A bamboo fungus egg coat polysaccharide and its extraction method and immune-promoting active product based thereon

Abstract

The invention discloses a bamboo fungus egg outer coat polysaccharide and an extraction method thereof and an immunostimulating active product based thereon. The invention extracts and separates a new bamboo fungus egg outer coat and middle layer colloid polysaccharide, which is named as bamboo fungus egg outer coat polysaccharide (Dictyophora indusiatapileus volva polysaccharide, DIVP). The bamboo fungus egg outer coat polysaccharide is composed of eight monosaccharides, namely glucose, glucuronic acid, galactose, mannose, xylose, galacturonic acid, fucose and glucosamine hydrochloride. The invention finds that the bamboo fungus egg outer coat polysaccharide has significant immunomodulatory activity on macrophages, and therefore has the potential to become an immune adjuvant. Based on the significant effect of regulating human immunity of the bamboo fungus egg outer coat polysaccharide obtained from the above research, the invention develops the bamboo fungus egg outer coat polysaccharide into a variety of immunostimulating related foods, which are palatable and convenient to eat, and various natural synergistic raw materials are compounded in the formula, which conforms to the diet and consumption habits of the general public.

Description

Technical Field

[0001] The invention belongs to the technical field of biopharmaceutical products, and specifically relates to a bamboo fungus egg coat polysaccharide and an extraction method thereof, and an immunomodulatory active product based thereon. Background Art

[0002] Dictyophora indusiata belongs to the Phallales, Phallaceae, and genus Dictyophora. It is one of the precious edible fungi. It is called the "veiled mushroom lady" or "mushroom queen" because of its white net-like veil that hangs down like a skirt and its delicious taste. It is very popular in China and other Asian countries.

[0003] There are two main types of products sold on the market: mature bamboo fungus (developed from bamboo fungus embryos) and bamboo fungus eggs. Bamboo fungus eggs are the young basidiosperm buds of bamboo fungus. They are spherical and have three layers of coating. The outer coating is thin, smooth, grayish white or light brown red; the middle layer is gelatinous; the inner coating is tough and fleshy, and all parts are edible. When the bamboo fungus eggs are mature, the coating cracks, and the bamboo fungus embryo develops into a stipe to push out the cap, and the coating remains at the bottom of the stipe to form a stipe. The young basidiosperm buds of bamboo fungus and mature bamboo fungus have different growth periods, which will lead to different primary structures (such as monosaccharide composition, amino acid type, oligopeptide type) and higher-level structures (such as sugar chain connection methods, peptide chain folding and spatial polymerization methods, etc.) of polysaccharides, which in turn lead to differences in polysaccharide functions. Some researchers have isolated, purified and characterized some water-soluble and water-insoluble polysaccharides from mature bamboo fungus, among which branched (1-3)-β-D-glucan has attracted much attention due to its anti-tumor potential and triple helical conformation. However, there are no reports on the immunomodulatory and anti-inflammatory mechanisms of polysaccharides from young basidiosperm fruit buds of bamboo fungus. Summary of the invention

[0004] The purpose of the present invention is to provide a bamboo fungus egg outer coat polysaccharide and an extraction method thereof and an immunomodulatory active product based thereon. The bamboo fungus egg outer coat polysaccharide is a new bamboo fungus egg outer coat polysaccharide and has significant immunomodulatory activity.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The invention discloses a bamboo fungus egg outer coat polysaccharide. The bamboo fungus egg outer coat polysaccharide is composed of eight monosaccharides, namely, glucose, glucuronic acid, galactose, mannose, xylose, galacturonic acid, fucose and glucosamine hydrochloride, with a molar ratio of 0.535:0.308:0.06:0.047:0.02:0.013:0.009:0.008.

[0007] Preferably, the bamboo fungus egg coat polysaccharide is a uniform polysaccharide with a weight average molecular weight of 96.6 kDa and a narrow molecular weight distribution range.

[0008] The present invention also discloses a method for preparing the bamboo fungus egg coat polysaccharide, comprising:

[0009] 1) sorting the outer coat of bamboo fungus eggs, crushing them, and soaking them in water overnight to obtain a raw material;

[0010] 2) treating the raw material with a composite enzyme at 40°C for 1 hour, extracting with hot water for 3 hours, filtering, concentrating the filtrate, and then adding 3 times the volume of ethanol and standing overnight;

[0011] 3) dissolving the precipitate obtained after the centrifugation in step 2), deproteinizing, shaking, centrifuging and taking the supernatant, repeating the shaking and centrifugation 3-5 times, and ethanol precipitation overnight;

[0012] 4) taking out the precipitate by centrifugation and freeze-drying to obtain the polysaccharide of the outer coat of the bamboo fungus egg.

[0013] Preferably, in step 1), the material-liquid ratio of the bamboo fungus egg coat to the water for soaking is 1 g: (20-50) mL.

[0014] Preferably, in step 2), the complex enzyme is cellulase: pectinase: neutral protease: acid protease prepared in a mass ratio of (1-3): (2-3): (1-2): (0.5-1), the dosage ratio of the complex enzyme to the raw material is (5-10) g: 1 kg, and hot water extraction is adopted at 75-85°C; the filtrate is concentrated to a solid-liquid ratio of 1 g: 3 mL.

[0015] Preferably, in step 3), the deproteinization treatment uses Sevag reagent, shakes for 20 minutes, and the centrifugation treatment is carried out at a speed of 3500 r / min for 10 minutes.

[0016] The invention also discloses the application of the bamboo fungus egg outer coat polysaccharide in the preparation of immune adjuvants. The bamboo fungus egg outer coat polysaccharide is a macrophage immunoregulatory substance.

[0017] The present invention also discloses related immune-promoting active products developed based on the above-mentioned bamboo fungus egg outer coat polysaccharide, including bamboo fungus egg outer coat polysaccharide immune-promoting oral liquid, bamboo fungus egg outer coat polysaccharide immune-promoting effervescent tablets, bamboo fungus egg outer coat polysaccharide immune-promoting lozenges for relieving pharyngitis, bamboo fungus egg outer coat polysaccharide immune-promoting instant tea and bamboo fungus egg outer coat polysaccharide immune-promoting solid freeze-dried soup. Specifically, it includes:

[0018] The invention discloses an oral liquid for promoting immunity of polysaccharide from the outer coat of bamboo fungus egg. The raw materials thereof include, by mass percentage, 60% to 80% of polysaccharide aqueous solution from the outer coat of bamboo fungus egg, 8% to 16% of white sugar, 2% to 2.5% of honey, 0.5% to 1.0% of carrageenan and 0.3% to 0.5% of citric acid.

[0019] The invention discloses an immune-boosting effervescent tablet of bamboo fungus egg outer coat polysaccharide, which comprises, by mass percentage, 13.3% of bamboo fungus egg outer coat polysaccharide, 50.6% of an effervescent disintegrant, 1.5% of aspartame, 6% of PEG 6000, 26.1% of lactose and 2.5% of PVPK30 (mass fraction is 5%).

[0020] A bamboo fungus egg outer coat polysaccharide lozenge for promoting immunity and relieving pharyngitis, wherein the raw materials include, by mass percentage: 10% to 15% of bamboo fungus egg outer coat polysaccharide powder, 8% to 10% of dextrin, 35% to 45% of mannitol, 3% to 5% of sodium carboxymethyl cellulose, 8% to 13% of sucrose, 5% to 10% of magnesium stearate and 1% to 2% of menthol;

[0021] The bamboo fungus egg outer coat polysaccharide powder is prepared by freeze-drying the bamboo fungus egg outer coat polysaccharide aqueous solution and then passing it through an 80-mesh sieve. The mass concentration of the bamboo fungus egg outer coat polysaccharide aqueous solution is 60% to 80%.

[0022] The invention discloses an instant tea for promoting immunity from the outer coat polysaccharide of bamboo fungus egg. The raw materials include, by weight percentage, 40% to 60% of the outer coat polysaccharide of bamboo fungus egg, 10% to 15% of tea leaves, 5% to 6% of red dates, 5% to 7% of honeysuckle, 3% to 5% of liquorice, 3% to 5% of wolfberry, 2% of hawthorn, 0.3% to 1.5% of xylitol, 0.004% to 0.02% of stevioside, 0.01% to 0.05% of aspartame and 0.1% to 0.5% of citric acid.

[0023] A bamboo fungus egg outer coat polysaccharide immune-promoting solid freeze-dried soup, comprising, by mass percentage: 30% to 40% bamboo fungus egg outer coat polysaccharide powder, 10% to 20% dried fungus, 2% to 10% dried bamboo fungus, 2% to 10% dried spinach, 2% to 10% dried egg drop, 2% to 5% dried scallion, 20% laver, 1% wolfberry, 0.5% to 1.5% pepper extract, 1% salt, and 0.1% to 0.5% citric acid;

[0024] The Zanthoxylum bungeanum extract was prepared as follows: 1 kg of Dahongpao Zanthoxylum bungeanum was crushed, passed through a 20-mesh sieve, and extracted with a 30% ethanol solution at 60°C for 4 hours, the supernatant was filtered, the ethanol was recovered, and the mixture was concentrated to a solid-liquid ratio of 1 g: 2 mL, and freeze-dried to obtain the Zanthoxylum bungeanum extract;

[0025] The bamboo fungus egg outer coat polysaccharide powder is prepared by freeze-drying the bamboo fungus egg outer coat polysaccharide aqueous solution and then passing it through an 80-mesh sieve. The mass concentration of the bamboo fungus egg outer coat polysaccharide aqueous solution is 60% to 80%.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] This study extracted and separated a new polysaccharide from the outer coating and middle layer of bamboo fungus eggs, named as Dictyophora indusiatapileus volva polysaccharide (DIVP). The present invention clarified the composition of the bamboo fungus egg outer coating polysaccharide through research, which is composed of 8 monosaccharides: glucose, glucuronic acid, galactose, mannose, xylose, galacturonic acid, fucose and glucosamine hydrochloride. At the same time, the present invention found that the bamboo fungus egg outer coating polysaccharide has significant immunomodulatory activity on macrophages, so it has the potential to become an immune adjuvant.

[0028] Based on the significant effect of regulating human immunity possessed by the bamboo fungus egg coat polysaccharide obtained in the above research, the present invention develops the bamboo fungus egg coat polysaccharide into a variety of immune-promoting related foods, which are palatable and easy to eat, and various natural synergistic raw materials are compounded in the formula, which conforms to the diet and consumption habits of the general public. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 For SEM analysis;

[0030] Figure 2 is Zeta potential;

[0031] Figure 3 is the molecular weight and purity of the polysaccharide;

[0032] Figure 4 It is composed of monosaccharides;

[0033] Figure 5 This is the result of FT-IR analysis;

[0034] Figure 6 This is the NMR analysis result diagram;

[0035] Figure 7 DIVP promotes macrophage proliferation;

[0036] Figure 8 The effect of DIVP on the phagocytic ability of RAW264.7 cells;

[0037] Fig. 9 The effect of DIVP on the NO release ability of RAW264.7 cells;

[0038] Fig.10 The results of DIVP intervention on the expression of iNOS (a), IL-1 (b), IL-6 (c) and TNF-αm (d) RNA in macrophages;

[0039] Fig.11 For antibody factor chip detection;

[0040] Fig.12DIVP activates the PI3K-AKT and ERK1 / 2 signaling pathways. DETAILED DESCRIPTION

[0041] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not 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 should fall within the scope of protection of the present invention.

[0042] It should be noted that the terms "first", "second", etc. in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged where appropriate, so that the embodiments of the present invention described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units that are clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0043] The present invention is further described in detail below in conjunction with the accompanying drawings:

[0044] 1. Analysis of the polysaccharide structure of bamboo shoot egg coat

[0045] 1. SEM analysis

[0046] See also Figure 1 , Figure 1 It can be observed that DIVP has a distinct honeycomb structure, and there are obvious gaps inside the polysaccharide, indicating that there are a large number of cavities inside DIVP. The surface of DIVP is extremely rough, with protrusions similar to spheres and chains, and the layers are tightly packed, indicating that DIVP may have a complex structure.

[0047] 2. Zeta potential

[0048] The stability of the solution depends largely on the attraction and repulsion between molecules; the size and change of the Zeta potential value can reflect the stability change of the polysaccharide solution; the larger the absolute value of the potential value, the greater the electrostatic interaction between the particles, and the more stable the solution tends to be. Figure 2As shown in the figure, when the pH value is greater than 4, the absolute value of the DIVP potential is greater than 15; as the pH increases, the absolute value of the potential gradually increases, and when pH = 8, the absolute value of the DIVP potential can reach a maximum of 28.2; then it gradually decreases, and when pH = 10, the absolute value of the potential still reaches 20, indicating that DIVP is a polysaccharide with strong stability.

[0049] 3. DIVP molecular weight

[0050] The molecular weight and purity of polysaccharides were determined by HPGPC. Figure 3 As shown. The resulting graph is a single symmetrical peak, indicating a uniform polysaccharide. The molecular weight of DIVP is mainly distributed at 96 kDa, and the molecular weight distribution range is narrow. The degree of polymerization and purity of the polysaccharide are good. The molecular weight of a polysaccharide is usually expressed by the average distribution of a series of polysaccharides with similar molecular weights, that is, the weight-average molecular weight (Mw). As can be seen from Table 1, the Mw of EPS1 is 96.6 kDa. The Mw / Mn value represents the polydispersity of the sample, and Mw / Mn is 1.56, indicating that the molecular weight distribution of DIVP is narrow.

[0051] Table 1

[0052]

[0053] 4. DIVP monosaccharide composition

[0054] The monosaccharide composition was determined by ion chromatography. Figure 4 As shown in the figure, DIVP is composed of eight monosaccharides, namely glucose, glucuronic acid, galactose, mannose, xylose, galacturonic acid, fucose, and glucosamine hydrochloride, with a molar ratio of 0.535:0.308:0.06:0.047:0.02:0.013:0.009:0.008. Glucose and glucuronic acid are the main monosaccharides that make up DIVP, which contains a small amount of uronic acid and is an acidic polysaccharide. The remaining galactose, mannose, xylose, and uronic acid are key functional molecules with strong biological activity.

[0055] 5. FI-IR analysis

[0056] like Figure 5 As shown in the figure, the infrared spectrum of DIVP has a typical polysaccharide characteristic absorption peak at 3272 cm -1 The broad absorption peak near 2905cm -1 The peak at 1720cm comes from CH stretching vibration. -1 The strong peak at 1404cm can be attributed to the absorption peak of bound water and the stretching vibration of C=O. -1 The absorption peak of 1200-800cm is attributed to the stretching vibration of the carboxyl group (COO-), indicating the presence of uronic acid. -1The range is the fingerprint region of polysaccharides, between 1193 and 1020 cm -1 The peak at 920.64cm is caused by the stretching vibration of COC and COH in the pyranose ring, indicating that the sugar chain of the sample contains a pyranose ring structure. -1 and 784.42cm -1 The characteristic peaks that appeared indicate that DIVP has α and β glycosidic bonds. -1 The small absorption peak at is the characteristic absorption peak of mannose residues, indicating that DIVP contains mannose, which is consistent with the results of monosaccharide composition determination.

[0057] 6. NMR analysis

[0058] 1 H NMR spectrum, Figure 6 As shown in (a), the four signals δ5.24 ppm, δ5.11 ppm, δ4.68 ppm and δ4.55 ppm are attributed to the terminal group signals of α-L-Manp, α-D-GalpA, β-D-Glup and β-D-GlcpA, respectively. 13 C NMR spectrum, Figure 6 As shown in (b), four characteristic signals appear in the resonance region of the anomeric carbon (δ95~110ppm), namely δ103.38ppm, δ102.24ppm, δ97.84ppm and δ95.84ppm. The DIVP glycosidic bond signal has a characteristic chemical shift in the range of δ68~69.5ppm. In the DEPT-135 spectrum, there is a negative methylene signal near 60.97, which can be judged that the C6 position is substituted; there is a characteristic signal with a chemical shift of δ176.02, indicating the presence of uronic acid.

[0059] Further studies were performed including H HCOSY, HSQC, NOSY and 1 H– 13According to the chemical shift signals and related literature, the H1 (δ4.68ppm) signal of 4)-β-D-Glcp-(1→ sugar residue and the C4 (δ80.38ppm) signal of →4)-β-D-GlcpA-(1→ sugar residue are related, indicating that 4)-β-D-Glcp-(1→ sugar residue is connected to the C4 of →4)-β-D-GlcpA sugar residue; the H1 (δ5.11ppm) signal of →1)-β-D-Galp-(→ sugar residue and the C4 (δ80.38ppm) signal of →4)-α-D-GlcpA-(1→ sugar residue are related, indicating that 4)-β-D-Glcp-(1→ sugar residue is connected to the C4 of →4)-β-D-GlcpA sugar residue; The C3 (δ72.62 ppm) signal of the up-(1→) sugar residue was correlated, indicating that the →1)-β-D-Galp-sugar residue was connected to the C3 of the →4)-α-D-Glup-(1→) sugar residue; the H4 (δ3.68 ppm) signal of the →4)-α-D-Manp sugar residue was correlated with the C2 (δ70.90 ppm) signal of the →4)-β-D-GlcpA sugar residue, indicating that the →4)-α-D-Manp sugar residue was connected to the C2 of the →4)-β-D-GlcpA sugar residue.

[0060] 2. Immunomodulatory effect of DIVP

[0061] 1. Effects on RAW264.7 cells

[0062] The basis for macrophages to exert their immune function is cell survival, and the state of survival directly affects their function and the strength of their response. The MTT method was used to determine the effect of DIVP on cell proliferation. Figure 7 It can be seen that DIVP has the activity of promoting macrophage proliferation in the mass concentration range of 25-200 μg / mL and has no cytotoxicity. The effect of neutral red test on the phagocytic ability of macrophages was determined by Figure 8 It can be seen that DIVP has a significant inhibitory effect on the phagocytic ability of LPS-activated macrophages. The Gress experiment determined the effect on the NO release ability of macrophages. Fig. 9 It can be seen that DIVP significantly increased the ability of RAW264.7 cells to release NO (P < 0.01), but it was weaker than the LPS group. It can be determined that DIVP is an immunomodulatory substance of macrophages and has immunomodulatory activity. The RT-PCR experiment determined the effect on the transcription level of cytokines. Fig.10 It can be seen that the expression levels of iNOS, IL-1, IL-6, and TNF-α mRNA increased to varying degrees after DIVP intervention in macrophages with different mass concentrations.

[0063] 2. Antibody factor chip detection of the effect of DIVP on U937 and THP-1 cytokines

[0064] Antibody chip scanning results are shown in Fig.11Compared with the blank group, the seven cytokines MMP-8, IL-1β, IL-4, IL-10, Fas Ligand, CINC-3, and CNTF in the LPS-induced inflammatory model group were significantly increased (P < 0.05), among which the expression levels of MMP-8 and IL-4 were the most significant (P < 0.002). Compared with the blank group, except for NEG, Agrin, LIX, and L-selectin, the other cytokines in the DIVP treatment group were significantly increased (P < 0.05), among which Fas Ligand and IL-1α were the most significant (P < 0.002).

[0065] The results of this experiment showed that DIVP had a significant regulatory effect on chemokines, interleukins, matrix metalloproteinases and endothelin in U937. Among them, M-CSF is an important factor in promoting the recruitment of macrophages in tissues, and plays an important role in the proliferation, differentiation and activity maintenance of monocytes. After DIVP intervention, the levels of chemokines CCL11 and CXCL11 were significantly reduced, indicating that the level of inflammatory response was reduced. Matrix metalloproteinases are the main factors that promote tissue remodeling during inflammatory response. The reduction of MMP-2, MMP-3 and MMP-9 by DIVP means that it is beneficial to extracellular matrix remodeling. After DIVP intervention, the levels of four interleukins (IL-10, IL-12, IL-22, IL-23) were reduced, and IL-17A was also slightly reduced, indicating that the overall level of inflammatory response was reduced.

[0066] 3. Signaling pathways by which DIVP affects immune cell function

[0067] Western Blot experiments were performed to study changes in inflammatory signaling pathways. Fig.12 It was shown that DIVP could activate the PI3K-AKT and ERK1 / 2 signaling pathways, thereby promoting the expression of inflammatory genes.

[0068] In summary, this study extracted a new acidic polysaccharide from the outer coat of Dictyophora indica, with an average molecular weight of 96 kDa. This polysaccharide has a good ability to promote immune cell proliferation and immune cytokine secretion, and can reduce LPS-induced macrophage inflammatory response. At the same time, the study found that DIVP may mediate the immune regulation process through the PI3K / AKT and MAPK signaling pathways.

[0069] 3. Several DIVP immune-boosting foods

[0070] 1. Immunity-boosting oral solution

[0071] Raw material: DIVP polysaccharide solution

[0072] Formula: DIVP aqueous solution (60%-80%), white sugar (8%-16%), honey 2%, carrageenan (0.5%-0.9%), citric acid (0.3%).

[0073] Process: DIVP polysaccharide solution, white sugar, citric acid, carrageenan and honey are mixed and homogenized once at 65℃ and 40MPa; canned and sealed at 85℃; autoclaved at 121℃ for 20min; cooled in stages (60℃-40℃-20℃); labeled and stored.

[0074] 2. Immunity-boosting effervescent tablets

[0075] Raw materials: DIVP aqueous solution (mass concentration of 60% to 80%) was freeze-dried and passed through an 80-mesh sieve to obtain DIVP powder.

[0076] Formula: 13.3% DIVP powder, 50.6% effervescent disintegrant (acid-base ratio 1.2:1), 1.5% aspartame, 6% PEG6000 and 26.1% lactose 5% PVPK30 2.5%.

[0077] Wet granulation process: DIVP powder is mixed with an acid source (citric acid and tartaric acid 1:1), a sweetener (aspartame 1-3), and a filler (lactose), mixed well, and a wetting agent (75% ethanol) is added to make a soft material, and then dried at 0°C to obtain granules A. DIVP powder is taken, mixed with an alkali source (sodium bicarbonate), a sweetener (aspartame), and a filler (lactose), mixed with a binder (5% PVPK30) to make a soft material, and then dried at 50°C to obtain granules B. Granules A and B are sieved through a 14-mesh sieve, and an appropriate amount of lubricant (PEG 6000) is added. After mixing, they are pressed into 0.5g effervescent tablets.

[0078] Direct mixing process: citric acid and polyethylene glycol are crushed by a grinder, passed through an 80-mesh sieve, DIVP powder (10%-20%), disintegrant (acid-base ratio 1.2:1, accounting for 40%-50% by mass), aspartame (1%-2%), and polyethylene glycol 6000 (1%) are mixed evenly and tableted.

[0079] 3. Lozenges to boost immunity and relieve throat symptoms

[0080] Raw materials: DIVP aqueous solution (mass concentration of 60% to 80%) was freeze-dried and passed through an 80-mesh sieve to obtain DIVP powder.

[0081] Formula: DIVP powder 15%, dextrin 10%, mannitol 40%, sodium carboxymethyl cellulose 4%, sucrose 19%, magnesium stearate 10%, menthol 2%.

[0082] Process:

[0083] 1) Mix the raw materials and auxiliary materials (except lubricant and menthol);

[0084] 2) Making soft materials: Add pure water, add pure water in small amounts several times while stirring continuously, adjust the humidity of the material, avoid adding too much pure water, and make soft materials;

[0085] 3) Drying: Spread the granular soft material evenly on a baking tray and dry it at 70℃, turning it over every 0.5h. After drying, the dry granules will not stick to your hands and can be crushed by squeezing them hard.

[0086] 4) Add lubricant (magnesium stearate 10%) and menthol (2%) and mix well;

[0087] 5) Tablet pressing machine.

[0088] 4. Immunity-boosting instant tea

[0089] Raw materials: DIVP aqueous solution (mass concentration of 60% to 80%) was freeze-dried and passed through an 80-mesh sieve to obtain DIVP powder.

[0090] Formula: DIVP powder (40%-60%), tea (10%-15%), red dates (5%-6%), honeysuckle (5%-7%), licorice (3%-5%), wolfberry (3%-5%), hawthorn (2%), sweetener (xylitol, 0.3%-1.5%), stevioside (0.004%-0.02%), aspartame (0.01%-0.05%), acidulant (citric acid, 0.1%-0.5%).

[0091] Process: Red dates, licorice, honeysuckle, wolfberry, and hawthorn are freeze-dried and then ultra-finely ground, and all are sterilized (microwave or steam high temperature); the components are mixed evenly; microwave sterilization (80-140s, 430-460W); cooled and bagged and sealed.

[0092] 5. Immunity-boosting solid freeze-dried soup

[0093] Raw materials: DIVP aqueous solution (mass concentration of 60% to 80%) was freeze-dried and passed through an 80-mesh sieve to obtain DIVP powder.

[0094] Formula: DIVP powder (30%-40%), dried fungus (10%-20%), dried bamboo fungus (2%-10%), dried spinach (2%-10%), dried egg drop (2%-10%), dried scallion (2%-5%), seaweed (20%), wolfberry (1%), pepper extract (0.5%-1.5%), 1% salt, acidulant (citric acid (0.1%-0.5%).

[0095] The Zanthoxylum bungeanum extract was prepared as follows: 1 kg of Dahongpao Zanthoxylum bungeanum was crushed, passed through a 20-mesh sieve, and extracted with a 30% ethanol solution at 60°C for 4 hours, the supernatant was filtered, the ethanol was recovered, and the mixture was concentrated to a solid-liquid ratio of 1 g: 2 mL, and freeze-dried to obtain the Zanthoxylum bungeanum extract;

[0096] Process: Mix the components according to the proportion; sterilize by microwave (80-140s, 430-460W); cool and bag and seal.

[0097] The above contents are only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. A bamboo fungus egg coat polysaccharide, characterized in that: The bamboo fungus egg outer coat polysaccharide is composed of eight monosaccharides, namely glucose, glucuronic acid, galactose, mannose, xylose, galacturonic acid, fucose and glucosamine hydrochloride, with a molar ratio of 0.535:0.308:0.06:0.047:0.02:0.013:0.009:0.008; The polysaccharide in the outer coat of the bamboo fungus egg showed a single symmetrical peak shape and was a uniform polysaccharide with a weight average molecular weight of 96.6 kDa.

2. The method for preparing the bamboo fungus egg coat polysaccharide according to claim 1, characterized in that: include: 1) sorting the outer coat of bamboo fungus eggs, crushing them, and soaking them in water overnight to obtain the raw material; 2) The raw material was treated with a composite enzyme for 1 hour at 40°C, extracted with hot water for 3 hours, filtered, the filtrate was concentrated, and then 3 times the volume of ethanol was added and allowed to stand overnight; 3) Dissolve the precipitate obtained after centrifugation in step 2), deproteinize, shake and centrifuge to obtain the supernatant, repeat shaking and centrifugation 3-5 times, and precipitate with ethanol overnight; 4) Centrifuge and take out the precipitate, freeze-dry it, and obtain the polysaccharide of the outer coat of bamboo fungus egg.

3. The method for preparing the bamboo fungus egg coat polysaccharide according to claim 2, characterized in that: In step 1), the solid-liquid ratio of the outer layer of bamboo fungus eggs to the water used for soaking is 1 g: (20 ~ 50) mL; in step 2), the complex enzyme is prepared by cellulase: pectinase: neutral protease: acidic protease according to the mass ratio of (1~3): (2~3): (1~2): (0.5~1), the dosage ratio of the complex enzyme to the raw material is (5~10) g: 1 kg, and hot water extraction is adopted at 75~85°C; the filtrate is concentrated to a solid-liquid ratio of 1 g: 3 mL; in step 3), the deproteinization treatment adopts Sevag reagent, shakes for 20 minutes, and the centrifugal treatment is processed at a speed of 3500r / min for 10 minutes.

4. The use of the bamboo fungus egg coat polysaccharide according to claim 1 in the preparation of an immune adjuvant, characterized in that: The bamboo fungus egg coat polysaccharide is a macrophage immunomodulatory substance.

5. An oral liquid for promoting immunity containing polysaccharides from the outer coat of bamboo fungus eggs, characterized in that: In terms of mass percentage, the raw materials include: 60% to 80% aqueous solution of polysaccharide from the outer coat of bamboo fungus eggs, 8% to 16% white sugar, 2% to 2.5% honey, 0.5% to 1.0% carrageenan, and 0.3% to 0.5% citric acid; The bamboo fungus egg outer coat polysaccharide aqueous solution is prepared from the bamboo fungus egg outer coat polysaccharide described in claim 1.

6. A bamboo fungus egg coat polysaccharide immune-boosting effervescent tablet, characterized in that: In terms of mass percentage, it includes: 13.3% of bamboo fungus egg outer coat polysaccharide, 50.6% of effervescent disintegrant, 1.5% of aspartame, 6% of PEG 6000, 26.1% of lactose and 2.5% of 5% PVPK30; The bamboo fungus egg outer coat polysaccharide is the bamboo fungus egg outer coat polysaccharide described in claim 1.

7. A lozenge containing polysaccharides from the outer coat of bamboo fungus eggs for promoting immunity and relieving pharyngitis, characterized in that: The raw materials include, by mass percentage: 10% to 15% of bamboo fungus egg coat polysaccharide powder, 8% to 10% of dextrin, 35% to 45% of mannitol, 3% to 5% of sodium carboxymethyl cellulose, 8% to 13% of sucrose, 5% to 10% of magnesium stearate and 1% to 2% of menthol; The bamboo fungus egg outer coat polysaccharide powder is prepared by freeze-drying the bamboo fungus egg outer coat polysaccharide aqueous solution and then passing it through an 80-mesh sieve, and the mass concentration of the bamboo fungus egg outer coat polysaccharide aqueous solution is 60% to 80%; The bamboo fungus egg outer coat polysaccharide aqueous solution is prepared from the bamboo fungus egg outer coat polysaccharide described in claim 1.

8. A bamboo fungus egg coat polysaccharide immune-boosting instant tea, characterized in that: In terms of percentage by mass, the raw materials include: 40% to 60% of bamboo fungus egg coat polysaccharide, 10% to 15% of tea leaves, 5% to 6% of red dates, 5% to 7% of honeysuckle, 3% to 5% of licorice, 3% to 5% of wolfberry, 2% of hawthorn, 0.3% to 1.5% of xylitol, 0.004% to 0.02% of stevioside, 0.01% to 0.05% of aspartame, and 0.1% to 0.5% of citric acid; The bamboo fungus egg outer coat polysaccharide is the bamboo fungus egg outer coat polysaccharide described in claim 1.

9. A solid freeze-dried soup containing polysaccharides from bamboo fungus eggs to promote immunity, characterized in that: Calculated by mass percentage, it includes: 30% to 40% of bamboo fungus egg coat polysaccharide powder, 10% to 20% of dried fungus, 2% to 10% of dried bamboo fungus, 2% to 10% of dried spinach, 2% to 10% of dried egg drop, 2% to 10% of dried scallion, 20% of seaweed, 1% of wolfberry, 0.5% to 1.5% of pepper extract, 1% of salt, and 0.1% to 0.5% of citric acid; The Zanthoxylum bungeanum extract was prepared as follows: 1 kg of Dahongpao Zanthoxylum bungeanum was crushed, passed through a 20-mesh sieve, and extracted with a 30% ethanol solution at 60°C for 4 hours, the supernatant was filtered, the ethanol was recovered, and the mixture was concentrated to a solid-liquid ratio of 1 g:2 mL, and freeze-dried to obtain the Zanthoxylum bungeanum extract; Among them, the bamboo fungus egg outer coat polysaccharide powder is obtained by freeze-drying the bamboo fungus egg outer coat polysaccharide aqueous solution and then passing it through an 80-mesh sieve. The mass concentration of the bamboo fungus egg outer coat polysaccharide aqueous solution is 60%~80%; the bamboo fungus egg outer coat polysaccharide aqueous solution is prepared from the bamboo fungus egg outer coat polysaccharide described in claim 1.

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