Compound probiotic and nerve-calming sleep-aiding fermentation composition as well as preparation method and application of compound probiotic and nerve-calming sleep-aiding fermentation composition

Abstract

The invention provides a compound probiotic and nerve soothing and sleep aiding fermentation composition and a preparation method and application thereof, and relates to the technical field of microorganisms, the compound probiotic comprises plant lactobacillus KMHD-1 and lactobacillus gasseri KMBGI 03, the compound probiotic is used in a compound mode, the fermentation effect of the compound probiotic can be enhanced, and the nerve soothing and sleep aiding effect of the compound probiotic can be improved. By adopting the compound probiotics fermentation, the contents of jujuboside, rare ginsenoside and polysaccharide in the product can be effectively increased, and the effects of soothing the nerves and helping sleep of the product can be effectively enhanced.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and in particular to a composite probiotic and nerve-soothing and sleep-inducing fermentation composition, and a preparation method and application thereof. Background Art

[0002] In the modern fast-paced and stressful living environment, people's anxiety and insomnia symptoms are common and on the rise. Severe anxiety and insomnia can cause serious physical and mental damage. Traditional Chinese medicine generally uses a variety of Chinese medicinal materials to mix, decoct and prepare medicinal soup for consumption, and also uses refined methods to prepare tablets or capsules for consumption. However, the efficacy of traditional Chinese medicine generally cannot achieve fast and obvious effects, and requires long-term use, a long course of treatment, and no obvious effect. Although Western medicine treatment is fast and obvious, it generally has serious side effects and dependence, which is not advisable for patients with mild anxiety and insomnia, and the gains outweigh the losses.

[0003] CN 117562958 A discloses a composition of sophora japonica seed and tangerine peel for improving sleep and relieving anxiety and a preparation method thereof. The composition contains tangerine peel extract, sophora japonica seed extract, banana powder, day lily powder, jujube seed powder, perilla powder, white lentil seed coat extract, modified Lactobacillus reuteri and modified Bifidobacterium animalis. The specially treated components are combined to have the effects of synergistically promoting sleep and improving anxiety. However, the preparation thereof requires a strict and cumbersome enzymolysis process, and is difficult to be industrially produced.

[0004] CN 116570662 A provides a fermented composition for calming the nerves and promoting sleep, and a preparation method and application thereof: the fermented composition comprises the following components by weight: 80-160 parts of spiny jujube seeds, 1-20 parts of pleurotus eryngii, 20-60 parts of notoginseng flowers, 1-10 parts of valerian, 0.3-1.5 parts of fermented bacterial powder, 5-30 parts of enzyme preparation and 1-10 parts of acid-base regulator; the fermented composition is prepared by enzymolysis, extraction and fermentation processes; the compatibility of the Chinese medicinal ingredients in the invention plays a good role in improving sleep quality, but its preparation requires strict and cumbersome enzymolysis, is difficult to be industrialized, and has not been subjected to sensory evaluation and human clinical verification.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] One of the purposes of the present invention is to provide a composite probiotic to at least solve one of the technical problems existing in the prior art. The composite probiotic comprises Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03. The composite probiotic can enhance its fermentation efficacy when used in combination.

[0007] The second object of the present invention is to provide an application of the composite probiotics in preparing a fermented composition for calming nerves and promoting sleep. The composite probiotics are used for fermentation to prepare a fermented composition with calming nerves and promoting sleep.

[0008] The third purpose of the present invention is to provide a fermented composition for calming nerves and helping sleep. The fermented composition for calming nerves and helping sleep includes a fermentation substrate and optional auxiliary materials; the fermented composition for calming nerves and helping sleep is prepared by fermenting the fermented substrate with the composite probiotics, and the components of the fermented substrate include: Chinese date seed and ginseng. The use of composite probiotics to ferment the fermented substrate can effectively increase the content of Chinese date seed saponin, ginseng rare saponin and polysaccharide, thereby enhancing the product's efficacy in calming nerves and helping sleep.

[0009] The fourth object of the present invention is to provide a method for preparing a fermented composition for calming the nerves and promoting sleep. The method comprises the following steps: mixing, extracting and concentrating the raw material components of the fermentation substrate in the formula amount in sequence to obtain a concentrated solution, then mixing the seed solution of the composite probiotics with the concentrated solution, and fermenting to obtain a fermented product. The preparation method can effectively increase the content of the active ingredient in the product, and the preparation method has a simple process, high efficiency and feasibility.

[0010] The fifth object of the present invention is to provide an application of the calming and sleep-inducing fermented composition in the preparation of a calming and sleep-inducing product.

[0011] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted: In a first aspect, the present invention provides a composite probiotic, comprising: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Lactobacillus gasseri KMBGI 03 ( Lactobacillus gas station KMBGI 03); The plant lactobacillus KMHD-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on August 28, 2023, with the deposit number CGMCC No.28269, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; The Lactobacillus gasseri KMBGI 03 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17177, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0012] In an optional embodiment, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Lactobacillus gasseri KMBGI 03 is (0.5-1.5):1.

[0013] In a second aspect, the present invention provides a use of the composite probiotics in preparing a fermented composition for calming nerves and promoting sleep.

[0014] In a third aspect, the present invention provides a fermented composition for calming the nerves and promoting sleep, wherein the fermented composition for calming the nerves and promoting sleep comprises a fermentation substrate and optional auxiliary materials; The calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with the composite probiotics, and the components of the fermentation substrate include: spiny jujube seeds and ginseng.

[0015] In an optional embodiment, the fermentation substrate further comprises at least one of Poria cocos and Longan pulp; Preferably, the fermentation substrate comprises the following components by weight: 12-15 parts of ginseng, 12-15 parts of spiny jujube seeds, 20-26 parts of tuckahoe and 30-35 parts of longan pulp; Preferably, the auxiliary material comprises at least one of blueberry concentrated juice, γ-aminobutyric acid, casein peptide and sweetener; Preferably, the sweetener comprises erythritol and natural sweetener; Preferably, the auxiliary materials include the following components by weight: 50-55 parts of blueberry concentrated juice, 12-16 parts of γ-aminobutyric acid, 5-7 parts of casein peptide, 60-70 parts of erythritol and 0.2-0.6 parts of natural sweetener.

[0016] In a fourth aspect, the present invention provides a method for preparing the fermented composition for calming nerves and promoting sleep, comprising the following steps: The raw material components of the fermentation substrate in the formula amount are mixed, extracted and concentrated in sequence to obtain a concentrated solution, and then the composite probiotics are mixed with the concentrated solution and fermented to obtain a fermentation product.

[0017] In an optional embodiment, the extraction includes a primary extraction and a secondary extraction; Preferably, the primary extraction comprises: adding water to the mixed components of the fermentation substrate, performing a boiling extraction and a filtering in sequence to obtain a primary extract; Preferably, the amount of water added for the extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time for the boiling extraction is 1-2 h; Preferably, the secondary extraction comprises: adding water, performing secondary boiling extraction and secondary filtration on the components of the fermentation substrate after the primary extraction in sequence to obtain a secondary extract; Preferably, the amount of water added in the secondary extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the secondary boiling extraction is 1-2 h; Preferably, the extraction further comprises: mixing the primary extract and the secondary extract to obtain a total extract.

[0018] In an optional embodiment, the weight of the concentrated liquid is 4-7 times the weight of the fermentation substrate raw material component; Preferably, the concentration temperature is 75-85°C; Preferably, after the concentration and before the fermentation, the concentrated liquid is sterilized for the first time; Preferably, the first sterilization adopts pasteurization; Preferably, the mixing of the composite probiotics with the concentrated solution comprises: mixing Lactobacillus plantarum KMHD-1 powder and Lactobacillus gasseri KMBGI 03 powder to obtain composite probiotic powder, then dissolving the composite probiotics in water and inoculating into the concentrated solution; Preferably, the added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution; Preferably, the fermentation time is 24-72 h.

[0019] In an optional embodiment, the method for preparing the fermented composition for calming the nerves and promoting sleep further comprises: mixing the auxiliary materials in the formula amount with the fermented product to prepare the fermented composition for calming the nerves and promoting sleep; Preferably, the mixing process comprises: dissolving the casein peptide and the sweetener separately, then mixing them to obtain a premix, mixing γ-aminobutyric acid and erythritol, and then adding the fermentation product, blueberry concentrate and the premix.

[0020] In a fifth aspect, the present invention provides a use of the aforementioned calming and sleep-inducing fermented composition in the preparation of a calming and sleep-inducing product.

[0021] Compared with the prior art, the present invention has the following beneficial effects: The composite probiotics provided by the present invention include Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03. During the fermentation process of the composite probiotics, the composite probiotics act synergistically to effectively increase the content of jujube seed saponin, ginseng rare saponin and polysaccharide in the product, and can effectively enhance the tranquilizing and sleep-inducing effect of the product. DETAILED DESCRIPTION

[0022] Unless otherwise defined herein, scientific and technical terms used in conjunction with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear, however, in the case of any potential ambiguity, the definitions provided herein take precedence over any dictionary or external definitions. In this application, unless otherwise stated, the use of "or" means "and / or". In addition, the use of the term "including" and other forms is non-limiting.

[0023] The technical solution of the present invention will be clearly and completely described below in conjunction with the 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.

[0024] The first aspect of the present invention provides a composite probiotic, the composite probiotic comprising: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Lactobacillus gasseri KMBGI 03 ( Lactobacillus gas station KMBGI 03); The plant lactobacillus KMHD-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on August 28, 2023, with the deposit number CGMCC No.28269, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; The Lactobacillus gasseri KMBGI 03 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17177, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0025] The composite probiotics provided by the present invention include Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03. During the fermentation process of the composite probiotics, the composite probiotics act synergistically to effectively increase the content of jujube seed saponin, ginseng rare saponin and polysaccharide in the product, and can effectively enhance the tranquilizing and sleep-inducing effect of the product.

[0026] Furthermore, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Lactobacillus gasseri KMBGI 03 is (0.5-1.5):1, for example, it can be 0.5:1, 1:1, 1.5:1, etc.

[0027] The second aspect of the present invention provides a use of the composite probiotics in preparing a fermented composition for calming nerves and promoting sleep.

[0028] In the present invention, the composite probiotics are used for fermentation to prepare a fermentation composition having a calming and sleep-inducing effect.

[0029] The third aspect of the present invention provides a fermented composition for calming the nerves and promoting sleep, wherein the fermented composition for calming the nerves and promoting sleep comprises a fermentation substrate and optional auxiliary materials; The calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with the composite probiotics, and the components of the fermentation substrate include: spiny jujube seeds and ginseng.

[0030] In the present invention, the components of the fermentation substrate include Chinese jujube seeds and ginseng. One of the main active ingredients in Chinese jujube seeds is Chinese jujube seed saponin, which has significant sedative and tranquilizing effects. The compound probiotic fermentation can increase the content of Chinese jujube seed saponin; the main active ingredient of ginseng, ginsenoside, can improve the body's tolerance and has an anti-fatigue effect. The compound probiotic fermentation can promote the generation or release of rare saponins in ginseng through specific enzymatic reactions; Chinese jujube seeds and ginseng are both rich in polysaccharide components, and these polysaccharides are further improved and activated by the action of the compound probiotics during the fermentation process, and these polysaccharides jointly enhance the tranquilizing and sleep-inducing effects of the product through multiple mechanisms such as antioxidant, immunomodulatory and anti-inflammatory effects.

[0031] Furthermore, the components of the fermentation substrate also include at least one of Poria cocos and longan pulp; In the present invention, the components of the fermentation substrate also include Poria cocos and longan pulp. The effective ingredients of longan include longan polysaccharide, and the main components of Poria cocos include Poria cocos polysaccharide. Both Poria cocos and longan pulp have sedative and anti-anxiety effects and can also improve matrix immunity. The effective ingredients of Poria cocos and longan pulp are further activated during the fermentation process, and together enhance the product's tranquilizing and sleep-inducing effects, thereby providing better sleep quality and overall health status.

[0032] Preferably, the fermentation substrate comprises the following components by weight: 12-15 parts of ginseng, 12-15 parts of spiny jujube seeds, 20-26 parts of tuckahoe and 30-35 parts of longan pulp; Wherein, in the fermentation substrate, the amount of ginseng added is 12 parts, 13 parts, 14 parts, 15 parts, etc.; Among them, in the fermentation substrate, the amount of sour jujube seeds added is 12 parts, 13 parts, 14 parts, 15 parts, etc.; Among them, in the fermentation substrate, the amount of Poria cocos added is 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, etc.; Among them, the added amount of longan pulp in the fermentation substrate is 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, etc.

[0033] Preferably, the auxiliary material comprises at least one of blueberry concentrated juice, γ-aminobutyric acid, casein peptide and sweetener; Preferably, the sweetener comprises erythritol and natural sweetener; In the present invention, the blueberry concentrated juice contains effective ingredients such as anthocyanins, γ-aminobutyric acid has a neuromodulatory effect, casein peptide has the effects of relaxing the nervous system, improving sleep and regulating immunity, erythritol and natural sweeteners are used to improve the taste of the product, and the natural sweeteners include mogroside and stevioside.

[0034] Preferably, the auxiliary materials include the following components by weight: 50-55 parts of blueberry concentrated juice, 12-16 parts of γ-aminobutyric acid, 5-7 parts of casein peptide, 60-70 parts of erythritol and 0.2-0.6 parts of natural sweetener.

[0035] Among them, the amount of blueberry concentrated juice added in the auxiliary materials is 50 parts, 51 parts, 52 parts, 53 parts, 54 parts, 55 parts, etc.; Among them, the amount of γ-aminobutyric acid added in the auxiliary material is 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, etc.; Among them, the amount of casein peptide added in the auxiliary materials is 5 parts, 6 parts, 7 parts, etc.; Among them, in the auxiliary materials, the added amount of erythritol is 60 parts, 61 parts, 62 parts, 63 parts, 64 parts, 65 parts, 66 parts, 67 parts, 68 parts, 69 parts, 70 parts, etc.; Among them, the added amount of natural sweetener in the auxiliary materials is 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.6 parts, etc.

[0036] The fourth aspect of the present invention provides a method for preparing the fermented composition for calming the nerves and promoting sleep, the method comprising the following steps: The raw material components of the fermentation substrate in the formula amount are mixed, extracted and concentrated in sequence to obtain a concentrated solution, and then the composite probiotics are mixed with the concentrated solution and fermented to obtain a fermentation product.

[0037] In the present invention, the preparation method is adopted to prepare the fermented composition for calming nerves and promoting sleep, which can effectively increase the content of effective ingredients in the product, and the preparation method has a simple process, high efficiency and feasibility.

[0038] Further, the extraction includes primary extraction and secondary extraction; In the present invention, a two-time extraction method is adopted to improve the extraction rate, avoid the waste of effective ingredients in the product, ensure thorough extraction, and thus improve product quality.

[0039] Preferably, the primary extraction comprises: adding water to the mixed components of the fermentation substrate, performing a boiling extraction and a filtering in sequence to obtain a primary extract; Preferably, the amount of water added for one extraction is 10-20 times the weight of the fermentation substrate raw material component, for example, it can be 10 times, 11 times, 12 times, 13 times, 14 times, 15 times, 16 times, 17 times, 18 times, 19 times, 20 times, etc., and the time for one boiling extraction is 1-2 h, for example, it can be 1 h, 1.5 h, 2 h, etc.; Preferably, the secondary extraction comprises: adding water, performing secondary boiling extraction and secondary filtration on the components of the fermentation substrate after the primary extraction in sequence to obtain a secondary extract; In the present invention, a double-layer filtration method of silk cloth is adopted in both the primary filtration and the secondary filtration.

[0040] Preferably, the amount of water added for the secondary extraction is 10-20 times the weight of the fermentation substrate raw material component, for example, it can be 10 times, 11 times, 12 times, 13 times, 14 times, 15 times, 16 times, 17 times, 18 times, 19 times, 20 times, etc., and the time for the secondary boiling extraction is 1-2 h, for example, it can be 1 h, 1.5 h, 2 h, etc.; Preferably, the extraction further comprises: mixing the primary extract and the secondary extract to obtain a total extract.

[0041] Further, the weight of the concentrated liquid is 4-7 times the weight of the fermentation substrate raw material component, for example, it can be 4 times, 5 times, 6 times, 7 times, etc.; Preferably, the concentration temperature is 75-85°C, for example, 75°C, 76°C, 77°C, 78°C, 79°C, 80°C, 81°C, 82°C, 83°C, 84°C, 85°C, etc.; In the present invention, a double-effect external circulation evaporator is preferably used for heating and concentration, and the vacuum degree is set to -0.08--0.03Mpa.

[0042] Preferably, after the concentration and before the fermentation, the concentrated liquid is sterilized for the first time, preferably by pasteurization; In the present invention, the concentrated liquid is poured into a fermentation tank without adding additional carbon and nitrogen sources, and then pasteurized and cooled to about 37°C.

[0043] Preferably, the mixing of the composite probiotics with the concentrated solution comprises: mixing Lactobacillus plantarum KMHD-1 powder and Lactobacillus gasseri KMBGI 03 powder to obtain composite probiotic powder, then dissolving the composite probiotics in water and inoculating into the concentrated solution; Preferably, the added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution; Preferably, the fermentation time is 24-72 h, for example, 24 h, 48 h, 72 h, etc.

[0044] In the present invention, the fermentation substrate is fermented with composite probiotics, stirred and fermented at 37° C. for 24-72 hours, and then pasteurized and refrigerated overnight to remove precipitation to prepare a fermentation liquid for later use.

[0045] Furthermore, the preparation method of the fermented composition for calming the nerves and promoting sleep also includes: mixing the auxiliary materials in the formula amount with the fermented product to prepare the fermented composition for calming the nerves and promoting sleep; Preferably, the mixing process comprises: dissolving casein peptide and natural sweetener separately, then mixing to obtain a premix, mixing γ-aminobutyric acid and erythritol, and then adding the fermentation product, blueberry concentrate and the premix.

[0046] In the present invention, casein peptide and natural sweetener are fully dissolved with appropriate amount of purified water to obtain pre-solution; add appropriate amount of purified water to the pre-dissolution tank, turn on stirring and steam heating, heat to 60-70°C, add γ-aminobutyric acid and erythritol, stir and dissolve, add fermentation concentrate, blueberry concentrate and premixed liquid, and stir and mix evenly. Use purified water to weigh to 1 ton, keep warm and stir, and mix evenly. The prepared liquid is filtered while hot through a plate and frame and then filled, and the filled semi-finished product is placed in a sterilization tray and placed in a sterilizer for wet heat sterilization. The sterilization conditions are as follows: sterilization temperature 121°C, sterilization time 20 min.

[0047] The fifth aspect of the present invention provides a use of the calming and sleep-inducing fermented composition in the preparation of a sleep-inducing product.

[0048] In the present invention, composite probiotics are used to ferment Chinese medicinal and edible Chinese medicines such as Chinese jujube seeds and ginseng to prepare a Chinese medicinal and edible Chinese medicine fermentation liquid that can calm the nerves and help sleep, as well as to prepare products that can calm the nerves and help sleep, such as products such as calming nerves and helping sleep beverages.

[0049] The present invention is further described below by way of examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or directly purchased from the market.

[0050] Example 1 This embodiment provides a calming and sleep-inducing fermented composition, which is prepared by fermenting composite probiotics (Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03), and the calming and sleep-inducing fermented composition comprises a fermentation substrate and auxiliary materials; Wherein, the weight ratio of Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03 is 1:1; The fermentation substrate includes the following components by weight: 14 parts of ginseng, 14 parts of spinach seeds, 23 parts of poria cocos and 34 parts of longan pulp; Among them, the auxiliary materials include the following components by weight: 53 parts of blueberry concentrated juice, 14 parts of γ-aminobutyric acid, 6 parts of casein peptide, 65 parts of erythritol, 0.2 parts of stevioside and 0.2 parts of mogroside.

[0051] Example 2 This embodiment provides a calming and sleep-inducing fermented composition, which is prepared by fermenting composite probiotics (Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03), and the calming and sleep-inducing fermented composition comprises a fermentation substrate and auxiliary materials; Wherein, the weight ratio of Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03 is 0.5:1; The fermentation substrate includes the following components by weight: 12 parts of ginseng, 15 parts of spinach seeds, 20 parts of poria cocos and 35 parts of longan pulp; Among them, the auxiliary materials include the following components by weight: 50 parts of blueberry concentrated juice, 16 parts of γ-aminobutyric acid, 5 parts of casein peptide, 70 parts of erythritol, 0.1 parts of stevioside and 0.1 parts of mogroside.

[0052] Example 3 This embodiment provides a calming and sleep-inducing fermented composition, which is prepared by fermenting composite probiotics (Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03), and the calming and sleep-inducing fermented composition comprises a fermentation substrate and auxiliary materials; Wherein, the weight ratio of Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03 is 1.5:1; The fermentation substrate includes the following components by weight: 15 parts of ginseng, 12 parts of spinach seeds, 26 parts of poria cocos and 30 parts of longan pulp; Among them, the auxiliary materials include the following components by weight: 55 parts of blueberry concentrated juice, 12 parts of γ-aminobutyric acid, 7 parts of casein peptide, 60 parts of erythritol, 0.3 parts of stevioside and 0.3 parts of mogroside.

[0053] Embodiment 4-6 Example 4-6 provides a method for preparing a fermented composition for calming the nerves and promoting sleep, comprising the following steps: Step 1: Extraction, according to the formula amount of the fermentation substrate components in Examples 1-3, respectively, the formula amount of the fermentation substrate components is put into a multifunctional extraction tank, 15 times the amount of purified water is added for the first extraction, the extraction is boiled for 1.5 hours, and then double-layer filtration is performed with silk cloth; 15 times the amount of purified water is added for the second extraction, the extraction is boiled for 1.5 hours, and then double-layer filtration is performed with silk cloth, and the two filtrates are combined to obtain the total extract.

[0054] Step 2: Concentrate, pump the total extract into a double-effect external circulation evaporator, start heating and concentrating, set the vacuum degree to -0.08--0.03Mpa, the temperature to 80°C, and concentrate to 6 times the weight of the raw material. The concentrate is the fermentation substrate.

[0055] Step 3: Examples 4-6 are fermented according to the formula amount of the powder ratio in Examples 1-3, and the concentrated solution is poured into the fermentation tank without adding additional carbon and nitrogen sources. Then pasteurize and cool to about 37°C. Mix the plant lactobacillus KMHD-1 powder and the gasseri lactobacillus KMBGI 03 powder, and the addition amount of the composite probiotic powder is 0.1% of the weight of the concentrated solution. Then, the composite probiotic powder is fully dissolved with sterile purified water at room temperature (wherein, the amount of sterile purified water used is 7-10 times the weight of the composite probiotic powder, which can achieve the purpose of fully dissolving the composite probiotic powder), and stir evenly. After stirring and fermenting at 37°C for 48 hours, pasteurize, refrigerate and stand overnight to remove precipitation, and set aside.

[0056] Step 4: In Examples 4-6, according to the formula amount of the auxiliary material components in Examples 1-3, casein peptides and natural sweeteners are fully dissolved with appropriate amount of purified water to obtain pre-solutions; add appropriate amount of purified water to the pre-dissolution tank, turn on stirring and steam heating, heat to 60-70°C, add γ-aminobutyric acid and erythritol, stir to dissolve, add fermentation concentrate, blueberry concentrate and premixed liquid, and stir and mix evenly. Use purified water to weigh to 1 ton, keep warm and stir, and mix evenly. The prepared liquid is filtered while hot through the plate and frame and then filled. The filled semi-finished product is placed in a sterilization tray and placed in a sterilizer for wet heat sterilization. The sterilization conditions are 121°C for 20 min.

[0057] Example 7 This embodiment provides a method for preparing a fermented composition for calming the nerves and promoting sleep, comprising the following steps: Step 1: Extraction. According to the formula amount of the fermentation substrate component in Example 1, the formula amount of the fermentation substrate component is put into a multifunctional extraction tank. For the first extraction, 10 times the amount of purified water is added, and the mixture is boiled and extracted for 2 hours, and then double-filtered with silk cloth; for the second extraction, 20 times the amount of purified water is added, and the mixture is boiled and extracted for 1 hour, and then double-filtered with silk cloth, and the two filtrates are combined to obtain the total extract.

[0058] Step 2: Concentrate, pump the total extract into a double-effect external circulation evaporator, start heating and concentrating, set the vacuum degree to -0.08--0.03Mpa, the temperature to 85°C, and concentrate to 4 times the weight of the raw material. The concentrate is the fermentation substrate.

[0059] Step 3: Fermentation, pour the concentrated solution into the fermentation tank without adding additional carbon and nitrogen sources. Then pasteurize and cool to about 37°C. Mix the plant lactobacillus KMHD-1 powder and the gasseri Lactobacillus KMBGI 03 powder, and add the composite probiotic powder in an amount of 0.12% of the weight of the concentrated solution. Then fully dissolve the composite probiotic powder with sterile purified water at room temperature (wherein, the amount of sterile purified water used is 7-10 times the weight of the composite probiotic powder, which can achieve the purpose of fully dissolving the composite probiotic powder), and stir evenly. After stirring and fermenting at 37°C for 72 hours, pasteurize, refrigerate and stand overnight to remove the precipitate, and set aside.

[0060] Step 4: According to the formula amount of the auxiliary material components in Example 1, Step 4 of Example 7 is consistent with Step 4 of Example 4.

[0061] Example 8 This embodiment provides a method for preparing a fermented composition for calming the nerves and promoting sleep, comprising the following steps: Step 1: Extraction. According to the formula amount of the fermentation substrate component in Example 1, the formula amount of the fermentation substrate component is put into a multifunctional extraction tank. For the first extraction, 20 times the amount of purified water is added, and the extraction is boiled for 1 hour, and then double-layer filtration is performed with silk cloth; for the second extraction, 10 times the amount of purified water is added, and the extraction is boiled for 2 hours, and then double-layer filtration is performed with silk cloth. The two filtrates are combined to obtain the total extract.

[0062] Step 2: Concentrate. Pump the total extract into a double-effect external circulation evaporator and start heating and concentrating. Set the vacuum degree to -0.08-0.03Mpa and the temperature to 75°C. Concentrate to 7 times the weight of the raw material. The concentrate is the fermentation substrate.

[0063] Step 3: Fermentation, pour the concentrated solution into the fermentation tank without adding additional carbon and nitrogen sources. Then pasteurize and cool to about 37°C. Lactobacillus plantarum KMHD-1 powder and Lactobacillus gasseri KMBGI 03 powder are mixed, and the addition amount of the composite probiotic powder is 0.07% of the weight of the concentrated solution. Then, the composite probiotic powder is fully dissolved with sterile purified water at room temperature (wherein, the amount of sterile purified water used is 7-10 times the weight of the composite probiotic powder, which can achieve the purpose of fully dissolving the composite probiotic powder), and stir evenly. After stirring and fermenting at 37°C for 24 hours, pasteurize, refrigerate and stand overnight to remove the precipitate, and set aside.

[0064] Step 4: According to the formula amount of the auxiliary material components in Example 1, Step 4 of Example 8 is consistent with Step 4 of Example 4.

[0065] Comparative Example 1 This comparative example provides a calming and sleep-inducing fermented composition, which is different from Example 1 in that the calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with Lactobacillus plantarum KMHD-1, and the rest is consistent with Example 1.

[0066] Comparative Example 2 This comparative example provides a calming and sleep-inducing fermented composition, which is different from Example 1 in that the calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with Lactobacillus gasseri KMBGI 03, and the rest is consistent with Example 1.

[0067] Comparative Example 3 This comparative example provides a calming and sleep-inducing fermented composition, which is different from Example 1 in that the calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with a composite probiotic (commercial bacteria Lactobacillus plantarum and commercial bacteria Lactobacillus gasseri), and the rest is consistent with Example 1.

[0068] Comparative Example 4 This comparative example provides a calming and sleep-inducing fermented composition, which is different from Example 1 in that the calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with a composite probiotic (Lactobacillus plantarum KMHD-1 and commercial bacteria Lactobacillus gasseri), and the rest is consistent with Example 1.

[0069] Comparative Example 5 This comparative example provides a calming and sleep-inducing fermented composition, which is different from Example 1 in that the calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with a composite probiotic (commercial bacteria Lactobacillus plantarum and Lactobacillus gasseri KMBGI 03), and the rest is consistent with Example 1.

[0070] Comparative Examples 6-10 Comparative Examples 6-10 provide a method for preparing a fermented composition for calming the nerves and promoting sleep. The preparation method of Example 4 is used to prepare the fermented compositions for calming the nerves and promoting sleep of Comparative Examples 1-5, respectively.

[0071] Test example 1. Active ingredient detection.

[0072] (1) Detection method: Rare ginsenosides of ginseng were separated by high performance liquid chromatography: acetonitrile containing 0.1% formic acid was used as mobile phase B, and water containing 0.1% formic acid was used as mobile phase C for gradient elution; the chromatographic column was ACQUITY UPLC BEH Shield RP18 (2.1 mm×100mm, 1.7 μm); the flow rate was 0.4 mL / min; and the column temperature was 30°C.

[0073] Jujube seed saponins were analyzed by high performance liquid chromatography: the chromatographic column was Ultimate AQ-C18 (4.6mm × 250mm, 5μm), the mobile phase was acetonitrile-water (33:77), the flow rate was 1.0mL·min-1, and the column temperature was 30℃.

[0074] The polysaccharide was determined by the phenol-sulfuric acid colorimetric method: accurately measure 0.0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of glucose reference solution and place them in 20 mL stoppered test tubes, and fill to 1.0 mL with water. Add 1.0 mL of 5% phenol solution, then quickly add 5.0 mL of sulfuric acid and let stand for 10 minutes. Mix the reaction solution thoroughly, react the test tube in a 30°C water bath for 20 minutes, and measure the absorbance at 490 nm. Draw a standard curve with glucose concentration as the horizontal axis and absorbance value as the vertical axis. The standard curve is: y=0.008x+0.0885, R 2 =0.9901.

[0075] (2) Test samples: The same method was used to test the sedative and sleep-inducing fermented compositions prepared in Examples 4-8 and Comparative Examples 6-10, and the results are shown in Table 1. Among them, there are 12 types of rare ginsenosides tested: Rh1, 20(R)-Rg2, 20(S)-Rg3, 20(R)-Rg3, 20(S)-PPT, 20(R)-PPT, CK, Rk1, Rg5, 20(S)-Rh2, 20(R)-Rh2, and 20(S)-PPD.

[0076] Table 1

[0077] As can be seen from Table 1, the tranquilizing and sleeping-aiding products prepared in the embodiments of the present invention are rich in rare ginseng saponins, jujube seed saponins, and polysaccharides; the rare ginseng saponins in Comparative Example 6 are higher than those in Comparative Example 8, indicating that the effect of fermenting and converting saponins by self-screened bacteria Lactobacillus plantarum is significant; compared with Comparative Example 6 and Comparative Example 7, Comparative Examples 9 and 10 use self-screened bacteria and commercial bacteria for mixed fermentation, and the contents of rare ginseng saponins, jujube seed saponins, and polysaccharides are improved to varying degrees, indicating that the mixed bacteria fermentation effect is better than that of single bacteria fermentation; the contents of rare saponins, jujube seed saponins, and polysaccharides in the product of Example 4 are significantly higher than those in Comparative Examples 8, 9, and 10, which shows that the specific interaction produced by the fermentation of the two self-screened strains affects the conversion process of rare ginseng saponins. In addition, the mixed fermentation of the two self-screened strains also significantly improves the contents of jujube seed saponins and polysaccharides, indicating that the mixed fermentation of the two self-screened strains has a more significant sleep-aiding effect.

[0078] Test case 2. Population test evaluation.

[0079] (1) Test method: According to the Guiding Principles for Clinical Research of New Chinese Medicines, patients who meet both the main and secondary symptoms at the stage of recruiting research subjects are diagnosed with insomnia and included in the study. The main symptoms are: restlessness and insomnia, or light sleep and easy awakening, or waking up and difficult to fall asleep again; the secondary symptoms are: five-heart heat, hot flashes and night sweats, dry mouth and throat, palpitations, and sore waist and knees.

[0080] The subjects were divided into 4 groups, each with 30 people. After completing the TCM symptom scoring form, the subjects received oral treatment with the product of the present invention. The main symptoms and secondary symptoms of the 4 groups of subjects before and after treatment were scored to obtain symptom scores. Each of the main symptoms was scored as 0, 2, 4, and 6 points according to the levels of none, mild, moderate, and severe; each of the secondary symptoms was scored as 0, 1, 2, and 3 points according to the levels of none, mild, moderate, and severe; and the scores of each item were added together to obtain the symptom score; and the clinical efficacy of the subjects was analyzed and judged based on the symptom score. According to the "Guidelines for Clinical Research of New Chinese Medicines", the clinical efficacy is divided into the following four levels: Recovery: Symptom score reduction is greater than or equal to 95%, and TCM clinical symptoms and signs are completely relieved or disappear; Markedly effective: The symptom score is reduced by 70% or more, and the clinical symptoms and signs of traditional Chinese medicine are significantly improved; Effective: The symptom score is reduced by 30% or more, and both clinical symptoms and signs of TCM show signs of improvement; Ineffective: The symptom score decreases by less than or equal to 30%, and the TCM clinical symptoms and signs do not improve, or even show signs of worsening.

[0081] The number of subjects who were cured, markedly effective, effective and ineffective was obtained, and the total effective number and total effective rate were calculated and recorded in Table 2 below; where, the total effective number = the number of cured cases + the number of markedly effective cases + the number of effective cases, and the total effective rate = (the number of cured cases + the number of markedly effective cases + the number of effective cases) / the total number of cases × 100%; (2) Test samples: The same method was used to test the calming and sleep-inducing fermented compositions prepared in Example 4 and Comparative Examples 6-10. The results are shown in Table 2 below.

[0082] Table 2. Clinical efficacy analysis

[0083] As shown in Table 2, after the subjects took the above 6 products for 1 month, the symptoms were relieved to varying degrees. The symptoms were scored, and the effective rate of improving insomnia of the volunteers by taking the product of Example 4 was 90%, the effective rate of improving insomnia of the subjects taking Comparative Example 6 was 76.7%, the effective rate of improving insomnia of the subjects taking Comparative Example 7 was 73.3%, the effective rate of improving insomnia of the subjects taking Comparative Example 8 was 76.7%, the effective rate of improving insomnia of the subjects taking Comparative Example 9 was 83.3%, and the effective rate of improving insomnia of the subjects taking Comparative Example 10 was 80.8%, which can prove that the product prepared in Example 4 of the present invention has significant therapeutic effect on insomnia subjects.

[0084] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composite probiotic, characterized in that: include: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Lactobacillus gasseri KMBGI 03 ( Lactobacillus gasseri KMBGI 03); The plant lactobacillus KMHD-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on August 28, 2023, with the deposit number CGMCC No.28269, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; The Lactobacillus gasseri KMBGI 03 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17177, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. The composite probiotic according to claim 1, characterized in that The weight ratio of the Lactobacillus plantarum KMHD-1 to the Lactobacillus gasseri KMBGI 03 is (0.5-1.5):

1.

3. Use of the composite probiotics as claimed in claim 1 in preparing a fermented composition for calming nerves and promoting sleep.

4. A fermented composition for calming nerves and promoting sleep, characterized in that: The calming and sleep-inducing fermented composition comprises a fermentation substrate and optional auxiliary materials; The calming and sleep-inducing fermented composition is prepared by fermenting a fermentation substrate with the composite probiotics according to claim 1 or 2, and the components of the fermentation substrate include: spiny jujube seeds and ginseng.

5. The fermented composition for calming nerves and promoting sleep according to claim 4, characterized in that: The components of the fermentation substrate also include at least one of Poria cocos and longan pulp; Preferably, the fermentation substrate comprises the following components by weight: 12-15 parts of ginseng, 12-15 parts of spiny jujube seeds, 20-26 parts of tuckahoe and 30-35 parts of longan pulp; Preferably, the auxiliary material comprises at least one of blueberry concentrated juice, γ-aminobutyric acid, casein peptide and sweetener; Preferably, the sweetener comprises erythritol and natural sweetener; Preferably, the auxiliary materials include the following components by weight: 50-55 parts of blueberry concentrated juice, 12-16 parts of γ-aminobutyric acid, 5-7 parts of casein peptide, 60-70 parts of erythritol and 0.2-0.6 parts of natural sweetener.

6. The method for preparing the fermented composition for calming nerves and promoting sleep as claimed in claim 4 or 5, characterized in that: The following steps are involved: The raw material components of the fermentation substrate in the formula amount are mixed, extracted and concentrated in sequence to obtain a concentrated solution, and then the composite probiotics are mixed with the concentrated solution and fermented to obtain a fermentation product.

7. The preparation method according to claim 6, characterized in that: The extraction includes primary extraction and secondary extraction; Preferably, the primary extraction comprises: adding water to the mixed components of the fermentation substrate, performing a boiling extraction and a filtering in sequence to obtain a primary extract; Preferably, the amount of water added for the extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time for the boiling extraction is 1-2 h; Preferably, the secondary extraction comprises: adding water, performing secondary boiling extraction and secondary filtration on the components of the fermentation substrate after the primary extraction in sequence to obtain a secondary extract; Preferably, the amount of water added in the secondary extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the secondary boiling extraction is 1-2 h; Preferably, the extraction further comprises: mixing the primary extract and the secondary extract to obtain a total extract.

8. The preparation method according to claim 6, characterized in that: The weight of the concentrated liquid is 4-7 times the weight of the fermentation substrate raw material component; Preferably, the concentration temperature is 75-85°C; Preferably, after the concentration and before the fermentation, the concentrated liquid is sterilized for the first time; Preferably, the first sterilization adopts pasteurization; Preferably, the mixing of the composite probiotics with the concentrated solution comprises: mixing Lactobacillus plantarum KMHD-1 powder and Lactobacillus gasseri KMBGI 03 powder to obtain composite probiotic powder, then dissolving the composite probiotics in water and inoculating into the concentrated solution; Preferably, the added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution; Preferably, the fermentation time is 24-72 h.

9. The preparation method according to claim 6, characterized in that: The preparation method of the fermented composition for calming the nerves and promoting sleep also includes: mixing the auxiliary materials in the formula amount with the fermented product to prepare the fermented composition for calming the nerves and promoting sleep; Preferably, the mixing process comprises: dissolving casein peptide and natural sweetener separately, then mixing to obtain a premix, mixing γ-aminobutyric acid and erythritol, and then adding the fermentation product, blueberry concentrate and the premix.

10. Use of the calming and sleep-inducing fermented composition according to claim 4 or 5 in the preparation of a sleep-inducing product.

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