A composite probiotic and qi-blood-replenishing fermentation composition and its preparation method and application

Abstract

The present invention provides a composite probiotic and a Qi and blood invigorating fermentation composition, and a preparation method and application thereof, and relates to the technical field of microorganisms. The composite probiotic comprises Lactobacillus plantarum KMHD‑1 and Bifidobacterium longum KMBGI 01. By fermenting the composite probiotic, a fermentation composition capable of invigorating Qi and blood, improving immunity and enhancing energy can be prepared. In addition, by fermenting a fermentation substrate with the composite probiotic, the types and contents of rare ginsenosides of ginseng can be effectively increased, as well as the contents of astragaloside I and polysaccharides can be increased, thereby enhancing the efficacy of the product.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to a composite probiotic and qi and blood invigorating fermentation composition, and a preparation method and application thereof. Background Art

[0002] In TCM theory, Qi and blood are related to human functions, so Qi and blood deficiency can easily lead to reduced human resistance and fatigue. Qi and blood deficiency refers to Qi deficiency and blood deficiency in TCM. Modern people often suffer from Qi and blood imbalance due to irregular diet and work and rest, excessive thinking or long-term illness, and symptoms of Qi and blood deficiency appear, mainly manifested as fatigue, palpitations, shortness of breath, night sweats and insomnia, pale or sallow complexion. Qi and blood deficiency is a common and frequent disease. Red ginseng, astragalus, wolfberry, polygonatum, longan meat, and jujube are all Chinese medicines that are both medicinal and edible. They contain special physiologically active substances and their many benefits to human health have been widely proven.

[0003] At present, there are many Chinese patent medicines used clinically for qi and blood deficiency, but most of them have a series of disadvantages such as unsatisfactory efficacy, bitter taste, adverse reactions, etc. For example, CN117085086A discloses a qi and blood replenishing fermented product and its preparation process and application: the qi and blood replenishing fermented product is prepared by fermenting a fermented substrate with fermented lactobacillus SF-L35, and the fermented substrate includes the following components in parts by weight: 1-2 parts of codonopsis extract, 1-3 parts of tuckahoe extract, 1-4 parts of atractylodes extract, 1-4 parts of licorice extract, 1-3 parts of astragalus extract, 1-4 parts of wolfberry extract, 1-4 parts of red date powder, 0.5-2 parts of polygonatum extract, and 2-6 parts of brown sugar; the fermented product has the effect of replenishing qi and blood. However, the preparation method of the extract in this patent is relatively complicated, and a single strain of bacteria is used for fermentation, the fermentation time is short, and thorough fermentation cannot be ensured; CN117224614A relates to a composite fermentation product with the effects of invigorating qi and nourishing blood, and its preparation method and application: the raw materials for preparing the composite fermentation product include: 100-300 parts of ginseng, 20-300 parts of donkey-hide gelatin, 300-1000 parts of mulberry, 250-2000 parts of fruit concentrate juice, 100-300 parts of longan powder, and 20-100 parts of inulin. This invention creatively uses probiotics to ferment traditional medicine and food together with natural fruit juice using lactic acid bacteria to obtain a composite fermentation product with the effects of invigorating qi and nourishing blood, and beautifying and nourishing the skin. However, the fermentation process of this patent is complicated, time-consuming and labor-intensive.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] 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 Bifidobacterium longum KMBGI 01. The composite probiotic can enhance its fermentation efficacy when used in combination.

[0006] The second purpose of the present invention is to provide an application of the composite probiotics in preparing a fermented composition for replenishing qi and blood. The composite probiotics are used for fermentation to prepare a fermented composition for replenishing qi and blood, improving immunity and enhancing energy.

[0007] The third object of the present invention is to provide a Qi and blood tonifying fermentation composition. The Qi and blood tonifying fermentation composition is prepared by fermenting a fermentation substrate with the composite probiotics, and the components of the fermentation substrate include: red ginseng and astragalus. The use of composite probiotics to ferment the fermentation substrate can effectively increase the types and contents of rare ginseng saponins, as well as the contents of astragaloside IV and polysaccharides, thereby enhancing the efficacy of the product.

[0008] The fourth object of the present invention is to provide a method for preparing a fermented composition for replenishing qi and blood. The method for preparing the fermented composition for replenishing qi and blood 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 is simple in process, efficient and feasible.

[0009] The fifth purpose of the present invention is to provide an application of the Qi and blood tonifying fermentation composition in the preparation of Qi and blood tonifying products.

[0010] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0011] In a first aspect, the present invention provides a composite probiotic, comprising: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Bifidobacterium longum KMBGI 01 ( Bifidobacterium long KMBGI 01);

[0012] 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;

[0013] The Bifidobacterium longum KMBGI 01 was deposited in the General Microbiology Center of the China Microorganism Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17175, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0014] Furthermore, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Bifidobacterium longum KMBGI 01 is (0.5-1.5):1.

[0015] In a second aspect, the present invention provides a use of the composite probiotics in preparing a Qi and blood-invigorating fermentation composition.

[0016] In a third aspect, the present invention provides a Qi and blood tonifying fermentation composition, the Qi and blood tonifying fermentation composition comprising a fermentation substrate and optional auxiliary materials;

[0017] The Qi and blood-replenishing fermentation composition is prepared by fermenting a fermentation substrate with composite probiotics, and the components of the fermentation substrate include red ginseng and astragalus.

[0018] Furthermore, the components of the fermentation substrate also include at least one of Polygonatum odoratum, Longan pulp and jujube;

[0019] Preferably, the fermentation substrate comprises the following components by weight: 16-20 parts of red ginseng, 16-20 parts of astragalus, 12-14 parts of polygonatum, 16-20 parts of longan pulp, and 20-24 parts of jujube;

[0020] Preferably, the auxiliary material includes at least one of wolfberry juice, pomegranate concentrated juice, donkey-hide gelatin peptide, inulin and sweetener;

[0021] Preferably, the sweetener comprises at least one of xylo-oligosaccharide, erythritol and natural sweetener;

[0022] Preferably, the auxiliary materials include the following components by weight: 100-110 parts of wolfberry juice, 60-70 parts of pomegranate concentrated juice, 5-8 parts of donkey-hide gelatin peptide, 30-40 parts of inulin, 10-15 parts of xylo-oligosaccharides, 30-40 parts of erythritol, and 0.1-0.3 parts of natural sweetener.

[0023] Furthermore, the Qi and blood-replenishing fermented composition contains ginsenosides, and the ginsenosides include ginsenoside Rk1, and at least one of ginsenoside Rh1, ginsenoside 20(R)-Rg2, ginsenoside 20(S)-Rg3, ginsenoside 20(R)-Rg3, ginsenoside Rg5 and ginsenoside Rh2.

[0024] In a fourth aspect, the present invention provides a method for preparing the Qi and blood-invigorating fermentation composition, comprising the following steps:

[0025] 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.

[0026] Further, the extraction includes a first extraction and a second extraction;

[0027] Preferably, the first extraction comprises: adding water to the mixed components of the fermentation substrate, performing a first boiling extraction and a first filtering in sequence to obtain a first extract;

[0028] Preferably, the amount of water added in the first extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the first boiling extraction is 1-2 h;

[0029] Preferably, the second extraction comprises: adding water, boiling extraction for the second time and filtering for the second time to the components of the fermentation substrate after the first extraction in sequence to obtain a second extract;

[0030] Preferably, the amount of water added in the second extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the second boiling extraction is 1-2 h;

[0031] Preferably, the extracting further comprises: mixing the first extract and the second extract to obtain a total extract.

[0032] Further, the weight of the concentrated liquid is 4-7 times the weight of the fermentation substrate raw material component;

[0033] Preferably, the concentration temperature is 75-85°C;

[0034] Preferably, after the concentration and before the fermentation, the concentrated liquid is sterilized for the first time;

[0035] Preferably, the first sterilization adopts pasteurization;

[0036] Preferably, the mixing of the composite probiotics with the concentrated solution comprises: mixing Lactobacillus plantarum KMHD-1 powder and Bifidobacterium longum KMBGI 01 powder to obtain composite probiotic powder, then dissolving the composite probiotic powder in water and inoculating it into the concentrated solution;

[0037] Preferably, the added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution;

[0038] Preferably, the fermentation time is 24-72 h.

[0039] Furthermore, the preparation method of the Qi-invigorating and blood-invigorating fermentation composition further comprises: mixing the auxiliary materials in the formula amount with the fermentation product to prepare the Qi-invigorating and blood-invigorating fermentation composition;

[0040] Preferably, the mixing process includes: dissolving donkey-hide gelatin peptide and natural sweetener separately, then mixing to obtain a premix, mixing inulin and erythritol, and then adding the fermentation product, wolfberry juice, pomegranate concentrate, xylo-oligosaccharide solution and the premix.

[0041] In a fifth aspect, the present invention provides an application of the Qi and blood tonifying fermentation composition in the preparation of Qi and blood tonifying products.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The composite probiotics provided by the present invention include Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01. Fermentation with the composite probiotics can effectively increase the types and contents of rare ginsenosides in the product, as well as the contents of astragaloside IV and polysaccharides, thereby enhancing the efficacy of the product. Moreover, compared with single-bacteria fermentation, the composite probiotics are used to ferment the fermentation substrate, and the types of rare ginsenosides in the obtained product are more than ginseng rare saponins Rk1, which can effectively enhance the product's effects of replenishing qi and blood, improving immunity, and the like. DETAILED DESCRIPTION

[0044] 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.

[0045] Generally, the nomenclature and its technology used in conjunction with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. Unless otherwise indicated, the methods and techniques of the present invention are generally carried out according to conventional methods as well-known in the art, and as described in various general and more specific references, which are cited and discussed throughout this specification. Enzymatic reactions and purification techniques are carried out according to the manufacturer's specifications, as generally achieved in the art, or as described herein. The nomenclature and its laboratory procedures and technology used in conjunction with analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein are those well-known and commonly used in the art.

[0046] 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.

[0047] The first aspect of the present invention provides a composite probiotic, comprising: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Bifidobacterium longum KMBGI 01 ( Bifidobacterium long KMBGI 01);

[0048] 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;

[0049] The Bifidobacterium longum KMBGI 01 was deposited in the General Microbiology Center of the China Microorganism Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17175, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0050] In the present invention, the composite probiotics are used in combination to enhance the fermentation efficacy thereof.

[0051] Furthermore, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Bifidobacterium longum KMBGI 01 is (0.5-1.5):1, for example, it can be 0.5:1, 1:1, 1.5:1, etc.

[0052] The second aspect of the present invention provides an application of the composite probiotics in preparing a Qi and blood-invigorating fermentation composition.

[0053] In the present invention, Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01 are used as composite probiotics for fermentation, which can improve the product efficacy of the qi and blood replenishing fermented composition in replenishing qi and blood, improving immunity, and enhancing energy.

[0054] The third aspect of the present invention provides a Qi and blood tonifying fermentation composition, the Qi and blood tonifying fermentation composition comprising a fermentation substrate and optional auxiliary materials;

[0055] The Qi and blood-replenishing fermentation composition is prepared by fermenting a fermentation substrate with the composite probiotics, and the components of the fermentation substrate include: red ginseng and astragalus.

[0056] In the present invention, the components of the fermentation substrate include red ginseng and astragalus, the main effective ingredients of red ginseng include rare ginsenosides and polysaccharides, etc., ginseng rare saponins have the effects of enhancing immune function, improving cardiovascular health, improving memory and cognitive function, and the polysaccharides in red ginseng help to enhance the body's immunity and promote the generation of white blood cells, thereby indirectly supporting the generation of qi and blood; the main effective ingredients of astragalus include astragaloside IV and polysaccharides, etc., astragaloside IV has strong antioxidant and anti-inflammatory effects, and the polysaccharides in astragalus can promote the activity of immune cells and enhance the body's resistance. The present invention adopts plant lactobacillus KMHD-1 and bifidobacterium longum KMBGI 01 as composite probiotic fermentation substrates, which can effectively improve the types and contents of rare ginsenosides in the product, as well as improve the content of astragaloside IV, thereby enhancing the product's effects of replenishing qi and blood and enhancing immunity.

[0057] In some preferred embodiments, the components of the fermentation substrate further include at least one of Polygonatum odoratum, Longan pulp and jujube.

[0058] In the present invention, the components of the fermentation substrate also include Polygonatum odoratum, which nourishes Yin fluid, balances Yin and Yang in the body, and indirectly contributes to the reconciliation of Qi and blood; the components of the fermentation substrate also include Longan meat, which is rich in vitamin C and other nutrients, can enhance spleen and stomach function, promote the digestion and absorption of food, and then is conducive to the generation of Qi and blood; the components of the fermentation substrate also include Chinese jujube, which is rich in various nutrients such as iron and vitamins, can promote blood production, increase the number of red blood cells, and thus directly improve the Qi and blood level of human body. At the same time, Chinese jujube also contributes to the recovery of spleen and stomach function, enhances digestion and absorption capacity, and makes Qi and blood sufficient. It can be seen that the Polygonatum odoratum, Longan meat and Chinese jujube in the fermentation substrate all have the effect of replenishing Qi and blood, can assist red ginseng and astragalus to further enhance the Qi and blood replenishing effect of the product, and by the fermentation of composite probiotics, the total polysaccharide content in the product is further increased.

[0059] Preferably, the fermentation substrate comprises the following components by weight: 16-20 parts of red ginseng, 16-20 parts of astragalus, 12-14 parts of polygonatum, 16-20 parts of longan pulp, and 20-24 parts of jujube;

[0060] Wherein, in the fermentation substrate, the amount of red ginseng added is 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, etc.;

[0061] Among them, in the fermentation substrate, the amount of astragalus added is 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, etc.;

[0062] Wherein, in the fermentation substrate, the amount of Polygonatum odoratum added is 12 parts, 13 parts, 14 parts, etc.;

[0063] Wherein, in the fermentation substrate, the amount of longan pulp added is 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, etc.;

[0064] Among them, the added amount of jujube in the fermentation substrate is 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, etc.

[0065] Preferably, the auxiliary material includes at least one of wolfberry juice, pomegranate concentrated juice, donkey-hide gelatin peptide, inulin and sweetener;

[0066] Preferably, the sweetener comprises at least one of xylo-oligosaccharide, erythritol and a natural sweetener.

[0067] In the present invention, wolfberry juice, pomegranate concentrated juice and donkey-hide gelatin peptides are all helpful to enhance qi and blood and enhance immunity, inulin and oligoxylose can optimize the intestinal environment, enhance the absorption efficiency of nutrients, and indirectly support the fullness of qi and blood, erythritol and sweeteners are mainly used for seasoning, and do not directly affect qi and blood, but can avoid physical discomfort caused by blood sugar fluctuations while controlling blood sugar stability, thereby indirectly maintaining qi and blood balance. Among them, natural sweeteners are preferably stevioside and mogroside. Different sweeteners can be mixed to improve the overall sweetness, and can fully simulate the sweetness of sucrose as much as possible, including the maintenance time and aftertaste of sweetness, etc., and the use of multiple sweeteners can improve the stability of beverages under different conditions.

[0068] Preferably, the auxiliary materials include the following components by weight: 100-110 parts of wolfberry juice, 60-70 parts of pomegranate concentrated juice, 5-8 parts of donkey-hide gelatin peptide, 30-40 parts of inulin, 10-15 parts of xylo-oligosaccharides, 30-40 parts of erythritol, and 0.1-0.3 parts of natural sweetener.

[0069] Among them, the amount of wolfberry juice added in the auxiliary materials is 100 parts, 101 parts, 102 parts, 103 parts, 104 parts, 105 parts, 106 parts, 107 parts, 108 parts, 109 parts, 110 parts, etc.;

[0070] Among them, the amount of pomegranate concentrated juice added in the auxiliary materials is 60 parts, 61 parts, 62 parts, 63 parts, 64 parts, 65 parts, 66 parts, 67 parts, 68 parts, 69 parts, 70 parts, etc.;

[0071] Among them, the amount of donkey-hide gelatin peptide added in the auxiliary materials is 5 parts, 6 parts, 7 parts, 8 parts, etc.;

[0072] Among them, the amount of inulin added in the auxiliary materials is 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, 40 parts, etc.;

[0073] Among them, the amount of xylo-oligosaccharide added in the auxiliary materials is 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, etc.;

[0074] Among them, in the auxiliary materials, the added amount of erythritol is 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, 40 parts, etc.;

[0075] Among them, the amount of natural sweetener added in the auxiliary materials is 0.1 part, 0.2 part, 0.3 part, etc.

[0076] In some preferred embodiments, the Qi and blood-replenishing fermented composition contains ginsenosides, and the ginsenosides include ginsenoside Rk1, and at least one of ginsenoside Rh1, ginsenoside 20(R)-Rg2, ginsenoside 20(S)-Rg3, ginsenoside 20(R)-Rg3, ginsenoside Rg5 and ginsenoside Rh2.

[0077] In the present invention, the product of qi and blood invigorating fermentation composition has a variety of ginseng rare saponins, which can effectively enhance the effects of qi and blood invigorating, improving immunity, etc. of the product. Moreover, compared with single bacteria fermentation, the composite probiotic fermentation substrate is used, and the ginseng rare saponin types in the obtained product are more than ginseng rare saponin Rk1, and ginseng rare saponin Rk1 has a strong cardiovascular protective effect, for example, it can reduce myocardial ischemia reperfusion injury, improve blood circulation, ginseng rare saponin Rk1 can provide effective support for qi and blood circulation, ginseng rare saponin Rk1 can also regulate the immune system, enhance the body's resistance, and ginseng rare saponin Rk1 can also help reduce cell damage caused by oxidative stress through its antioxidant effect, and then through qi invigoration, achieve physical strength enhancement and fatigue relief. Therefore, compared with single bacteria fermentation, the product obtained by the composite probiotic fermentation has more ginseng rare saponin Rk1, which effectively improves the efficacy of the product.

[0078] The fourth aspect of the present invention provides a method for preparing the Qi and blood invigorating fermentation composition, characterized in that it comprises the following steps:

[0079] 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.

[0080] In the present invention, the preparation method can effectively increase the content of active ingredients in the product, and the preparation method has a simple process, high efficiency and feasibility.

[0081] In some preferred embodiments, the extraction includes a first extraction and a second extraction.

[0082] In the present invention, two extractions are adopted to ensure thorough extraction, thereby improving product quality.

[0083] Preferably, the first extraction comprises: adding water to the mixed components of the fermentation substrate, performing a first boiling extraction and a first filtering in sequence to obtain a first extract.

[0084] In the present invention, a double-layer filtering method of silk cloth is adopted for the first filtering.

[0085] Preferably, the amount of water added in the first extraction is 10-20 times the weight of the fermentation substrate raw material component, for example, it can be 10 times, 15 times, 20 times, etc., and the time of the first boiling extraction is 1-2 h, for example, it can be 1 h, 1.5 h, 2 h, etc.;

[0086] Preferably, the second extraction comprises: adding water to the components of the fermentation substrate after the first extraction, performing a second boiling extraction and a second filtration in sequence to obtain a second extract.

[0087] In the present invention, a silk cloth double-layer filtering method is adopted to carry out the second filtering.

[0088] Preferably, the amount of water added for the second extraction is 10-20 times the weight of the fermentation substrate raw material component, for example, it can be 10 times, 15 times, 20 times, etc., and the time for the second boiling extraction is 1-2 h, for example, it can be 1 h, 1.5 h, 2 h, etc.;

[0089] Preferably, the extracting further comprises: mixing the first extract and the second extract to obtain a total extract.

[0090] In some preferred embodiments, 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.;

[0091] Preferably, the concentration temperature is 75-85°C, for example, 75°C, 80°C, 85°C, etc.

[0092] 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.

[0093] Preferably, after the concentration and before the fermentation, the concentrated liquid is sterilized for the first time;

[0094] Preferably, the first sterilization is performed by pasteurization.

[0095] 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.

[0096] Preferably, the mixing of the composite probiotics with the concentrated solution comprises: mixing Lactobacillus plantarum KMHD-1 powder and Bifidobacterium longum KMBGI 01 powder to obtain composite probiotic powder, then dissolving the composite probiotic powder in water and inoculating it into the concentrated solution;

[0097] Preferably, the added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution;

[0098] Preferably, the fermentation time is 24-72 h, for example, it can be 24 h, 30 h, 35 h, 40 h, 45 h, 50 h, 55 h, 60 h, 65 h, 72 h, etc.

[0099] In the present invention, the fermentation substrate is fermented with composite probiotics at 37°C for 24-72 hours under stirring, then pasteurized, refrigerated and left to stand overnight (the temperature is maintained at 2-4°C) to remove precipitation, and then used to obtain a fermentation liquid.

[0100] In some preferred embodiments, the method for preparing the Qi-invigorating and blood-invigorating fermentation composition further comprises: mixing the auxiliary materials in the formula amount with the fermentation product to prepare the Qi-invigorating and blood-invigorating fermentation composition;

[0101] Preferably, the mixing process comprises:

[0102] Dissolving donkey-hide gelatin peptide and natural sweetener separately, and then mixing to obtain a premixed solution;

[0103] After inulin and erythritol are mixed, the fermented product, wolfberry juice, pomegranate concentrate, xylo-oligosaccharide solution and the premixed solution are added.

[0104] In the present invention, appropriate amount of purified water is used to fully dissolve donkey-hide gelatin peptide and natural sweetener respectively to obtain premixed liquid, then appropriate amount of purified water is added to the predissolving tank, stirring and steam heating are turned on, heated to 60-70°C, inulin and erythritol are added, stirred and dissolved, fermentation concentrate, wolfberry juice, pomegranate concentrate, oligoxylose liquid and premixed liquid are added, stirred and mixed evenly, then purified water is used to weigh to 1 ton, heat preservation and stirring are carried out, and the prepared liquid is filled after hot filtration through a plate frame, and the filled semi-finished product is loaded into a sterilization tray and loaded into a sterilizing cabinet for wet heat sterilization, and the sterilization conditions are: temperature is 121°C and time is 20 min.

[0105] The fifth aspect of the present invention provides a use of the Qi and blood tonifying fermentation composition in the preparation of Qi and blood tonifying products.

[0106] In the present invention, composite probiotics are used to ferment red ginseng, astragalus, longan pulp and other Chinese medicinal and edible herbs, which can be used to prepare a Chinese medicinal and edible herbal fermentation liquid that can replenish qi and blood, improve immunity and enhance energy, as well as to prepare qi and blood replenishing products, such as qi and blood replenishing beverages and other products.

[0107] 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.

[0108] Example 1

[0109] This embodiment provides a Qi and blood tonifying fermentation composition, which is prepared by fermenting a fermentation substrate with a composite probiotic (Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01), and the components of the Qi and blood tonifying fermentation composition include a fermentation substrate and auxiliary materials;

[0110] The weight ratio of the plant lactobacillus KMHD-1 to the long bifidobacterium KMBGI 01 is 1:1; the fermentation substrate comprises the following components by weight: 18 parts of red ginseng, 18 parts of astragalus, 13 parts of polygonatum, 18 parts of longan pulp, and 22 parts of jujube;

[0111] Among them, the auxiliary materials include the following components by weight: 105 parts of wolfberry juice, 65 parts of pomegranate concentrated juice, 7 parts of donkey-hide gelatin peptide, 35 parts of inulin, 13 parts of xylo-oligosaccharides, 35 parts of erythritol, and 0.2 parts of natural sweetener (wherein the natural sweetener includes stevioside and mogroside in a weight ratio of 1:1).

[0112] Example 2

[0113] This embodiment provides a Qi and blood tonifying fermentation composition, which is prepared by fermenting a fermentation substrate with a composite probiotic (Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01), and the components of the Qi and blood tonifying fermentation composition include a fermentation substrate and auxiliary materials;

[0114] Wherein, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Bifidobacterium longum KMBGI 01 is 0.5:1;

[0115] The fermentation substrate includes the following components by weight: 16 parts of red ginseng, 20 parts of astragalus, 12 parts of polygonatum, 20 parts of longan pulp, and 20 parts of jujube;

[0116] Among them, the auxiliary materials include the following components by weight: 100 parts of wolfberry juice, 70 parts of pomegranate concentrated juice, 5 parts of donkey-hide gelatin peptide, 40 parts of inulin, 10 parts of xylo-oligosaccharides, 40 parts of erythritol, and 0.1 parts of sweetener (wherein the natural sweetener includes stevioside and mogroside in a weight ratio of 1:1).

[0117] Example 3

[0118] This embodiment provides a Qi and blood tonifying fermentation composition, which is prepared by fermenting a fermentation substrate with a composite probiotic (Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01), and the components of the Qi and blood tonifying fermentation composition include a fermentation substrate and auxiliary materials;

[0119] Wherein, the weight ratio of the Lactobacillus plantarum KMHD-1 to the Bifidobacterium longum KMBGI 01 is 1.5:1;

[0120] The fermentation substrate comprises the following components by weight: 20 parts of red ginseng, 16 parts of astragalus, 14 parts of polygonatum, 16 parts of longan pulp, and 24 parts of jujube;

[0121] Among them, the auxiliary materials include the following components by weight: 110 parts of wolfberry juice, 60 parts of pomegranate concentrated juice, 8 parts of donkey-hide gelatin peptide, 30 parts of inulin, 15 parts of xylo-oligosaccharides, 30 parts of erythritol, and 0.3 parts of sweetener (wherein the natural sweetener includes stevioside and mogroside in a weight ratio of 1:1).

[0122] Embodiment 4-6

[0123] Example 4-6 provides a method for preparing a Qi-replenishing and blood-replenishing fermentation composition, comprising the following steps:

[0124] (1) Extraction: The fermentation substrate components of Examples 1-3 were respectively put into a multifunctional extraction tank according to the formula amount of the fermentation substrate components. For the first extraction, 15 times the amount of purified water was added, and the mixture was boiled for 1.5 h, and then filtered through a double layer of silk cloth. For the second extraction, 15 times the amount of purified water was added, and the mixture was boiled for 1.5 h, and then filtered through a double layer of silk cloth. The two filtrates were combined to obtain the total extract.

[0125] (2) Concentration: The total extract is pumped into a double-effect external circulation evaporator and heated for concentration. The vacuum degree is set to -0.08-0.03Mpa and the temperature is 80°C. Concentrate to 6 times the weight of the raw material. The concentrate is the fermentation substrate.

[0126] (3) Examples 4-6 were fermented according to the formula amount of the powder ratio in Examples 1-3: the concentrated solution was poured into the fermentation tank without adding additional carbon and nitrogen sources. Then pasteurized and cooled to about 37°C. The plant lactobacillus KMHD-1 powder and the long bifidobacterium KMBGI 01 powder were mixed to obtain a composite probiotic powder, the addition amount of the composite probiotic powder was 0.1% of the weight of the concentrated solution, and then the composite probiotic powder was fully dissolved with sterile purified water at room temperature (wherein the amount of sterile purified water used was 7-10 times the weight of the composite probiotic powder, which can achieve the purpose of fully dissolving the composite probiotic powder), inoculated into the sterilized concentrated solution, added to the concentrated solution, and stirred evenly. After stirring and fermenting at 37°C for 48 hours, pasteurized, refrigerated and left to stand overnight (2-4°C) to remove the precipitate, and set aside.

[0127] (4) Example 4-6: According to the formula amount of the auxiliary material components in Example 1-3, use an appropriate amount of purified water to fully dissolve the donkey-hide gelatin peptide and natural sweetener to obtain a pre-solution; add an appropriate amount of purified water to the pre-dissolution tank, turn on the stirring and steam heating, heat to 60-70°C, add inulin and erythritol, stir and dissolve, add fermentation concentrate, wolfberry juice, pomegranate concentrate, oligoxylose liquid 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 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.

[0128] Example 7

[0129] This embodiment provides a method for preparing a Qi and blood-replenishing fermentation composition, comprising the following steps:

[0130] (1) Extraction: according to the formula amount of the fermentation substrate components in Example 1, the formula amount of the fermentation substrate components was put into a multifunctional extraction tank, and 10 times the amount of purified water was added for the first extraction, and the mixture was boiled for 2 h, and then filtered through a double layer of silk cloth; for the second extraction, 20 times the amount of purified water was added, and the mixture was boiled for 1 h, and then filtered through a double layer of silk cloth, and the two filtrates were combined to obtain the total extract.

[0131] (2) Concentration: The total extract is pumped into a double-effect external circulation evaporator and heated for concentration. The vacuum degree is set to -0.08-0.03Mpa and the temperature is 75°C. Concentrate to 7 times the weight of the raw material. The concentrate is the fermentation substrate.

[0132] (3) Fermentation: Pour the concentrated solution into a fermentation tank without adding additional carbon and nitrogen sources. Then pasteurize and cool to about 37°C. Mix the KMHD-1 powder of Lactobacillus plantarum and the KMBGI 01 powder of Bifidobacterium longum to obtain a composite probiotic powder. The amount of the composite probiotic powder added is 0.07% of the weight of the concentrated solution. Then, fully dissolve the composite probiotic powder with sterile purified water at room temperature and inoculate it into the sterilized concentrated solution (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 (2-4°C) to remove the precipitate, and set aside.

[0133] (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.

[0134] Example 8

[0135] (1) Extraction: according to the formula amount of the fermentation substrate components in Example 1, the formula amount of the fermentation substrate components was put into a multifunctional extraction tank, and 10 times the amount of purified water was added for the first extraction, and the mixture was boiled for 2 h, and then filtered through a double layer of silk cloth; for the second extraction, 20 times the amount of purified water was added, and the mixture was boiled for 1 h, and then filtered through a double layer of silk cloth, and the two filtrates were combined to obtain the total extract.

[0136] (2) Concentration: The total extract is pumped into a double-effect external circulation evaporator and heated for concentration. The vacuum degree is set to -0.08-0.03Mpa and the temperature is 85°C. Concentrate to 4 times the weight of the raw material. The concentrate is the fermentation substrate.

[0137] (3) Fermentation: Pour the concentrated solution into a fermentation tank without adding additional carbon and nitrogen sources. Then pasteurize and cool to about 37°C. Mix the KMHD-1 powder of Lactobacillus plantarum and the KMBGI 01 powder of Bifidobacterium longum to obtain a composite probiotic powder. The amount of the composite probiotic powder added is 0.12% of the weight of the concentrated solution. Then, fully dissolve the composite probiotic powder with sterile purified water at room temperature and inoculate it into the sterilized concentrated solution (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 (2-4°C) to remove the precipitate, and set aside.

[0138] (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.

[0139] Comparative Example 1

[0140] This comparative example provides a Qi and blood tonifying fermentation composition, which is different from Example 1 in that the Qi and blood tonifying fermentation composition is prepared by fermenting a fermentation substrate with Lactobacillus plantarum KMHD-1, and the rest is consistent with Example 1.

[0141] Comparative Example 2

[0142] This comparative example provides a Qi and blood invigorating fermentation composition, which is different from Example 1 in that the Qi and blood invigorating fermentation composition is prepared by fermenting a fermentation substrate with Bifidobacterium longum KMBGI 01, and the rest is consistent with Example 1.

[0143] Comparative Example 3

[0144] This comparative example provides a Qi and blood tonifying fermentation composition, which is different from Example 1 in that the Qi and blood tonifying fermentation composition is prepared by fermenting a fermentation substrate with a composite probiotic (commercial bacteria Lactobacillus plantarum and commercial bacteria Bifidobacterium longum), and the rest is consistent with Example 1.

[0145] Comparative Example 4

[0146] This comparative example provides a Qi and blood tonifying fermentation composition, which is different from Example 1 in that the Qi and blood tonifying fermentation composition is prepared by fermenting a fermentation substrate with a composite probiotic (Lactobacillus plantarum KMHD-1 and commercial bacteria Bifidobacterium longum), and the rest is consistent with Example 1.

[0147] Comparative Example 5

[0148] This comparative example provides a Qi and blood tonifying fermentation composition, which is different from Example 1 in that the Qi and blood tonifying fermentation composition is prepared by fermenting a fermentation substrate with a composite probiotic (commercial bacteria Lactobacillus plantarum and Bifidobacterium longum KMBGI 01), and the rest is consistent with Example 1.

[0149] Comparative Examples 6-10

[0150] Comparative Examples 6-10 provide a method for preparing a Qi and blood tonifying fermentation composition, and the Qi and blood tonifying fermentation compositions of Comparative Examples 1-5 are prepared respectively by the preparation method of Example 4.

[0151] Test Example 1

[0152] Active ingredient detection:

[0153] (1) Detection method:

[0154] Rare ginsenosides of Panax 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; the column temperature was 30°C; the injection volume was 3 μL; and the autosampler temperature was 4°C.

[0155] Astragaloside IV was analyzed by high performance liquid chromatography:

[0156] ACQUITY UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm), mobile phase acetonitrile-water (33:77), flow rate 1.0 mL min －1 , column temperature 30℃.

[0157] 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.

[0158] (2) Test samples: The Qi-blood-replenishing fermented compositions prepared in Examples 4-8 and Comparative Examples 6-10 were tested using the same method. The results are shown in Table 1 below.

[0159] Table 1

[0160]

[0161] It can be seen from the data in the table that according to the data of the comparative examples, compared with comparative examples 6 and 7, comparative examples 9 and 10 use self-screened bacteria and commercial bacteria for mixed fermentation, and the contents of ginseng rare saponins, astragaloside IV, and polysaccharides are all improved to varying degrees, which shows that the mixed bacteria fermentation effect is better than single bacteria fermentation, and when the two self-screened bacteria in Example 4 are compounded, the contents of ginseng rare saponins, astragaloside IV, and polysaccharides are further significantly improved, and the types of rare saponins are more than all comparative examples. Rk1 may be that the specific metabolites produced by the two strains interact with each other, affecting the transformation process of ginseng rare saponins, thereby increasing the types of ginseng rare saponins. Ginseng rare saponins, astragaloside IV, and polysaccharides are the main effective ingredients of the product, which are closely related to the effectiveness of the product, so the product of Example 4 has the best effect of replenishing qi and blood.

[0162] Test Example 2

[0163] Population test evaluation:

[0164] According to the Guiding Principles for Clinical Research of New Chinese Medicines, patients who meet two main symptoms and two secondary symptoms at the stage of recruiting research subjects were diagnosed with Qi and blood deficiency and included in the study. The main symptoms were pale or sallow complexion, dizziness, palpitations, shortness of breath, fatigue, and secondary symptoms were spontaneous sweating, laziness, insomnia, and numbness of hands and feet.

[0165] The subjects were divided into 4 groups, each with 30 people. After completing the TCM symptom scoring form, the subjects were treated with the product of the present invention orally. The main symptoms and secondary symptoms of the 4 groups of subjects before and after treatment were scored to obtain symptom scores, and then the clinical efficacy of the subjects was analyzed and judged based on the symptom scores. Each symptom was scored as 0, 1, 2, and 3 points according to the levels of none, mild, moderate, and severe; the scores of each item were added together to obtain the symptom score; according to the "Guidelines for Clinical Research of New Chinese Medicines", the clinical efficacy was divided into the following four levels:

[0166] Recovery: Symptom score reduction is greater than or equal to 95%, and TCM clinical symptoms and signs are completely relieved or disappear;

[0167] Markedly effective: The symptom score is reduced by 70% or more, and the clinical symptoms and signs of traditional Chinese medicine are significantly improved;

[0168] Effective: The symptom score is reduced by 30% or more, and both clinical symptoms and signs of TCM show signs of improvement;

[0169] 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.

[0170] The number of subjects who were cured, significantly effective, effective and ineffective was obtained, and the total effective number and total effective rate were calculated and recorded in the following Table 2; among which, the total effective number = the number of cured cases + the number of significantly effective cases + the number of effective cases, and the total effective rate = (the number of cured cases + the number of significantly effective cases + the number of effective cases) / the total number of cases × 100%.

[0171] Test samples: The same method was used to test the Qi and blood-replenishing fermented compositions prepared in Example 4 and Comparative Examples 6-10. The results are shown in Table 2 below.

[0172] The test results are shown in Table 2 (Clinical efficacy analysis table).

[0173] Table 2

[0174]

[0175] 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 product of Example 4 was 93.3% effective for improving the deficiency of qi and blood in volunteers, 70.0% effective for improving the deficiency of qi and blood in the subjects of Comparative Example 6, 63.3% effective for improving the deficiency of qi and blood in the subjects of Comparative Example 7, 76.7% effective for improving the deficiency of qi and blood in the subjects of Comparative Example 8, and 80.0% effective for improving the deficiency of qi and blood in the subjects of Comparative Example 9; the effective rate of improving the deficiency of qi and blood in the subjects of Comparative Example 10 was 73.3%, which can prove that the product prepared by Example 4 of the present invention has the most significant effect on the subjects of deficiency of qi and blood.

[0176] 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: Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1) and Bifidobacterium longum KMBGI 01 ( Bifidobacterium longum KMBGI 01) composition; 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 Bifidobacterium longum KMBGI 01 was deposited in the General Microbiological Center of China Microorganism Culture Collection Administration on January 14, 2019, with the deposit number CGMCC No.17175, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; The weight ratio of the Lactobacillus plantarum KMHD-1 to the Bifidobacterium longum KMBGI 01 is (0.5-1.5):

1.

2. A method for preparing a Qi and blood-replenishing fermentation composition, characterized in that: The Qi and blood tonifying fermentation composition comprises a fermentation substrate and auxiliary materials; the Qi and blood tonifying fermentation composition is prepared by fermenting the fermentation substrate with the composite probiotics according to claim 1, the components of the fermentation substrate include: red ginseng, astragalus, polygonatum, longan pulp and jujube; the auxiliary materials include wolfberry juice, pomegranate concentrated juice, donkey-hide gelatin peptide, inulin and sweetener; the sweetener includes xylo-oligosaccharide, erythritol and natural sweetener; The fermentation substrate comprises the following components by weight: 16-20 parts of red ginseng, 16-20 parts of astragalus, 12-14 parts of polygonatum, 16-20 parts of longan pulp and 20-24 parts of jujube; The auxiliary materials include the following components by weight: 100-110 parts of wolfberry juice, 60-70 parts of pomegranate concentrated juice, 5-8 parts of donkey-hide gelatin peptide, 30-40 parts of inulin, 10-15 parts of xylo-oligosaccharide, 30-40 parts of erythritol and 0.1-0.3 parts of natural sweetener; The preparation method of the Qi-replenishing and blood-invigorating fermentation composition comprises the following steps: The raw material components of the fermentation substrate in the formula amount are sequentially mixed, extracted and concentrated to obtain a concentrated solution, and then the composite probiotics are mixed with the concentrated solution and fermented to obtain a fermentation product; Wherein, the extraction includes a first extraction and a second extraction; The first extraction comprises: adding water to the mixed fermentation substrate components, performing a first boiling extraction and a first filtering in sequence to obtain a first extract; The amount of water added in the first extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the first boiling extraction is 1-2 h; The second extraction comprises: adding water, boiling extraction for the second time and filtering for the second time to the components of the fermentation substrate after the first extraction in sequence to obtain a second extract; The amount of water added in the second extraction is 10-20 times the weight of the fermentation substrate raw material component, and the time of the second boiling extraction is 1-2 h; The extracting further comprises: mixing the first extract and the second extract to obtain a total extract; The weight of the concentrated liquid is 4-7 times the weight of the fermentation substrate raw material component; The concentration temperature is 75-85°C; After the concentration, and before the fermentation, the concentrated liquid is sterilized for the first time; The first sterilization adopts pasteurization method; The added amount of the composite probiotic powder is 0.07-0.12% by weight of the concentrated solution; The fermentation time is 24-72 hours; The preparation method of the Qi-invigorating and blood-invigorating fermentation composition further comprises: mixing the auxiliary materials in the formula amount with the fermentation product to prepare the Qi-invigorating and blood-invigorating fermentation composition; The mixing process includes: dissolving donkey-hide gelatin peptide and natural sweetener respectively, then mixing to obtain a premixed solution, mixing inulin and erythritol, and then adding the fermented product, wolfberry juice, pomegranate concentrate, xylo-oligosaccharide solution and the premixed solution.

3. The method for preparing the Qi-invigorating and blood-invigorating fermentation composition according to claim 2, characterized in that: The Qi and blood-replenishing fermentation composition contains ginsenosides, which include ginsenoside Rk1 and at least one of ginsenoside Rh1, ginsenoside 20(R)-Rg2, ginsenoside 20(S)-Rg3, ginsenoside 20(R)-Rg3, ginsenoside Rg5 and ginsenoside Rh2.

4. The Qi-invigorating and blood-invigorating fermented composition prepared by the preparation method of the Qi-invigorating and blood-invigorating fermented composition according to claim 2 or 3.

5. Use of the Qi-blood-invigorating fermentation composition according to claim 4 in the preparation of Qi-blood-invigorating products.

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