize at 36°C and 30 MPa for 15 min, and then homogenize at the same temperature and 15 MPa for 8 min to obtain a composite bacterial emulsion; (2) Mix and dissolve the above-mentioned goat milk, food additives, oligofructose and lactose, homogenize at 75°C and 25 MPa for 25 minutes, and sterilize to obtain a composite emulsion; (3) The composite bacterial emulsion and the composite emulsion were mixed evenly, allowed to stand for 18 h, homogenized for 15 min at 36 °C and 30 MPa, and then homogenized for 8 min at 15 MPa at the same temperature. The mixture was spray-dried at low temperature to obtain selenium-enriched sleep-promoting goat milk powder.

[0046] Comparative Example 1 The difference from Example 1 is that Candida tropicalis (CICC 1779) is replaced by Saccharomyces cerevisiae (CICC1251), and the remaining steps are the same as those in Example 1.

[0047] Comparative Example 2 The difference from Example 1 is that Bifidobacterium animalis (CICC 6165) is replaced by Bifidobacterium longum (CICC6193), and the remaining steps are the same as those in Example 1.

[0048] Comparative Example 3 The difference from Example 1 is that Enterococcus duratus (CICC 20379) is replaced by Bacillus licheniformis (CICC10037), and the remaining steps are the same as those in Example 1.

[0049] Comparative Example 4 The difference from Example 1 is that the yam in step 2 is replaced by taro, and the remaining steps are the same as those in Example 1.

[0050] Comparative Example 5 The difference from Example 1 is that the wolfberry in step 2 is replaced by polygonatum, and the remaining steps are the same as those in Example 1.

[0051] Comparative Example 6 The difference from Example 1 is that the wheat in step 2 is replaced by hawthorn, and the remaining steps are the same as those in Example 1.

[0052] Comparative Example 7 The difference from Example 1 is that no food additives are used in the preparation of goat milk powder, and the remaining steps are the same as those in Example 1.

[0053] Comparative Example 8 The difference from Example 1 is that the walnut oil in step 3 is replaced by olive oil, and the remaining steps are the same as those in Example 1.

[0054] Comparative Example 9 The difference from Example 1 is that the lactose in step 3 is replaced by whey protein, and the remaining steps are the same as those in Example 1.

[0055] Comparative Example 10 The difference from Example 1 is that in step (1) of step 3, homogenization is replaced by homogenization at 32° C. and 20 MPa for 20 min, and the remaining steps are the same as those in Example 1.

[0056] Comparative Example 11 The difference from Example 1 is that the standing step in step (3) of step 3 is removed.

[0057] Experimental Example 1 1. Experimental animals: The animal experiments in this study were conducted in accordance with the Technical Specifications for Inspection and Evaluation of Health Food (2003 edition), and the implementation plan was reviewed and approved by the Ethics Committee. SPF mice were fed and adapted under laboratory conditions for 3 days, and the mice had free access to food and water.

[0058] 2, grouping, gavage and observation of mouse state: mice were randomly divided into 16 groups, namely normal control group, model group, embodiment 1 food additive group, embodiment 1-3 milk powder group and comparative example 1-11 milk powder group, 10 in each group. Daily gavage gave the test substance 0.03g / kg, the experimental time was 30 days, and the mouse state was observed during the experiment. Before the last administration, 12h fasting and water were not allowed. After gavage for 1h on the 30th day, except for the normal control group, the remaining groups were injected with sodium pentobarbital (30mg / kg) intraperitoneally at one time, and the administration time, the disappearance time of righting reflex and the recovery time of righting reflex were recorded, and the sleep latency and sleep duration of mice were calculated respectively (with the injection of sodium pentobarbital to the disappearance of righting reflex as the sleep latency, and the disappearance of righting reflex to the reappearance of righting reflex as the sleep duration), and the results are shown in Table 1.

[0059] After observing the status of each group of mice after gavage for 30 days, all groups of mice were in good condition. They were fed with the food additive prepared in Example 1 of the present invention and the milk powder prepared in Examples 1-3 and Comparative Examples 1-11, and had no adverse reactions within 30 days, and were in good mental state.

[0060] Table 1 Sleep latency and sleep time of mice in each group

[0061] As can be seen from Table 1, after a one-time intraperitoneal injection of sodium pentobarbital (30 mg / kg), mice were given food additives or milk powder mice in groups for 30 days, and the sleep latency was shortened, which was significantly different from the model group. The sleep latency of mice in the milk powder groups of Examples 1-3 was also shortened relative to the milk powder groups of Comparative Examples 1-11. And the sleep duration of food additives or milk powder mice in groups was significantly prolonged compared with the model group mice. The sleep duration of mice in the milk powder groups of Examples 1-3 was also prolonged relative to the milk powder groups of Comparative Examples 1-11. It shows that the milk powder prepared by Examples 1-3 can better improve the sleep condition of mice.

[0062] 3. Sampling and detection: Blood was collected from the eye socket 24 hours after the injection of sodium pentobarbital. The separated mouse whole blood was centrifuged at 4°C and 3500r / min for 8 minutes to collect the supernatant serum. The supernatant serum after centrifugation was taken and the levels of NOS and 5-HT in the serum were determined using the mouse nitric oxide synthase (NOS) ELISA kit and the mouse 5-hydroxytryptamine (5-HT) ELISA kit, respectively. The results are shown in Table 2.

[0063] Table 2 NOS and 5-HT levels in serum of mice in each group

[0064] The sleep condition of mice was verified by detecting the content of sleep-promoting factors 5-HT and NOS in mouse serum. As shown in Table 2, compared with the normal control group, the content of 5-HT and NOS in the mouse serum of the model group increased, indicating that the secretion of sleep-promoting factors in the mouse brain increased, and the model was successfully established; compared with the normal control group, the content of 5-HT and NOS in the serum of mice in the food additive, the milk powder group of Example 1-3 and the milk powder group of Comparative Example 1-11 increased, indicating that the food additive prepared by the present invention has a sleep-promoting effect. However, the content in the milk powder group of Example 1-3 increased significantly, indicating that the milk powder prepared by the technical solution of the present invention has a more significant sleep-promoting effect.

[0065] After blood was drawn, mice in each group were immediately decapitated and killed. The hippocampus of the mouse brain was immediately dissected and separated on an ice tray. The brain tissue of the same weight was weighed and chopped, and then placed in an ice-cold tissue grinder. 12 times the amount of pre-cooled PBS buffer was added to grind the homogenate on ice, centrifuged at 4°C and 20,000 rpm for 15 minutes, and the supernatant was collected to obtain a brain tissue homogenate sample. The contents of GABA, Glu, NE and SOD in mouse brain tissue were determined according to the instructions of the commercially available ELISA kit. The results are shown in Table 3.

[0066] Table 3 Contents of GABA, Glu, NE, SOD and MDA in brain tissues of mice in each group

[0067] As shown in Table 3, the food additive prepared in Example 1 of the present invention, the milk powder prepared in Examples 1-3, and the milk powder prepared in Comparative Examples 1-11 can increase the GABA content and SOD content in the brain tissue of mice, and reduce the Glu content and NE content. The milk powder prepared in Examples 1-3 has a more significant effect in increasing the GABA content and SOD content in the brain tissue of mice and reducing the Glu content and NE content, indicating that the goat milk powder prepared by the technical solution of the present invention has a significant improvement and sleep-aiding effect.

[0068] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A microbial composition, characterized in that The microbial composition comprises animal bifidobacterium, duractic enterococcus and tropical candida; the strain number of animal bifidobacterium is CICC 6165, the strain number of duractic enterococcus is CICC 20379, and the strain number of tropical candida is CICC 1779.

2. Use of the microbial composition according to claim 1 in the preparation of food additives.

3. A method for preparing a food additive using the microbial composition according to claim 1, characterized in that: The steps include: Mix 15-25 parts by weight of yam with water, heat until the yam is cooked, and then pulp it after cooling to obtain yam slurry; Mix 10-20 parts by weight of selenium-enriched black beans, 8-12 parts by weight of wolfberry, 1-4 parts by weight of liquorice, 5-11 parts by weight of wheat and 4-10 parts by weight of tremella with water, add the mixed bacterial liquid of animal Bifidobacterium, Enterococcus duracticus and Candida tropicalis, ferment, filter and collect the filtrate to obtain a fermentation liquid; The yam slurry is mixed with the fermentation liquid, left to stand for 8-12 hours, and dried to obtain a food additive.

4. The method according to claim 3, characterized in that The preparation of the mixed bacterial solution comprises the following steps: activating Bifidobacterium animalis, Enterococcus durophilus and Candida tropicalis respectively, and culturing at 32-37°C until the bacterial concentration reaches 1×10 8 -6×10 8 cfu / mL of bacterial solution, and then mixed with animal Bifidobacterium bacterial solution, durable Enterococcus bacterial solution and tropical Candida solution in a volume ratio of 1: (0.1-3.5): (0.1-3.5).

5. The method according to claim 3, characterized in that The fermentation temperature is 28-37° C., and the fermentation time is 12-84 hours.

6. A food additive prepared by the method according to any one of claims 3 to 5.

7. Use of the food additive prepared by the method according to any one of claims 3 to 5 or the food additive according to claim 6 in the preparation of a sleep-aiding product.

8. A selenium-enriched sleep-aiding goat milk powder, characterized in that: The invention comprises the following raw materials in parts by weight: 70-110 parts of goat milk, 3-15 parts of the food additive according to claim 6, 2-20 parts of oligofructose, 8-30 parts of lactose, 0.005-0.20 parts of linseed oil, 0.005-0.20 parts of walnut oil and 0.005-0.20 parts of compound bacterial agent.

9. The selenium-enriched sleep-aiding goat milk powder as claimed in claim 8, characterized in that: The composite bacterial agent is the mixed bacterial solution described in claim 4.

10. The method for preparing the selenium-enriched sleep-aiding goat milk powder as claimed in claim 8, characterized in that: The method comprises the following steps: mixing linseed oil, walnut oil and a composite bacterial agent, homogenizing to obtain a composite bacterial emulsion; mixing and dissolving goat milk, food additives, oligofructose and lactose, homogenizing to obtain a composite emulsion; mixing the composite bacterial emulsion and the composite emulsion, standing, homogenizing and drying to obtain selenium-enriched sleep-aiding goat milk powder.

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