Functional solid beverage obtained by edible fungus fermentation and preparation method thereof

Abstract

The present invention belongs to the field of solid beverage preparation, and specifically relates to a functional solid beverage obtained by fermenting edible fungi and a preparation method thereof. The present invention performs bidirectional liquid fermentation on shiitake mushrooms, caragana and apple powder, and the obtained substances are compounded with the polysaccharide components of Agaricus bisporus and Flammulina velutipes to obtain a composite extract of edible fungi. A liquid culture medium prepared with a marine source extract is used, and Grifola frondosa is used for two fermentations to obtain a multi-component fermentation product. The composite extract of edible fungi and the multi-component fermentation product have a strong synergistic antioxidant effect. The milk pulp liquid provides preliminary flavor and color, and maltitol and citric acid give the functional solid beverage appropriate sweetness and sourness. The functional solid beverage obtained by fermenting edible fungi obtained by the present invention has fine powder, is easy to brew, has uniform color, rich aroma and rich layered feeling, is safe to eat, and has been verified by animal experiments to have a certain effect of enhancing immunity.

Description

Technical Field

[0001] The invention belongs to the field of solid beverage preparation, and particularly relates to a functional solid beverage obtained by fermenting edible fungi and a preparation method thereof. Background Art

[0002] Edible fungi are not only widely used as a food source for their unique taste and subtle flavor, but are also an important source of medicine in many countries for treating cancer, inflammation, liver damage, diabetes, and neuropathy. The most commonly cultivated edible fungi are Agaricus bisporus, Shiitake mushrooms, Oyster mushrooms, and Flammulina velutipes. Considering their health-promoting effects, edible fungi including Ganoderma lucidum, Antrodia cinnamomea, Cordyceps sinensis, Poria cocos, and Trichoderma lucidum have significant medicinal value. Nucleosides, flavonoids, polysaccharides, sterols, and terpenes are the most extensively studied functional active substances in edible fungi. These active substances exhibit pharmacological effects such as immunomodulation, antioxidant effects, antiviral effects, hypoglycemic and lipid-lowering effects, and anti-tumor effects.

[0003] Fermentation technology is an effective way to enhance the nutritional and health benefits of food. Solid-state fermentation refers to microbial fermentation using a solid matrix as raw material. The culture substrate exists in solid form and is not easily soluble in water, providing the microorganisms with the nutrients and growth environment they need. Liquid fermentation, also known as deep culture, is a process in which the nutrients and growth environment required for mycelial growth are prepared into a liquid culture medium, which is then inoculated with the bacterial strain for cultivation. Edible fungi are widely used in the development of functional foods due to their rich nutrition and low calorie content. Their fermentation process is simple and has a short growth cycle. Therefore, fermentation technology can be used to efficiently, stably, and rapidly separate and obtain the bioactive components in edible fungi, effectively increasing the yield of bioactive components in fermentation products and production efficiency, and can realize the industrial production of edible fungi. Summary of the Invention

[0004] Based on the above problems, the present invention develops and utilizes a variety of edible fungi, performs bidirectional liquid fermentation on shiitake mushrooms, caragana and apple powder, and the obtained substance is compounded with the polysaccharide components of Agaricus bisporus and Flammulina velutipes to obtain a composite extract of edible fungi, and at the same time, grisea bulb is inoculated on wheat grains for preliminary fermentation. The secondary fermentation adopts a liquid culture medium prepared by a marine source extract. The multi-component fermentation product obtained after the two fermentations has no fishy smell, and both the edible fungi composite extract and the multi-component fermentation product have good antioxidant effects, and under the formula of functional solid beverages, have a strong synergistic antioxidant effect. The milky fungus liquid of the present invention provides the preliminary flavor and color of the functional solid beverage, and maltitol and citric acid give the functional solid beverage suitable sweetness and acidity. The functional solid beverage finally obtained by fermentation of edible fungi has fine powder, is easy to brew, has uniform color, rich fragrance and rich layered feeling, is safe to eat, and has been verified by animal experiments to have a certain effect of enhancing immunity.

[0005] The present invention provides a functional solid beverage obtained by fermenting edible fungi. The functional solid beverage comprises the following raw materials in parts by mass: 8-12 parts of a composite edible fungus extract, 6-10 parts of a multi-component fermentation product, 1-3 parts of milk mushroom liquid, 0.8-1.5 parts of maltitol, 0.6-1.2 parts of locust bean gum, and 0.4-0.8 parts of citric acid.

[0006] The preparation method of the edible fungus composite extract is as follows:

[0007] L1. Wash and drain fresh, mature shiitake mushrooms, disinfect, cut into chunks, and inoculate onto a slant culture medium. Incubate at 24-26°C for 7-10 days until mycelium covers the entire slant, obtaining a stock culture. Inoculate this stock culture onto a plate culture medium and incubate at 25-28°C for 6-8 days. Centrifuge at 5000-5500g for 20-30 minutes. Wash the pellet three times with distilled water and dry at 60-65°C to obtain an activated culture.

[0008] L2. The activated strain obtained in step L1 was inoculated into the seed culture medium and cultured at 25-28°C for 2-3 days to obtain a shiitake mushroom culture liquid. Caragana, apple powder, and talc were added to the shiitake mushroom culture liquid and cultured at 25-28°C for 6-10 days. After the culture was completed, the filtrate was filtered, sterilized at high temperature, and naturally cooled to obtain a fermentation liquid. The pH of the fermentation liquid was adjusted to 4.0 and stirred at a speed of 100-150 rpm for 8-10 minutes. After the fermentation, the precipitate was collected, dissolved in distilled water, adjusted to pH 7.0, dialyzed for 24-36 hours, and then freeze-dried to obtain substance 1;

[0009] L3. Grind Agaricus bisporus and Flammulina velutipes into powder, sieve, and weigh to obtain a mixed powder in a mass ratio of 1:3-5. Weigh the mixed powder and deionized water in a mass-to-volume ratio of 60-80 g:1 L, heat to boiling for 2-4 hours, filter to obtain a filtrate, centrifuge the filtrate at 5000-6000 g for 20-30 minutes, mix the supernatant with anhydrous ethanol in a volume ratio of 1:3-5, stir until uniformly dispersed, and then keep at 4°C for 10-15 hours. Centrifuge at 8000-10000 g for 8-10 minutes. After the precipitate is washed with 75% ethanol, dried, and dissolved in ultrapure water to obtain Solution 1.

[0010] L4. Solution 1 obtained in step L1 was frozen at -20°C 8-9 times, each time for 9-10 hours. After each freezing, it was thawed at 15°C. The final thawed solution was centrifuged at 6000-8000g for 25-30 minutes, filtered through a 0.45μm filter membrane, and the filtrate was recorded as solution 2;

[0011] L5. Mix the substance 1 obtained in step L2 with the solution 2 obtained in step L4 at a mass-to-volume ratio of 1 g: 60-100 mL, adjust the pH to 8.0, and incubate in a water bath at 80-90°C for 24-30 h. After incubation, cool naturally to room temperature to obtain solution 3. Add sodium alginate to solution 3, stir until uniformly dispersed, and freeze-dry to obtain a powder, which is the edible fungus composite extract.

[0012] Preferably, during inoculation in step L2, the mass volume ratio of the activated strain: the seed culture medium is 1 g: 100-150 mL, and the seed culture medium is prepared as follows: 100 g of glucose, 15 g of magnesium sulfate heptahydrate, 12 g of potassium dihydrogen phosphate, and 5 L of water. The mass percentage concentration of Caragana in the shiitake mushroom culture liquid is 8-10%, the mass percentage concentration of apple powder in the shiitake mushroom culture liquid is 4-6%, and the mass percentage concentration of talc in the shiitake mushroom culture liquid is 1-1.5%. The talc is preferably 800-1250 mesh, and dialysis preferably uses a dialysis bag with a molecular weight cutoff of 7 KD.

[0013] Preferably, the sieving in step L3 is preferably performed using a 10-mesh sieve, and the concentration of the precipitate in solution 1 is 5-8 g / L.

[0014] Preferably, the concentration of sodium alginate in solution 3 in step L5 is 0.4-0.6 g / L.

[0015] The preparation method of the multi-fermentation product is as follows:

[0016] S1. Wash kelp and mussel meat, dry them, grind them, sieve them, and weigh them in a mass ratio of 1:2-4 to obtain a mixed powder. Add 15-20 times the mass of water to the mixed powder, heat to 80-90°C for 5-8 hours, and filter to obtain a filtrate, which is designated as Solution 4. Control the temperature of Solution 4 to 30-40°C, add an aqueous solution of hydroxypropyl-β-cyclodextrin, and stir at 120-160 rpm for 3-4 hours. Centrifuge at 8000-10000 g for 15-20 minutes. The supernatant is used as a liquid culture medium.

[0017] S2. The wheat grains were cleaned and soaked for 12-15h, then evenly spread out and sprayed with a sucrose solution. After standing for 45-60s, the wheat grains were evenly sprayed with a Grifola frondosa solution and incubated at 26-28°C for 3-4 days to obtain a preliminary fermentation product.

[0018] S3. The primary fermentation product obtained in step S2 is added to the liquid culture medium obtained in step S1. The amount of primary fermentation product added is calculated as 1200-1500 wheat grains per liter of liquid culture medium. The fermentation is stirred at 26-28°C and 80-100 rpm for 48-54 hours. After fermentation, steam sterilize the fermentation product, cool it naturally to room temperature, and filter it. The filtrate is decolorized to obtain a secondary fermentation product. The secondary fermentation product is concentrated to 0.3-0.4 times the original volume, which is recorded as solution 5. Solution 5 is freeze-dried to obtain a powdered multi-fermentation product.

[0019] Preferably, the sieving in step S1 is preferably a 2-mesh sieve, the volume ratio of solution 4: hydroxypropyl-β-cyclodextrin aqueous solution is 20-25:1, the mass percentage concentration of hydroxypropyl-β-cyclodextrin aqueous solution is 6-10%, and the liquid culture medium is prepared as follows: 10L supernatant, 300-350g soluble starch, 50-60g sucrose, 5-7g magnesium sulfate heptahydrate, 3-4g dipotassium hydrogen phosphate, 0.5-1g vitamin B1, and the pH is adjusted to 6.0.

[0020] Preferably, the mass percentage concentration of the sucrose aqueous solution in step S2 is 5-8%, and the method for obtaining the Grifola frondosa liquid is the same as that for obtaining the Lentinus edodes liquid.

[0021] Preferably, the decolorization and concentration in step S3 are performed using methods acceptable in the food preparation process.

[0022] The preparation method of the milk pulp bacteria liquid is as follows:

[0023] The milk fungus is cleaned, drained, and weighed and water is added according to a mass volume ratio of 1g:40-50mL. The mixture is processed by colloid mill to obtain a mixture with a fineness of 30-40μm. The mixture is heated to boiling and maintained for 10-15min. After the boiling, the mixture is centrifuged at a centrifugal force of 8000-10000g for 30-40min to obtain the supernatant, which is the milk fungus liquid.

[0024] The present invention provides a method for preparing a functional solid beverage obtained by fermenting edible fungi, the specific steps of which are as follows:

[0025] V1. The edible fungus composite extract and multi-fermentation mixture was mixed, 5-10 times the mass of the edible fungus composite extract was added to water, stirred until uniformly dispersed, the milk pulp liquid, maltitol, locust bean gum, citric acid was added to obtain a solution 6;

[0026] V2. The solution 6 obtained in step V1 is spray-dried. The centrifugal speed of the spray drying is 30,000-35,000 rpm, the inlet temperature is 150-160°C, and the outlet temperature is 70-80°C. After the end, a functional solid beverage is obtained.

[0027] The beneficial effects of the present invention are as follows:

[0028] The present invention develops and utilizes a variety of edible fungi, performs bidirectional liquid fermentation on shiitake mushrooms, caragana, and apple powder, obtains a substance 1 rich in small molecule proteins after treatment, adds talcum powder during the process to avoid excessive growth of shiitake mushroom mycelium, affects material transfer, adopts water extraction and alcohol precipitation combined with multiple freeze-thaw methods to obtain the polysaccharide components of Agaricus bisporus and Flammulina velutipes, and finally uses the wet Maillard reaction to compound the substance 1 rich in small molecule proteins with solution 2 to obtain an edible fungus composite extract, and adds sodium alginate to enhance the stability of the edible fungus composite extract. The present invention inoculates maitake mushrooms on wheat grains for primary fermentation, and the secondary fermentation uses a liquid culture medium prepared with a marine source extract. The multi-fermentation obtained after the two fermentations has no fishy smell, has certain antioxidant and anti-inflammatory effects, and can enhance the edible effect of the final functional solid beverage. The milk pulp fungus liquid of the present invention provides the initial flavor and color of the functional solid beverage. With the brewing of hot water, the edible fungus composite extract and the multi-fermentation will further release different levels of flavor, giving the functional solid beverage a better sensory experience. Experiments have verified that the edible fungus composite extract and multi-fermentation product of the present invention both have good antioxidant effects, and under the formula of the functional solid beverage provided by the present invention, have a strong synergistic antioxidant effect.

[0029] The functional solid beverage obtained by fermenting edible fungi obtained by the present invention has fine powder, is easy to brew, has uniform color, rich aroma and rich layered feeling, appropriate sweetness and sourness, is safe for consumption, and has been verified by animal experiments to have a certain effect of enhancing immunity. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0031] Figure 1 This is the result of DPPH free radical scavenging test;

[0032] Figure 2 ABTS + Free radical scavenging test results diagram;

[0033] Figure 3 This is an analysis chart of the antioxidant test of edible fungus compound extracts and multi-fermentation products;

[0034] Figure 4 This is a graph showing the results of the NK cell activity assay. DETAILED DESCRIPTION

[0035] In order to more clearly illustrate the overall concept of the present application, the following is a detailed description of the embodiments in conjunction with the accompanying drawings. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0036] Example 1: This example provides a method for preparing a composite extract of edible fungi, and the specific steps are as follows:

[0037] L1. Wash and drain fresh, mature shiitake mushrooms. Disinfect with 75% ethanol, then cut into 0.5 cm long and wide pieces. Inoculate the pieces onto a slant culture medium and incubate at 24°C for 10 days until mycelium has fully grown on the slant. Obtain the mother culture. Cut a 1 cm x 1 cm piece of the mother culture uniformly grown on the slant culture medium and inoculate it onto a plate culture medium (90 mm Petri dish). Incubate uniformly at 25°C for 8 days. Centrifuge at 5000 g for 20 minutes. Wash the pellet three times with distilled water and dry at 60°C to obtain the activated culture.

[0038] L2. Preparation of seed culture medium: 100 g of glucose, 15 g of magnesium sulfate heptahydrate, 12 g of potassium dihydrogen phosphate, and 5 L of water. The activated strain obtained in step L1 was inoculated into the seed culture medium. The mass volume ratio of the activated strain: the seed culture medium was 1 g: 100 mL. The culture was cultured at 25 ° C for 3 days to obtain a shiitake mushroom solution. The caragana, apple powder, and talc were added to the shiitake mushroom solution. The mass percentage concentrations of caragana, apple powder, and talc (800 mesh) in the shiitake mushroom solution were 8%, 4%, and 1%, respectively. The solution was cultured at 25 ° C for 10 days, filtered, sterilized at high temperature, and cooled naturally to obtain a fermentation broth. The fermentation broth was adjusted to pH 4.0 and stirred at 100 rpm for 8 min. Afterwards, the solution was centrifuged at 8000 g for 20 min. The precipitate was collected, dissolved in distilled water, adjusted to pH 7.0, and dialyzed for 24 h using a dialysis bag with a molecular weight cutoff of 7 KD and then freeze-dried to obtain substance 1.

[0039] L3. Grind Agaricus bisporus and Flammulina velutipes into powder, pass through a 10-mesh sieve, and weigh the mixture in a mass ratio of 1:3 to obtain a mixed powder. The mixed powder was then weighed with deionized water in a mass-to-volume ratio of 60 g:1 L. The mixture was heated to boiling for 2 h, filtered, and the filtrate obtained was centrifuged at 5000 g for 20 min. The supernatant was mixed with anhydrous ethanol in a volume ratio of 1:3, stirred until uniformly dispersed, and then placed at 4°C for 10 h. The mixture was then centrifuged at 8000 g for 8 min. The precipitate was washed with 75% ethanol, dried, and dissolved in ultrapure water to obtain Solution 1. The concentration of the precipitate in Solution 1 was 5 g / L.

[0040] Solution 1 obtained in step L1 was frozen at -20°C eight times, each time for 9 hours. After each freezing, it was thawed at 15°C. The final thawed solution was centrifuged at 6000 g for 25 minutes and filtered through a 0.45 μm filter. The filtrate was designated Solution 2.

[0041] L5. Mix the substance 1 obtained in step L2 with the solution 2 obtained in step L4 at a mass-to-volume ratio of 1 g:60 mL, adjust the pH to 8.0, and incubate in an 80°C water bath for 24 h. After incubation, naturally cool to room temperature to obtain solution 3. Add sodium alginate to solution 3 at a concentration of 0.4 g / L. Stir until uniformly dispersed, and freeze-dry to obtain a powder, which is the composite edible fungus extract.

[0042] Example 2: This example provides a method for preparing a composite extract of edible fungi, and the specific steps are as follows:

[0043] L1. Wash and drain fresh, mature shiitake mushrooms. Disinfect with 75% ethanol, cut into 0.5 cm long and wide blocks, and inoculate them onto a slant culture medium. Incubate at 25°C for 8 days until mycelium has completely grown on the slant, obtaining the mother culture. Cut a 1 cm x 1 cm piece of the mother culture uniformly grown on the slant culture medium and inoculate it onto a plate culture medium (90 mm Petri dish). Incubate uniformly at 26°C for 7 days. Centrifuge at 5200 g for 25 minutes. Wash the pellet three times with distilled water and dry at 62°C to obtain the activated culture.

[0044] L2. Preparation of seed culture medium: 100 g glucose, 15 g magnesium sulfate heptahydrate, 12 g potassium dihydrogen phosphate, 5 L water, the activated strain obtained in step L1 was inoculated into the seed culture medium, the activated strain: seed culture mass volume ratio was 1 g: 120 mL, cultured at 27 ° C for 2 days to obtain shiitake mushroom liquid, the caragana, apple powder, talc was added to the shiitake mushroom liquid, the mass percentage concentration of caragana, apple powder, talc (1000 mesh) in the shiitake mushroom liquid was 9%, 5%, 1.2%, respectively, cultured at 27 ° C for 8 days, filtered, sterilized at high temperature, and cooled naturally to obtain a fermentation broth, the fermentation broth was adjusted to pH 4.0, stirred at 120 rpm for 9 min, and then centrifuged at 9000 g for 25 min, the precipitate was collected, dissolved in distilled water, adjusted to pH 7.0, dialyzed for 30 h using a dialysis bag with a molecular weight cutoff of 7 KD, and then freeze-dried to obtain substance 1;

[0045] L3. Grind Agaricus bisporus and Flammulina velutipes into powder, pass through a 10-mesh sieve, and weigh the mixture in a mass ratio of 1:4 to obtain a mixed powder. The mixed powder was then weighed with deionized water in a mass-to-volume ratio of 70 g:1 L. The mixture was heated to boiling for 3 hours and filtered to obtain a filtrate. The filtrate was centrifuged at 5500 g for 24 minutes. The supernatant was mixed with anhydrous ethanol in a volume ratio of 1:4, stirred until uniformly dispersed, and then kept at 4°C for 12 hours. The mixture was then centrifuged at 9000 g for 9 minutes. The precipitate was washed with 75% ethanol, dried, and dissolved in ultrapure water to obtain Solution 1. The concentration of the precipitate in Solution 1 was 6 g / L.

[0046] Solution 1 obtained in step L1 was frozen at -20°C nine times for 9 h each time. After each freezing, it was thawed at 15°C. The final thawed solution was centrifuged at 7000 g for 28 min and filtered through a 0.45 μm filter. The filtrate was designated Solution 2.

[0047] L5. Mix the substance 1 obtained in step L2 with the solution 2 obtained in step L4 at a mass-to-volume ratio of 1 g:80 mL, adjust the pH to 8.0, and incubate in an 85°C water bath for 27 h. After incubation, naturally cool to room temperature to obtain solution 3. Add sodium alginate to solution 3 at a concentration of 0.5 g / L. Stir until uniformly dispersed, and freeze-dry to obtain a powder, which is the composite edible fungus extract.

[0048] Example 3: This example provides a method for preparing a composite extract of edible fungi, and the specific steps are as follows:

[0049] L1. Wash and drain fresh, mature shiitake mushrooms. Disinfect with 75% ethanol, then cut into 0.5 cm long and wide pieces. Inoculate the pieces onto a slant culture medium and incubate at 26°C for 7 days until mycelium covers the entire slant. Obtain the mother culture. Cut a 1 cm x 1 cm piece of the mother culture uniformly grown on the slant culture medium and inoculate it onto a plate culture medium (90 mm Petri dish). Incubate uniformly at 28°C for 6 days. Centrifuge at 5500 g for 30 minutes. Wash the pellet three times with distilled water and dry at 65°C to obtain the activated culture.

[0050] L2. Preparation of seed culture medium: 100 g glucose, 15 g magnesium sulfate heptahydrate, 12 g potassium dihydrogen phosphate, 5 L water, the activated strain obtained in step L1 was inoculated into the seed culture medium, the activated strain: seed culture medium mass volume ratio was 1 g: 150 mL, cultured at 28 ° C for 2 days to obtain shiitake mushroom liquid, the caragana, apple powder, talc was added to the shiitake mushroom liquid, the mass percentage concentrations of caragana, apple powder, and talc (1250 mesh) in the shiitake mushroom liquid were 10%, 6%, and 1.5%, respectively, cultured at 28 ° C for 6 days, filtered, sterilized at high temperature, and cooled naturally to obtain a fermentation broth, the fermentation broth was adjusted to pH 4.0, stirred at 150 rpm for 10 min, and then centrifuged at 10,000 g for 30 min. The precipitate was collected, dissolved in distilled water, adjusted to pH 7.0, dialyzed for 36 h using a dialysis bag with a molecular weight cutoff of 7 KD, and then freeze-dried to obtain substance 1;

[0051] L3. Grind Agaricus bisporus and Flammulina velutipes into powder, pass through a 10-mesh sieve, and weigh the mixture in a mass ratio of 1:5 to obtain a mixed powder. The mixed powder was then weighed with deionized water in a mass-to-volume ratio of 80 g:1 L, heated to boiling for 4 h, and filtered to obtain a filtrate. The filtrate was centrifuged at 6000 g for 30 min. The supernatant was mixed with anhydrous ethanol in a volume ratio of 1:5, stirred until uniformly dispersed, and then kept at 4°C for 15 h. Centrifuged at 10000 g for 10 min, the precipitate was washed with 75% ethanol, dried, and dissolved in ultrapure water to obtain Solution 1. The concentration of the precipitate in Solution 1 was 8 g / L.

[0052] Solution 1 obtained in step L1 was frozen at -20°C nine times, each time for 10 h. After each freezing, it was thawed at 15°C. The final thawed solution was centrifuged at 8000 g for 30 min and filtered through a 0.45 μm filter. The filtrate was designated Solution 2.

[0053] L5. Mix the substance 1 obtained in step L2 with the solution 2 obtained in step L4 at a mass-to-volume ratio of 1 g:100 mL, adjust the pH to 8.0, and incubate in a 90°C water bath for 30 h. After incubation, naturally cool to room temperature to obtain solution 3. Add sodium alginate to solution 3 to a concentration of 0.6 g / L. Stir until uniformly dispersed, and freeze-dry to obtain a powder, which is the composite edible fungus extract.

[0054] The slant culture medium involved in Examples 1-3 was prepared as follows: 200 g peeled potatoes, 20 g glucose, 20 g agar, 3 g potassium dihydrogen phosphate, 1.5 g magnesium sulfate heptahydrate, and 1 L water, and the pH was adjusted to 6.0.

[0055] The plate culture medium involved in Examples 1-3 was prepared as follows: 240 g potato extract, 35 g glucose, 1 g anhydrous magnesium sulfate, 2 g potassium dihydrogen phosphate, 15 g agar, and 1 L water, and the pH was adjusted to 6.5.

[0056] Example 4: This example provides a method for preparing a multi-component fermentation product, and the specific steps are as follows:

[0057] S1. Clean kelp and mussel meat, dry them, and grind them into powder. Pass them through a 2-mesh sieve and weigh them in a mass ratio of 1:2 to obtain a mixed powder. Add 15 times the mass of water to the mixed powder, heat to 80°C and maintain for 5 hours, and filter to obtain a filtrate. The filtrate is recorded as Solution 4. The temperature of Solution 4 is controlled to 30°C, and a 6% (by mass) hydroxypropyl-β-cyclodextrin aqueous solution is added. The volume ratio of Solution 4 to hydroxypropyl-β-cyclodextrin aqueous solution is 20:1. Stir at 120 rpm for 3 hours, and then centrifuge at 8000 g for 15 minutes. The supernatant is prepared into a liquid culture medium: 10 L of supernatant, 300 g of soluble starch, 50 g of sucrose, 5 g of magnesium sulfate heptahydrate, 3 g of potassium dihydrogen phosphate, and 0.5 g of vitamin B1. The pH is adjusted to 6.0.

[0058] S2. Wash and dry fresh, mature maitake mushrooms, sterilize with 75% ethanol, and cut into 0.5 cm long and wide blocks. Inoculate the blocks onto a slant medium and incubate at 24°C for 10 days. When the mycelium has fully grown on the slant, obtain the mother culture. Cut a 1 cm × 1 cm piece of the mother culture uniformly grown on the slant medium and inoculate it onto a plate medium (90 mm Petri dish). Incubate the slant medium uniformly at 25°C for 8 days. Centrifuge at 5000 g for 20 min. Wash the precipitate three times with distilled water and centrifuge at 60°C. Dry to obtain activated bacteria, inoculate the activated bacteria into seed culture medium at a mass volume ratio of 1 g of activated bacteria to 100 mL of seed culture medium, and culture at 25°C for 3 days to obtain a Grifola frondosa solution. Wash the wheat grains and soak them for 12 hours. Remove and evenly spread them, slightly separating them so that every 1,200 grains occupy a layer, and evenly spray them with a 5% by mass concentration of sucrose aqueous solution. After standing for 45 seconds, evenly spray them with Grifola frondosa solution and culture them at 26°C for 4 days to obtain a preliminary fermentation product.

[0059] S3. The primary fermentation product obtained in step S2 is added to the liquid culture medium obtained in step S1. The amount of primary fermentation product added is calculated as 1,200 wheat grains per liter of liquid culture medium. The fermentation is stirred at 26°C and 80 rpm for 54 hours. After fermentation, steam sterilization is performed, and the product is naturally cooled to room temperature and filtered. The filtrate is decolorized to obtain a secondary fermentation product. The secondary fermentation product is concentrated to 0.3 times the original volume, recorded as solution 5, and solution 5 is freeze-dried to obtain a powdered multi-fermentation product.

[0060] Example 5: This example provides a method for preparing a multi-component fermentation product, and the specific steps are as follows:

[0061] S1. Clean kelp and mussel meat, dry them, and grind them. After passing through a 2-mesh sieve, weigh them in a mass ratio of 1:3 to obtain a mixed powder. Add 18 times the mass of water to the mixed powder, heat to 85°C and maintain for 6 hours, and filter to obtain a filtrate. The filtrate is recorded as Solution 4. The temperature of Solution 4 is controlled to 35°C, and an 8% by mass concentration of hydroxypropyl-β-cyclodextrin aqueous solution is added. The volume ratio of Solution 4 to hydroxypropyl-β-cyclodextrin aqueous solution is 22:1. Stir at 140 rpm for 3.5 hours, and then centrifuge at 9000 g for 18 minutes. The supernatant is prepared into a liquid culture medium: 10 L of supernatant, 320 g of soluble starch, 55 g of sucrose, 6 g of magnesium sulfate heptahydrate, 3.5 g of potassium hydrogen phosphate, and 0.8 g of vitamin B1. The pH is adjusted to 6.0.

[0062] S2. Wash and dry fresh, mature maitake mushrooms, sterilize with 75% ethanol, and cut into 0.5 cm long and wide blocks. Inoculate the blocks onto a slant medium and incubate at 25°C for 8 days. When the mycelium has fully grown on the slant, obtain the mother culture. Cut a 1 cm × 1 cm piece of the mother culture uniformly grown on the slant medium and inoculate it onto a plate medium (90 mm Petri dish). Incubate the slant medium uniformly at 26°C for 7 days. Centrifuge at 5200 g for 25 minutes. Wash the precipitate three times with distilled water and incubate at 62°C. Dry to obtain activated bacteria, inoculate the activated bacteria into seed culture medium at a mass volume ratio of 1 g of activated bacteria to 120 mL of seed culture medium, and culture at 27°C for 2 days to obtain a Grifola frondosa solution. Wash the wheat grains and soak them for 14 hours. Remove and evenly spread them, slightly separating them so that every 1,400 grains occupy a layer, and evenly spray them with a 6% mass percentage concentration of sucrose aqueous solution. After standing for 50 seconds, evenly spray them with Grifola frondosa solution and culture them at 27°C for 4 days to obtain a preliminary fermentation product.

[0063] S3. The primary fermentation product obtained in step S2 is added to the liquid culture medium obtained in step S1. The amount of primary fermentation product added is calculated as wheat grains, with 1,400 wheat grains added per liter of liquid culture medium. The fermentation is stirred at 27°C and 90 rpm for 52 hours. After fermentation, steam sterilization is performed, and the product is naturally cooled to room temperature and filtered. The filtrate is decolorized to obtain a secondary fermentation product. The secondary fermentation product is concentrated to 0.35 times the original volume, recorded as solution 5, and solution 5 is freeze-dried to obtain a powdered multi-fermentation product.

[0064] Example 6: This example provides a method for preparing a multi-component fermentation product, and the specific steps are as follows:

[0065] S1. Clean kelp and mussel meat, dry them, and grind them into powder. After passing through a 2-mesh sieve, weigh them in a mass ratio of 1:4 to obtain a mixed powder. Add 20 times the mass of water to the mixed powder, heat to 90°C and maintain for 8 hours, and filter to obtain a filtrate. The filtrate is recorded as solution 4. The temperature of solution 4 is controlled to 40°C, and a 10% mass concentration of hydroxypropyl-β-cyclodextrin aqueous solution is added. The volume ratio of solution 4 to hydroxypropyl-β-cyclodextrin aqueous solution is 25:1. Stir at 160 rpm for 4 hours, and then centrifuge at 10,000 g for 20 minutes. The supernatant is prepared into a liquid culture medium: 10 L of supernatant, 350 g of soluble starch, 60 g of sucrose, 7 g of magnesium sulfate heptahydrate, 4 g of potassium dihydrogen phosphate, and 1 g of vitamin B1. The pH is adjusted to 6.0.

[0066] S2. Wash and dry fresh, mature maitake mushrooms, sterilize with 75% ethanol, cut into 0.5 cm long and wide pieces, and inoculate them into slant culture medium. Incubate at 26°C for 7 days until mycelium has completely covered the slant. Obtain the mother culture. Cut 1 cm × 1 cm pieces of the mother culture uniformly grown on the slant culture medium and inoculate them into plate culture medium (90 mm Petri dish). Incubate uniformly at 28°C for 6 days. Centrifuge at 5500 g for 30 min. Wash the precipitate three times with distilled water and centrifuge at 65°C. Dry to obtain activated bacteria, inoculate the activated bacteria into seed culture medium at a mass volume ratio of 1 g of activated bacteria to 150 mL of seed culture medium, and culture at 28°C for 2 days to obtain a Grifola frondosa solution. Wash the wheat grains and soak them for 15 hours. Remove and evenly spread them, slightly separating them so that every 1,500 grains occupy a layer, and evenly spray them with an 8% by mass concentration of sucrose aqueous solution. After standing for 60 seconds, evenly spray them with the Grifola frondosa solution and culture them at 28°C for 3 days to obtain a preliminary fermentation product.

[0067] S3. The primary fermentation product obtained in step S2 is added to the liquid culture medium obtained in step S1. The amount of primary fermentation product added is calculated as wheat grains, with 1,500 wheat grains added per liter of liquid culture medium. The fermentation is stirred at 28°C and 100 rpm for 48 hours. After fermentation, steam sterilization is performed, and the product is naturally cooled to room temperature and filtered. The filtrate is decolorized to obtain a secondary fermentation product. The secondary fermentation product is concentrated to 0.4 times the original volume, recorded as solution 5, and solution 5 is freeze-dried to obtain a powdered multi-fermentation product.

[0068] The preparation of the slant culture medium, plate culture medium, and seed culture medium in Examples 4-6 is the same as that in Examples 1-3.

[0069] Example 7: This example provides a method for preparing a functional solid beverage obtained by fermentation of edible fungi, the specific contents of which are as follows:

[0070] The functional solid beverage obtained by edible fungus fermentation in this embodiment comprises the following raw materials in parts by weight: 8 parts of a composite edible fungus extract, 6 parts of a multi-component fermentation product, 1 part of a milk fungus liquid, 0.8 parts of maltitol, 0.6 parts of locust bean gum, and 0.4 parts of citric acid. The composite edible fungus extract is prepared using the method of Example 1, and the multi-component fermentation product is prepared using the method of Example 4.

[0071] The preparation method of the milk pulp liquid is as follows: the milk pulp is cleaned, drained, and then water is weighed and added according to a mass volume ratio of 1g:40mL, and the mixture is processed by colloid mill to obtain a mixture with a fineness of 30μm. The mixture is heated to boiling and maintained for 10 minutes, and then centrifuged at a centrifugal force of 8000g for 30 minutes to obtain a supernatant, which is the milk pulp liquid;

[0072] The preparation method of the functional solid beverage obtained by edible fungus fermentation comprises the following specific steps:

[0073] V1. The edible fungus composite extract and multi-fermentation product were mixed, 5 times the mass of the edible fungus composite extract was added to water, stirred until uniformly dispersed, the milk pulp liquid, maltitol, locust bean gum, citric acid was added to obtain a solution 6;

[0074] V2. The solution 6 obtained in step V1 was spray-dried at a centrifugal speed of 30,000 rpm, an inlet temperature of 150°C, and an outlet temperature of 70°C. After the spray drying, a functional solid beverage was obtained.

[0075] Example 8: This example provides a method for preparing a functional solid beverage obtained by fermentation of edible fungi, the specific contents of which are as follows:

[0076] The functional solid beverage obtained by edible fungus fermentation in this embodiment comprises the following raw materials in parts by mass: 10 parts of a composite edible fungus extract, 8 parts of a multi-component fermentation product, 2 parts of milk fungus liquid, 1 part of maltitol, 0.8 parts of locust bean gum, and 0.5 parts of citric acid. The composite edible fungus extract is prepared using the method of Example 2, and the multi-component fermentation product is prepared using the method of Example 5.

[0077] The preparation method of the milk pulp liquid is as follows: the milk pulp is cleaned, drained, and then water is weighed and added according to a mass volume ratio of 1g:45mL, and the mixture is processed by colloid mill to obtain a mixture with a fineness of 35μm. The mixture is heated to boiling and maintained for 12 minutes, and then centrifuged at a centrifugal force of 9000g for 34 minutes to obtain a supernatant, which is the milk pulp liquid;

[0078] The preparation method of the functional solid beverage obtained by edible fungus fermentation comprises the following specific steps:

[0079] V1. The edible fungus composite extract and multi-fermentation product were mixed, 8 times the mass of the edible fungus composite extract was added to water, stirred until uniformly dispersed, the milk pulp liquid, maltitol, locust bean gum, citric acid was added to obtain a solution 6;

[0080] V2. The solution 6 obtained in step V1 was spray-dried at a centrifugal speed of 32,000 rpm, an inlet temperature of 155°C, and an outlet temperature of 72°C. After the spray drying, a functional solid beverage was obtained.

[0081] Example 9: This example provides a method for preparing a functional solid beverage obtained by fermentation of edible fungi, the specific contents of which are as follows:

[0082] The functional solid beverage obtained by edible fungus fermentation in this embodiment comprises the following raw materials in parts by weight: 12 parts of a composite edible fungus extract, 10 parts of a multi-component fermentation product, 3 parts of milk fungus liquid, 1.5 parts of maltitol, 1.2 parts of locust bean gum, and 0.8 parts of citric acid. The composite edible fungus extract is prepared using the method of Example 3, and the multi-component fermentation product is prepared using the method of Example 6.

[0083] The preparation method of the milk pulp liquid is as follows: the milk pulp is cleaned, drained, and then water is weighed and added according to a mass volume ratio of 1g:50mL, and the mixture is processed by colloid mill to obtain a mixture with a fineness of 40μm. The mixture is heated to boiling and maintained for 15 minutes, and then centrifuged at a centrifugal force of 10000g for 40 minutes to obtain a supernatant, which is the milk pulp liquid;

[0084] The preparation method of the functional solid beverage obtained by edible fungus fermentation comprises the following specific steps:

[0085] V1. The edible fungus composite extract and multi-fermentation product were mixed, 10 times the mass of the edible fungus composite extract was added to water, stirred until uniformly dispersed, the milk pulp liquid, maltitol, locust bean gum, citric acid was added to obtain a solution 6;

[0086] V2. The solution 6 obtained in step V1 was spray-dried at a centrifugal speed of 35,000 rpm, an inlet temperature of 160°C, and an outlet temperature of 80°C. After the spray drying, a functional solid beverage was obtained.

[0087] The maltitol, locust bean gum, and citric acid used in Examples 7-9 are all food grade and purchased from Zhengzhou Kunli Food Additive Co., Ltd., and the shiitake mushrooms, Agaricus bisporus, Flammulina velutipes, Grifola frondosa, and Milk Fungus used in Examples 1-9 are all edible fungi produced by Dalian Kangzhiyuan Biotechnology Co., Ltd.

[0088] Experimental test:

[0089] Solid beverage sensory scoring rules:

[0090] With reference to the national standard GB / T 29602-2013, "Solid Beverages," the sensory evaluation criteria for solid beverages shown in Table 1 were developed. The functional solid beverages obtained by fermenting edible fungi in Examples 7-9 were evaluated. Taking Example 7 as an example, 3 g of the functional solid beverage obtained by fermenting edible fungi was added to 350 mL of hot water at temperatures of 60, 70, 80, 90, and 100°C, respectively. After brewing, the beverage was briefly stirred until evenly dispersed. The brewing procedures for Examples 8 and 9 were the same as those for Example 7. Ten food professionals were invited to rate the functional solid beverages obtained by fermenting edible fungi based on Table 1 for appearance, color, flavor, and mouthfeel. The flavor score was limited to within five minutes of infusion, with a total score of 100.

[0091] Table 1 Sensory scoring criteria for solid beverages

[0092]

[0093] Table 2 Sensory evaluation results of functional solid beverages obtained in Examples 7-9 at different temperatures

[0094]

[0095] The sensory scoring results of the functional solid beverages obtained by fermenting edible fungi in Examples 7-9 are shown in Table 2. As can be seen from Table 2, the functional solid beverages obtained by fermenting edible fungi in Examples 7-9 performed best in the four scoring items of appearance, color, flavor and taste when brewed in hot water at 80°C or 90°C. When brewed in hot water at 60°C, the powder cannot dissolve quickly in a short time, but more stirring can alleviate this problem.

[0096] Antioxidant test:

[0097] The antioxidant test was carried out using the edible fungus composite extract obtained in Examples 1-3 and the multi-component fermentation products obtained in Examples 4-6. The antioxidant test mainly involved DPPH free radical scavenging test and ABTS + Free radical scavenging test, the specific contents are as follows:

[0098] 1) DPPH free radical scavenging test:

[0099] Use anhydrous ethanol to dissolve DPPH to obtain a mother solution with a concentration of 0.2 mM, store in the dark, and use within 24 hours. Prepare solutions of edible fungi composite extract / multi-fermentation product with concentrations of 2, 4, 8, 16, and 32 mg / mL. Pipette 2 mL of each solution into a centrifuge tube, add 2 mL of DPPH mother solution to each centrifuge tube, mix thoroughly, and react in the dark for 25 minutes. Measure the absorbance at 517 nm using a UV spectrophotometer, and calculate the DPPH free radical scavenging rate according to the following formula:

[0100] DPPH free radical scavenging rate = (OD0-(OD1-OD2)) / OD0×100%;

[0101] OD0 is the absorbance value of the control group, OD1 is the absorbance value of the sample group, OD2 is the absorbance value of the blank group, the control group is mother solution + anhydrous ethanol, and the blank group is sample solution + anhydrous ethanol.

[0102] 2) ABTS + Free radical scavenging test:

[0103] ABTS+ (7mM, 5mL) was mixed with potassium persulfate solution (140mM, 80μL) to obtain a mother solution, which was stored in the dark at room temperature. Before the test, the mother solution was diluted with phosphate buffer to an absorbance of 0.6 to obtain a mother solution dilution. The edible fungus composite extract / multi-fermentation product was prepared into solutions with concentrations of 0.05, 0.1, 0.2, 0.4, and 0.8mg / mL. 0.4mL of the solution at each concentration was aspirated and 3.6mL of the mother solution dilution was added. The mixture was thoroughly mixed and stored in the dark at room temperature for 6min. The absorbance at 734nm was measured and the ABTS was calculated according to the following formula: + Free radical scavenging rate:

[0104] ABTS + Free radical scavenging rate = (OD0-(OD1-OD2)) / OD0×100%;

[0105] OD0 is the absorbance value of the control group, OD1 is the absorbance value of the sample group, OD2 is the absorbance value of the blank group, the control group is the mother solution diluent + water, and the blank group is the sample solution + anhydrous ethanol. Parallel groups have been set up for the above experiments.

[0106] The results of the antioxidant test were as follows Figure 1 and Figure 2 As shown by Figure 1 and Figure 2 It can be seen that the edible fungus composite extracts obtained in Examples 1-3 and the multi-component fermentation products obtained in Examples 4-6 have high DPPH free radical scavenging rates and ABTS + The free radical scavenging rate proves that the edible fungus composite extracts obtained in Examples 1-3 and the multi-component fermentation products obtained in Examples 4-6 have strong antioxidant capacity.

[0107] The DPPH free radical scavenging test was used to study the antioxidant effects of different concentrations of edible fungus composite extracts (Example 2) and multi-fermentation products (Example 4). The experimental results were analyzed using the SynergyFinder program. Figure 3 As shown, Figure 3The 2D synergy diagram obtained by the ZIP model is displayed. In the 2D synergy diagram, red indicates the synergistic dose area and green indicates the antagonistic dose area. According to experience, a synergy score <-10 means that the interaction between the two substances is antagonistic, a synergy score between -10 and 10 means that the interaction between the two substances may be additive, and a synergy score >10 means that the interaction between the two substances is synergistic. Figure 3 It can be seen that the edible fungus compound extract is at 1-6 mg / mL, and the multi-fermentation product is at 1-5 mg / mL, and the two have a better synergistic antioxidant effect when paired with each other. Further considering the overall effect of the edible fungus compound extract and the multi-fermentation product after adding them to the functional solid beverage, as well as the concentration after brewing, the present invention clarifies the mass fraction of the edible fungus compound extract and the multi-fermentation product in the functional solid beverage, and recommends brewing the functional solid beverage with 3g:300-350mL hot water. At this time, not only is the synergistic antioxidant effect of the edible fungus compound extract and the multi-fermentation product better, but the flavor of the functional solid beverage is also better, with rich layers and a suitable taste.

[0108] 3. Verification of immunity-related effects:

[0109] SPF-grade ICR mice weighing 18-22g were housed in a clean environment, maintained at 22±2°C, and maintained on a 12-hour day / night cycle. After a week of adaptive feeding, immunity-related tests were performed. The mice were randomly divided into four groups: a control group (sterile water), a low-dose group (29.2mL / kg BW), a medium-dose group (58.3mL / kg BW), and a high-dose group (166.7mL / kg BW). These doses correspond to 5, 10, and 20 times the recommended human dose, respectively, of 350mL (based on a 60kg adult). The functional solid beverage obtained by fermenting the edible fungi obtained in Examples 7-9 was added to water (3g:350mL) to prepare a beverage. The beverage was then administered orally to the mice and concentrated using a rotary evaporator. The dose per mouse was 0.4mL.

[0110] The test validation projects involved mouse thymus and spleen index, thymus and spleen morphological observation, antibody-producing cell detection, serum hemolysin determination, delayed-type hypersensitivity test, mouse carbon clearance test and NK cell activity assay. The test methods and results of the delayed-type hypersensitivity test and NK cell activity assay are shown below:

[0111] 1) Delayed hypersensitivity test:

[0112] Mice were intraperitoneally injected with 2% (V / V) SRBC (0.2 mL / mouse). Four days after sensitization, the thickness of the left hind paw plantar was measured. Then, 20% (V / V) SRBC (20 μL / mouse) was subcutaneously injected at the measurement site. The thickness of the left hind paw plantar was measured 24 h after injection. The same site was measured three times and the average value was taken.

[0113] Table 3 Results of delayed-type hypersensitivity test in mice

[0114]

[0115] The results of the delayed-type hypersensitivity test in mice are shown in Table 3. As can be seen from Table 3, the paw swelling of mice in the low-, medium-, and high-dose groups of the functional solid beverage obtained by fermentation of edible fungi in Example 7 was greater than that in the control group, and the difference was significant (P<0.001 (***)). Similarly, the paw swelling of mice in the low-, medium-, and high-dose groups of the functional solid beverage obtained by fermentation of edible fungi in Examples 8 and 9 was also significantly different from that in the control group (P<0.001, or P<0.05 (**)).

[0116] 2) NK cell activity assay:

[0117] Mouse lymphoma cells (YAC-1 cells) were used as target cells. They were subcultured 24 h before the experiment and washed three times with Hank's solution before use. The cell number was adjusted to 4 × 10 cells using RPMI1640 complete culture medium. 5 / mL. The effector cells are spleen cells. Specifically, the spleen is obtained aseptically and placed in a small dish containing an appropriate amount of sterile Hank's solution. The spleen is gently ground with tweezers to make a single-cell suspension, and washed twice with Hank's solution, centrifuged at 1000rpm for 10min each time. The supernatant is discarded and the cell slurry is flicked off. 0.5mL of sterile water is added for 20s. After the red blood cells are lysed, 0.5mL of 2x Hank's solution and 8mL of Hanks solution are added, and centrifuged at 1000rpm for 10min. Resuspend with 1mL of RPMI1640 complete culture medium containing 10% calf serum, dilute with 1% glacial acetic acid and count, and count the number of living cells (more than 95%) with trypan blue staining. Finally, the cell concentration is adjusted to 2×10 with RPMI1640 complete culture medium. 7 pieces / mL.

[0118] In a U-shaped 96-well plate, 100 μL of target cells and 100 μL of effector cells were added as reaction wells. A target cell natural release well (100 μL of target cells and culture medium) and a target cell maximum release well (100 μL of target cells and 1% NP40) were also set up. Three replicates were set up for each of these three wells. The plates were incubated at 37°C, 5% CO2 for 4 hours. Afterward, the 96-well plate was centrifuged at 1500 rpm for 5 minutes. 100 μL of the supernatant was aspirated from each well and placed in a flat-bottomed 96-well plate. 100 μL of LDH matrix solution was added and the reaction was continued at room temperature (25°C) for 8 minutes. 30 μL of 1 mol / L hydrochloric acid was added to each well. The OD value was measured at 490 nm, and NK cell activity was calculated according to the following formula:

[0119] NK cell activity = (OD1-OD2) / (OD3-OD2) × 100%;

[0120] OD1 is the OD value of the natural pore, OD2 is the OD value of the natural release pore, and OD3 is the OD value of the maximum release pore.

[0121] NK cell activity assay results Figure 4 As shown by Figure 4 It can be seen that the NK cell activity of the medium and high dose groups of the functional solid beverage obtained by fermentation of edible fungi in Example 7 was significantly higher than that of the control group. The same pattern was also seen in the functional solid beverages obtained by fermentation of edible fungi in Examples 8 and 9. Therefore, the functional solid beverages obtained by fermentation of edible fungi in Examples 7-9 have a positive effect on the nonspecific immunity of mice.

[0122] The results of the mouse delayed-type hypersensitivity test and the NK cell activity assay test were both positive, which preliminarily indicates that the functional solid beverages obtained by fermenting edible fungi obtained in Examples 7-9 have a certain effect of enhancing immunity.

[0123] The food safety of the functional solid beverages obtained by fermentation of edible fungi in Examples 7-9 has been verified by acute toxicity tests and short-term feeding tests. The mice did not show any abnormalities during the test period, and there was no difference in body weight and organs such as heart, liver, and kidney compared with the control group.

[0124] The above descriptions are merely several embodiments of the present invention and do not constitute any form of limitation to the present invention. Although the present invention is disclosed as above in terms of preferred embodiments, they are not intended to limit the present invention. Any technician familiar with the present profession who, without departing from the scope of the technical solution of the present invention, makes slight changes or modifications using the technical contents disclosed above are equivalent to equivalent implementation cases and fall within the scope of protection of the technical solution of the present invention.

Claims

1. A functional solid beverage obtained by fermentation of edible fungi, characterized in that: The functional solid beverage obtained by edible fungus fermentation comprises the following raw materials in parts by mass: 8-12 parts of edible fungus composite extract, 6-10 parts of multi-fermentation product, 1-3 parts of milk pulp fungus liquid, 0.8-1.5 parts of maltitol, 0.6-1.2 parts of locust bean gum, and 0.4-0.8 parts of citric acid; The preparation method of the edible fungus composite extract is as follows: L1. Wash and drain fresh, mature shiitake mushrooms, disinfect, and cut into chunks. Inoculate the slant medium until mycelium grows all over the slant to obtain a stock culture. Inoculate the stock culture onto a plate medium. After incubation, centrifuge, wash the precipitate with distilled water, and dry at 60-65°C to obtain an activated culture. L2. The activated strain obtained in step L1 was inoculated into a seed culture medium to obtain a shiitake mushroom culture solution. Caragana, apple powder, and talc were added to the shiitake mushroom culture solution and the culture was continued. After the culture was completed, the solution was filtered, sterilized at high temperature, and naturally cooled to obtain a fermentation broth. The pH of the fermentation broth was adjusted to 4.0 and stirred. After the fermentation was completed, the precipitate was collected and dissolved in distilled water, the pH was adjusted, and the solution was dialyzed and freeze-dried to obtain substance 1. L3. Grind Agaricus bisporus and Flammulina velutipes into powder, sieve, and weigh to obtain a mixed powder in a mass ratio of 1:3-5. The mixed powder and deionized water are weighed in a mass-to-volume ratio of 60-80 g:1 L. The mixture is heated to boiling and maintained. Filter to obtain a filtrate, centrifuge the filtrate, and mix the supernatant with anhydrous ethanol in a volume ratio of 1:3-5. Stir until uniformly dispersed, then maintain at 4°C and centrifuge. The precipitate is washed with 75% ethanol, dried, and dissolved in ultrapure water to obtain Solution 1. L4. Solution 1 obtained in step L1 was frozen at -20°C 8-9 times, each time for 9-10 hours, and thawed at 15°C after each freezing. The final thawed solution was centrifuged and filtered through a 0.45 μm filter membrane. The filtrate was recorded as solution 2; L5. The substance 1 obtained in step L2 was mixed with the solution 2 obtained in step L4 at a mass volume ratio of 1 g: 60-100 mL, the pH was adjusted to 8.0, and the mixture was incubated in a water bath. After the mixture was cooled naturally to room temperature, a solution 3 was obtained. Sodium alginate was added to the solution 3, stirred until uniformly dispersed, and freeze-dried to obtain a powder which was the composite edible fungus extract. The preparation method of the multi-fermentation product is as follows: S1. Wash kelp and mussel meat, dry them, grind them, sieve them, and weigh them in a mass ratio of 1:2-4 to obtain a mixed powder. Add 15-20 times the mass of the mixed powder with water, heat and maintain the mixture, and filter to obtain a filtrate, which is designated as Solution 4. Control the temperature of Solution 4 to 30-40°C, add a hydroxypropyl-β-cyclodextrin aqueous solution, stir, and centrifuge. The supernatant is used to prepare the liquid culture medium. S2. The wheat grains were cleaned and soaked, and then evenly spread out after being removed, and evenly sprayed with a sucrose aqueous solution. After standing for 45-60s, the Grifola frondosa solution was evenly sprayed, cultured, and a preliminary fermentation product was obtained; S3. The primary fermentation product obtained in step S2 is added to the liquid culture medium obtained in step S1. The amount of primary fermentation product added is calculated as 1200-1500 wheat grains per liter of liquid culture medium. The fermentation is then steam sterilized, cooled naturally to room temperature, and filtered. The filtrate is decolorized to obtain a secondary fermentation product. The secondary fermentation product is concentrated to 0.3-0.4 times its original volume, designated as solution 5. Solution 5 is freeze-dried to obtain a powdered multi-component fermentation product. The preparation method of the milk pulp bacteria liquid is as follows: The milk fungus was cleaned, drained, and then weighed and added with water according to a mass volume ratio of 1g:40-50mL. The mixture was processed by colloid mill to obtain a mixture with a fineness of 30-40μm. The mixture was heated to boiling and maintained, and then centrifuged to obtain the supernatant, which is the milk fungus liquid.

2. The functional solid beverage obtained by fermentation of edible fungi according to claim 1, characterized in that: During inoculation in step L2, the mass volume ratio of activated bacteria to seed culture medium is 1 g: 100-150 mL, and the mass percentage concentrations of caragana, apple powder, and talc in the shiitake mushroom culture liquid are 8-10%, 4-6%, and 1-1.5%, respectively.

3. The functional solid beverage obtained by fermentation of edible fungi according to claim 2, characterized in that: The concentration of the precipitate in solution 1 in step L3 is 5-8 g / L, and the concentration of sodium alginate in solution 3 in step L5 is 0.4-0.6 g / L.

4. The functional solid beverage obtained by fermentation of edible fungi according to claim 3, characterized in that: In step S1, the volume ratio of solution 4 to the hydroxypropyl-β-cyclodextrin aqueous solution is 20-25:1, and the mass percentage concentration of the hydroxypropyl-β-cyclodextrin aqueous solution is 6-10%.

5. The functional solid beverage obtained by fermentation of edible fungi according to claim 4, characterized in that: The mass percentage concentration of the sucrose aqueous solution in step S2 is 5-8%.

6. A method for preparing a functional solid beverage obtained by fermentation of edible fungi according to any one of claims 1 to 5, characterized in that: The specific steps are as follows: V1. The edible fungus composite extract and multi-fermentation mixture was mixed, 5-10 times the mass of the edible fungus composite extract was added to water, stirred until uniformly dispersed, the milk pulp liquid, maltitol, locust bean gum, citric acid was added to obtain a solution 6; V2. The solution 6 obtained in step V1 is spray-dried to obtain a functional solid beverage.

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