Ganoderma lucidum peptide composition as well as preparation method and application thereof

Abstract

The invention relates to the technical field of ganoderma lucidum peptides, in particular to a ganoderma lucidum peptide composition as well as a preparation method and application thereof. The composition comprises the following components in parts by mass: 30-50 parts of ganoderma lucidum peptide, 10-20 parts of echinacea polysaccharide, 8-18 parts of polygonatum polysaccharide, 5-25 parts of dendrobium polysaccharide, 9-16 parts of astragalus polysaccharide, 12-24 parts of acanthopanax polysaccharide and 20-50 parts of probiotic viable bacteria powder, wherein the viable bacteria count in the probiotic viable bacteria powder is 4 * 10 < 10 > CFU / g-6 * 10 < 20 > CFU / g. The ganoderma lucidum peptide composition provided by the invention has an excellent immune regulation function, and can be used for improving the immunity of the organism, enhancing the physical quality and resisting pathogen invasion.

Description

Technical Field

[0001] The present invention belongs to the technical field of Ganoderma lucidum peptides, and particularly relates to a Ganoderma lucidum peptide composition, a preparation method thereof, and an application thereof. Background Art

[0002] Ganoderma lucidum peptides are a class of bioactive peptides extracted, separated from Ganoderma lucidum, or obtained by means of biotechnology enzymatic hydrolysis, etc. They are formed by connecting multiple amino acids with peptide bonds and have a relatively small molecular mass. This characteristic enables them to be more efficiently absorbed by the human gastrointestinal tract, avoiding problems such as difficult absorption and incomplete decomposition during digestion like some macromolecular proteins, and can quickly enter the blood circulation to exert their effects. Ganoderma lucidum peptides contain a variety of essential amino acids for the human body, and the reasonable combination of these amino acids is the material basis for their numerous physiological activities. For example, branched-chain amino acids such as leucine and isoleucine among them are of great significance for muscle repair and energy metabolism regulation. At present, as an immune regulator, Ganoderma lucidum peptides can activate immune cells such as macrophages, NK cells, T cells, and B cells in the human immune system, prompting them to release cytokines, enhancing the body's resistance to foreign pathogens, and helping to prevent various infectious diseases such as colds and influenza. Therefore, they are often made into health products for daily use by people with low immunity. However, the effect of taking Ganoderma lucidum peptides alone is limited, and the function of improving immunity is limited.

[0003] For example, CN110522901A discloses a composition of lamb bone marrow peptides and Ganoderma lucidum peptides. Its raw materials include, by weight parts: 10 - 20 parts of lamb bone marrow peptides, 10 - 20 parts of Ganoderma lucidum peptide polysaccharides, 0.5 - 1.6 parts of SGF hydrolase, 5 - 10 parts of animal protein extract, 5 - 10 parts of natural vegetable extract, 5 - 10 parts of seaweed extract, 3 - 5 parts of milk, 5 - 10 parts of wheat extract, 1 - 3 parts of thickener, 1 - 3 parts of sweetener, and the balance of purified water. This application enhances the hematopoietic function of the human bone marrow and improves immunity through the synergistic effect of lamb bone marrow peptides and Ganoderma lucidum peptides. However, when two kinds of peptide components are superimposed, the body's metabolic burden will increase during gastrointestinal digestion and absorption. Especially for the elderly and patients with gastrointestinal diseases whose digestive function is inherently poor, such as slow gastrointestinal motility and insufficient secretion of digestive enzymes, a large amount of peptide substances rush into the digestive system at once, making it more difficult to be fully decomposed and absorbed, and easily causing symptoms such as abdominal distension, belching, and indigestion.

[0004] As disclosed in CN104522461A, a ganoderma lucidum peptide polygonatum polysaccharide and its preparation process are provided. It consists of ganoderma lucidum peptide polysaccharide, polygonatum sibiricum peptide polysaccharide, and urtica polysaccharide, and their weight percentages are: ganoderma lucidum peptide polysaccharide 20 - 60%, polygonatum polysaccharide 10 - 50%, and urtica polysaccharide 10 - 50%. It is prepared by cultivation, filtration, purification, drying, pulverization, compounding, and filling. The health food provided by this application has obvious detoxifying, anti-inflammatory, and analgesic effects. In addition, it also has the effect of expelling wind and dredging collaterals and regulating the nerve center. However, the composition of this application does not have the effect of improving immunity.

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

[0006] The object of the present invention is to provide a ganoderma lucidum peptide composition, its preparation method, and application. The ganoderma lucidum peptide composition provided by the present invention has excellent immune regulation function, can improve the body's immunity, enhance physical fitness, and resist the invasion of pathogens.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] In the first aspect, an embodiment of the present invention provides a ganoderma lucidum peptide composition, including the following components by mass parts: 30 - 50 parts of ganoderma lucidum peptide, 10 - 20 parts of echinacea polysaccharide, 8 - 18 parts of polygonatum polysaccharide, 5 - 25 parts of dendrobium polysaccharide, 9 - 16 parts of astragalus polysaccharide, 12 - 24 parts of acanthopanax senticosus polysaccharide, and 20 - 50 parts of probiotic live bacteria powder, and the number of live bacteria in the probiotic live bacteria powder is 4×10 10 CFU / g - 6×10 20 CFU / g.

[0009] In a preferred embodiment, the composition includes the following components by mass parts: 33 - 46 parts of ganoderma lucidum peptide, 12 - 18 parts of echinacea polysaccharide, 10 - 16 parts of polygonatum polysaccharide, 10 - 18 parts of dendrobium polysaccharide, 11 - 14 parts of astragalus polysaccharide, 15 - 22 parts of acanthopanax senticosus polysaccharide, and 25 - 40 parts of probiotic live bacteria powder.

[0010] The present invention can synergistically improve immunity from the aspects of immune cell activation, immune factor regulation, antioxidant and anti-stress, and intestinal flora optimization by compounding ganoderma lucidum peptide and polysaccharides of traditional Chinese medicine extracts.

[0011] At the level of immune cell activation, echinacea polysaccharide can activate macrophages and natural killer cells, enhancing their ability to phagocytose and kill pathogens; astragalus polysaccharide can promote the proliferation and differentiation of lymphocytes and enhance the activity of T cells and B cells. When they act together, different polysaccharides target different types of immune cells they are good at, synergistically stimulating the awakening and activation of more types of immune cells. Ganoderma lucidum peptide can also help regulate the signal transduction of immune cells, making the activation process smoother and more efficient, and comprehensively improving immunity. At the same time, eleutherococcus senticosus polysaccharide helps to strengthen the memory function of immune cells. When the same pathogen invades again, the immune response can be quickly and strongly initiated. In combination with dendrobium officinale polysaccharide, dendrobium officinale polysaccharide provides energy for the metabolism and proliferation of immune cells, stabilizes the intracellular environment, enabling immune memory cells to survive longer and function stronger. Ganoderma lucidum peptide then assists in correcting the recognition tags of immune memory cells, further consolidating the body's memory of pathogens.

[0012] At the level of immune factor regulation, polygonatum sibiricum polysaccharide induces the body to produce immune factors such as interferon and interleukin, and astragalus polysaccharide also stimulates the release of various immune regulatory factors. Multiple polysaccharides work together to significantly increase the concentration of immune factors in the body in a short period of time. Ganoderma lucidum peptide acts like a regulatory switch, precisely controlling the appropriate secretion amount and action duration of various immune factors, preventing over-immune responses, and constructing a balanced and active immune regulation network. At the same time, these polysaccharides and Ganoderma lucidum peptide jointly create a stable in vivo microenvironment, preventing immune factors from being inactivated due to temperature, pH changes or enzymatic hydrolysis, and ensuring that immune factors continuously exert pressure on pathogens for a long time.

[0013] At the level of antioxidant and anti-stress, echinacea polysaccharide, polygonatum sibiricum polysaccharide, etc. all have antioxidant properties. Working together, they can efficiently scavenge free radicals generated by body metabolism and pathogen invasion, reducing the damage of free radicals to immune cells and immune factors. Ganoderma lucidum peptide repairs cell membranes and organelles damaged by free radicals, restoring the vitality and function of immune cells. At the same time, in the face of external stressors such as external pressure and bacterial infection, eleutherococcus senticosus polysaccharide and astragalus polysaccharide regulate the secretion of body stress hormones, weakening the inhibition of stress on the immune system. Coupled with Ganoderma lucidum peptide to enhance the stress tolerance of immune cells, the immune system can still maintain a high combat effectiveness in an adverse environment.

[0014] At the level of intestinal flora optimization, polysaccharide components such as dendrobium officinale polysaccharide and polygonatum sibiricum polysaccharide are food for beneficial intestinal bacteria, which can promote the reproduction of probiotics such as bifidobacteria and lactobacilli. Eleutherococcus senticosus polysaccharide regulates the distribution of intestinal flora, constructing a healthy intestinal microecology. At the same time, probiotics ferment polysaccharides to produce metabolites such as short-chain fatty acids, some of which can be absorbed and indirectly stimulate the immune system. Ganoderma lucidum peptide assists in the repair of the intestinal barrier, preventing pathogens from invading through the intestine, and strengthening the immune defense starting from the intestine.

[0015] Furthermore, the present invention simultaneously adds probiotic live bacteria powder, which synergistically improves the body's immunity with various traditional Chinese medicine extracts and simultaneously solves the problem that traditional Chinese medicine extracts are difficult to be fully decomposed and absorbed.

[0016] In a preferred embodiment, the probiotic live bacteria powder is a mixture of Bifidobacterium live bacteria powder and Lactobacillus rhamnosus live bacteria powder, and the mass ratio of the Bifidobacterium live bacteria powder to the Lactobacillus rhamnosus live bacteria powder is 1:1.

[0017] In a preferred embodiment, the viable count of the Bifidobacterium live bacteria powder is 2×10 10 CFU / g - 3×10 20 CFU / g.

[0018] In a preferred embodiment, the viable count of the Lactobacillus rhamnosus live bacteria powder is 2×10 10 CFU / g - 3×10 20 CFU / g.

[0019] The present invention further defines the probiotics as Bifidobacterium and Lactobacillus rhamnosus, and the two synergistically improve the function of the composition in regulating immunity in terms of strengthening the intestinal barrier, activating immune cells, regulating immune factors, and enhancing metabolites.

[0020] In terms of strengthening the intestinal barrier, Bifidobacterium can promote the intestinal epithelial cells to secrete mucus, which coats the inner wall of the intestine with a protective layer to block the adhesion and invasion of harmful bacteria; Lactobacillus rhamnosus stimulates the proliferation of intestinal epithelial cells and accelerates the repair of damaged intestinal mucosa, making the intestinal barrier more solid and complete, reducing the risk of pathogens taking advantage of the weakness to cause diseases, and reducing the burden on the immune system. At the same time, the combined action of the two can regulate the expression of tight junction proteins between intestinal epithelial cells, making the cell connections more tight, preventing harmful substances and allergens in the intestine from penetrating the intestinal wall into the blood, reducing the frequency of abnormal activation of immune cells, and maintaining the stability of the intestinal immune system.

[0021] In terms of activating immune cells, both Bifidobacterium and Lactobacillus rhamnosus have unique cell wall components, such as peptidoglycan, teichoic acid, etc. These components are recognized as "danger signals" by intestinal immune cells, and then activate innate immune cells such as macrophages and dendritic cells, prompting them to start the phagocytosis and bactericidal mode; the two bacteria can also synergistically induce these immune cells to release immune regulatory factors to summon more immune forces. They interact with gut-associated lymphoid tissue, stimulate the differentiation and maturation of T lymphocytes and B lymphocytes, and help the body build a precise adaptive immune response, enabling the immune system to respond more efficiently to various specific pathogens. For example, when infected with a virus, it helps the body produce specific antibodies.

[0022] In terms of immune factor regulation, Bifidobacterium and Lactobacillus rhamnosus can promote intestinal epithelial cells and immune cells to secrete a variety of immune regulatory factors, including interleukins, interferons, etc. The combined action of the two bacteria can significantly increase the secretion of these immune factors in a short period of time, regulate the immune response in all directions, and enhance the body's anti-inflammatory and antiviral capabilities. At the same time, after the immune response ends, they will cooperate tacitly to inhibit overactive immune factors, avoid persistent inflammation, guide the immune system back to a stable state, and prevent autoimmune diseases caused by immune imbalance.

[0023] In terms of metabolite enhancement, Bifidobacterium and Lactobacillus rhamnosus ferment dietary fiber in the intestines to produce a large number of short-chain fatty acids, such as acetic acid, propionic acid, and butyric acid. These short-chain fatty acids not only provide energy for intestinal epithelial cells and maintain cell health, but can also be absorbed into the blood and act on immune cells throughout the body, further activating immune function and strengthening the body's ability to resist pathogens. At the same time, the two synergistically participate in the synthesis of B vitamins, vitamin K, etc. These vitamins are necessary nutrients for the metabolism and proliferation of immune cells. Adequate supply helps maintain the activity and number of immune cells, indirectly enhancing immunity.

[0024] Further, the compounded live bacteria powder of the present invention and the Chinese medicine extract provided by the present invention can synergistically improve the body's immunity. Specifically, bifidobacteria and Lactobacillus rhamnosus are good at colonizing the intestine. They promote intestinal epithelial cells to secrete mucus and strengthen the physical barrier of the intestine; at the same time, they regulate tight junction proteins between cells to prevent pathogen leakage. Echinacea polysaccharides and dendrobium polysaccharides can provide nutrition for intestinal epithelial cells and accelerate the repair of damaged cells. Astragalus polysaccharides also help maintain the metabolic stability of epithelial cells. A multi-pronged approach makes the intestinal barrier indestructible, blocks the invasion of most pathogens, and reduces the stress burden of the immune system. And the cell wall components of probiotics act as natural immune signals to activate macrophages, dendritic cells, etc. in intestinal-associated lymphoid tissues. Polygonatum sibiricum polysaccharides and Acanthopanax senticosus polysaccharides work synergistically to enhance the sensitivity of such immune cells to probiotic signals, prompting them to release immune regulatory factors faster, recruit lymphocytes, etc., and Ganoderma lucidum peptides regulate the efficiency of signal transmission between immune cells, allowing the intestinal immune response to unfold rapidly and orderly.

[0025] It is further explained that the combined stimulation of Bifidobacterium and Lactobacillus rhamnosus causes the phagocytic activity of innate immune cells such as macrophages to soar; Echinacea polysaccharides replenish energy for phagocytes, enhance their continuous combat capability, and enable them to clear pathogens more efficiently. Astragalus polysaccharides can also induce the proliferation of natural killer cells and broaden the scope of innate immunity. At the same time, probiotics interact with intestinal lymphoid tissue to promote the maturation of T and B lymphocytes. A variety of polysaccharides provide a material basis for the proliferation and differentiation of lymphocytes, helping them to develop into fully functional effector cells. Ganoderma lucidum peptides participate in regulating cell differentiation signals, allowing adaptive immunity to accurately respond to different pathogens and produce corresponding antibodies and cellular immune responses.

[0026] In addition, Bifidobacterium and Lactobacillus rhamnosus induce intestinal cells to secrete immune factors such as interleukins and interferons. Echinacea polysaccharides and Dendrobium polysaccharides follow closely, greatly increasing the secretion volume, increasing the concentration of immune factors in the body in a short period of time, and enhancing the body's anti-inflammatory and antiviral abilities. In this process, Astragalus polysaccharides can also stabilize the cell's endocrine environment to ensure continuous and smooth secretion. At the same time, Acanthopanax polysaccharides and Ganoderma lucidum peptides work together to accurately regulate the activity of immune factors at the peak and trough of the immune response, avoiding excessive secretion in the early stage and causing excessive inflammation, and preventing the inactivation of immune factors in the later stage from being too fast, making the immune defense long-term stable and efficient.

[0027] Echinacea polysaccharides and Polygonatum sibiricum polysaccharides have antioxidant properties and can eliminate free radicals produced by infection and metabolism in the body; the antioxidant enzymes synthesized by Bifidobacterium and Lactobacillus rhamnosus can also reduce the damage of free radicals to immune cells and immune factors, maintain the stability of immune function, and Ganoderma lucidum peptides repair cell membranes damaged by free radicals, thereby helping immune cells to reproduce.

[0028] In a second aspect, an embodiment of the present invention provides a method for preparing a Ganoderma lucidum peptide composition, comprising the following steps:

[0029] (1) Ganoderma lucidum is crushed and sieved, mixed with water, inactivated by enzymatic hydrolysis, centrifuged and dried to obtain Ganoderma lucidum peptide:

[0030] (2) crushing and sieving the Dendrobium officinale, mixing with water, soaking, heating and extracting, centrifuging to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve, standing, centrifuging and drying to obtain a Dendrobium polysaccharide;

[0031] (3) crushing and sieving polygonatum, mixing with water, soaking, heating and extracting, centrifuging to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve, standing, centrifuging and drying to obtain polygonatum polysaccharide;

[0032] (4) crushing and sieving the astragalus, mixing it with water, soaking it, heating it for extraction, and centrifuging it to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve it, standing it, centrifuging it and drying it to obtain astragalus polysaccharide;

[0033] (5) crushing and sieving the roots and rhizomes of Acanthopanax senticosus, mixing them with water, soaking them, heating them for extraction, and centrifuging them to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve it, standing it, centrifuging and drying it to obtain Acanthopanax senticosus polysaccharide;

[0034] (6) mixing Echinacea and water, adjusting the pH to 3-5, inactivating after enzymatic hydrolysis, and centrifuging and drying to obtain Echinacea polysaccharide;

[0035] (7) Activate and culture the freeze-dried powder of Bifidobacterium to a bacterial density of 2×10 10CFU / ml - 3×10 20 After reaching 3×10 CFU / ml, continue culturing for 2 - 3 generations, and lyophilize the obtained bacterial suspension to obtain viable Bifidobacterium powder:

[0036] (8) Activate the freeze-dried strain powder of Lactobacillus rhamnosus until the bacterial density reaches 2×10 10 CFU / ml - 3×10 20 CFU / ml, then continue culturing for 2 - 3 generations, and lyophilize the obtained bacterial suspension to obtain viable Lactobacillus rhamnosus powder;

[0037] (9) Mix the viable Lactobacillus rhamnosus powder, viable Bifidobacterium powder, ganoderma peptide, dendrobium polysaccharide, polygonatum polysaccharide, astragalus polysaccharide, acanthopanax senticosus polysaccharide, and echinacea polysaccharide to obtain a ganoderma peptide composition.

[0038] In a preferred embodiment, in step (1), the sieve used for sieving has a mesh size of 200 - 400 meshes.

[0039] In a preferred embodiment, in step (1), the mass ratio of Ganoderma sinense to water is 1:(50 - 100).

[0040] In a preferred embodiment, in step (1), the conditions for enzymatic hydrolysis are:

[0041] Perform enzymatic hydrolysis using a composite enzyme composed of hemicellulase and papain with a weight ratio of 1:1. The enzymatic hydrolysis time is 60 - 100 min, the enzymatic hydrolysis temperature is 40 - 50 °C, and the addition amount of the composite enzyme is 1 wt% - 3 wt% of the Ganoderma sinense powder.

[0042] In a preferred embodiment, in step (2), the sieve used for sieving has a mesh size of 200 - 400 meshes.

[0043] In a preferred embodiment, in step (2), the mass ratio of Dendrobium officinale to water is 1:(50 - 100).

[0044] In a preferred embodiment, in step (2), the soaking time is 2 - 4 h.

[0045] In a preferred embodiment, in step (2), the conditions for heating extraction are: heat to 60 - 100 °C and then keep warm for 1.5 - 3 h.

[0046] In a preferred embodiment, in step (2), the volume of ethanol added is 2 - 4 times the volume of water.

[0047] In a preferred embodiment, in step (3), the sieve used for sieving has a mesh size of 200 - 400 meshes.

[0048] In a preferred embodiment, in step (3), the mass ratio of polygonatum sibiricum to water is 1:(50 - 100).

[0049] In a preferred embodiment, in step (3), the soaking time is 2 - 4 h.

[0050] In a preferred embodiment, in step (3), the conditions for heating and extraction are: heating to 60 - 100 °C and then holding for 1.5 - 3 h.

[0051] In a preferred embodiment, in step (3), the volume of ethanol added is 2 - 4 times the volume of water.

[0052] In a preferred embodiment, in step (4), the sieve used for sieving is 200 - 400 mesh.

[0053] In a preferred embodiment, in step (4), the mass ratio of astragalus membranaceus to water is 1:(50 - 100).

[0054] In a preferred embodiment, in step (4), the soaking time is 2 - 4 h.

[0055] In a preferred embodiment, in step (4), the conditions for heating and extraction are: heating to 60 - 100 °C and then holding for 1.5 - 3 h.

[0056] In a preferred embodiment, in step (4), the volume of ethanol added is 2 - 4 times the volume of water.

[0057] In a preferred embodiment, in step (5), the sieve used for sieving is 200 - 400 mesh.

[0058] In a preferred embodiment, in step (5), the mass ratio of the roots and rhizomes of acanthopanax senticosus to water is 1:(50 - 100).

[0059] In a preferred embodiment, in step (5), the soaking time is 2 - 4 h.

[0060] In a preferred embodiment, in step (5), the conditions for heating and extraction are: heating to 60 - 100 °C and then holding for 1.5 - 3 h.

[0061] In a preferred embodiment, in step (5), the volume of ethanol added is 2 - 4 times the volume of water.

[0062] In a preferred embodiment, in step (6), the mass ratio of echinacea purpurea to water is 1:(50 - 100).

[0063] In a preferred embodiment, in step (6), the conditions for enzymatic hydrolysis are as follows:

[0064] Perform enzymatic hydrolysis using pectinase. The addition amount of pectinase is 1 wt% - 4 wt% of echinacea, the enzymatic hydrolysis temperature is 40 - 60 °C, and the enzymatic hydrolysis time is 2 - 4 h.

[0065] In a preferred embodiment, in steps (7) and (8), the culture medium used for cultivation is MRS medium. Taking 1000 mL of water as the relative amount, the raw materials in the MRS medium include: peptone 5 - 15 g; yeast extract 2 - 5 g; beef extract 8 - 15 g; glucose 15 - 20 g; sodium acetate 2 - 4 g; diammonium citrate 1 - 2 g; Tween 80 1 - 2 mL; magnesium sulfate 0.2 - 0.4 g; manganese sulfate 0.01 - 0.04 g; dipotassium hydrogen phosphate 2 - 4 g; agar 15 - 20 g; ganoderma peptide 3 - 5 g, echinacea polysaccharide 1 - 2 g, polygonatum polysaccharide 0.8 - 1.8 g, dendrobium polysaccharide 0.5 - 2.5 g, astragalus polysaccharide 0.9 - 1.6 g, acanthopanax senticosus polysaccharide 1.2 - 2.4 g.

[0066] In a third aspect, the present invention provides an application of the ganoderma peptide composition as described above in the preparation of a drug for enhancing immunity and / or an adjuvant drug for enhancing immunity.

[0067] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0068] 1. The present invention can synergistically enhance immunity from aspects such as immune cell activation, immune factor regulation, antioxidant and anti-stress, and intestinal flora optimization by compounding ganoderma peptide and traditional Chinese medicine extract polysaccharide.

[0069] 2. The present invention simultaneously adds probiotic live powder, which synergistically enhances the body's immunity with various traditional Chinese medicine extracts and simultaneously solves the problem that traditional Chinese medicine extracts are difficult to be fully decomposed and absorbed.

[0070] 3. The present invention further defines the probiotics as bifidobacterium and lactobacillus rhamnosus, and the two synergistically enhance the function of the composition in regulating immunity in terms of intestinal barrier reinforcement, immune cell activation, immune factor regulation, and metabolite synergistic effect.

[0071] 4. The compounded live bacteria powder of the present invention and the Chinese medicine extract provided by the present invention can synergistically improve the body's immunity. Specifically, bifidobacteria and Lactobacillus rhamnosus are good at colonizing the intestine. They promote intestinal epithelial cells to secrete mucus and strengthen the physical barrier of the intestine; at the same time, they regulate tight junction proteins between cells to prevent pathogen leakage. Echinacea polysaccharides and dendrobium polysaccharides can provide nutrition for intestinal epithelial cells and accelerate the repair of damaged cells. Astragalus polysaccharides also help maintain the metabolic stability of epithelial cells. A multi-pronged approach makes the intestinal barrier indestructible, blocks the invasion of most pathogens, and reduces the stress burden of the immune system. And the cell wall components of probiotics act as natural immune signals to activate macrophages, dendritic cells, etc. in intestinal-associated lymphoid tissues. Polygonatum sibiricum polysaccharides and Acanthopanax senticosus polysaccharides work synergistically to enhance the sensitivity of such immune cells to probiotic signals, prompting them to release immune regulatory factors faster, recruit lymphocytes, etc., and Ganoderma lucidum peptides regulate the efficiency of signal transmission between immune cells, allowing the intestinal immune response to unfold rapidly and orderly. DETAILED DESCRIPTION

[0072] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0073] Unless otherwise specified, the reagents and raw materials used in the present invention are commercially available.

[0074] Lactobacillus rhamnosus, MP108, was purchased from Jinqiao Biotechnology Co., Ltd.

[0075] Bifidobacterium, BB-12, was purchased from Chr. Hansen, Denmark.

[0076] The culture medium used for culture is MRS culture medium, with 1000 mL of water as the relative amount. The raw materials in the MRS culture medium include: 10 g of peptone; 5 g of yeast extract; 12 g of beef extract; 18 g of glucose; 3 g of sodium acetate; 1.5 g of diammonium citrate; 1.5 ml of Tween 80; 0.3 g of magnesium sulfate; 0.02 g of manganese sulfate; 3 g of dipotassium hydrogen phosphate; 18 g of agar; 5 g of Ganoderma lucidum peptide, 2 g of Echinacea polysaccharide, 1.8 g of Polygonatum sibiricum polysaccharide, 2.5 g of Dendrobium polysaccharide, 1.6 g of Astragalus polysaccharide, and 2.4 g of Acanthopanax senticosus polysaccharide.

[0077] Example 1

[0078] This embodiment provides a method for preparing a Ganoderma lucidum peptide composition, comprising the following steps:

[0079] (1) By mass fraction, 1 part of Ganoderma sinense is pulverized through a 200-mesh sieve and then mixed with 70 parts of water. It is enzymatically hydrolyzed using a composite enzyme composed of hemicellulase and papain with a weight ratio of 1:1. The enzymatic hydrolysis time is 80 min, the enzymatic hydrolysis temperature is 45 °C, and the addition amount of the composite enzyme is 1.5 wt% of the Ganoderma sinense powder. Then, it is inactivated, centrifuged, separated, and dried to obtain Ganoderma peptide:

[0080] (2) By mass fraction, 1 part of Dendrobium officinale is pulverized through a 200-mesh sieve and then mixed with 70 parts of water. It is soaked for 2.5 h, heated for extraction, centrifuged and filtered to obtain a liquid phase. The liquid phase is vacuum concentrated, dissolved in ethanol, allowed to stand, centrifuged, separated, and dried to obtain Dendrobium officinale polysaccharide

[0081] (3) By mass fraction, 1 part of Polygonatum sibiricum is pulverized through a 200-mesh sieve and then mixed with 70 parts of water. It is soaked for 2.5 h, heated to 80 °C and then kept warm for 3 h, centrifuged and filtered to obtain a liquid phase. The liquid phase is vacuum concentrated, dissolved in 150 parts of ethanol, allowed to stand, centrifuged, separated, and dried to obtain Polygonatum sibiricum polysaccharide;

[0082] (4) By mass fraction, 1 part of Astragalus membranaceus is pulverized through a 200-mesh sieve and then mixed with 70 parts of water. It is soaked for 2.5 h, heated to 80 °C and then kept warm for 3 h, centrifuged and filtered to obtain a liquid phase. The liquid phase is vacuum concentrated, dissolved in 150 parts of ethanol, allowed to stand, centrifuged, separated, and dried to obtain Astragalus membranaceus polysaccharide;

[0083] (5) By mass fraction, 1 part of the roots and rhizomes of Acanthopanax senticosus is pulverized through a 200-mesh sieve and then mixed with 70 parts of water. It is soaked for 2.5 h, heated to 80 °C and then kept warm for 3 h, centrifuged and filtered to obtain a liquid phase. The liquid phase is vacuum concentrated, dissolved in 150 parts of ethanol, allowed to stand, centrifuged, separated, and dried to obtain Acanthopanax senticosus polysaccharide;

[0084] (6) By mass fraction, 1 part of Echinacea purpurea is mixed with 70 parts of water, the pH is adjusted to 3.5, and it is enzymatically hydrolyzed using pectinase. The addition amount of pectinase is 2 wt% of Echinacea purpurea. The enzymatic hydrolysis temperature is 50 °C, and the enzymatic hydrolysis time is 2.5 h. It is inactivated, centrifuged, separated, and dried to obtain Echinacea purpurea polysaccharide;

[0085] (7) After the freeze-dried powder of the Bifidobacterium strain is activated and cultured to a cell density of 2.5 × 10 20 CFU / ml, it is continuously cultured for 2 generations, and the obtained bacterial suspension is freeze-dried to obtain Bifidobacterium viable powder:

[0086] (8) After the freeze-dried powder of the Lactobacillus rhamnosus strain is activated and cultured to a cell density of 2.5 × 10 20 CFU / ml, it is continuously cultured for 2 generations, and the obtained bacterial suspension is freeze-dried to obtain Lactobacillus rhamnosus viable powder;

[0087] (9) Mix 13 parts by mass of live Lactobacillus rhamnosus powder, 13 parts by mass of live Bifidobacterium powder, 33 parts by mass of ganoderma peptide, 10 parts by mass of dendrobium polysaccharide, 10 parts by mass of polygonatum polysaccharide, 11 parts by mass of astragalus polysaccharide, 15 parts by mass of eleutherococcus senticosus polysaccharide, and 12 parts by mass of echinacea polysaccharide to obtain a ganoderma peptide composition.

[0088] Example 2

[0089] This example provides a preparation method of a ganoderma peptide composition, which includes the following steps:

[0090] (1) Mix 1 part of Ganoderma sinense powder that has been ground through a 200-mesh sieve with 90 parts of water by mass, and perform enzymatic hydrolysis using a composite enzyme composed of hemicellulase and papain with a weight ratio of 1:1. The enzymatic hydrolysis time is 80 min, the enzymatic hydrolysis temperature is 45 °C, and the addition amount of the composite enzyme is 1.5 wt% of the Ganoderma sinense powder. Then inactivate, centrifuge and separate, and dry to obtain ganoderma peptide:

[0091] (2) Mix 1 part of Dendrobium officinale powder that has been ground through a 200-mesh sieve with 90 parts of water by mass, soak for 2.5 h, raise the temperature for extraction, centrifuge and filter to obtain a liquid phase. Vacuum concentrate the liquid phase, add ethanol for dissolution, let stand, centrifuge and separate, and dry to obtain dendrobium polysaccharide

[0092] (3) Mix 1 part of Polygonatum sibiricum powder that has been ground through a 200-mesh sieve with 90 parts of water by mass, soak for 2.5 h, raise the temperature to 80 °C and keep warm for 3 h, centrifuge and filter to obtain a liquid phase. Vacuum concentrate the liquid phase, add 150 parts of ethanol for dissolution, let stand, centrifuge and separate, and dry to obtain polygonatum polysaccharide;

[0093] (4) Mix 1 part of Astragalus membranaceus powder that has been ground through a 200-mesh sieve with 90 parts of water by mass, soak for 2.5 h, raise the temperature to 80 °C and keep warm for 3 h, centrifuge and filter to obtain a liquid phase. Vacuum concentrate the liquid phase, add 150 parts of ethanol for dissolution, let stand, centrifuge and separate, and dry to obtain astragalus polysaccharide;

[0094] (5) Mix 1 part of the roots and rhizomes of Eleutherococcus senticosus that have been ground through a 200-mesh sieve with 90 parts of water by mass, soak for 2.5 h, raise the temperature to 80 °C and keep warm for 3 h, centrifuge and filter to obtain a liquid phase. Vacuum concentrate the liquid phase, add 150 parts of ethanol for dissolution, let stand, centrifuge and separate, and dry to obtain eleutherococcus senticosus polysaccharide;

[0095] (6) Mix 1 part of Echinacea purpurea and 90 parts of water by mass, adjust the pH to 3.5, perform enzymatic hydrolysis using pectinase, the addition amount of pectinase is 2 wt% of Echinacea purpurea, the enzymatic hydrolysis temperature is 50 °C, the enzymatic hydrolysis time is 2.5 h, inactivate, centrifuge and separate, and dry to obtain echinacea polysaccharide;

[0096] (7) Activate and culture the freeze-dried powder of Bifidobacterium to a cell density of 2.5ⅹ10 20After reaching 2.5×10 CFU / ml, continue culturing for 2 generations, and freeze-dry the obtained bacterial suspension to obtain live Bifidobacterium powder:

[0097] (8) Activate the freeze-dried strain of Lactobacillus rhamnosus until the bacterial density reaches 2.5×10 20 CFU / ml, continue culturing for 2 generations, and freeze-dry the obtained bacterial suspension to obtain live Lactobacillus rhamnosus powder;

[0098] (9) By mass, mix 20 parts of live Lactobacillus rhamnosus powder, 20 parts of live Bifidobacterium powder, 46 parts of Ganoderma lucidum peptide, 18 parts of Dendrobium officinale polysaccharide, 16 parts of Polygonatum sibiricum polysaccharide, 14 parts of Astragalus membranaceus polysaccharide, 22 parts of Eleutherococcus senticosus polysaccharide, and 18 parts of Echinacea purpurea polysaccharide to obtain a Ganoderma lucidum peptide composition.

[0099] Comparative Example 1

[0100] The difference between this comparative example and Example 1 is that no live Lactobacillus rhamnosus powder and live Bifidobacterium powder are added.

[0101] Comparative Example 2

[0102] The difference between this comparative example and Example 1 is that only 26 parts of live Lactobacillus rhamnosus powder are added to the probiotic powder.

[0103] Comparative Example 3

[0104] The difference between this comparative example and Example 1 is that only 26 parts of live Bifidobacterium longum powder are added to the probiotic powder.

[0105] Comparative Example 4

[0106] The difference between this comparative example and Example 1 is that Dendrobium officinale polysaccharide is not added, and the mass of Ganoderma lucidum peptide is changed to 43 parts.

[0107] Comparative Example 5

[0108] The difference between this comparative example and Example 1 is that Polygonatum sibiricum polysaccharide is not added, and the mass of Ganoderma lucidum peptide is changed to 43 parts.

[0109] Comparative Example 6

[0110] The difference between this comparative example and Example 1 is that Astragalus membranaceus polysaccharide is not added, and the mass of Ganoderma lucidum peptide is changed to 44 parts.

[0111] Comparative Example 7

[0112] The difference between this comparative example and Example 1 is that Eleutherococcus senticosus polysaccharide is not added, and the mass of Ganoderma lucidum peptide is changed to 48 parts.

[0113] Comparative Example 8

[0114] The difference between this comparative example and Example 1 is that Echinacea purpurea polysaccharide is not added, and the mass of Ganoderma lucidum peptide is changed to 45 parts.

[0115] Animal experiment:

[0116] 1. Animal grouping: After adaptively raising 200 SPF-grade SD rats (weighing 160 ± 10 g) at 6 - 8 weeks of age for 7 days, they were randomly divided into a normal group, an Example 1 group, and a Comparative Example 8 group, with 20 rats in each group.

[0117] 2. Animal modeling and drug administration: The Example 1 group was correspondingly given the Ganoderma lucidum peptide composition prepared in Example 1 (0.1 g each time, twice a day), the Comparative Example 8 group was correspondingly given the composition obtained in Comparative Examples 1 - 8 (0.1 g each time, twice a day), and no operation was performed on the normal group. This was continuously carried out for 21 days. On the 22nd day, except for the normal group, the rats in each group were intraperitoneally injected with cyclophosphamide at 60 mg / kg for 4 consecutive days, and the normal group was given an equal amount of normal saline. On the 35th day, all the rats were anesthetized and sacrificed by carbon dioxide asphyxiation, and then blood was taken.

[0118] 3. Testing: The blood of the rats in each test group was centrifuged at 13400 rpm for 10 minutes at 4°C, and the supernatant was taken for immune index analysis. Through an enzyme-linked immunosorbent assay kit (ELISA), the concentrations of interferon-γ (IFN-γ), interleukin-4 (IL-4), immunoglobulin M (IgM), and immunoglobulin G (IgG) in the rat serum were measured, and the results are shown in Table 1.

[0119] Table 1 Analysis table of rat immune indexes

[0120]

[0121]

[0122] It can be seen from the above performance test results that Example 1 has a better improvement on the immune function of rats. This is mainly because the compound traditional Chinese medicine extract and probiotic live bacteria powder in the present invention synergistically improve the effect of the composition on immunity.

[0123] In the comparative example, because the necessary technical solutions were not adopted, its performance in the corresponding performance tests was significantly worse than that of the example, which better proves the irreplaceability of the specific technical solution of this application for achieving the technical effect and solving the technical problem.

[0124] The above is the preferred implementation manner of the present invention. It should be noted that for those of ordinary skill in the art in this technical field, without departing from the principle described in the present invention, several improvements and refinements can still be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A Ganoderma lucidum peptide composition, characterized in that, Comprising the following components by mass parts: Ganoderma lucidum peptide 30 - 50 parts, Echinacea purpurea polysaccharide 10 - 20 parts, Polygonatum sibiricum polysaccharide 8 - 18 parts, Dendrobium officinale polysaccharide 5 - 25 parts, Astragalus membranaceus polysaccharide 9 - 16 parts, Acanthopanax senticosus polysaccharide 12 - 24 parts, 20 - 50 parts of probiotic live bacteria powder, the viable bacteria count in the probiotic live bacteria powder being 4ⅹ10 10 CFU / g - 6ⅹ10 20 CFU / g.

2. The Ganoderma lucidum peptide composition according to claim 1, wherein The probiotic live bacteria powder is a mixture of bifidobacterium live bacteria powder and Lactobacillus rhamnosus live bacteria powder, and the mass ratio of the bifidobacterium live bacteria powder to the Lactobacillus rhamnosus live bacteria powder is 1:1; The viable count of the Bifidobacterium viable bacteria powder is 2×10 10 CFU / g - 3×10 20 CFU / g; The viable count of the Lactobacillus rhamnosus viable bacteria powder is 2×10 10 CFU / g - 3×10 20 CFU / g.

3. A method for preparing the Ganoderma lucidum peptide composition according to claim 1 or 2, characterized in that, The following steps are involved: (1) Ganoderma lucidum is crushed and sieved, mixed with water, inactivated by enzymatic hydrolysis, centrifuged and dried to obtain Ganoderma lucidum peptide: (2) crushing and sieving the Dendrobium officinale, mixing with water, soaking, heating and extracting, centrifuging to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve, standing, centrifuging and drying to obtain a Dendrobium polysaccharide; (3) crushing and sieving polygonatum, mixing with water, soaking, heating and extracting, centrifuging to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve, standing, centrifuging and drying to obtain polygonatum polysaccharide; (4) crushing and sieving the astragalus, mixing it with water, soaking it, heating it for extraction, and centrifuging it to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve it, standing it, centrifuging it and drying it to obtain astragalus polysaccharide; (5) crushing and sieving the roots and rhizomes of Acanthopanax senticosus, mixing them with water, soaking them, heating them for extraction, and centrifuging them to obtain a liquid phase, vacuum concentrating the liquid phase, adding ethanol to dissolve it, standing it, centrifuging and drying it to obtain Acanthopanax senticosus polysaccharide; (6) mixing Echinacea and water, adjusting the pH to 3-5, inactivating after enzymatic hydrolysis, and centrifuging and drying to obtain Echinacea polysaccharide; (7) Activate the freeze-dried powder of the Bifidobacterium strain and culture it until the bacterial density reaches 2×10 10 CFU / ml - 3×10 20 CFU / ml, then continue to culture for 2 - 3 generations, and freeze-dry the obtained bacterial suspension to obtain the Bifidobacterium live bacteria powder: (8) Activate the freeze-dried powder of Lactobacillus rhamnosus until the bacterial density reaches 2×10 10 CFU / ml - 3×10 20 CFU / ml, then continue to culture for 2 - 3 generations, and freeze-dry the obtained bacterial suspension to obtain the live powder of Lactobacillus rhamnosus; (9) Mixing live bacteria powder of Lactobacillus rhamnosus, live bacteria powder of Bifidobacterium, Ganoderma lucidum peptide, Dendrobium officinale polysaccharide, Polygonatum sibiricum polysaccharide, Astragalus polysaccharide, Acanthopanax senticosus polysaccharide, and Echinacea purpurea polysaccharide to obtain a Ganoderma lucidum peptide composition.

4. The preparation method of the ganoderma lucidum peptide composition according to claim 3, characterized in that, In step (1), the sieving uses a sieve with a mesh size of 200-400; And / or, in step (1), the mass ratio of the Ganoderma lucidum to water is 1:(50-100); And / or, in step (1), the enzymatic hydrolysis conditions are: enzymatic hydrolysis using a composite enzyme consisting of hemicellulase and papain in a weight ratio of 1:1, the enzymatic hydrolysis time is 60-100 min, the enzymatic hydrolysis temperature is 40-50° C., and the amount of the composite enzyme added is 1wt%-3wt% of the ganoderma lucidum powder.

5. The preparation method of the Ganoderma lucidum peptide composition according to claim 3, characterized in that, In step (2), the sieving uses a sieve with a mesh size of 200-400; And / or, in step (2), the mass ratio of the Dendrobium officinale to water is 1:(50-100). And / or, in step (2), the soaking time is 2-4 hours; And / or, in step (2), the temperature-raising extraction conditions are: heating to 60-100° C. and then keeping the temperature for 1.5-3 h; And / or, in step (2), relative to the volume of the water, the volume of the added ethanol is 2-4 times that of the water.

6. The preparation method of the ganoderma lucidum peptide composition according to claim 3, wherein, In step (3), the sieving uses a sieve with a mesh size of 200-400; And / or, in step (3), the mass ratio of polygonatum to water is 1:(50-100); And / or, in step (3), the soaking time is 2-4 hours; And / or, in step (3), the conditions for the temperature extraction are: heating to 60-100° C. and then keeping the temperature for 1.5-3 h; And / or, in step (3), relative to the volume of the water, the volume of the added ethanol is 2-4 times that of the water.

7. The preparation method of the ganoderma lucidum peptide composition according to claim 3, wherein, In step (4), the sieving uses a sieve with a mesh size of 200-400; And / or, in step (4), the mass ratio of the astragalus membranaceus to water is 1:(50 - 100); And / or, in step (4), the soaking time is 2 - 4 h; And / or, in step (4), the conditions for heating extraction are: heating to 60 - 100 °C and then holding for 1.5 - 3 h; And / or, in step (4), the volume of ethanol added is 2 - 4 times the volume of water.

8. The preparation method of the ganoderma lucidum peptide composition according to claim 3, wherein, In step (5), the sieve used for sieving is 200 - 400 mesh. And / or, in step (5), the mass ratio of the acanthopanax senticosus roots and rhizomes to water is 1:(50 - 100); And / or, in step (5), the soaking time is 2 - 4 h; And / or, in step (5), the conditions for heating extraction are: heating to 60 - 100 °C and then holding for 1.5 - 3 h; And / or, in step (5), the volume of ethanol added is 2 - 4 times the volume of water; And / or, in step (6), the mass ratio of echinacea purpurea to water is 1:(50 - 100); And / or, in step (6), the conditions for enzymatic hydrolysis are: enzymatic hydrolysis is carried out using pectinase, the addition amount of pectinase is 1 wt% - 4 wt% of echinacea purpurea, the enzymatic hydrolysis temperature is 40 - 60 °C, and the enzymatic hydrolysis time is 2 - 4 h.

9. The preparation method of the Ganoderma lucidum peptide composition according to claim 3, characterized in that, In steps (7) and (8), the medium used for cultivation is MRS medium. Taking 1000 mL of water as the relative amount, the raw materials in the MRS medium include: peptone 5 - 15 g; yeast extract 2 - 5 g; beef extract 8 - 15 g; glucose 15 - 20 g; sodium acetate 2 - 4 g; diammonium citrate 1 - 2 g; Tween 80 1 - 2 mL; magnesium sulfate 0.2 - 0.4 g; manganese sulfate 0.01 - 0.04 g; dipotassium hydrogen phosphate 2 - 4 g; agar 15 - 20 g; ganoderma lucidum peptide 3 - 5 g, echinacea purpurea polysaccharide 1 - 2 g, polygonatum sibiricum polysaccharide 0.8 - 1.8 g, dendrobium officinale polysaccharide 0.5 - 2.5 g, astragalus membranaceus polysaccharide 0.9 - 1.6 g, acanthopanax senticosus polysaccharide 1.2 - 2.4 g.

10. Use of a ganoderma lucidum peptide composition as claimed in claim 1 or 2 in the preparation of a drug for enhancing immunity and / or an adjuvant drug for enhancing immunity.

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