Method for fermenting cynanchum auriculatum by using lactic acid bacteria and application of product in spleen-tonifying stomach-benefiting digestion-promoting product

Abstract

The invention belongs to the technical field of bioengineering, and particularly relates to a method for fermenting cynanchum auriculatum with lactic acid bacteria and application of a product in a product for strengthening the spleen, benefiting the stomach and promoting digestion. The method specifically comprises the following steps: (1) pretreatment: adding cynanchum auriculatum powder into water, and performing high-temperature stewing to obtain a pretreatment solution; (2) enzymolysis: continuously adding water into the pretreatment solution, and then adding an enzyme preparation for enzymolysis to obtain an enzymolysis solution; (3) enzyme deactivation and fermentation: sterilizing the enzymatic hydrolysate, adding plant lactobacillus at the temperature of 25-30 DEG C, and fermenting for 1-3 hours to obtain fermentation liquor A; and (4) secondary fermentation: adding a compound strain into the fermentation liquor A, fermenting for 14-18 hours to obtain fermentation liquor, and carrying out solid-liquid separation on the fermentation liquor to obtain an upper layer which is fermentation clear liquor. The content of C21 steroid saponin in the fermentation product is high, and the fermentation product has the effects of strengthening the spleen, benefiting the stomach, promoting digestion and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of bioengineering, and specifically relates to a method for fermenting ear leaf cow peel shavings with lactic acid bacteria and application of the product in spleen and stomach invigorating and digestion-aiding products. Background Art

[0002] Cynanchumanriculatum Royle ex Wight is generally referred to as Cynanchumanriculatum Royle ex Wight, which is the dried root of Cynanchumanriculatum Royle ex Wight, a plant of the genus Cynanchus of the family Apocynaceae. It is a common food in the traditional eating habits of Jiangsu Province. In the theory of traditional Chinese medicine, Cynanchumanriculatum Royle ex Wight is sweet and bitter in nature, slightly warm in taste, and enters the spleen, stomach, and lung meridians. It has the effects of strengthening the stomach and removing stagnation, detoxifying and removing dampness, and is used in the treatment of diseases such as indigestion, abdominal pain, bloating, and ascites. Modern research shows that Cynanchumanriculatum Royle ex Wight mainly contains acetophenones, steroidal glycosides, organic acids, and contains polysaccharides, phospholipids, amino acids and other nutrients. The C21 steroidal saponin component contained therein is the largest class of compounds isolated from Cynanchumanriculatum so far, including caudatin, kidjolanin, deacymetaplexigenin and gagamine.

[0003] There are many methods for extracting C21 steroidal saponins, for example, Chinese patent CN 1660865A discloses a Radix Cynomorii saponin compound, total saponins and their use in drugs for treating gastrointestinal diseases. This method uses an organic solvent to extract C21 saponins from Radix Cynomorii, but alcohol solvents are expensive and the extraction cost is high. Secondly, alcohols are flammable and explosive, and high safety requirements are placed on the production process.

[0004] Another example is Chinese patent CN 116585366A, which discloses a Radix Cynomorii extract and a method for its use, and its use in regulating intestinal motility and treating hemorrhoids. The patent uses a water extraction method to extract the active ingredients in Radix Cynomorii. However, the extraction rate is low when water is used as a solvent, and repeated extraction is required, which takes a long time to operate.

[0005] Modern Chinese medicine fermentation is a new process that combines modern processes such as fermentation with traditional Chinese medicine fermentation. One or more beneficial bacteria are added during the fermentation of Chinese medicinal materials, and the enzymes produced during the growth and metabolism of microorganisms react with the complex active ingredients in Chinese medicine. This can not only increase the content of active ingredients, but also accelerate the completion of the reaction by utilizing the catalytic effect of enzymes. It has been widely used to improve the medicinal properties of Chinese medicine and produce new drug varieties. No literature has reported the relevant fermentation process of Erye Niupixiao. Summary of the invention

[0006] To this end, the present invention provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which innovatively uses lactic acid bacteria to ferment ear leaf cow skin shavings, significantly increasing the content of C21 steroidal saponin in the fermentation product. The obtained fermentation product has a better effect in promoting appetite and improving indigestion symptoms such as abdominal pain, belching, pantothenic acid and abdominal distension.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] First, the present invention provides a method for fermenting ear leaf cow hide shavings with lactic acid bacteria, which specifically comprises the steps of:

[0009] (1) Pretreatment: Add the ear leaf cow skin powder into water and simmer at high temperature to obtain a pretreatment solution;

[0010] (2) enzymolysis: adding water to the pretreatment solution, and then adding an enzyme preparation to perform enzymolysis to obtain an enzymolysis solution;

[0011] (3) enzyme inactivation and fermentation: after sterilizing the enzymatic hydrolysate, adding Lactobacillus plantarum and fermenting for 1-3 hours at a temperature of 25-30° C. to obtain fermentation liquid A;

[0012] (4) Secondary fermentation: adding the composite strain to the fermentation liquid A and fermenting for 14-18 hours to obtain a fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0013] The enzyme preparation comprises medium-temperature alpha-amylase, saccharifying enzyme and acid protease; and the composite strain comprises Lactobacillus acidophilus and Lactobacillus fermentans.

[0014] Preferably, the weight ratio of the Bacillus subtilis powder to the Lactobacillus plantarum is (500-800):1, and the weight ratio of the Bacillus subtilis powder to the composite strain is (500-1000):1.

[0015] Further preferably, the weight ratio of the Bacillus subtilis powder to the Lactobacillus plantarum is 500:1; the weight ratio of the Bacillus subtilis powder to the composite strain is 500:1.

[0016] More preferably, the weight ratio of Lactobacillus acidophilus to Lactobacillus mucosa fermentans is (1-3):(2-5).

[0017] More preferably, the weight ratio of Lactobacillus acidophilus to Lactobacillus mucofermentans is 3:2.

[0018] Preferably, the weight ratio of the Auricularia aurantifolia powder to the enzyme preparation is (30-50):1.

[0019] Further preferably, the weight ratio of the Auricularia aurantifolia powder to the enzyme preparation is 32:1.

[0020] More preferably, the weight ratio of the medium-temperature α-amylase, saccharifying enzyme and acid protease is (2-10): (2-7): (3-8).

[0021] More preferably, the weight ratio of the medium-temperature α-amylase, saccharifying enzyme and acid protease is 2:3:3.

[0022] Preferably, the amount of water added in step (1) is based on the amount of water added to submerge the ear leaf cowpea powder by 3-5 cm.

[0023] Preferably, in step (1), the weight ratio of water to the Auricularia aurantifolia powder is (2-5):1.

[0024] Further preferably, the weight ratio of water to the Auricularia aurantifolia powder in step (1) is 4:1.

[0025] Preferably, the weight ratio of water to the Auricularia aurantifolia powder in step (2) is (5-15):1.

[0026] Further preferably, the weight ratio of water to the Auricularia aurantifolia powder is 10:1.

[0027] Preferably, the high temperature stewing in step (1) is at a temperature of 80-100° C. and a time of 30-60 min.

[0028] More preferably, the high temperature stewing in step (1) is at 90° C. and the time is 50 minutes.

[0029] Preferably, the enzymatic hydrolysis temperature in step (2) is 40-60° C., and the enzymatic hydrolysis time is 30-60 min.

[0030] Further preferably, the enzymatic hydrolysis temperature in step (2) is 45° C. and the enzymatic hydrolysis time is 45 min.

[0031] Preferably, the enzymatic solution in step (3) is sterilized for 30-90 minutes at a temperature of 90-120°C.

[0032] Preferably, the fermentation time in step (3) is 2 hours; and the fermentation time in step (4) is 16 hours.

[0033] Further preferably, the enzymatic activity of the medium-temperature α-amylase is 1000-10000 U / g, the enzymatic activity of the saccharifying enzyme is 50000-150000 U / g, and the enzymatic activity of the acid protease is 50000-100000 U / g.

[0034] Further preferably, the viable bacteria count of Lactobacillus plantarum is 100-300 billion CFU / g, the viable bacteria count of Lactobacillus acidophilus is 100-300 billion CFU / g, and the viable bacteria count of Lactobacillus mucosa fermentans is 100-300 billion CFU / g.

[0035] Preferably, the method further comprises the step of concentrating the fermentation supernatant and then spray-drying it to obtain a fermentation product.

[0036] More preferably, the concentration is carried out by rotary evaporation.

[0037] Further preferably, the air inlet temperature of the spray drying is 110-150°C, and the air outlet temperature is 80-90°C.

[0038] The fermented product of ear leaf cow hide is obtained according to the above preparation method.

[0039] The fermentation product obtained according to the above preparation method is used in the preparation of digestion-aiding, spleen-strengthening and stomach-benefiting products.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] 1. The preparation method of the present invention has the problem that the effective ingredients are difficult to extract because the root tissue of the ear-leaf cow skin xiao is hard and has a high degree of lignification. However, the high-temperature stewing treatment of the ear-leaf cow skin xiao is conducive to the dissolution of the effective ingredients and shortens the subsequent process time.

[0042] 2. The preparation method of the present invention comprises the following processes: high-temperature stewing followed by enzymatic hydrolysis, enzyme inactivation fermentation, and a second fermentation. The fermentation product obtained after enzymatic hydrolysis and fermentation by specific enzymes (medium-temperature α-amylase, saccharifying enzyme and acid protease) and strains (Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus mucosa fermentation) has a high content of C21 steroidal saponins and has the effects of strengthening the spleen and stomach and aiding digestion. DETAILED DESCRIPTION

[0043] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0044] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those generally understood by those of ordinary skill in the art to which the present invention belongs.

[0045] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Products from different manufacturers have no significant effect on the effects.

[0046] In the following embodiments, ear leaf cow hide is purchased from Haozhou Shishi Pharmaceutical Co., Ltd.; medium temperature α-amylase, saccharifying enzyme, acid protease are purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd.; plant lactobacillus, acidophilus, fermented mucus lactobacillus are purchased from Jiangsu Xinshenao Biotechnology Co., Ltd. plant lactobacillus YS4, acidophilus LA16 and fermented mucus lactobacillus CQPC07.

[0047] Example 1

[0048] This embodiment provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0049] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 960 mL of water, heat to 80° C. and stew for 60 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0050] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 60°C, 1600 mL of water is added, followed by 3.2 g of medium-temperature α-amylase, 2.24 g of saccharifying enzyme and 2.56 g of acid protease, and the mixture is stirred and enzymolyzed at 60°C for 50 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0051] (3) Inactivation of enzymes and fermentation: the enzymatic solution was inactivated at 90°C for 60 min, then cooled to 30°C, 0.53 g of Lactobacillus plantarum was added, and the solution was fermented at 30°C for 1 h to obtain fermentation solution A;

[0052] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 30° C., adding 0.1 g of Lactobacillus acidophilus and 0.3 g of Lactobacillus mucilaginosus, and continuing fermentation at 30° C. for 14 h to obtain fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0053] (5) Separation and drying: The fermentation supernatant was concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 110°C and an air outlet temperature of 90°C to obtain the fermentation product of ear leaf cow hide.

[0054] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0055] Example 2

[0056] This embodiment provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0057] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 640 mL of water, heat to 100° C. and stew for 30 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0058] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 40°C, 4800 mL of water is added, followed by 2.56 g of medium-temperature α-amylase, 1.92 g of saccharifying enzyme and 1.92 g of acid protease, and the mixture is stirred and enzymolyzed at 40°C for 60 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0059] (3) Inactivation of enzymes and fermentation: The enzymatic hydrolysate was inactivated at 120°C for 30 min, then cooled to 25°C, 0.4 g of Lactobacillus plantarum was added, and the mixture was fermented at 25°C for 3 h to obtain fermentation liquid A;

[0060] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 25° C., adding 0.137 g of Lactobacillus acidophilus and 0.183 g of fermented Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 18 h to obtain fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0061] (5) Separation and drying: The fermentation supernatant was concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 150°C and an air outlet temperature of 80°C to obtain the fermentation product of ear leaf cow hide.

[0062] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0063] Example 3

[0064] This embodiment provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0065] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1280 mL of water, heat to 90° C. and stew for 50 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0066] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 45°C, 3200 mL of water is added, followed by 2.5 g of medium-temperature α-amylase, 3.75 g of saccharifying enzyme and 3.75 g of acid protease, and the mixture is stirred and enzymolyzed at 45°C for 45 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0067] (3) Inactivation of enzyme and fermentation: the enzymatic solution was inactivated at 100°C for 50 min, then cooled to 25°C, 0.64 g of Lactobacillus plantarum was added, and the solution was fermented at 25°C for 2 h to obtain fermentation solution A;

[0068] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 25° C., adding 0.384 g of Lactobacillus acidophilus and 0.256 g of fermented Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 16 h to obtain a fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0069] (5) Separation and drying: The fermentation supernatant was concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the fermentation product of ear leaf cow hide.

[0070] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0071] Example 4

[0072] This embodiment provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0073] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1600 mL of water, heat to 95° C. and stew for 45 minutes, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0074] (2) Enzymolysis: After the temperature of the pretreatment solution dropped to 50°C, 2560 mL of water was added, followed by 3.2 g of medium-temperature α-amylase, 1.6 g of saccharifying enzyme and 2.4 g of acid protease, and the mixture was stirred and enzymolyzed at 50°C for 30 min. Sodium bicarbonate was added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0075] (3) Inactivation of enzyme and fermentation: the enzymatic solution was inactivated at 110° C. for 90 min, then cooled to 25° C., 0.46 g of Lactobacillus plantarum was added, and the solution was fermented at 25° C. for 1.5 h to obtain fermentation solution A;

[0076] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 25° C., adding 0.16 g of Lactobacillus acidophilus and 0.4 g of fermented Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 15 h to obtain fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0077] (5) Separation and drying: After the fermentation supernatant was concentrated by rotary evaporation, it was spray-dried at an air inlet temperature of 140°C and an air outlet temperature of 80°C to obtain the fermentation product of ear leaf cow hide.

[0078] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0079] Comparative Example 1

[0080] This comparative example provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0081] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot, add 1280 mL of water, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0082] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 45°C, 3200 mL of water is added, followed by 2.5 g of medium-temperature α-amylase, 3.75 g of saccharifying enzyme and 3.75 g of acid protease, and the mixture is stirred and enzymolyzed at 45°C for 45 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0083] (3) Inactivation of enzyme and fermentation: the enzymatic solution was inactivated at 100°C for 50 min, then cooled to 25°C, 0.64 g of Lactobacillus plantarum was added, and the solution was fermented at 25°C for 2 h to obtain fermentation solution A;

[0084] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 25° C., adding 0.384 g of Lactobacillus acidophilus and 0.256 g of fermented Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 16 h to obtain a fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid;

[0085] (5) Separation and drying: The fermentation supernatant was concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the fermentation product of ear leaf cow hide.

[0086] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0087] The difference from Example 3 is that there is no stewing process.

[0088] Comparative Example 2

[0089] This comparative example provides a method for fermenting ear leaf cow hide shavings with lactic acid bacteria, which is different from Example 3 in that the enzyme preparation used for enzymolysis is different, specifically: (2) enzymolysis: after the temperature of the pretreatment liquid drops to 45°C, 3200 mL of water is added, followed by adding 4 g of medium-temperature α-amylase and 6 g of saccharifying enzyme, and stirring the enzymolysis at a temperature of 45°C for 45 min, and adding sodium bicarbonate to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0090] The activity of medium-temperature α-amylase is 4000U / g, and the activity of saccharifying enzyme is 100000U / g.

[0091] The remaining steps are the same as in Example 3.

[0092] Comparative Example 3

[0093] This comparative example provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0094] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1280 mL of water, heat to 90° C. and stew for 50 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0095] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 45°C, 3200 mL of water is added, followed by 2.5 g of medium-temperature α-amylase, 3.75 g of saccharifying enzyme and 3.75 g of acid protease, and the mixture is stirred and enzymolyzed at 45°C for 45 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0096] (3) Inactivation of enzymes and fermentation: After inactivating the enzymes in the enzymatic solution at 100° C. for 50 min, the temperature was lowered to 25° C., 0.64 g of plant lactobacillus, 0.384 g of lactobacillus acidophilus and 0.256 g of fermented lactobacillus mucilaginosus were added, and the mixture was fermented at 25° C. for 18 h to obtain a fermentation solution;

[0097] (4) Separation and drying: The fermentation liquid is separated into solid and liquid, and the upper layer is the fermentation clear liquid. The fermentation clear liquid is concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the ear leaf cow hide fermentation product.

[0098] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0099] The difference from Example 3 is that there is no separate fermentation.

[0100] Comparative Example 4

[0101] This comparative example provides a method for fermenting ear leaf cow hide shavings with lactic acid bacteria, which differs from Example 3 in that the strains selected during the secondary fermentation are different, specifically: (4) Secondary fermentation: maintain the temperature of the fermentation liquid A at 25°C, add 0.64 g of Lactobacillus acidophilus and continue fermenting at 25°C for 16 hours to obtain a fermentation liquid.

[0102] The number of live bacteria of Lactobacillus acidophilus is 200 billion CFU / g.

[0103] The remaining steps are the same as in Example 3.

[0104] Comparative Example 5

[0105] This comparative example provides a method for fermenting ear leaf cow hide shavings with lactic acid bacteria, which differs from Example 3 in that the strain selection during the secondary fermentation process is different, specifically: (4) Secondary fermentation: Maintain the temperature of the fermentation liquid A at 25°C, add 0.64g of fermented mucus lactobacillus and continue fermenting at 25°C for 16 hours to obtain a fermentation liquid.

[0106] The number of viable bacteria in fermented mucus lactobacillus is 200 billion CFU / g.

[0107] The remaining steps are the same as in Example 3.

[0108] Comparative Example 6

[0109] This comparative example provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0110] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1280 mL of water, heat to 90° C. and stew for 50 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0111] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 45°C, 3200 mL of water is added, followed by 2.5 g of medium-temperature α-amylase, 3.75 g of saccharifying enzyme and 3.75 g of acid protease, and the mixture is stirred and enzymolyzed at 45°C for 45 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0112] (3) Inactivation of enzyme and fermentation: the enzymatic solution was inactivated at 100°C for 50 min, then cooled to 25°C, 0.384 g of Lactobacillus acidophilus was added, and the solution was fermented at 25°C for 2 h to obtain fermentation solution A;

[0113] (4) Secondary fermentation: maintaining the temperature of the fermentation liquid A at 25° C., adding 0.64 g of Lactobacillus plantarum and 0.256 g of Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 16 h to obtain a fermentation liquid;

[0114] (5) Separation and drying: The fermentation liquid is separated into solid and liquid, and the upper layer is the fermentation clear liquid. The fermentation clear liquid is concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the fermentation product of ear leaf cow hide.

[0115] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0116] The difference from Example 3 is that the order of adding Lactobacillus plantarum and Lactobacillus acidophilus is different.

[0117] Comparative Example 7

[0118] This comparative example provides a method for fermenting ear leaf cow skin shavings with lactic acid bacteria, which specifically comprises the following steps:

[0119] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1280 mL of water, heat to 90° C. and stew for 50 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0120] (2) Enzymolysis: After the temperature of the pretreatment solution drops to 45°C, 3200 mL of water is added, followed by 2.5 g of medium-temperature α-amylase, 3.75 g of saccharifying enzyme and 3.75 g of acid protease, and the mixture is stirred and enzymolyzed at 45°C for 45 min, and sodium bicarbonate is added to adjust the pH to 6.0 to obtain an enzymolysis solution;

[0121] (3) Inactivation of enzymes and fermentation: The enzymatic hydrolysate was inactivated at 100° C. for 50 min, then cooled to 25° C., 0.64 g of Lactobacillus plantarum was added, and the mixture was fermented at 25° C. for 5 h to obtain fermentation liquid A;

[0122] (4) Secondary fermentation: maintaining the temperature of fermentation liquid A at 25° C., adding 0.384 g of Lactobacillus acidophilus and 0.256 g of fermented Lactobacillus mucilaginosus, and continuing fermentation at 25° C. for 13 h to obtain a fermentation liquid;

[0123] (5) Separation and drying: The fermentation liquid is separated into solid and liquid, and the upper layer is the fermentation clear liquid. The fermentation clear liquid is concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the fermentation product of ear leaf cow hide.

[0124] Among them, the enzyme activity of mesophilic α-amylase is 4000U / g, the enzyme activity of saccharifying enzyme is 100000U / g, the enzyme activity of acid protease is 50000U / g; the live bacteria count of Lactobacillus plantarum is 200 billion CFU / g, the live bacteria count of Lactobacillus acidophilus is 200 billion CFU / g, and the live bacteria count of Lactobacillus fermentative mucus is 200 billion CFU / g.

[0125] The difference from Example 3 is that the first fermentation time is extended and the second fermentation time is shortened.

[0126] Comparative Example 8

[0127] This comparative example provides a method for preparing Psoralea corylifolia, which specifically comprises the following steps:

[0128] (1) Pretreatment: grind the ear leaf cow hide shavings and pass it through a 200 mesh sieve to obtain ear leaf cow hide shavings powder, take 320 g of the ear leaf cow hide shavings powder and add it to a stewing pot and add 1280 mL of water, heat to 90° C. and stew for 50 min, add anhydrous citric acid to adjust the pH to 5.0, and obtain a pretreatment solution;

[0129] (2) Separation and drying: The pre-treated liquid is separated into solid and liquid, and the upper clear liquid is concentrated by rotary evaporation and then spray-dried at an air inlet temperature of 130°C and an air outlet temperature of 85°C to obtain the ear leaf cow hide product.

[0130] The difference from Example 3 is that only boiling is performed without enzymolysis or fermentation.

[0131] Experimental Example 1

[0132] This experimental example is the content detection and analysis of C21 steroidal saponins in the fermentation product of Erye Niupixiao. The detection was carried out by high performance liquid chromatography, and the specific method is as follows.

[0133] 1.1 Preparation of the standard product of Auricularia auricula-leaf saponin

[0134] Use an analytical balance to accurately weigh 10 mg each of calcidin, kadogenin, deacetyl rutinin and gacaminin in a 50 mL beaker, and ultrasonically dissolve with an appropriate amount of chromatographic grade 65% ethanol. After complete dissolution, aspirate into a 10 mL volumetric flask and dilute to the scale to obtain a mixed stock solution containing calcidin, kadogenin, deacetyl rutinin and gacaminin at a concentration of 1 mg / mL.

[0135] The mixed solution was then gradiently diluted into mixed standard solutions with concentrations of 200, 150, 100, 50, 10, 5, and 1 μg / mL, and sterilized by filtration using a 0.22 μm filter membrane.

[0136] Among them, gadatenin aglycone was purchased from Sinopharm Chemical Reagent Co., Ltd., model TargetMol-T3150 (HPLC≥98%); kaidenin aglycone was purchased from Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd., model TZT310 / (HPLC≥98%); deacetyl rutin aglycone was purchased from Bio-Tech Biotechnology Co., Ltd., model BCC8163 / (HPLC≥98%); gagamin aglycone was purchased from Bio-Tech Biotechnology Co., Ltd., model BCC8164 / (HPLC≥98%).

[0137] 1.2 Chromatographic conditions

[0138] Chromatographic column model: Shimadzu C18 column (4.6mm×250mm, 5μm); mobile phase: acetonitrile-water; flow rate: 1mL / min; column temperature: 25℃; detection wavelength: 220nm; injection volume: 20μL; detection method: 0-10min, 20%-30% acetonitrile; 10-20min, 30%-40% acetonitrile; 20-30min, 40%-50% acetonitrile; 30-50min, 50%-70% acetonitrile; 50-60min, 70%-20% acetonitrile.

[0139] 1.3 Drawing of standard curve

[0140] The standard curve was drawn with the peak area of ​​the standard as the ordinate and the concentration of each monomer saponin as the abscissa, and the regression line equations of the four C21 steroidal aglycone standards were calculated, as shown in Table 1.

[0141] Table 1 Linear regression equations for each C21 steroid

[0142] Element Regression equation Linear range (μg / mL) <![CDATA[R 2 ]]> Gaudatin Aglycone Y=10.3221X+2.4306 1.00～50.00 0.9997 Kaide Aglycone Y=12.4501X+0.6846 1.00～10.00 0.9996 Deacetyl rutin Y=15.2165X+0.8652 0.10～5.00 0.9999 Gagamin Aglycone Y=24.4874X+1.7478 0.01～0.10 0.9998

[0143] 1.4 Determination of C21 steroid content in Rhizoma Auriculariae

[0144] Take 1.00 g of the fermented product of ear leaf cow hide after separation and drying from Examples 1-4 and Comparative Examples 1-8, respectively, extract it with 65% ethanol ultrasonically (250 W, 50 kHz) for 2 h, collect the supernatant, and extract the precipitate twice under the same conditions, collect the supernatant, make the volume to 1 mL with 65% ethanol, and filter it with a 0.22 μm microporous filter membrane for testing.

[0145] 1.5 Analysis results of C21 steroid content in the fermentation products of Niupixiao

[0146] The saponin content analysis of the fermentation products of Erye Niupixiao of Examples 1-4 and Comparative Examples 1-8 is shown in Table 2.

[0147] Data analysis: The experimental data were statistically analyzed using Excel and SPSS19.0 software. The results were recorded as "mean ± standard deviation". The variance homogeneity test was used for significance analysis, and P < 0.05 was considered to be significantly different.

[0148] Table 2 Analysis results of C21 steroid content in the fermentation products of Auricularia niupixiao (μg / mL, n=3)

[0149]

[0150]

[0151] Note: In the same column of the table, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0152] Result analysis:

[0153] (1) The contents of gaudatin aglycone, kaidengenin, deacetylated rutingenin and gamagnin in the fermented products of the Auricularia sutchuenensis in Examples 1-4 were significantly higher than those in Comparative Examples 1-8, indicating that the preparation method of the Auricularia sutchuenensis provided by the present invention can significantly increase the content of C21 steroid components in the Auricularia sutchuenensis.

[0154] (2) Comparative Example 1 is significantly different from Example 3, indicating that the stewing process before fermentation can significantly increase the content of C21 steroid components in the ear leaf cow skin shavings.

[0155] (3) Comparative Example 2 is significantly different from Example 3, indicating that the selection of enzyme preparations for enzymatic hydrolysis has a significant effect on the content of C21 steroid components in Auricularia auricula, and the selected combination of medium-temperature α-amylase, saccharifying enzyme and acid protease is a better choice.

[0156] (4) Comparative Examples 3-7 groups have significant differences compared with Example 3. This means that the number of fermentations, the selection of strains in the first and second fermentation processes, and the fermentation time have a significant impact on the content of C21 steroidal components in the ear leaf cow hide shavings. The first fermentation (Lactobacillus plantarum) and the second fermentation (Lactobacillus acidophilus and fermented Lactobacillus mucilaginosus) are the better combinations, and increasing the first fermentation time and reducing the second fermentation time cannot significantly increase the content of C21 steroids in the fermentation product. The better fermentation time of the first fermentation is 2h, and the optimal time of the second fermentation is 16h.

[0157] (5) There are significant differences between Comparative Example 8 and Example 3, indicating that enzymatic hydrolysis and fermentation are also effective steps for maximizing the content of C21 steroid components in Rhizoma Auriculariae Niupixiao.

[0158] Experimental Example 2: Experiment on improving digestion: Animal experiment on body weight, weight gain, food intake and food utilization

[0159] 2.1 Experimental animals: Male adult SD rats were used in the experiment. The rat weight was 120-150 g / rat, 10 rats in each group. The difference in animal weight at the beginning of the experiment should not exceed 10% of the average body weight.

[0160] 2.2 Experimental groups: The experiment set up a normal group, embodiment 1 group, embodiment 2 group, embodiment 3 group, embodiment 4 group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, comparative example 5 group, comparative example 6 group, comparative example 7 group, and comparative example 8 group.

[0161] 2.3 Experimental method: Oral test samples were taken for 30 days, and the corresponding fermented products of ear leaf cow peel were fed at a dose of 40 mg / kg in Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5, Comparative Example 6, Comparative Example 7, and Comparative Example 8, and the normal group was fed with an equal amount of physiological saline. Body weight and food intake were measured twice a week. At the end of the experiment, body weight, weight gain, food intake and food utilization were calculated. The food utilization calculation formula is:

[0162]

[0163] 2.4 Data analysis: The experimental data were statistically analyzed using Excel and SPSS19.0 software. The results were recorded as “mean ± standard deviation” and the Decan test was used for significance analysis. P < 0.05 was considered to be significantly different.

[0164] 2.5 Test results:

[0165] After the experiment, the data of rats' weight gain, food intake and food utilization are shown in Table 3.

[0166] Table 3 Effects of fermented products of Auricularia australis on body weight, weight gain, food intake and food utilization of rats (Mean±SD, n=10)

[0167]

[0168]

[0169] Note: In the two columns of weight gain and food utilization, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0170] Result analysis:

[0171] In the embodiment and comparative example, when compared with the normal group, any one of the three indicators of the last body weight, body weight gain and food intake increased, and the difference was significant after statistical processing, and the food utilization rate was not significantly reduced compared with the normal group, the experimental result can be judged to be positive.

[0172] Regarding the weight gain index: compared with the normal group, the weight gain of Example 1-4 groups and Comparative Example 1-8 groups was significantly higher than that of the normal group, indicating that the fermentation product of Auricularia australis can significantly increase the weight gain of rats; compared with the Comparative Example 1-8 groups, the weight gain of rats in Example 1-4 groups was significantly higher than that of rats in Comparative Example 1-8 groups, indicating that the fermentation method selected by the present invention can increase the digestion and absorption effect of the fermentation product of Auricularia australis in rats.

[0173] Food utilization index: compared with the normal group, the food utilization rates of Example 1-4 groups and Comparative Example 1-8 groups were significantly higher than those of the normal group, indicating that the fermentation products of Coix lacryma-vomica can significantly improve the food utilization rate of rats; compared with Comparative Example 1-8 groups, the food utilization rates of Example 1-4 groups were significantly improved, indicating that the fermentation method selected by the present invention can improve the food utilization rate of Coix lacryma-vomica in rats.

[0174] Combining the two indicators of weight gain and food utilization, it can be concluded that the fermentation products of Rhizoma Corydalis can increase the food intake and food utilization of rats.

[0175] Experimental Example 3: Small Intestinal Motility Animal Test

[0176] 3.1 Experimental Grouping: BALB / c mice were adaptively fed for one week and randomly divided into blank control group, model control group, Example 1 group, Example 2 group, Example 3 group, Example 4 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, Comparative Example 5 group, Comparative Example 6 group, Comparative Example 7 group and Comparative Example 8 group. The model control group, Example 1-4 groups and Comparative Example 1-8 groups were modeled with loperamide 2-4 mg / kg.

[0177] 3.2 Test samples: fermented products of Rhizoma Auriculariae Niupixiao obtained from Example 1-4 and Comparative Example 1-8.

[0178] 3.3 Experimental method: The mice in the blank control group and the model control group were given 5 mL of distilled water daily, and the other groups were gavaged with 80 mg / kg of the fermentation product of ear leaf cow hide, for 20 days. Before the end of the experiment, they were fasted but not watered for 16 hours. On the day of the measurement, each experimental group was given the corresponding test sample again, and the blank control group and the model control group were given distilled water. After 30 minutes, except for the blank control group, which was given distilled water, the other groups were given loperamide (3 mg / kg). After 30 minutes, each group was given 0.5 mL of ink as an indicator, and the animals were killed after 30 minutes. The entire section of the small intestine from the pylorus to the cecum was laid straight on white paper, the total length of the small intestine and the distance from the pylorus to the front of the ink were measured and recorded, and the small intestinal propulsion rate was calculated. The specific results are shown in Table 4, and the calculation formula for the small intestinal propulsion rate is:

[0179]

[0180] 3.4 Test results:

[0181] Table 4 Results of mouse small intestinal motility test

[0182]

[0183]

[0184] Note: In the ink advancement rate column, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0185] Result analysis:

[0186] Compared with the blank control group, the ink propulsion rate of the model control group decreased by 11.24%, which was significantly different, indicating that the indigestion model of this experiment was successfully established.

[0187] Compared with the model control group, the ink propulsion rate of each group of Examples 1-4 and Comparative Examples 1-8 was significantly higher than that of the model control group, which indicates that the intestinal propulsion capacity of mice is related to the fermented products of Auricularia nigra ingested by them; compared with the Comparative Examples 1-8, the intestinal ink propulsion rate of mice in Example 1-4 was significantly higher than that of Comparative Examples 1-8, which indicates that the fermentation method selected by the present invention can improve the intestinal propulsion capacity of mice that ingest the fermented products of Auricularia nigra. The fermented products of Auricularia nigra ingested by the present invention can significantly improve the motility of the small intestine of mice.

[0188] Experimental Example 4 Human Food Tasting Test (Children)

[0189] 4.1 Basic information of the subjects: The subjects were all children aged 4-10 years old with indigestion symptoms such as loss of appetite, reduced food intake, and partial eating, and their weight caused by poor diet alone was less than 1 standard deviation from the average normal weight of the same age. The subjects included in the trial did not have the following diseases: acute or chronic diarrhea, positive worm eggs in routine stool examination, combined with systemic diseases such as cardiovascular, liver, kidney and hematopoietic system, taking items related to the test function in a short period of time, affecting the judgment of the results, and not taking the test samples consistently.

[0190] 4.2 Grouping: There are 50 people in both the trial group and the control group, with 25 men and 25 women in each group. The trial group took the fermented product of Auricularia auricula obtained in Example 3 at a dose of 560 mg within 1 hour after lunch and dinner every day during the drinking period. The control group took an equal amount of purified water. The test lasted for 30 days, including a 5-day emptying period and a 25-day drinking period. The trial test was conducted in a blind manner. During the test, the original eating habits were not changed and a normal diet was followed. The efficacy indicators that need to be observed during the test are shown in Table 5.

[0191] Table 5 Observation indicators of human food testing (children)

[0192]

[0193] 4.3 Data processing and result determination:

[0194] (1) Improved appetite: The appetite score of the test group increased significantly when compared with that of the control group before and after the test. The appetite score of the test group or the difference between the two scores before and after the test increased after the test. The difference was statistically significant, and the indicator was judged to be positive.

[0195] (2) Improvement in food intake: The food intake of the test group increased significantly before and after the test. After the test, the food intake of the test group increased compared with that of the control group. The difference between the food intake of the test group and that of the control group increased. The difference was statistically significant and the indicator was judged to be positive.

[0196] (3) Improvement of partial eclipse: The partial eclipse score of the trial group increased significantly when compared with the control group before and after the trial. The partial eclipse score of the trial group or the difference between the two scores before and after the trial increased after the trial. The difference was statistically significant, and the indicator was judged to be positive.

[0197] (4) Body weight measurement and hemoglobin content determination: before and after the tasting, the body weight or hemoglobin content of the tasting group was significantly increased compared with that of the control group. After the tasting, the body weight or hemoglobin content of the tasting group was significantly increased compared with that of the control group. The difference was statistically significant, and the body weight or hemoglobin index was determined to be positive.

[0198] 4.4 Test results

[0199] (1) Appetite: As can be seen from Table 6, the appetite score of the test group increased significantly when compared with itself before and after the test. The difference between the test group and the control group increased after the test. The difference was statistically significant, indicating that the fermentation product of Auricularia auricula can significantly improve the appetite of the test children.

[0200] Table 6 Statistical analysis results of functional index appetite of children before and after trial use

[0201]

[0202] (2) Picky eating: As can be seen from Table 7, the picky eating score of the trial group increased significantly when compared with itself before and after the trial. The difference between the trial group and the control group increased after the trial. The difference was statistically significant, indicating that the fermentation product of ear leaf cow peel can significantly improve the picky eating of the test children.

[0203] Table 7 Statistical analysis results of functional index partial eclipse in children before and after trial use

[0204]

[0205] (3) Food intake: As can be seen from Table 8, the food intake of the test group increased significantly when compared with itself before and after the trial. The difference between the test group and the control group increased after the trial. The difference was statistically significant, indicating that the fermentation product of Auricularia auricula-Coprinus can significantly increase the food intake of the test children.

[0206] Table 8 Statistical analysis results of functional index food intake of children before and after trial use

[0207]

[0208]

[0209] (4) Body weight: As can be seen from Table 9, the weight of the test children increased significantly when the test group was compared with itself before and after the test. The difference between the test group and the control group increased after the test. The difference was significant after statistical processing, indicating that the fermentation product of Auricularia auricula can significantly increase the weight of the test children.

[0210] Table 9 Statistical analysis results of functional indicators and weight of children before and after the trial

[0211]

[0212] (5) Hemoglobin: As can be seen from Table 10, the hemoglobin content of the test children increased significantly when the test group was compared with itself before and after the test. The difference between the test group and the control group increased after the test. The difference was significant after statistical processing, indicating that the fermentation product of ear leaf cow hide can significantly increase the hemoglobin content of the test children.

[0213] Table 10 Statistical analysis results of functional index hemoglobin in children before and after trial use

[0214]

[0215] To sum up: the results for improving children's digestive function, appetite, food intake, and picky eating were positive, and the results for weight and hemoglobin were positive. It can be confirmed that the fermentation products of Auricularia auricula have a digestive effect.

[0216] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. The present invention provides a method for fermenting ear leaf cow skin Xiao with lactic acid bacteria, characterized in that: The specific steps include: (1) Pretreatment: Add the ear leaf cow skin powder into water and simmer at high temperature to obtain a pretreatment solution; (2) enzymolysis: adding water to the pretreatment solution, and then adding an enzyme preparation to perform enzymolysis to obtain an enzymolysis solution; (3) enzyme inactivation and fermentation: after sterilizing the enzymatic hydrolysate, adding Lactobacillus plantarum and fermenting for 1-3 hours at a temperature of 25-30° C. to obtain fermentation liquid A; (4) Secondary fermentation: adding the composite strain to the fermentation liquid A and fermenting for 14-18 hours to obtain a fermentation liquid, separating the fermentation liquid into solid and liquid, and the upper layer is the fermentation clear liquid; The composite bacterial strain comprises Lactobacillus acidophilus and Lactobacillus mucosa fermentans.

2. The preparation method according to claim 1, characterized in that: The weight ratio of the Auricularia auriculariae powder to the Lactobacillus plantarum is (500-800):1, and the weight ratio of the Auricularia auriculariae powder to the composite strain is (500-1000):

1.

3. The preparation method according to claim 1, characterized in that: The weight ratio of the Lactobacillus acidophilus to the Lactobacillus mucilaginosus fermentation is (1-3): (2-5).

4. The preparation method according to claim 1, characterized in that: The weight ratio of the ear leaf cow peel powder to the enzyme preparation is (30-50):

1.

5. The preparation method according to claim 1, characterized in that: The enzyme preparation comprises medium-temperature α-amylase, saccharifying enzyme and acidic protease; the weight ratio of the medium-temperature α-amylase, saccharifying enzyme and acidic protease is (2-10): (2-7): (3-8).

6. The preparation method according to claim 1, characterized in that: In the step (1), the weight ratio of water to the Auricularia australis powder is (2-5):1; and in the step (2), the weight ratio of water to the Auricularia australis powder is (5-15):

1.

7. The preparation method according to claim 1, characterized in that: In the step (1), the high temperature stewing temperature is 80-100° C. and the time is 30-60 minutes.

8. The preparation method according to claim 1, characterized in that: In step (2), the enzymatic hydrolysis temperature is 40-60°C and the enzymatic hydrolysis time is 30-60 min; in step (3), the enzymatic hydrolysis solution is sterilized for 30-90 min at a temperature of 90-120°C.

9. The preparation method according to claim 1, characterized in that: The fermentation time in step (3) is 2 hours; the fermentation time in step (4) is 16 hours.

10. The preparation method according to claim 1, characterized in that: The method also includes the step of concentrating the fermentation clear liquid and then spray-drying it to obtain a fermentation product.

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