TW202007404A - Method for stimulating activity of natural killer cells or formation of IgA or IgM antibody using glucan and mushroom polysaccharide composition capable of strengthening immune capability - Google Patents

Abstract

The present invention provides a glucan and mushroom polysaccharide composition, which contains 20-50 wt% of beta-glucan concentrate, 20-50 wt% of ganoderma lucidum mycelium extract, 1-5 wt% of antrodia camphorate mycelium extract, 1-5 wt% of cordyceps sinensis mycelium extract, 1-5 wt% of phellinus linteus fruiting body extract, 1-5 wt% of agaricus blazei mrill fruiting body extract, 1-5 wt% of grifola frondosa fruiting body extract, and 1-5 wt% of hericium erinaceus fruiting body extract.

Description

Dextran and mushroom polysaccharide composition

The invention relates to a polysaccharide composition; in particular, it relates to a glucan and mushroom polysaccharide composition that can enhance immune function.

Polysaccharides present in mushrooms are known to have an immune-enhancing effect. Taking Ganoderma as an example, it has long been widely used as a raw material for nutrition and health food. β-Glucan (β-Glucan) is a kind of polysaccharide, which is extremely related to the immune effect of polysaccharide.

In the past, polysaccharide health care products have both single-component products and compound products, and compound products have better immune regulation effects and are more popular with consumers. With the increase in consumer demand for health food in modern society, it is urgent to develop compound polysaccharide products with both flavor and health benefits.

In view of this, the object of the present invention is to provide a glucan and lentinan composition that can improve immune function.

In order to achieve the above object, the present invention provides a glucan and lentinan composition comprising 20 to 50% by weight of β-polyglucose concentrate and 20 to 50% by weight of Ganoderma lucidum Mycelium extract, 1 to 5% by weight of Antrodia camphorata mycelium extract, 1 to 5% by weight of Cordyceps mycelium extract, 1 to 5% by weight of mulberry fruit body extract , 1 to 5% by weight of Brazilian mushroom fruit body extract, 1 to 5% by weight of Maitake mushroom body extract and 1 to 5% by weight Hericium erinaceus fruit body extract.

In order to achieve the above object, the present invention also provides a method for manufacturing a glucan and mushroom mushroom polysaccharide composition, which comprises a black yeast cell fermentation product, a ganoderma fungus cell fermentation product, and antrodia The fungus fermentation product of a fungus, a fungus fermentation product of Mortierella sinensis, a fruiting body powder of Phellinus igniarius, a fruiting body powder of Brazil mushroom, a fruiting body powder of Maitake mushroom and a fruiting body powder of Hericium erinaceus, respectively Water is mixed in a ratio of 10:1 to 40:1 by weight, and then the mixture is stirred at 70 to 100°C for 2 to 6 hours, filtered and solids are removed, and the filtrate is taken and heated and sterilized. Obtain a β-polyglucose concentrate, a Ganoderma lucidum mycelium extract, an Antrodia camphorata mycelium extract, a Cordyceps mycelium extract, a mulberry fruit body extract, a Brazilian mushroom fruit body extract, a Maitake fruit body extract and a hericium erinaceus fruit body extract, and then all the aforementioned extracts are mixed and concentrated to obtain the glucan and mushroom mushroom polysaccharide body composition.

In order to explain the present invention more clearly, the preferred embodiments are described in detail below with reference to the drawings.

The glucan and mushroom mushroom polysaccharide composition of the present invention is an extract obtained from a variety of mushroom fungi. The main formula includes a β-polyglucose concentrate, a ganoderma lucidum mycelium extract, and an Antrodia camphorata mycelium extract , A Cordyceps sinensis mycelium extract, a Phellinus linteus fruit body extract, a Brazilian mushroom fruit body extract, a Maitake fruit body extract and a Hericium erinaceus fruit body extract.

其中，黑酵母葡聚醣（單方組）係作為本實例中之對照組配方，其中，此單方組係將β-聚葡萄糖濃縮液濃縮至規格：多醣體＞6 g/L、葡聚醣＞5 g/L。黑酵母葡聚醣及菇蕈多醣體組成物（複方組）係作為本實例中之實驗組配方，其中，此複方組係先將牛樟芝菌絲體萃取液、冬蟲夏草菌絲體萃取液、桑黃子實體萃取液及巴西蘑菇子實體萃取液以1：1：1：1之比例進行混合後濃縮至多醣體＞5 g/L，得到一混合液A，再另外將舞茸子實體萃取液及猴頭菇子實體萃取液以1：1之比例進行混合後濃縮至多醣體＞5 g/L，得到一混合液B，最後添加β-聚葡萄糖濃縮液及靈芝菌絲體萃取液至前述混合液A及混合液B中，再進行混合後濃縮至多醣體＞6 g/L，即為本實例之黑酵母葡聚醣及菇蕈多醣體組成物（複方組）。The preparation method of the glucan and lentinan composition of the present invention will be described below. (1) The method of obtaining polysaccharides from the fermentation product of the bacterial cell 1.1 Preparation of the fermentation broth of the bacterial cell Inoculate each bacterial species in a carbon source (such as glucose, sucrose or mannitol) 0.5 to 5.0 weight percent, a nitrogen source (such as yeast extract) , Yeast protein cocoon or soy protein cocoon) 0.1 to 1.5 weight percent and other trace substances (such as trace elements and inorganics), a medium with a pH value between 5.0 and 6.5, aerated and stirred at a temperature of 20 to 30°C for 2 to After 7 days, the bacterial fermentation product can be obtained. 1.2 Obtain the polysaccharide extract and mix the fermentation product of the cells with water at a weight ratio of 10:1 to 40:1 to form a mixture, stir the mixture at 70 to 100°C for 2 to 6 hours, and press the mixture Filter, remove solids, take the filtrate and heat sterilize to obtain polysaccharide extract. (2) How to obtain polysaccharides from fruit bodies 2.1 Preparation of fruit body mixed solution After mixing fruit body powder and water in a weight ratio of 10:1 to 40:1 to form a mixture, stir the mixture at 70 to 100°C 2 to 6 hours. 2.2 Obtaining polysaccharide extracts The respective mixtures were pressure filtered to remove solids, and the filtrate was taken and sterilized by heating to obtain polysaccharide extracts. (3) Determination method of polysaccharide specifications 3.1 Calculation of β-glucan content After adding β-polyglucose concentrate to an appropriate volume of buffer and mixing well, add α-amylase and proteolytic enzyme in sequence (Protease) and Amyloglucosidase (Amyloglucosidase) three enzyme treatment, precipitation with 4 times the volume of alcohol, β-glucan sedimentation and separation from the solution, the precipitate was collected and washed with alcohol and dried, dried The precipitate was hydrolyzed with strong acid and high temperature. After acid-base neutralization, glucose was measured, and β-glucan content was calculated. 3.2 Detection of polysaccharide concentration: β-polyglucose concentrate, Ganoderma lucidum mycelium extract, Antrodia camphorata mycelium extract, Cordyceps sinensis mycelium extract, Phellinus igniarius fruit body extract, Brazil mushroom fruit body extract, Maitake The fruit body extract and Hericium erinaceus fruit body extract are respectively diluted to appropriate concentrations, injected into the dialysis membrane (specification: MW6000～8000), and dialysis with flowing water for 48 hours (flow rate: 0.2L/min). The system uses a phenol-sulfuric acid assay to determine the polysaccharide concentration. When the sugar encounters a strong acid, the hydroxyl group in the structural formula combines with the phenol to produce an orange-yellow liquid, so the absorbance value can be measured by colorimetry to detect the concentration of its polysaccharide. (4) Examples This example uses strains of black yeast ( Aureobasidium pullulans , BCRC No. 930184), Ganoderma lucidum ( Ganoderma lucidum ), Antrodia camphorate ( Antrodia camphorate ), and Hirsutella sinensis ( Hirsutella sinensis ), and mulberry ( Phellinus linteus), Agaricus (Agaricus blazei murill), Maitake (Grifola frondosa) and Hericium (Hericium erinaceus) fruiting bodies of polysaccharide extracts were prepared in accordance with the preceding embodiment, respectively, and comprises the following formulation:

Among them, the black yeast glucan (unilateral group) is used as the control group formulation in this example, wherein the unilateral group concentrates the β-polydextrose concentrate to specifications: polysaccharide>6 g/L, dextran> 5 g/L. The black yeast glucan and mushroom mushroom polysaccharide composition (composite group) is used as the experimental group formula in this example, in which the compound group consists of the Antrodia cinnamomea mycelium extract, Cordyceps sinensis mycelium extract, and Phellinus linteus. The fruit body extract and the Brazilian mushroom fruit body extract are mixed in a ratio of 1:1:1:1 and concentrated to polysaccharides> 5 g/L to obtain a mixed liquid A, and then the Maitake fruit body extract and Hericium erinaceus fruit body extract is mixed in a ratio of 1:1, then concentrated to a polysaccharide> 5 g/L to obtain a mixed solution B, and finally β-polyglucose concentrated solution and Ganoderma lucidum mycelium extract are added to the foregoing mixture The liquid A and the mixed liquid B are mixed again and concentrated to a polysaccharide> 6 g/L, which is the composition of the black yeast glucan and lentinan in this example (composite group).

The glucan and mushroom polysaccharide composition of the present invention comprises: 44% by weight of β-polyglucose concentrate, 44% by weight of Ganoderma lucidum mycelium extract, and 2% by weight of Antrodia cinnamomea mycelium Body extract, Cordyceps sinensis mycelium extract at 2% by weight, Phellinus linteus fruit body extract at 2% by weight, Brazil mushroom fruit body extract at 2% by weight, Maitake fruit body extract at 2% by weight Liquid and 2% by weight of Hericium erinaceus fruit body extract.

In order to evaluate the ability of the glucan and lentinan composition of the present invention to promote the proliferation of immune cells, the following animal experiments were evaluated on specific and non-specific immunomodulatory effects, respectively. (1) Design of test group and dosage of test substance administration: 6-week-old BALB/c female mice were used for animal testing. As shown in Table 1, the test group was designed with a water control group and a low-dose glucan of black yeast Group (hereinafter referred to as the unilateral low-dose group), black yeast glucan high-dose group (hereinafter referred to as the unilateral high-dose group), black yeast glucan and the compound mushroom mushroom polysaccharide low-dose group (hereinafter referred to as the compound low-dose group) and The high-dose group of black yeast glucan and compound mushroom mushroom polysaccharide (hereinafter referred to as the compound high-dose group), the number of experimental animals in each group is 8 animals, of which the water control group is only administered with deionized water, single and compound low dose groups The dosage of the test substance is 1 times the human recommended dose, the single and compound high-dose groups are 3 times the recommended human dose, of which the recommended dose for the human body is 60 kg for a person 30 mL/day, according to the animal and human body weight per kg The dose conversion factor 12.3 converts the dose administered to mice in each group. The test animals were given the test substance and the negative control solution (ie deionized water) by oral administration through a gastric tube daily for 9 weeks, and the administration volume was 20 mL/kg.

(2) 致敏小鼠 如圖1所示，於投予試驗物質3週後，以卵白蛋白（Ovalbumin, OVA）做為免疫原致敏試驗小鼠，以皮下注射方式給予注射體積100 μL之使用劑量3 μg OVA及佛氏完全輔劑(complete Freund’s adjuvant)混合液，2週後再次皮下注射100 μL之使用劑量12 μg OVA及佛氏不完全輔劑(incomplete Freund’s adjuvant)混合液為第二次致敏，2週後再次皮下注射100 μL之使用劑量12 μg OVA及佛氏不完全輔劑(incomplete Freund’s adjuvant)混合液為第三次致敏。分別於致敏前、後自眼窩採血，進行總抗體及OVA特異性抗體的分析。試驗結束後犧牲小鼠，採取血液以及脾臟細胞進行免疫功能評估分析，包括測定免疫細胞增生、免疫細胞激素分泌、免疫細胞種類以及血清抗體濃度等，藉此評估本發明之葡聚醣及菇蕈多醣體組成物特異性及非特異性免疫調節之功效。 (3) 試驗動物觀察 在餵食9週後，各組間之體重、脾臟絕對及相對重以及脾臟細胞數量並無明顯差異，顯示二試驗物質對於小鼠之生長無不良影響，如表2、表3及圖2所示。Table 1. Design of test group and dosage of test substance

(2) The sensitized mice are shown in Figure 1. Three weeks after the administration of the test substance, ovalbumin (OVA) is used as the immunogen sensitization test mice, and the injection volume is 100 μL by subcutaneous injection. Use a dose of 3 μg of OVA and complete Freund's adjuvant mixed solution, and 2 weeks later, subcutaneously inject 100 μL of the used dose of 12 μg OVA and incomplete Freund's adjuvant mixed solution as the second Second sensitization. After 2 weeks, 100 μL of subcutaneous injection of 12 μg OVA and incomplete Freund's adjuvant mixed solution was injected subcutaneously for the third sensitization. Blood was collected from the eye socket before and after sensitization to analyze total antibodies and OVA-specific antibodies. After the test, the mice were sacrificed, and blood and spleen cells were taken for immune function evaluation and analysis, including the determination of immune cell proliferation, immune cell hormone secretion, immune cell type, and serum antibody concentration, etc., thereby evaluating the glucan and mushroom of the present invention The efficacy of specific and non-specific immunomodulation of polysaccharide composition. (3) Test animals observed that after 9 weeks of feeding, there was no significant difference in body weight, absolute and relative weight of the spleen, and the number of spleen cells between the groups, showing that the two test substances had no adverse effects on the growth of mice, as shown in Table 2 and Table 2. 3 and Figure 2.

Table 2. Weekly average weight change of test animals

(4) 非特異性免疫調節功效 4.1 吞噬細胞活性試驗 利用Phagotest kit檢測血液中吞噬細胞的吞噬活性。於第4週，自採集的血液取100 μl抗凝之全血與20 μl opsonized FITC-labelledE. coli 混合，置於37℃水浴槽作為實驗組或置於冰上作為陰性對照組作用10分鐘，然後置於冰上以終止吞噬作用，加入100 μl冰冷之quenching solution，混合均勻。離心、清洗。加入200 μl之DNA staining solution，置於冰上，避光反應10分鐘，而後在60分鐘內以流式細胞儀進行分析。吞噬作用活性（%）=實驗組之吞噬作用（%）－陰性對照組之吞噬作用（%）。 如表4及圖3所示，單方低劑量組之血液中吞噬細胞活性顯著高於水對照組（p＜0.05），單方高劑量組平均值雖亦高於水對照組，但未達統計上顯著差異。複方低劑量組及高劑量組與水對照組之間皆無明顯差異。Table 3. Test animal spleen weight and cell number

(4) Non-specific immunomodulatory effects 4.1 Phagocytic activity test Phagotest kit was used to detect the phagocytic activity of phagocytes in the blood. At week 4, 100 μl of anticoagulated whole blood was mixed with 20 μl of opsonized FITC-labelled E. coli from the collected blood and placed in a 37°C water bath as an experimental group or placed on ice as a negative control group for 10 minutes. , And then placed on ice to stop the phagocytosis, add 100 μl ice cold quenching solution, mix well. Centrifuge and wash. 200 μl of DNA staining solution was added, placed on ice, and protected from light for 10 minutes, and then analyzed by flow cytometry within 60 minutes. Phagocytosis activity (%) = phagocytosis of experimental group (%)-phagocytosis of negative control group (%). As shown in Table 4 and Figure 3, the blood phagocytic activity of the unilateral low-dose group was significantly higher than that of the water control group (p<0.05). Although the average of the unilateral high-dose group was also higher than that of the water control group, it was not statistically significant. Significant differences. There was no significant difference between the compound low-dose group and the high-dose group and the water control group.

4.2 自然殺手細胞活性試驗 利用LIVE/DEAD cell-mediated cytotoxicity kit檢測自然殺手細胞毒殺標的細胞YAC-1的能力。先以染劑DIOC₁₈ 標示YAC-1細胞，於37℃、5%CO₂ 培養箱內，作用4小時，再以標的細胞：脾臟細胞=1：100或200之比例混合作為實驗組，於37℃、5%CO₂ 培養箱內，作用4小時。陰性對照組則以RPMI 1640+10%FBS取代脾臟細胞。以流式細胞儀進行分析。自然殺手細胞活性（%）=（實驗組之細胞死亡率）－（陰性對照組之細胞死亡率）。 如表5及圖4所示，複方低劑量組、高劑量組均可顯著促進自然殺手細胞之活性（p＜0.05）。Table 4. Phagocytic cell activity

4.2 Natural killer cell activity test The LIVE/DEAD cell-mediated cytotoxicity kit was used to detect the ability of natural killer cytotoxicity to kill the target cell YAC-1. First mark the YAC-1 cells with the dye DIOC ₁₈ , and incubate at 37°C in a 5% CO ₂ incubator for 4 hours. Then mix the target cells: spleen cells = 1:100 or 200 as the experimental group at 37 ℃, 5% CO ₂ incubator for 4 hours. The negative control group replaced spleen cells with RPMI 1640+10% FBS. Analyze by flow cytometry. Natural killer cell activity (%) = (cell death rate of experimental group)-(cell death rate of negative control group). As shown in Table 5 and Figure 4, the compound low-dose group and high-dose group can significantly promote the activity of natural killer cells (p<0.05).

4.3 免疫細胞增生試驗 96孔培養盤內分別加入100 μl/well之培養基或含2.5 μg/ml Concanavalin A (Con A)、或10 μg/ml lipopolysaccharide (LPS)、或200 μg/ml OVA之培養基，每一處理三重複。然後加入100 μl/well之4×10⁶ cells/ml脾臟細胞懸浮液，置於37℃、5%CO₂ 培養箱內，培養48或72小時。之後每孔洞中加入20μl之MTT試劑（5 mg/ml），繼續培養4至6小時。離心250×g、10分鐘。吸棄200 μl/well上清液。加入200 μl/well之dimethyl sulfoxid（DMSO），震盪5分鐘後，用ELISA reader測A₅₇₀ nm吸光值。刺激指數（Stimulation index, S.I.）=（A₅₇₀ nm treatment－A₅₇₀ nm medium control）/（A₅₇₀ nm cell only－A₅₇₀ nm medium control）。 如表6所示，從實驗結果發現，不論是在Con A或LPS處理下，各組之間並無差異。Table 5. Activity of natural killer cells

4.3 Immune cell proliferation test 96-well culture plates were added with 100 μl/well medium or medium containing 2.5 μg/ml Concanavalin A (Con A), or 10 μg/ml lipopolysaccharide (LPS), or 200 μg/ml OVA, Repeat three times for each treatment. Then add 100 μl/well of 4×10 ⁶ cells/ml spleen cell suspension and place in a 37°C, 5% CO ₂ incubator for 48 or 72 hours. After that, add 20 μl of MTT reagent (5 mg/ml) to each hole, and continue culturing for 4 to 6 hours. Centrifuge at 250×g for 10 minutes. Aspirate 200 μl/well supernatant. Add 200 μl/well of dimethyl sulfoxid (DMSO), shake for 5 minutes, and measure the absorbance of A ₅₇₀ nm with ELISA reader. Stimulation index (SI) = (A ₅₇₀ nm treatment-A ₅₇₀ nm medium control)/(A ₅₇₀ nm cell only-A ₅₇₀ nm medium control). As shown in Table 6, from the experimental results, it was found that there was no difference between the groups, whether under Con A or LPS treatment.

4.4 細胞激素分析試驗 24孔培養盤上分別加入培養基、Con A（最終濃度：2.5 μg/ml）或OVA（最終濃度：100 μg/ml），加入脾臟細胞，使每孔細胞總數為5×10⁶ cells/well。分別於24及48小時後收集細胞培養上清液。之後以三明治型酵素連結免疫分析法（sandwich enzyme-linked immunosorbent assay, sandwich-ELISA）測試細胞培養上清液中干擾素-γ（Interferon γ, IFN-γ）以及細胞白介素-5（Interleukin-5, IL-5）含量。 如表7所示，除複方低劑量組之Con A-stimulated IL-5顯著低於水對照組（p＜0.05）外，其餘各組間並無明顯差異。Table 6. Spleen immune cell proliferation ability

4.4 Cytokine analysis test Add medium, Con A (final concentration: 2.5 μg/ml) or OVA (final concentration: 100 μg/ml) to the 24-well culture plate, add spleen cells to make the total number of cells per well 5×10 ⁶ cells/well. The cell culture supernatant was collected after 24 and 48 hours, respectively. Then, sandwich enzyme-linked immunosorbent assay (sandwich enzyme-linked immunosorbent assay, sandwich-ELISA) was used to test interferon-γ (Interferon γ, IFN-γ) and cell interleukin-5 (Interleukin-5, IL-5) content. As shown in Table 7, except that Con A-stimulated IL-5 in the compound low-dose group was significantly lower than that in the water control group (p<0.05), there was no significant difference between the remaining groups.

4.5 血液中抗體含量試驗 將保存於4℃之抗凝全血以sandwich-ELISA測定其中所含IgG、IgM、IgA總抗體。如表8及圖5所示，第8週之血液抗體檢測結果顯示，複方高劑量組可顯著促進IgA及IgM抗體的生成（p＜0.05）。Table 7. Content of cytokine secretion

4.5 Test of antibody content in blood Anticoagulated whole blood stored at 4°C was tested by sandwich-ELISA for the total antibodies contained in IgG, IgM and IgA. As shown in Table 8 and Figure 5, the blood antibody test results at week 8 show that the compound high-dose group can significantly promote the production of IgA and IgM antibodies (p<0.05).

(5) 特異性免疫調節功效 將保存於4℃之抗凝全血以sandwich-ELISA測定其中所含OVA特異性IgG抗體的濃度，96孔盤中被覆的是10 μg/ml的OVA（溶於pH9.6之0.05M Sodium carbonate緩衝液）。 再參照表8，在特異性抗體anti-OVA IgG生成方面，單方低劑量組及高劑量組、複方低劑量組及高劑量組均與水對照組之間沒有差異。 另再參照表6及表7，在OVA專一性淋巴細胞增生以及OVA專一性細胞激素兩項指標，單方低劑量組及高劑量組、複方低劑量組及高劑量組均與水對照組之間沒有差異。Table 8. Serum antibody production

(5) Specific immunomodulation effect: The concentration of OVA-specific IgG antibody contained in anticoagulated whole blood stored at 4℃ was determined by sandwich-ELISA. The 96-well plate was coated with 10 μg/ml OVA (dissolved in 0.05M Sodium carbonate buffer pH 9.6). Referring again to Table 8, regarding the production of specific antibody anti-OVA IgG, there was no difference between the unilateral low-dose group and the high-dose group, the compound low-dose group and the high-dose group from the water control group. Also refer to Table 6 and Table 7, in the two indicators of OVA-specific lymphocyte hyperplasia and OVA-specific cytokine, the unilateral low-dose group and the high-dose group, the compound low-dose group and the high-dose group are all between the water control group no difference.

According to the above, saccharomyces cerevisiae glucan, saccharomyces cerevisiae glucan and complex mushroom mushroom polysaccharides all have a regulating effect on non-specific immune responses.

The above is only the preferred and feasible embodiments of the present invention, and any equivalent changes in applying the description of the present invention and the scope of patent application should be included in the patent scope of the present invention.

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Figure 1 is the sensitization process of the test animals; Figure 2 is the weekly average body weight change chart of the test animals; Figure 3 is the test animal phagocytic cell activity chart; Figure 4 is the test animal natural killer cell activity chart; and Figure 5 is the test animal blood The content of non-specific antibodies IgA and IgM.

Claims (5)

A glucan and mushroom mushroom polysaccharide composition, comprising 20 to 50% by weight of β-polydextrose concentrate, 20 to 50% by weight of Ganoderma lucidum mycelium extract, 1 to 1% by weight 5% of Antrodia camphorata mycelium extract, 1 to 5% by weight of Cordyceps sinensis mycelium extract, 1 to 5% by weight of Mulberry fruit body extract, 1 to 5% by weight of Brazil Mushroom fruit body extract, 1 to 5% by weight of Maitake mushroom fruit body extract and 1 to 5% by weight of Hericium erinaceus fruit body extract. The glucan and mushroom mushroom polysaccharide composition according to claim 1, wherein the β-polydextrose concentrate contains a β-polydextrose concentrate of black yeast. A method for manufacturing a glucan and mushroom mushroom polysaccharide composition, which comprises a black yeast yeast cell fermentation product, a Ganoderma lucidum bacterial cell fermentation product, an Antrodia cinnamomea bacterial cell fermentation product, and a Chinese porangia spore The fermentation product of the fungus, the fruit body powder of a mulberry, the fruit body powder of a brazilian mushroom, the fruit body powder of a Maitake mushroom, and the fruit body powder of a hericium erinaceus, respectively, in a weight ratio of 10:1 to 40:1 with water Mix each proportion into a mixture, and then stir the aforementioned mixture at 70 to 100°C for 2 to 6 hours, perform pressure filtration to remove solids, and take the filtrate to heat and sterilize to obtain a β-polyglucose concentrate and a Ganoderma lucidum mycelium Body extract, an Antrodia camphorata mycelium extract, a Cordyceps sinensis mycelium extract, a mulberry fruit body extract, a Brazilian mushroom fruit body extract, a Maitake fruit body extract and a hericium erinaceus fruit body The extract, and then all the aforementioned extracts are blended and concentrated to obtain the glucan and lentinan composition. The method for manufacturing the glucan and mushroom mushroom polysaccharide composition according to claim 3, wherein the bacterial fermentation product of the black yeast, the bacterial fermentation product of the Ganoderma lucidum, the bacterial fermentation product of the Antrodia camphorata or The preparation step of the bacterial fermentation product of Mortierella sinensis consists of inoculating a strain of black yeast, a strain of Ganoderma lucidum, a strain of Antrodia cinnamomea or a strain of Mortierella chinensis respectively The carbon source is 0.5 to 5.0% by weight, the nitrogen source is 0.1 to 1.5% by weight, other trace substances, and the pH value is 5.0 to 6.5 in a medium, and the culture is carried out at a temperature of 20 to 30°C with aeration and stirring for 2 to 7 days. The method for manufacturing the glucan and mushroom mushroom polysaccharide composition according to claim 3, further comprising 44% by weight of the β-polydextrose concentrate and 44% by weight of the Ganoderma lucidum mycelium extraction Liquid, 2% by weight of the Antrodia camphorata mycelium extract, 2% by weight of the Cordyceps sinensis mycelium extract, 2% by weight of the Phellinus linteus fruit body extract, 2% by weight of the Brazilian mushroom seed The steps of reconciling and concentrating the physical extract, the 2% by weight of the Maitake mushroom fruit body extract and the 2% by weight of the Hericium erinaceus fruit body extract.

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