Preparation method of edible fungus mycelium fermentation liquid, fermentation liquid and application thereof

Abstract

The present application relates to the field of fermentation technology, and specifically discloses a method for preparing a mycelium fermentation liquid of edible fungi, the fermentation liquid and an application thereof. The method for preparing the mycelium fermentation liquid of edible fungi comprises taking mycelium of edible fungi and inoculating it on a PDA culture medium under a sterile environment, culturing until the mycelium grows to more than 3 / 5 of the plate, and obtaining mycelium seeds of edible fungi for standby use; preparing a fermentation medium and sterilizing it, taking mycelium seeds of edible fungi and inoculating them into the fermentation medium under a sterile environment, culturing with shaking for 4-8 days, treating the fermentation liquid and collecting it, and obtaining the mycelium fermentation liquid of edible fungi; the method for preparing the mycelium fermentation liquid of edible fungi is simple and convenient to operate, safe and controllable, and a large amount of mycelium of edible fungi can be obtained in a short time. The content of adenine nucleotide, an effective ingredient, in the metabolites of the fermentation liquid is high and the quality is stable. The obtained mycelium fermentation liquid of edible fungi is used in the preparation of food, medicine, condiment and feed to cover up the bitter taste.

Description

Technical Field

[0001] The present application relates to the field of fermentation technology, and more specifically, to a method for preparing edible fungus mycelium fermentation liquid, the fermentation liquid and applications thereof. Background Art

[0002] Bitterness itself is an unpleasant taste sensation, which can be tasted at a very low threshold (quinine 0.005%), and bitterness lasts longer than other taste sensations. Pure bitterness is the fundamental reason why consumers refuse to eat bitter substances. Usually, bitterness masking is divided into three steps, including "bitterness suppression", "sweetening" and "flavoring", among which "bitterness suppression" is the fundamental and key, and "sweetening" and "flavoring" play an auxiliary and reinforcing role. At present, the main method used at home and abroad to mask bitterness is to add sweeteners. Although the bitterness is reduced to a certain extent, it cannot meet the consumer's demand for green and healthy food. In addition, traditional sweeteners such as acesulfame potassium and aspartame, while providing sweetness to food, will bring obvious bitterness and metallic taste to the food. Therefore, sweeteners or fragrances alone cannot fundamentally achieve the purpose of masking bitterness.

[0003] Edible fungi are a type of edible large fungi with fleshy or gelatinous fruiting bodies. Edible fungi have high edible and medicinal value and are rich in amino acids, proteins, sugars, lipids, vitamins, mineral elements, nucleotides and other nutrients. Among them, adenine nucleotide (AMP for short) in nucleotides has the property of inhibiting bitterness after testing on humans. AMP can compete with bitter substances for bitter receptors or block the transmission of bitter signals to the brain, thereby weakening the perception of bitterness and effectively masking the bitterness of food.

[0004] However, the current extraction of active ingredients of edible fungi mainly relies on artificially cultivated fruiting bodies as raw materials. The cultivation process of edible fungi fruiting bodies is cumbersome and time-consuming, with high environmental requirements and harsh management requirements. It is difficult to control product quality and difficult to mass produce. In addition, the composition and content of active ingredients extracted from different batches vary greatly. Cultivating edible fungi mycelium using liquid fermentation technology can not only increase its biomass, but also make the cultivation method more convenient, safe and controllable, and can also obtain a large amount of mycelium and valuable metabolites in a short time. However, the AMP content in edible fungi mycelium cultivated by existing liquid fermentation technology is low, and the quality stability is poor. It needs to be used with sweeteners at the same time to achieve a better effect of enhancing or improving the flavor of food. Although it can reduce the amount of sweeteners to a certain extent, it still cannot meet consumers' requirements for green and healthy food. Therefore, in order to obtain a safe, efficient, universal, stable and inexpensive bitterness blocker, the present application provides a method for preparing an edible fungus mycelium fermentation liquid, a fermentation liquid and its application. Summary of the invention

[0005] In order to solve the problems of low AMP content and poor quality stability in edible fungus mycelium cultured by liquid fermentation technology in related technologies, the present application provides a method for preparing edible fungus mycelium fermentation liquid, fermentation liquid and application thereof.

[0006] In a first aspect, the present application provides a method for preparing edible fungus mycelium fermentation liquid, using the following technical scheme:

[0007] A method for preparing edible fungus mycelium fermentation liquid comprises the following preparation steps:

[0008] S1. Preparation of edible fungus mycelium seeds: inoculate edible fungus mycelium onto PDA medium under a sterile environment, and culture at 25-28° C. until mycelium grows to more than 3 / 5 of the plate, and obtain edible fungus mycelium seeds for later use;

[0009] S2, preparing edible fungus mycelium fermentation liquid: preparing a fermentation medium and sterilizing it, taking the edible fungus mycelium seeds prepared in step S1 and inoculating them into the fermentation medium under a sterile environment, shaking and culturing at 24-30° C. for 4-8 days, treating the fermentation liquid and collecting it, and obtaining the edible fungus mycelium fermentation liquid.

[0010] By adopting the above technical scheme, the preparation method of the edible fungus mycelium fermentation liquid of the present application is simple and convenient to operate, safe and controllable, easy to scale up production, and a large amount of edible fungus mycelium can be obtained in a short time. The fermentation cycle is short, and the content of the effective ingredient adenine nucleotide in the metabolites of the treated fermentation liquid is high and the quality is stable, which significantly reduces the industrial production cost of the bitterness blocker adenine nucleotide (AMP) and improves the economic benefits of the enterprise. The obtained edible fungus mycelium fermentation liquid is safe and natural, and can be directly used in food, medicine, and feed, which reduces or even eliminates the perception of bitterness, effectively masks the bitterness, avoids the large-scale use of sweeteners, and meets consumers' requirements for green and healthy food.

[0011] The fermentation broth prepared in the present application also contains bioactive substances such as high-molecular polysaccharides, β-glucans, high-molecular RNA complexes, nucleic acid degradation products, cAMP and triterpenoid compounds, which can play anti-cancer, antibacterial and antiviral, blood pressure lowering, blood lipid lowering, stomachic, digestion promoting, cough and asthma relieving, blood sugar lowering, urination and defecation, immune regulation and the like effects, play an important role in maintaining human health, and greatly promote the in-depth utilization of edible fungus resources.

[0012] Preferably, the edible fungi in step S1 include at least one of shiitake mushrooms, king oyster mushrooms, enoki mushrooms, ganoderma lucidum, truffles, cordyceps militaris, and hericium erinaceus.

[0013] By adopting the above technical scheme, the above edible fungi have high nutritional value, fast mycelium growth rate, high strain purity, high content of effective ingredient adenine nucleotide in its fermentation liquid metabolites, high quality and good stability.

[0014] Preferably, the PDA culture medium formula in step S1 is: 200 g potato, 20 g glucose, 20 g agar, and 1000 mL water.

[0015] Preferably, the shaking culture speed in step S2 is 100-180 rpm / min.

[0016] Preferably, the preparation conditions of the fermentation medium in step S2 are as follows: 20-25 g of carbon source, 4-6 g of nitrogen source, 2-3 g of inorganic salt, 1000 mL of water, 60-140 mL of medium, and the pH is adjusted to 4-8.

[0017] Preferably, the carbon source is selected from at least one of glucose, sucrose, corn flour, soluble starch and malt extract powder.

[0018] Preferably, the nitrogen source is selected from at least one of tryptone, ammonium sulfate, soybean powder, corn steep liquor powder and yeast extract powder.

[0019] Preferably, the inorganic salt is selected from at least one of MnSO4·4H2O, KH2PO4, C3H3NaO3, MgSO4, and (NaPO3)6.

[0020] Preferably, the preparation conditions of the fermentation medium are as follows: 20-25 g corn flour, 4-6 g yeast extract powder, 2-3 g KH2PO4, 1000 mL water, 80-120 mL culture medium in a 250 mL conical flask, and the pH is adjusted to 4-8.

[0021] Preferably, the preparation conditions of the fermentation medium are as follows: 20-25 g glucose, 4-6 g tryptone, 2-3 g MgSO4, 1000 mL water, 80-120 mL culture medium in a 250 mL conical flask, and the pH is adjusted to 4-7.

[0022] By adopting the above technical scheme, the fermentation medium has safe ingredients and rich nutrition, which can meet the growth requirements of edible fungus mycelium, and is beneficial to improving the fermentation efficiency and fermentation quality. By optimizing the fermentation conditions of edible fungus mycelium, including optimizing the carbon source, carbon source addition, nitrogen source, nitrogen source addition, inorganic salt, inorganic salt addition and pH value, liquid volume and fermentation days in the fermentation medium, the content of adenine nucleotide, an effective ingredient in the metabolites of the fermentation liquid, can be effectively increased, and the quality stability of adenine nucleotide can be improved.

[0023] Preferably, the fermentation liquid treatment method in step S2 is one of freeze-thaw, enzymatic hydrolysis, high temperature or ultrasound.

[0024] Preferably, the freeze-thaw operation is as follows: the fermented edible fungus mycelium fermentation liquid is placed in a -20°C refrigerator, and after 2 hours, it is taken out and thawed to room temperature, which is freeze-thaw once, and the freeze-thaw is repeated 3 times.

[0025] Preferably, the enzymatic hydrolysis is specifically performed as follows: the pH value of the fermented edible fungus mycelium fermentation liquid is adjusted to 7, and snail enzyme and 5′-phosphodiesterase are added for enzymatic hydrolysis for 1 hour in a water bath at 50° C. The total mass of snail enzyme and 5′-phosphodiesterase added is 0.1% of the mass of the fermentation liquid, and the mass ratio of snail enzyme to 5′-phosphodiesterase is 1:1.

[0026] Preferably, the high temperature is specifically operated as follows: the fermented edible fungus mycelium fermentation liquid is maintained at 120° C. for 30 minutes.

[0027] Preferably, the ultrasonic treatment is specifically performed as follows: the fermented edible fungus mycelium fermentation liquid is placed in an ultrasonic cleaning machine for 1 hour, with the ultrasonic power set to 240W and the ultrasonic temperature set to 35°C.

[0028] By adopting the above technical scheme, the fermentation liquid of edible fungi mycelium at the end of fermentation is treated, so that the intracellular substances in the fermented mycelium can be released, thereby promoting the production and release of adenine nucleotides, and further increasing the content of adenine nucleotides, the effective ingredient, in the metabolites of the fermentation liquid, and improving the quality stability of adenine nucleotides.

[0029] Preferably, a release promoter is added during the fermentation broth treatment process, and the release promoter is a compound of spermidine and riboflavin.

[0030] Preferably, the mass ratio of spermidine to riboflavin is 1:4-6.

[0031] Preferably, the mass ratio of spermidine to riboflavin is 1:4.8.

[0032] By adopting the above technical scheme, when the fermentation liquid of edible fungi mycelium is treated, the release promoter is added, which can further promote the release of intracellular adenine nucleotides, significantly increase the content of effective ingredient adenine nucleotides in the fermentation liquid metabolites, and improve the quality stability of adenine nucleotides.

[0033] In a second aspect, the present application provides an edible fungus mycelium fermentation liquid, which adopts the following technical solution:

[0034] An edible fungus mycelium fermentation liquid is prepared by the following method:

[0035] (1) Preparation of PDA culture medium:

[0036] Peel the potatoes, cut them into small pieces and put them into a pot, add 1000 mL of distilled water, boil for 20 minutes, then filter with 4 layers of gauze, take 200 g of potato filtrate, add 20 g of glucose and 20 g of agar, heat and stir with a glass rod to dissolve, and add water to 1000 mL, divide the culture medium into Erlenmeyer flasks and seal them, sterilize at 121°C for 30 minutes, pour the culture medium in the Erlenmeyer flask into a culture dish in a sterile room, and invert the culture dish after the culture medium condenses to obtain PDA culture medium;

[0037] (2) Preparation of edible mushroom mycelium seeds:

[0038] Take 1cm 2 The mycelium of edible fungi is inoculated on PDA medium under a sterile environment and cultured at 25-28°C until the mycelium grows to more than 3 / 5 of the plate. The mycelium morphology is observed to be uniform, indicating that the strain is relatively pure and the next step of the experiment can be carried out to obtain the mycelium seeds of edible fungi for future use;

[0039] (3) Preparation of fermentation medium:

[0040] Heat 20 g of glucose, 4 g of tryptone, and 3 g of MgSO4 and stir them with a glass rod to dissolve them, and add water to 1000 mL. Pour 100 mL of the culture medium into 250 mL conical flasks and seal them, sterilize at 121°C for 30 min to obtain a fermentation medium.

[0041] (4) preparing edible fungus mycelium fermentation liquid:

[0042] Take a piece of 0.5cm 2 The edible fungus mycelium seeds are inoculated into the fermentation medium under a sterile environment, and cultured with shaking at 24-30°C for 4-8 days, with a shaking culture speed of 100-180rpm / min. The edible fungus mycelium fermentation liquid after the fermentation is placed in a -20°C refrigerator, and after being placed for 2 hours, taken out and thawed to room temperature, i.e., freeze-thawed once. The freeze-thawed method is repeated 3 times, and the processed liquid is collected to obtain the edible fungus mycelium fermentation liquid.

[0043] By adopting the above technical scheme, the fermentation liquid metabolites finally obtained in the present application have a high content of the effective ingredient adenine nucleotide and good quality stability; the present application prepares a PDA culture medium, cultivates edible fungus mycelium seeds on the PDA culture medium, and then further ferments and cultivates the edible fungus mycelium seeds on the prepared fermentation medium, the fermentation cycle is short, and the edible fungus mycelium balls obtained are small and large in number; by treating the fermentation liquid, the intracellular substances in the fermented mycelium can be released, thereby promoting the production and release of adenine nucleotides.

[0044] In a third aspect, the present application provides an application of edible fungus mycelium fermentation liquid, adopting the following technical scheme: an application of edible fungus mycelium fermentation liquid, wherein the edible fungus mycelium fermentation liquid is used in the preparation of food, medicine, condiment, and feed to mask the bitter taste.

[0045] By adopting the above technical scheme, the edible fungus mycelium fermentation liquid prepared in the present application can be directly applied to food, medicine, condiments, and feed, and can significantly mask the bitterness and improve the taste without being used together with a sweetener; when used together with a sweetener, it can also mask the aftertaste and metallic taste of the sweetener.

[0046] In summary, this application has the following beneficial effects:

[0047] 1. The preparation method of the edible fungus mycelium fermentation liquid of the present application is simple and convenient to operate, safe and controllable, easy to scale up production, and can obtain a large amount of edible fungus mycelium in a short time. The fermentation cycle is short, and the content of the effective ingredient adenine nucleotide in the metabolites of the treated fermentation liquid is high and the quality is stable, which significantly reduces the industrial production cost of the bitterness blocker adenine nucleotide (AMP) and improves the economic benefits of the enterprise. The obtained edible fungus mycelium fermentation liquid is safe and natural, and can be directly used in food, medicine, and feed. It can reduce or even eliminate the perception of bitterness, effectively mask the bitterness, avoid the large-scale use of sweeteners, and meet consumers' requirements for green and healthy food.

[0048] 2. The fermentation broth prepared in the present application also contains bioactive substances such as high-molecular polysaccharides, β-glucans, high-molecular RNA complexes, nucleic acid degradation products, cAMP and triterpenoid compounds, which can play an anti-cancer, antibacterial and antiviral role, lower blood pressure, lower blood lipids, strengthen the stomach, promote digestion, relieve cough and asthma, lower blood sugar, promote urination and defecation, and regulate immunity. It plays an important role in maintaining human health and greatly promotes the in-depth utilization of edible fungus resources.

[0049] 3. The fermentation medium of the present application has safe ingredients and rich nutrition, which can meet the growth requirements of edible fungus mycelium, and is beneficial to improving the fermentation efficiency and fermentation quality. By optimizing the fermentation conditions of edible fungus mycelium, the content of adenine nucleotide, an effective ingredient in the metabolites of the fermentation liquid, can be effectively increased, and the quality stability of adenine nucleotide can be improved.

[0050] 4. The fermentation liquid treatment process in the present application can release the intracellular substances in the mycelium obtained by fermentation, thereby promoting the production and release of adenine nucleotides, and further increasing the content of adenine nucleotides, the effective ingredient, in the fermentation liquid metabolites, and improving the quality stability of adenine nucleotides.

[0051] 5. The edible fungus mycelium fermentation liquid prepared in the present application can be directly applied to food, medicine, condiments, and feed. It does not need to be used together with a sweetener, and can significantly mask the bitterness and improve the taste. When used together with a sweetener, it can also mask the aftertaste of the sweetener, such as the metallic taste and other undesirable tastes. DETAILED DESCRIPTION

[0052] The present application is further described in detail below with reference to the embodiments.

[0053] Examples 1-5 and Comparative Examples 1-6 provide methods for preparing edible fungus mycelium fermentation broth, which are described below using Example 1 as an example.

[0054] Example 1

[0055] An edible fungus mycelium fermentation liquid is prepared by the following method:

[0056] (1) Preparation of PDA culture medium:

[0057] Peel the potatoes, cut them into small pieces and put them into a pot, add 1000 mL of distilled water, boil for 20 minutes, then filter with 4 layers of gauze, take 200 g of potato filtrate, add 20 g of glucose and 20 g of agar, heat and stir with a glass rod to dissolve, and add water to 1000 mL, divide the culture medium into Erlenmeyer flasks and seal them, sterilize at 121°C for 30 minutes, pour the culture medium in the Erlenmeyer flask into a culture dish in a sterile room, and invert the culture dish after the culture medium condenses to obtain PDA culture medium;

[0058] (2) Preparation of Lentinus edodes mycelium seeds:

[0059] Take 1cm 2 The mycelium of Lentinus edodes was inoculated on PDA medium under a sterile environment and cultured at 28°C until the mycelium grew to 4 / 5 of the plate. The mycelium morphology was observed to be uniform, indicating that the strain was relatively pure and the next step of the experiment could be carried out to obtain the mycelium seeds for later use.

[0060] (3) Preparation of fermentation medium:

[0061] Heat 20 g of glucose, 4 g of tryptone, and 3 g of MgSO4 and stir them with a glass rod to dissolve them, add water to 1000 mL, adjust the pH to 6, and then divide 100 mL of the culture medium into 250 mL conical flasks and seal them, sterilize them at 121°C for 30 min, and obtain a fermentation medium;

[0062] (4) Preparation of Lentinus edodes mycelium fermentation liquid:

[0063] Take a piece of 0.5cm 2The shiitake mycelium seeds were inoculated into the fermentation medium under a sterile environment, and cultured at 28°C with shaking for 5 days at a shaking culture speed of 150 rpm / min. The shiitake mycelium fermentation liquid after fermentation was placed in a -20°C refrigerator, and after being placed for 2 hours, it was taken out and thawed to room temperature, i.e., freeze-thawed once. The freeze-thawed method was repeated 3 times, and the treated liquid was collected to obtain the shiitake mycelium fermentation liquid.

[0064] Embodiment 2-10 is the same as embodiment 1, except that different edible fungus mycelia are used in preparing edible fungus mycelia seeds, as shown in Table 1.

[0065] Table 1:

[0066]

[0067]

[0068] Example 11-35 is the same as Example 1, except that the formula for preparing the fermentation medium is different, as shown in Table 2 below.

[0069] Table 2:

[0070]

[0071]

[0072] Examples 36-51 are the same as Example 1, except that the conditions for preparing the edible fungus mycelium fermentation liquid are different, as shown in Table 3 below.

[0073] Table 3:

[0074]

[0075] Examples 52-55 are the same as Example 1, except that the processing method of the fermented shiitake mycelium fermentation liquid after fermentation is different, as follows:

[0076] Example 52 is the same as Example 1, except that the step (4) of "placing the fermented shiitake mycelium fermentation liquid in a -20°C refrigerator for 2 hours and then taking it out to thaw at room temperature, i.e., performing freeze-thaw once, repeating the freeze-thaw process three times and collecting the treated liquid" is replaced with "adjusting the pH value of the fermented shiitake mycelium fermentation liquid to 7, adding snail enzyme and 5'-phosphodiesterase for enzymatic hydrolysis for 1 hour in a water bath at 50°C, wherein the total mass of snail enzyme and 5'-phosphodiesterase added is 0.1% of the mass of the fermentation liquid, and the mass ratio of snail enzyme to 5'-phosphodiesterase is 1:1".

[0077] Example 53 is the same as Example 1, except that the step (4) of "placing the fermented shiitake mycelium fermentation liquid in a -20°C refrigerator for 2 hours and then taking it out to thaw at room temperature, i.e., performing freeze-thaw once, repeating the freeze-thaw process three times, and collecting the treated liquid" is replaced with "keeping the fermented shiitake mycelium fermentation liquid at 120°C for 30 minutes and collecting the treated liquid".

[0078] Example 54 is the same as Example 1, except that the step (4) of "placing the fermented shiitake mycelium fermentation liquid in a -20°C refrigerator for 2 hours and then taking it out to thaw at room temperature, i.e., performing freeze-thaw once, repeating the freeze-thaw process three times, and collecting the treated liquid" is replaced with "placing the fermented shiitake mycelium fermentation liquid in an ultrasonic cleaner for 1 hour, setting the ultrasonic power to 240 W and the ultrasonic temperature to 35°C, and collecting the treated liquid".

[0079] Example 55 is the same as Example 1, except that, in step (4), after "placing the fermented shiitake mycelium fermentation liquid in a -20°C refrigerator for 2 hours, taking it out and thawing it to room temperature, i.e., performing freeze-thaw once, repeating the freeze-thaw process three times, and collecting the treated liquid", the pH value of the collected treated liquid is adjusted to 7, and snail enzyme and 5'-phosphodiesterase are added to perform enzymatic hydrolysis for 1 hour in a water bath at 50°C. The total mass of snail enzyme and 5'-phosphodiesterase added is 0.1% of the mass of the treated liquid, and the mass ratio of snail enzyme to 5'-phosphodiesterase is 1:1.

[0080] In Examples 56-62, a release-promoting agent is added during the fermentation broth treatment process, as follows:

[0081] Example 56 is the same as Example 1, except that 0.3% of the total mass of a release promoter is added to the fermentation liquid of Lentinus edodes mycelium after fermentation in step (4), the mixture is stirred and evenly mixed, and then placed in a -20°C refrigerator. After 2 hours, the mixture is taken out and thawed to room temperature, i.e., freeze-thawed once. The freeze-thaw process is repeated three times, and the treated liquid is collected. The release promoter is a compound of spermidine and riboflavin in a mass ratio of 1:4.

[0082] Example 57 is the same as Example 56, except that the release-enhancing agent is prepared by mixing spermidine and riboflavin in a mass ratio of 1:4.8.

[0083] Example 58 is the same as Example 56, except that the release-enhancing agent is prepared by mixing spermidine and riboflavin in a mass ratio of 1:5.

[0084] Example 59 is the same as Example 56, except that the release-enhancing agent is prepared by mixing spermidine and riboflavin in a mass ratio of 1:6.

[0085] Example 60 is the same as Example 52, except that 0.3% of the total mass of the release promoter is added to the fermentation liquid of Lentinus edodes mycelium after fermentation in step (4), the mixture is stirred and mixed, the pH value is adjusted to 7, snail enzyme and 5′-phosphodiesterase are added for enzymatic hydrolysis for 1 hour in a water bath at 50° C., the treated liquid is collected, the total mass of the snail enzyme and 5′-phosphodiesterase added is 0.1% of the mass of the fermentation liquid, and the mass ratio of the snail enzyme to the 5′-phosphodiesterase is 1:1; the release promoter is obtained by compounding spermidine and riboflavin in a mass ratio of 1:4.

[0086] Example 61 is the same as Example 53, except that 0.3% of the total mass of a release promoter is added to the fermentation liquid of the shiitake mushroom mycelium after the fermentation in step (4), the mixture is stirred and mixed evenly, and then maintained at 120° C. for 30 minutes, and the treated liquid is collected; the release promoter is a compound of spermidine and riboflavin in a mass ratio of 1:4.

[0087] Example 62 is the same as Example 54, except that 0.3% of the total mass of the release promoter is added to the fermentation liquid of the shiitake mushroom mycelium after the fermentation in step (4), the mixture is stirred and evenly mixed, and then ultrasonicated for 1 hour in an ultrasonic cleaning machine with the ultrasonic power set to 240 W and the ultrasonic temperature set to 35°C, and the treated liquid is collected; the release promoter is obtained by compounding spermidine and riboflavin in a mass ratio of 1:4.

[0088] In order to verify the inventiveness of Examples 1-62 of the present application, the applicant set up Comparative Examples 1-5, which are as follows:

[0089] Comparative Example 1 is the same as Example 1, except that in step (4), the fermentation liquid of edible fungi mycelium after fermentation is directly collected without freeze-thaw treatment.

[0090] Comparative Example 2 is the same as Example 1, except that, during the preparation of the fermentation medium in step (3), a release enhancer of 0.3% of the total mass of the medium is added, stirred and dissolved, and then water is added to 1000 mL, wherein the release enhancer is prepared by mixing spermidine and riboflavin in a mass ratio of 1:4.

[0091] Comparative Example 3 is the same as Example 56, except that the release-enhancing agent is prepared by mixing spermidine and riboflavin in a mass ratio of 1:3.

[0092] Comparative Example 4 is the same as Example 56, except that the release-enhancing agent is prepared by mixing spermidine and riboflavin in a mass ratio of 1:7.

[0093] Comparative Example 5 is the same as Example 56, except that the release-enhancing agent is prepared by compounding spermidine and vitamin C in a mass ratio of 1:4.

[0094] Performance Testing

[0095] In accordance with the national food safety standard GB5413.40-2016 "Determination of nucleotides in infant food and dairy products", the AMP content in the edible fungus fermentation broth prepared in Examples 1-62 of the present application and Comparative Examples 1-5 was determined, and the results are shown in Table 4 below.

[0096] The electronic tongue was used to detect the bitterness blocking effect of the edible fungus fermentation liquid prepared in Examples 1-62 and Comparative Examples 1-5 of the present application. The specific detection method is as follows:

[0097] 1. Preparation of electronic tongue cleaning solution: ① Preparation of positive cleaning solution: accurately weigh 7.46g of potassium chloride, stir and dissolve with 500mL of distilled water, then accurately add 300mL of anhydrous ethanol solution, add 0.56g of potassium hydroxide accurately weighed while stirring, after dissolution, transfer to a 1000mL volumetric flask, dilute to volume for use. ② Preparation of negative cleaning solution: accurately measure 300mL of anhydrous ethanol, shake and mix with 500mL of distilled water, then add 8.3mL of concentrated hydrochloric acid, stir and mix, transfer to a 1000mL volumetric flask, dilute to volume for use. ③ Preparation of reference solution: accurately weigh 2.24g of potassium chloride and 0.045g of tartaric acid, dissolve with 500mL of water, transfer to a 1000mL volumetric flask, dilute to volume for use.

[0098] 2. Sample preparation for electronic tongue test: Accurately weigh 50 mg of berberine and prepare a 0.025 mg / mL berberine standard solution with distilled water as the standard bitter solution. Take 50 mL of the prepared standard bitter solution and add 50 mL of edible fungus mycelium fermentation liquid as the sample to be tested.

[0099] 3. Electronic tongue test procedure: Before data collection, the electronic tongue is activated, calibrated, diagnosed and other operations are performed to ensure the reliability and stability of the test data. The sample to be tested is placed in a special beaker for the electronic tongue, and sample collection and cleaning are performed alternately. The sample collection time is 120s, and each sample data is collected 4 times. The minimum error value of the 3 times is taken as the measurement data of the sample.

[0100] 4. Establishment of evaluation indexes for the bitterness suppression effect of electronic tongue method:

[0101] Bitterness inhibition rate (BIR) is used as an evaluation index for the bitterness inhibition effect of fermentation broth. Bitterness inhibition rate represents the ratio of the reduction in the bitterness value of the sample solution after adding the fermentation broth to the bitterness value of the bitter sample solution before adding. The calculation formula of bitterness inhibition rate BIR is: BIR = (I0-I) / I0*100%

[0102] According to the test results of the electronic tongue sensor and the calculation formula, the BIR of the edible fungus mycelium fermentation broth was obtained and recorded in Table 4.

[0103] Table 4:

[0104]

[0105]

[0106]

[0107] Combined with the test data in Table 4, we can see that:

[0108] The difference between Examples 2-10 of the present application and Example 1 is that the types of edible fungi are different. It can be seen from Example 10 and Example 1 that the selected edible fungi have 0.5 cm 2 Ganoderma lucidum mycelium plus 0.5cm 2 Hericium erinaceus mycelium replacement 1cm 2 When Lentinus edodes mycelium is added, the AMP content and the bitterness blocking effect of the final fermentation liquid are significantly improved.

[0109] The only difference between Examples 11-35 of the present application and Example 1 is that the formula for preparing the fermentation medium is different. It can be seen from the above examples that by optimizing the raw materials and raw material dosage for preparing the fermentation medium, the AMP content in the metabolites of the fermentation broth can be effectively increased, and the bitterness blocking effect of the fermentation broth can be improved.

[0110] The only difference between Examples 36-51 of the present application and Example 1 is that the conditions for preparing the shiitake mushroom mycelium fermentation broth are different. It can be seen from the above examples that by optimizing the conditions for preparing the shiitake mushroom mycelium fermentation broth, the AMP content in the metabolites of the fermentation broth can be effectively increased, thereby improving the bitterness blocking effect of the fermentation broth.

[0111] The only difference between Examples 52-55 and Example 1 is that the treatment methods of the fermentation broth are different. From the above data, it can be seen that when only a single treatment method is used to treat the fermentation broth, the enzymatic treatment effect is the best, and the AMP content of the fermentation broth and the effect of blocking bitterness are better; when two treatment methods are used for superposition treatment, the treatment effect will be further improved, but the overall improvement effect cannot reach the superposition effect.

[0112] In Examples 56-59, a release promoter was added during the freeze-thaw treatment of the fermentation broth. From the above data, it can be seen that adding a release promoter during the freeze-thaw treatment of the fermentation broth can further improve the AMP content of the fermentation broth and the effect of blocking bitterness. In addition, when other factors remain unchanged, only changing the ratio of the release promoter raw materials will have a certain impact on the quality of the fermentation broth obtained by the final treatment. Among them, when the release promoter is obtained by compounding spermidine and riboflavin in a mass ratio of 1:4.8, the treatment effect obtained is better.

[0113] It can be seen from Examples 60-62 that when the same release-enhancing agent is added during the fermentation broth treatment process and different treatment methods are used, the release-enhancing effects achieved by the release-enhancing agent are not the same. It can be seen from the above data that, when other factors remain unchanged, the synergistic release effect of adding a release-enhancing agent to the enzymatic treatment is the best, the AMP content of the obtained fermentation broth is high, and the effect of blocking bitterness is the best.

[0114] It can be seen from Example 1 and Comparative Example 1 that by directly collecting the fermentation liquid of edible fungi mycelium after fermentation without treating the fermentation liquid, the AMP content in the fermentation liquid finally obtained is significantly reduced, and the effect of blocking bitterness is also average.

[0115] It can be seen from Example 56 and Comparative Example 2 that the comprehensive quality of the fermentation liquid finally obtained by adding the release promoter during the preparation of the fermentation medium in step (3) is obviously inferior to the comprehensive quality of the fermentation liquid obtained by adding the release promoter to the shiitake mycelium fermentation liquid after the fermentation in step (4).

[0116] It can be seen from Example 56 and Comparative Examples 3-4 that when the mass ratio of the release-enhancing agent raw materials spermidine and riboflavin is not within the range of 1:4-6 specified in the present application, the overall quality of the fermentation broth finally obtained will be significantly reduced.

[0117] It can be seen from Example 56 and Comparative Example 5 that when riboflavin in the release-promoting agent raw material is replaced by vitamin C, its release-promoting effect is significantly reduced, and the overall quality of the fermentation broth finally obtained will also be significantly reduced.

[0118] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A method for preparing edible fungus mycelium fermentation liquid, characterized in that: The method comprises the following preparation steps: S1. Preparation of edible fungus mycelium seeds: inoculate edible fungus mycelium onto PDA medium under a sterile environment, and culture at 25-28° C. until mycelium grows to more than 3 / 5 of the plate, and obtain edible fungus mycelium seeds for later use; S2, preparing edible fungus mycelium fermentation liquid: preparing a fermentation medium and sterilizing it, taking the edible fungus mycelium seeds prepared in step S1 and inoculating them into the fermentation medium under a sterile environment, shaking and culturing at 24-30° C. for 4-8 days, treating the fermentation liquid and collecting it, and obtaining the edible fungus mycelium fermentation liquid; The fermentation liquid treatment method in step S2 is one of freeze-thaw, enzymatic hydrolysis, high temperature or ultrasound; During the fermentation liquid treatment process, a release promoter is added, wherein the release promoter is a compound of spermidine and riboflavin, and the mass ratio of spermidine to riboflavin is 1:4-6.

2. The method for preparing edible fungus mycelium fermentation liquid according to claim 1, characterized in that: The edible fungi in step S1 include at least one of shiitake mushrooms, king oyster mushrooms, enoki mushrooms, ganoderma lucidum, truffles, cordyceps militaris, and hericium erinaceus.

3. The method for preparing edible fungus mycelium fermentation liquid according to claim 1, characterized in that: The PDA culture medium formula in step S1 is: 200 g potato, 20 g glucose, 20 g agar, and 1000 mL water.

4. The method for preparing edible fungus mycelium fermentation liquid according to claim 1, characterized in that: The preparation conditions of the fermentation medium in step S2 are as follows: 20-25 g of carbon source, 4-6 g of nitrogen source, 2-3 g of inorganic salt, 1000 mL of water, 60-140 mL of medium, and the pH is adjusted to 4-8.

5. The method for preparing edible fungus mycelium fermentation liquid according to claim 4, characterized in that: The carbon source is selected from at least one of glucose, sucrose, corn flour, soluble starch, and malt extract powder; the nitrogen source is selected from at least one of trypsin, ammonium sulfate, soybean flour, corn steep liquor powder, and yeast extract powder; the inorganic salt is selected from at least one of MnSO4·4H2O, KH2PO4, C3H3NaO3, MgSO4, and (NaPO3)6.

6. The method for preparing edible fungus mycelium fermentation liquid according to claim 5, characterized in that: The preparation conditions of the fermentation medium are as follows: 20-25 g corn flour, 4-6 g yeast extract powder, 2-3 g KH2PO4, 1000 mL water, 60-140 mL culture medium in a 250 mL conical flask, pH adjusted to 4-8, and fermentation for 3-6 days.

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