Sleep-aiding lactic acid bacteria afterbiogen preparation, composition, preparation method and application of lactic acid bacteria afterbiogen preparation

Abstract

The invention relates to a lactic acid bacteria afterbiogen preparation for assisting sleep, a composition, a preparation method and application of the lactic acid bacteria afterbiogen preparation, and the lactic acid bacteria afterbiogen preparation is obtained by fermenting and culturing Nanqu 4580 and then sterilizing and drying fermentation liquor; a culture medium for fermentation culture comprises the following components in parts by weight: 15-20 parts of milk powder, 1-4 parts of sodium glutamate and 40-80 parts of water; the temperature of the fermentation culture is 30-45 DEG C, and the time is 50-90 hours. And when the fermentation culture is performed for 0-15 hours, supplementing sodium glutamate into the fermentation culture system until the concentration of the sodium glutamate in the fermentation system is 10-30 g / L. In the fermentation process, active ingredients for improving sleep can be accumulated to the maximum extent, the application effect of the product in improving sleep is improved, and the product is good in taste and sour, sweet and palatable in taste.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial application, and in particular relates to a lactic acid bacteria postbiotic preparation for assisting sleep, a composition, a preparation method and an application thereof. Background Art

[0002] Sleep is a basic function of organisms and plays a very important role in maintaining normal physiological activities. High-quality sleep not only helps improve the quality of life, but also plays an important role in relieving fatigue, restoring physical strength, repairing neurons, consolidating memory, maintaining metabolic homeostasis, etc. Therefore, sleep quality has received more and more attention from the public. However, according to surveys, in today's fast-paced and stressful social life, nearly 60% of people believe that they have various problems with their sleep or even sleep disorders. Difficulty falling asleep, waking up easily, and difficulty falling asleep after waking up have become the three typical manifestations of sleep problems.

[0003] In order to solve the problem of poor sleep quality among the people, the Chinese invention patent "Application of Naqu 4580 and Preparations in Foods for Improving Sleep Quality" with application number ZL202010705870.4 discloses that Naqu 4580 can improve sleep quality. However, it needs to be taken in the form of live bacteria, and its colonization and metabolism in the body are used to produce metabolites that improve sleep, thereby achieving the effect of improving sleep. Therefore, in order to ensure the activity of Naqu 4580, the fermentation liquid can only be dried by freeze-drying, and the drying cost is extremely high. Therefore, it is necessary to develop a postbiotic preparation of lactic acid bacteria Naqu 4580 and its composition, and use its fermentation product to improve sleep. Compared with the prior art, the product sterilizes the bacteria of Naqu 4580, and does not need to retain the live bacteria of Naqu 4580, but there are the following problems: Since the postbiotic preparation is taken directly, its palatability must be ensured, which leads to some ingredients such as beef extract and other culture medium raw materials with poor palatability cannot be used for the fermentation culture of Naqu 4580. For Naqu 4580, the composition of its culture medium and the fermentation process are crucial for the accumulation of active ingredients for improving sleep and the effect of its postbiotic preparation on improving sleep. When the culture medium and fermentation conditions are not suitable, even if it is extremely beneficial to the growth of Naqu 4580, it cannot maximize the accumulation of active ingredients for improving sleep, resulting in its failure to achieve the application effect of improving sleep. Therefore, suitable culture medium composition and fermentation conditions are the key to ensuring the palatability of postbiotic preparations and the application effect of assisting sleep. Summary of the invention

[0004] The purpose of the present invention is to overcome the defects in the prior art and provide a lactic acid bacteria postbiotic preparation, composition, preparation method and application thereof for assisting sleep, which can maximize the accumulation of active ingredients for improving sleep during the fermentation process, thereby improving its application effect in improving sleep, and the product has a good taste and a sweet and sour taste.

[0005] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0006] A lactic acid bacteria postbiotic preparation capable of assisting sleep, wherein Naqu 4580 is fermented and cultured, and the fermented liquid is sterilized and dried to obtain the fermented liquid; the culture medium used for the fermentation culture comprises, by weight, 15-20 parts of milk powder, 1-4 parts of sodium glutamate, and 40-80 parts of water;

[0007] The fermentation culture temperature is 30-45°C and the fermentation time is 50-90h.

[0008] As a further technical solution, the Naqu 4580 comprises the following raw materials in percentage by weight: 10-20% of Lactobacillus paracasei L578; 75%-88% of Lactobacillus helveticus L551; and 2%-5% of Streptococcus salivarius subsp. thermophilus S709.

[0009] As a further technical solution, when the fermentation culture reaches 0-15h (preferably 10-15h), it is also necessary to feed sodium glutamate into the fermentation culture system until the concentration of sodium glutamate in the fermentation system is 10-30g / L.

[0010] The preparation method of the lactic acid bacteria postbiotic preparation comprises the following steps:

[0011] Step 1, mixing: add culture medium into the mixing tank and stir at 55-60℃ for 10-15min;

[0012] Step 2, sterilization: the liquid in the material tank is pumped into the fermentation tank and pasteurized at 90-110°C for 5-30 minutes;

[0013] Step 3, fermentation: after inoculating Naqu 4580 into a fermentation tank, ferment and culture at a fermentation temperature of 30-45° C. for 50-90 hours to obtain a fermentation liquid;

[0014] Step 4: sterilize the fermentation liquid at high temperature to inactivate the strain, dry it, and grind it to obtain the lactic acid bacteria postbiotic preparation.

[0015] As a further technical solution, the drying adopts vacuum belt drying.

[0016] A composition comprising the following raw materials in percentage by weight:

[0017] 20-60% of the lactic acid bacteria postbiotic preparation;

[0018] Fructo-oligosaccharides 2-15%;

[0019] Isomaltooligosaccharide 0-20%;

[0020] Xylitol 10-30%;

[0021] Erythritol 0-30% (preferably 5-30%).

[0022] Application of the lactic acid bacteria postbiotic preparation or the composition in preparing sleep-aiding food or medicine.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] In order to ensure the palatability of the postbiotic preparation of the present invention, the present invention uses milk powder to ferment and culture Naqu 4580, which not only provides a nitrogen source for Naqu 4580, but also has the characteristics of good palatability. The present invention feeds sodium glutamate during the fermentation process, which helps the production and accumulation of the metabolite γ-aminobutyric acid with the effect of improving sleep, thereby improving the application effect of the present invention in improving sleep.

[0025] By screening and optimizing the culture medium and fermentation conditions, the present invention makes the inactivated fermentation product of Naqu 4580 also have an excellent effect of improving sleep and better palatability. Compared with the prior art, since it is an inactivated product, it is not necessary to ensure the activity of the bacteria during drying. Therefore, more drying methods can be selected, thereby reducing the drying cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a venn diagram of the intersection target of postbiotic preparations and insomnia;

[0027] Figure 2 GO biological process diagram for the core targets of postbiotic preparations;

[0028] Figure 3 KEGG pathway enrichment bubble diagram for the core targets of postbiotic preparations;

[0029] Figure 4 A Sankey diagram for the classification of medicinal materials-ingredients-targets-pathways-pathways for postbiotic preparations;

[0030] Figure 5 It is a statistical diagram of sleep disorders among people in the application embodiment;

[0031] Figure 6 This is a statistical chart of people's reliance on drugs to assist sleep in the application examples. DETAILED DESCRIPTION

[0032] The technical solution of the present invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. 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.

[0033] In addition, it should be noted that the terms "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or apparatus.

[0034] In the present invention,

[0035] Naqu 4580 comprises the following raw materials in percentage by weight: 10-20% of Lactobacillus paracasei L578; 75%-88% of Lactobacillus helveticus L551; and 2%-5% of Streptococcus salivarius subsp. thermophilus S709.

[0036] Among them, Lacticaseibacillus paracasei L578 is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number of CGMCC NO.15603 and the deposit date of April 11, 2018;

[0037] Lactobacillus helveticus L551 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC NO.15604 and the deposit date of April 11, 2018;

[0038] Streptococcus salivarius subsp. thermophilus S709 is deposited in the General Microbiology Center of China Culture Collection Administration, with the deposit number of CGMCCNO.18028.

[0039] The above strains have all been disclosed in the prior art.

[0040] Unless otherwise specified, the raw materials used in the present invention are all commercially available.

[0041] Example 1

[0042] A composition capable of assisting sleep, comprising the following raw materials in percentage by weight: the same as Example 1, except that the preparation method of the lactic acid bacteria postbiotic preparation is different;

[0043] The preparation method of the lactic acid bacteria postbiotic preparation of this embodiment comprises the following steps:

[0044] Step 1, mixing: add culture medium into the mixing tank and stir at 58°C for 12 minutes;

[0045] The culture medium comprises, by weight: 20 parts of milk powder, 4 parts of sodium glutamate, and 80 parts of water;

[0046] Step 2, sterilization: the culture medium in the feed tank is pumped into the fermentation tank and pasteurized at 95°C for 20 minutes;

[0047] Step 3, fermentation: after inoculating Naqu 4580 into a fermentation tank, fermentation culture was carried out at a fermentation temperature of 38° C. for 60 hours to obtain a fermentation liquid;

[0048] During the fermentation process, when the fermentation culture reaches 12 hours, sterilized sodium glutamate is added until the concentration of sodium glutamate in the fermentation culture system reaches 20 g / L;

[0049] Step 4, sterilization and drying: sterilize the fermentation liquid at high temperature to inactivate the strain, vacuum belt dry it, and grind it to obtain the lactic acid bacteria postbiotic preparation.

[0050] Example 2

[0051] A composition capable of assisting sleep, comprising the following raw materials in percentage by weight: the same as Example 1, except that the preparation method of the lactic acid bacteria postbiotic preparation is different;

[0052] The preparation method of the lactic acid bacteria postbiotic preparation of this embodiment comprises the following steps:

[0053] Step 1, mixing: add culture medium into the mixing tank and stir at 60°C for 14 minutes;

[0054] The culture medium comprises, by weight: 15 parts of milk powder, 2 parts of sodium glutamate, and 70 parts of water;

[0055] Step 2, sterilization: the culture medium in the feed tank is pumped into the fermentation tank and pasteurized at 95°C for 20 minutes;

[0056] Step 3, fermentation: after inoculating Naqu 4580 into a fermentation tank, fermentation culture was carried out at a fermentation temperature of 40° C. for 58 hours to obtain a fermentation liquid;

[0057] During the fermentation process, when the fermentation culture reaches 10 hours, sterilized sodium glutamate is added until the concentration of sodium glutamate in the fermentation culture system reaches 22 g / L;

[0058] Step 4, sterilization and drying: sterilize the fermentation liquid at high temperature to inactivate the strain, vacuum belt dry it, and grind it to obtain the lactic acid bacteria postbiotic preparation.

[0059] Example 3

[0060] A composition capable of assisting sleep, comprising the following raw materials in percentage by weight: the same as Example 1, except that the preparation method of the lactic acid bacteria postbiotic preparation is different;

[0061] The preparation method of the lactic acid bacteria postbiotic preparation of this embodiment comprises the following steps:

[0062] Step 1, mixing: add culture medium into the mixing tank and stir at 55°C for 11 minutes;

[0063] The culture medium comprises, by weight: 18 parts of milk powder, 2 parts of sodium glutamate, and 75 parts of water;

[0064] Step 2, sterilization: the culture medium in the feed tank is pumped into the fermentation tank and pasteurized at 95°C for 20 minutes;

[0065] Step 3, fermentation: after inoculating Naqu 4580 into a fermentation tank, fermentation culture was carried out at a fermentation temperature of 39° C. for 65 hours to obtain a fermentation liquid;

[0066] During the fermentation process, when the fermentation culture reaches 15 hours, sterilized sodium glutamate is added until the concentration of sodium glutamate in the fermentation culture system reaches 25 g / L;

[0067] Step 4, sterilization and drying: sterilize the fermentation liquid at high temperature to inactivate the strain, vacuum belt dry it, and grind it to obtain the lactic acid bacteria postbiotic preparation.

[0068] Effect Example 1: Research on the targets and pathways of active ingredients in fermentation products

[0069] 1. Test methods

[0070] The following method was used to study the mechanism of sleep assistance of the active ingredients of the lactic acid bacteria postbiotic preparation obtained in Example 1. The results are shown in Figure 1-4 ;

[0071] The mechanism research method includes the following steps:

[0072] (1) Obtain the active ingredients of the postbiotic preparation Naqu 4580 and analyze the targets of the active ingredients;

[0073] (2) Obtain potential targets for insomnia;

[0074] (3) Intersection of potential targets for insomnia and targets of the active ingredients of Naqu 4580;

[0075] (4) PPI network was constructed for the obtained intersection targets, and visual screening was performed through software. The targets were screened for GO analysis and KEGG enrichment analysis, and the analysis results included signaling pathways;

[0076] (5) The obtained intersection targets and the active ingredients of the lactic acid bacteria postbiotic preparation (inactivated lactic acid bacteria powder Naqu 4580) were screened to screen out the effective ingredients, and a Sangkey diagram was constructed through the core targets and signal pathways to analyze the sleep-aiding mechanism of Naqu 4580.

[0077] 2. Results and Analysis

[0078] from Figure 1-4 It can be seen that the lactic acid bacteria postbiotic preparation of the present invention helps to relieve insomnia, has the characteristics of multiple targets and multiple pathways, and mainly exerts a sleep-promoting effect through γ-aminobutyric acid (GABA), tryptophan, indole-3-lactic acid (ILA), etc. There are 162 related targets for the treatment of insomnia, mainly GABBR1, GABBR2, GABBR1, GABRB2, HTR1A, HTR1B, MTNR1A, OPRM1, etc. These core targets act on biological processes such as calcium signaling pathways, cAMP signaling pathways, serotonin synaptic pathways, GABAergic synaptic pathways, melatonin action pathways (circadian rhythms), neuroactive ligand-receptor interaction signaling pathways, retrograde endocannabinoid signaling pathways, cholinergic synaptic signaling pathways, and dopaminergic synaptic pathways, inhibiting neural overexcitation and exerting a sleep-promoting effect.

[0079] Application Example 1:

[0080] 1. Test methods

[0081] (1) Scale survey description

[0082] The improvement of sleep-related symptoms of the test subjects after taking the composition of Example 1 was investigated to evaluate the improvement effect of the Naqu 4580 composition on sleep.

[0083] The Naqu 4580 composition was distributed to the trial group at a dosage of 2 to 4 pieces per day. After taking it continuously for 15 days, the effects of the product on the sleep-related symptoms and the improvement of sleep disorders in the trial group were statistically analyzed.

[0084] The design of the trial form is based on the Pittsburgh Sleep Quality Index (PSQI), which is simplified to a certain extent according to the actual situation. The Pittsburgh Sleep Quality Index (PSQI) has become a commonly used scale for international research and psychiatric clinical assessment due to its high reliability and validity, and high correlation with polysomnography test results. It is currently the most applicable and authoritative self-assessment scale for sleep quality.

[0085] On this basis, a statistical analysis of the sleep quality data of the trial population was conducted. Whether from the subjective perception of the trial population or from objective sleep condition indicators, the sleep-aiding effect of the product of the present invention can be well evaluated.

[0086] (2) Basic information of the population

[0087] The total number of people who participated in the trial service was 107, with a male-female ratio of 15:92, mostly female, and an average age of 65.5 years old. People over 50 years old accounted for more than 98%, and the majority of the population was middle-aged and elderly. The proportion of different ages is shown in Table 1:

[0088] Table 1

[0089] age Number of people Proportion <50 years old 2 1.87％ 50≤m<60 years old 15 14.02％ 60≤m<70 years old 60 56.07％ ≥70 years 30 28.04％ total 107 100％

[0090] The sleep disorders of the population were investigated, and the results are shown in Figure 5-6 ;

[0091] in, Figure 5 The data shows that only 27.10% of people think they do not have sleep disorders, and more than half of the people have insomnia or sleep disorders for half a year or more.

[0092] Figure 6 The results showed that 29 of the people who took the drug were dependent on drugs to help them sleep, accounting for 27.10%. Among them, 15 people were seriously dependent on drugs, that is, taking drugs for more than 3 days a week, accounting for 14.02%.

[0093] 2. Test results and analysis

[0094] The composition product of the present invention has a good somatic effect on most people with sleep disorders, with an improvement rate of more than 91%. The most obvious improvement symptoms are the time to fall asleep, the number of times you get up at night, the actual sleep time, the situation of falling asleep again after waking up, and dreaminess, mental state after waking up, etc. It has a more significant effect on improving sleep quality, as follows:

[0095] (1) The overall results of the composition on sleep improvement are shown in Table 2;

[0096] The survey results after taking the composition for trial showed that more than 91% of the people believed that the product of the present invention provided benefits to their sleep.

[0097] Table 2

[0098] Effect Number of people Proportion Very effective 14 13.08％ efficient 46 42.99％ Some effect 38 35.51％ Basically ineffective 9 8.41％ total 107 100％

[0099] (2) The improvement of the composition of the present invention on the sleep disorder group and the drug dependence group is statistically analyzed. The results are shown in Table 3;

[0100] Table 3

[0101]

[0102] From the data in Table 3, it can be seen that the present invention has a certain improvement effect on people with different sleep disorders. The improvement rate of people with sleep disorders for more than 5 years is more than 96%, and the improvement rate of people who rely on sedatives is more than 86%.

[0103] (3) The statistical data of the effect of the composition of the present invention on the sleep time of the test group are shown in Table 4;

[0104] Table 4

[0105]

[0106] From the data in Table 4, it can be seen that after intervention with the composition of the present invention, the number of people who could fall asleep within 30 minutes increased from 51.4% to 65.4%, and 39.3% of the people took less time to fall asleep than before taking the product.

[0107] (4) The Naqu 4580 composition was used to analyze the waking-up situation of the test group. The results are shown in Table 5.

[0108] Table 5

[0109]

[0110]

[0111] From the statistical results in Table 5, it can be seen that after the intervention of the composition of the present invention, the number of people who woke up 2 times or less in the middle of the night increased from 73.8% to 84.1%, the number of people who frequently woke up at night (5 times or more) decreased from 5.6% to 0, and the number of people who woke up at night decreased significantly in 21.5% of the people compared with before.

[0112] (5) The effect of the composition of the present invention on the actual sleep time of the test group is statistically analyzed. The results are shown in Table 6;

[0113] Table 6

[0114]

[0115] From the statistical results in Table 6, it can be seen that after the intervention of the composition of the present invention, the proportion of people whose actual average sleep time in a week was less than 5 hours decreased from 48.6% to 21.5%, and 41.1% of the people believed that their actual sleep time was extended.

[0116] (6) The statistical results of the effect of the composition of the present invention on the sleep after waking up of the test group are shown in Table 7;

[0117] Table 7

[0118]

[0119]

[0120] From the statistical results in Table 7, it can be seen that after the intervention of the composition of the present invention, the proportion of people who can fall asleep quickly after waking up at night increased from 33.7% to 77.6%, the proportion of people who have difficulty falling asleep after waking up decreased from 29.9% to 4.7%, and 55.1% of the test subjects believed that they could continue to fall asleep better after waking up at night than before the trial.

[0121] (7) The statistical results of the effect of the Naqu 4580 combination on the dreaminess and post-awakening state of the test group are shown in Table 8;

[0122] Table 8

[0123]

[0124] From the statistical results in Table 8, it can be seen that after the intervention of the composition of the present invention, the night dreaming of the test group was alleviated, and the proportion of the group decreased from the original 62.6% to 36.4%; at the same time, the proportion of people who felt tired after waking up in the morning decreased from the original 42.1% to 12.1%. After taking the Nagqu 4580 composition, 36.4% of the people felt that they woke up in the morning with better spirits than before.

[0125] The above-described embodiments are only preferred embodiments of the present invention, and are not exhaustive of the feasible implementations of the present invention. For those skilled in the art, any obvious changes made thereto without departing from the principles and spirit of the present invention should be considered to be included in the scope of protection of the claims of the present invention.

Claims

1. A lactic acid bacteria postbiotic preparation capable of assisting sleep, characterized in that: After the Naqu 4580 is fermented and cultured, the fermentation liquid is sterilized and dried to obtain; The culture medium used for the fermentation culture comprises, by weight: 15-20 parts of milk powder, 1-4 parts of sodium glutamate, and 40-80 parts of water; The fermentation culture temperature is 30-45°C and the fermentation time is 50-90h.

2. A lactic acid bacteria postbiotic preparation according to claim 1, characterized in that: The Naqu 4580 comprises the following raw materials in percentage by weight: 10-20% of Lactobacillus paracasei L578; 75%-88% of Lactobacillus helveticus L551; and 2%-5% of Streptococcus salivarius subsp. thermophilus S709.

3. A lactic acid bacteria postbiotic preparation according to claim 1, characterized in that: When the fermentation culture reaches 0-15h, it is also necessary to feed sodium glutamate into the fermentation culture system until the concentration of sodium glutamate in the fermentation system is 10-30g / L.

4. The method for preparing the lactic acid bacteria postbiotic preparation according to any one of claims 1 to 3, characterized in that: The steps include: Step 1, mixing: add culture medium into the mixing tank and stir at 55-60℃ for 10-15min; Step 2, sterilization: the liquid in the material tank is pumped into the fermentation tank and pasteurized at 90-110°C for 5-30 minutes; Step 3, fermentation: after inoculating Naqu 4580 into a fermentation tank, ferment and culture at a fermentation temperature of 30-45° C. for 50-90 hours to obtain a fermentation liquid; Step 4: sterilize the fermentation liquid at high temperature to inactivate the strain, dry it, and grind it to obtain the lactic acid bacteria postbiotic preparation.

5. The preparation method according to claim 4, characterized in that: The drying is performed by vacuum belt drying.

6. A composition, characterized in that The following raw materials are included in weight percentage: 20-60% of the lactic acid bacteria postbiotic preparation according to any one of claims 1 to 3; Fructo-oligosaccharides 2-15%; Isomaltooligosaccharide 0-20%; Xylitol 10-30%; Erythritol 0-30%.

7. Use of the lactic acid bacteria postbiotic preparation according to any one of claims 1 to 3 or the composition according to claim 6 in the preparation of sleep-aiding food or medicine.

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