Inonotus obliquus fermentation product and application thereof in hypoglycemic drugs

Abstract

The invention relates to an inonotus obliquus fermentation product and application of the inonotus obliquus fermentation product in hypoglycemic drugs, and belongs to the technical field of inonotus obliquus fermentation products. A component culture medium is prepared from 10-15 parts of plant flour, 3-6 parts of glucose, 0.8-1.2 parts of dipotassium phosphate, 1-2 parts of magnesium sulfate, 5-8 parts of protein peptide, 4-8 parts of agar and 60-80 parts of purified water; the component B culture medium is prepared from the following raw materials in parts by weight: 12-15 parts of plant starch, 2-4 parts of glucose, 1-3 parts of oxytetracycline, 1-3 parts of chloramphenicol, 8-16 parts of sodium hyaluronate and 60-80 parts of purified water. Through the multi-target synergistic effect of improving insulin resistance, protecting islet cells, inhibiting sugar absorption and the like, the hypoglycemic effect is obvious, and the prepared inonotus obliquus fermentation product is simple in process, low in operation difficulty, suitable for industrial production and capable of being applied to the fields of cosmetics, textile industry and medicine.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of Inonotus obliquus fermentation product, and particularly relates to an Inonotus obliquus fermentation product and application thereof in hypoglycemic drugs. BACKGROUND

[0002] At present, oral hypoglycemic drugs for type II diabetes mellitus include sulfonylureas, biguanides, thiazolidinediones and alpha-glucosidase inhibitors. The alpha-glucosidase inhibitors can delay the absorption of glucose in the human body without inhibiting the absorption of protein and fat, so they can effectively reduce postprandial blood glucose in clinical practice. The alpha-glucosidase inhibitors on the market mainly include acarbose, voglibose and miglitol. However, the price of the alpha-glucosidase inhibitors is relatively high, and most of them are synthetic drugs, which can cause some adverse effects on the body of patients, such as abdominal discomfort and bloating, when they are taken for a long time. SUMMARY

[0003] The present application provides an Inonotus obliquus fermentation product and application thereof in hypoglycemic drugs to solve the above problems.

[0004] The present application achieves the above-mentioned purpose through the following technical solutions: The present application provides an Inonotus obliquus fermentation product, which is obtained by fermenting Inonotus obliquus strains. The Inonotus obliquus fermentation product is prepared by the following method: (1) The Inonotus obliquus strains are transferred to the A component culture medium for primary fermentation culture (primary shake flask), and cultured at 23℃ for 14 days at a shaking speed of 185r / min. Then, the Inonotus obliquus strains are transferred to the A component culture medium at a 15% inoculation amount for secondary fermentation culture (secondary shake flask), and cultured at 28℃ for 14 days at a shaking speed of 175r / min. Finally, the Inonotus obliquus strains are transferred to the A component culture medium at a 10% inoculation amount for tertiary fermentation culture (tertiary shake flask), and cultured at 28℃ for 7 days at a shaking speed of 165r / min to obtain fermentation product A; (2) The Inonotus obliquus strains are transferred to the B component culture medium for primary fermentation culture (primary shake flask), and cultured at 23℃ for 14 days at a shaking speed of 185r / min. Then, the Inonotus obliquus strains are transferred to the B component culture medium at a 15% inoculation amount for secondary fermentation culture (secondary shake flask), and cultured at 28℃ for 14 days at a shaking speed of 175r / min. Finally, the Inonotus obliquus strains are transferred to the B component culture medium at a 10% inoculation amount for tertiary fermentation culture (tertiary shake flask), and cultured at 28℃ for 7 days at a shaking speed of 165r / min to obtain fermentation product B; (3) The fermentation product A and the fermentation product B are mixed and placed in a culture dish, and cultured at 28℃ for 14 days to obtain a mixed fermentation product; (four) the mixed fermentation is filtered, the mycelium and the bacterial liquid are separated, the mycelium is dried and crushed, the bacterial liquid is added to a macroporous resin column, and then the column is eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e., a water fraction, a 20% ethanol fraction, a 40% ethanol fraction, a 60% ethanol fraction and an 80% ethanol fraction; the five fractions are mixed to obtain the Inonotus obliquus fermentation product.

[0005] As a further optimization scheme of the present application, the preparation raw materials of the A component culture medium include, by weight: 10-15 parts of plant flour, 3-6 parts of glucose, 0.8-1.2 parts of dipotassium hydrogen phosphate, 1-2 parts of magnesium sulfate, 5-8 parts of protein peptide, 4-8 parts of agar, and 60-80 parts of purified water; and the preparation raw materials of the B component culture medium include: 12-15 parts of plant starch, 2-4 parts of glucose, 1-3 parts of oxytetracycline, 1-3 parts of chloramphenicol, 8-16 parts of sodium hyaluronate, and 60-80 parts of purified water.

[0006] As a further optimization scheme of the present application, the preparation process of the A component culture medium is as follows: (1) the purified water is divided into two parts with the same weight for standby; (2) one part of the purified water is heated to 35℃, and the plant flour is added and stirred at 25 r / min for 5-10 min to obtain a pre-preparation A; (3) the pre-preparation A is cooled to 25℃, and the glucose, dipotassium hydrogen phosphate and magnesium sulfate are added and stirred at 30 r / min for 20-25 min to obtain a solution A; (4) the other part of the purified water is heated to 65℃, and the agar is added and stirred at 65 r / min for 30-40 min to obtain a pre-preparation B; (5) the pre-preparation B is cooled to 25℃, and the protein peptide is added and stirred at 30 r / min for 20-25 min to obtain a solution B; and (6) the solution A and the solution B are mixed, uniformly distributed, and sterilized at 120℃ to obtain the A component culture medium.

[0007] As a further optimization scheme of the present application, the preparation process of the B component culture medium is as follows: (1) the purified water is divided into two parts with the same weight for standby; (2) one part of the purified water is heated to 35℃, and the plant flour is added and stirred at 25 r / min for 5-10 min to obtain a pre-preparation A; (3) the pre-preparation A is cooled to 25℃, and the glucose, dipotassium hydrogen phosphate and magnesium sulfate are added and stirred at 30 r / min for 20-25 min to obtain a solution A; (4) the other part of the purified water is heated to 65℃, and the agar is added and stirred at 65 r / min for 30-40 min to obtain a pre-preparation B; (5) the pre-preparation B is cooled to 25℃, and the protein peptide is added and stirred at 30 r / min for 20-25 min to obtain a solution B; and (6) the solution A and the solution B are mixed, uniformly distributed, and sterilized at 120℃ to obtain the A component culture medium.

[0008] As a further optimization scheme of the present application, the plant flour is mixed by wheat flour, mung bean flour and sorghum flour; the mass ratio of the wheat flour, mung bean flour and sorghum flour is 1:1:0.5.

[0009] As a further optimization scheme of the present application, the plant starch is mixed by potato starch, corn starch and cassava starch; the mass ratio of the potato starch, corn starch and cassava starch is 0.2:1:0.6.

[0010] The application of the Inonotus obliquus fermentation product in hypoglycemic drugs.

[0011] The application has the advantages that the Inonotus obliquus fermentation product prepared by the application has obvious hypoglycemic effect through the multi-target synergistic effect of improving insulin resistance, protecting islet cells and inhibiting sugar absorption, and the Inonotus obliquus fermentation product prepared by the application has simple process, low operation difficulty, is suitable for industrial production, and can be applied in the fields of cosmetics, textile industry and medicine. DETAILED DESCRIPTION

[0012] The application will be further described below, and it is necessary to point out here that the following detailed description is only used to further illustrate the application, and cannot be understood as limiting the protection scope of the application, and the skilled in the art can make some non-essential improvements and adjustments to the application according to the above application content.

[0013] The method used in the application is a conventional method known by the skilled in the art, and the reagents and other materials used are commercially available products, unless otherwise specified.

[0014] Example 1 The plant flour is mixed by wheat flour, mung bean flour and sorghum flour; the mass ratio of the wheat flour, mung bean flour and sorghum flour is 1:1:0.5. The plant starch is mixed by potato starch, corn starch and cassava starch; the mass ratio of the potato starch, corn starch and cassava starch is 0.2:1:0.6. Preparation process of A component medium: (1) 60 parts of purified water were divided into two equal parts, ready for use; (2) one part of the purified water was heated to 35℃, 10 parts of plant flour were added and stirred at 25 r / min for 5 min to obtain a pre-prepared solution A; (3) the pre-prepared solution A was cooled to 25℃, 3 parts of glucose, 0.8 parts of hydrogen phosphate and 1 part of magnesium sulfate were added at the same time, and stirred at 30 r / min for 20 min to obtain solution A; (4) the other part of the purified water was heated to 65℃, 4 parts of agar were added and stirred at 65 r / min for 30 min to obtain a pre-prepared solution B; (5) the pre-prepared solution B was cooled to 25℃, 5 parts of protein peptide were added and stirred at 30 r / min for 20 min to obtain solution B; (6) solution A and solution B were mixed and sterilized at 120℃ after being uniform to obtain A component medium; Preparation process of B component medium: (1) 60 parts of purified water were divided into two equal parts, ready for use; (2) one part of the purified water was heated to 35℃, 12 parts of plant starch were added and stirred at 25 r / min for 5 min to obtain a pre-prepared solution C; (3) the pre-prepared solution C was cooled to 25℃, 2 parts of glucose and 1 part of oxytetracycline were added at the same time, and stirred at 30 r / min for 20 min to obtain solution C; (4) the other part of the purified water was heated to 65℃, 8 parts of sodium hyaluronate were added and stirred at 65 r / min for 30 min to obtain a pre-prepared solution D; (5) the pre-prepared solution D was cooled to 25℃, 1 part of chloramphenicol was added and stirred at 30 r / min for 20 min to obtain solution D; (6) solution C and solution D were mixed and sterilized at 120℃ after being uniform to obtain B component medium; The Inonotus obliquus fermentation product is prepared by the following method: (1) The Inonotus obliquus strain was transferred to the A component medium for primary fermentation culture (primary shake flask), cultured at 23℃ for 14d, the shaking bed speed was 185r / min, then transferred to the A component medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultured at 28℃ for 14d, the shaking bed speed was 175r / min, finally transferred to the A component medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultured at 28℃ for 7d, the shaking bed speed was 165r / min, to obtain fermentation product A; (2) The Inonotus obliquus strain was transferred to the B component medium for primary fermentation culture (primary shake flask), cultured at 23℃ for 14d, the shaking bed speed was 185r / min, then transferred to the B component medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultured at 28℃ for 14d, the shaking bed speed was 175r / min, finally transferred to the B component medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultured at 28℃ for 7d, the shaking bed speed was 165r / min, to obtain fermentation product B; (Three) ferment A and ferment B are mixed and placed in a culture dish, and cultured at 28℃ for 14 days to obtain a mixed ferment; (Four) the mixed ferment is filtered, and the mycelium and the broth are separated. The mycelium is dried and crushed. The broth is loaded onto a macroporous resin column, and eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e. water fraction, 20% ethanol fraction, 40% ethanol fraction, 60% ethanol fraction and 80% ethanol fraction. The five fractions are mixed to obtain the ferment product of Inonotus obliquus.

[0015] Example 2 The plant flour is a mixture of wheat flour, mung bean flour and sorghum flour, and the mass ratio of the wheat flour, mung bean flour and sorghum flour is 1:1:0.5. The plant starch is a mixture of potato starch, corn starch and cassava starch, and the mass ratio of the potato starch, corn starch and cassava starch is 0.2:1:0.6. The preparation process of the A component culture medium is as follows: (One) 70 parts of purified water are divided into two equal parts, and are ready for use; (Two) one part of the purified water is heated to 35℃, 12 parts of plant flour are added and stirred at 25 r / min for 8 min to obtain a pre-preparation A; (Three) the pre-preparation A is cooled to 25℃, 4 parts of glucose, 1 part of dipotassium hydrogen phosphate and magnesium sulfate are added at the same time, and stirred at 30 r / min for 23 min to obtain a solution A; (Four) the other part of the purified water is heated to 65℃, 6 parts of agar are added and stirred at 65 r / min for 35 min to obtain a pre-preparation B; (Five) the pre-preparation B is cooled to 25℃, 6 parts of protein peptide are added and stirred at 30 r / min for 22 min to obtain a solution B; (Six) the solution A and the solution B are mixed, sterilized at 120℃ after being uniform, and the A component culture medium is obtained; The preparation process of the B component culture medium is as follows: (One) 70 parts of purified water are divided into two equal parts, and are ready for use; (Two) one part of the purified water is heated to 35℃, 13 parts of plant starch are added and stirred at 25 r / min for 7 min to obtain a pre-preparation C; (Three) the pre-preparation C is cooled to 25℃, 3 parts of glucose and 2 parts of terramycin are added at the same time, and stirred at 30 r / min for 23 min to obtain a solution C; (Four) the other part of the purified water is heated to 65℃, 12 parts of sodium hyaluronate are added and stirred at 65 r / min for 35 min to obtain a pre-preparation D; (Five) the pre-preparation D is cooled to 25℃, 2 parts of chloramphenicol are added and stirred at 30 r / min for 22 min to obtain a solution D; (Six) the solution C and the solution D are mixed, sterilized at 120℃ after being uniform, and the B component culture medium is obtained; The ferment product of Inonotus obliquus is prepared by the following method: (I) the Inonotus obliquus species is transferred to the A component culture medium for primary fermentation culture (primary shake flask), 23℃ environment, 185r / min rotation speed, 14d, then transferred to the A component culture medium for secondary fermentation culture (secondary shake flask) at 15% inoculation amount, 28℃ environment, 175r / min rotation speed, 14d, finally transferred to the A component culture medium for tertiary fermentation culture (tertiary shake flask) at 10% inoculation amount, 28℃ environment, 165r / min rotation speed, 7d, to obtain the fermentation product A; (II) the Inonotus obliquus species is transferred to the B component culture medium for primary fermentation culture (primary shake flask), 23℃ environment, 185r / min rotation speed, 14d, then transferred to the B component culture medium for secondary fermentation culture (secondary shake flask) at 15% inoculation amount, 28℃ environment, 175r / min rotation speed, 14d, finally transferred to the B component culture medium for tertiary fermentation culture (tertiary shake flask) at 10% inoculation amount, 28℃ environment, 165r / min rotation speed, 7d, to obtain the fermentation product B; (III) the fermentation product A and the fermentation product B are mixed and placed in a culture dish, 28℃ environment, 14d, to obtain the mixed fermentation product; (IV) the mixed fermentation product is filtered, the mycelium and the broth are separated, the mycelium is dried and crushed; the broth is subjected to macroporous resin column, and then eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e. water fraction, 20% ethanol fraction, 40% ethanol fraction, 60% ethanol fraction and 80% ethanol fraction; the five fractions are mixed to obtain the Inonotus obliquus fermentation product.

[0016] Example 3 The plant flour is a mixture of wheat flour, green bean flour and sorghum flour; the mass ratio of the wheat flour, the green bean flour and the sorghum flour is 1:1:0.5; The plant starch is a mixture of potato starch, corn starch and cassava starch; the mass ratio of the potato starch, the corn starch and the cassava starch is 0.2:1:0.6; Preparation process of A component medium: (1) 80 parts of purified water were divided into two equal parts, ready for use; (2) one part of the purified water was heated to 35℃, 15 parts of plant flour were added and stirred at 25 r / min for 10 min to obtain a pre-prepared solution A; (3) the pre-prepared solution A was cooled to 25℃, 6 parts of glucose, 1.2 parts of di-potassium hydrogen phosphate and magnesium sulfate were added at the same time, and stirred at 30 r / min for 25 min to obtain solution A; (4) the other part of the purified water was heated to 65℃, 8 parts of agar were added and stirred at 65 r / min for 40 min to obtain a pre-prepared solution B; (5) the pre-prepared solution B was cooled to 25℃, 8 parts of protein peptide were added and stirred at 30 r / min for 25 min to obtain solution B; (6) solution A and solution B were mixed and sterilized at 120℃ to obtain A component medium; Preparation process of B component medium: (1) 80 parts of purified water were divided into two equal parts, ready for use; (2) one part of the purified water was heated to 35℃, 12 parts of plant starch were added and stirred at 25 r / min for 10 min to obtain a pre-prepared solution C; (3) the pre-prepared solution C was cooled to 25℃, 4 parts of glucose and 3 parts of oxytetracycline were added at the same time, and stirred at 30 r / min for 25 min to obtain solution C; (4) the other part of the purified water was heated to 65℃, 16 parts of sodium hyaluronate were added and stirred at 65 r / min for 30 min to obtain a pre-prepared solution D; (5) the pre-prepared solution D was cooled to 25℃, 3 parts of chloramphenicol were added and stirred at 30 r / min for 25 min to obtain solution D; (6) solution C and solution D were mixed and sterilized at 120℃ to obtain B component medium; The Inonotus obliquus fermentation product is prepared by the following method: (1) The Inonotus obliquus strain was transferred to the A component medium for primary fermentation culture (primary shake flask), cultured at 23℃ for 14d, the shaking bed speed was 185r / min, then transferred to the A component medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultured at 28℃ for 14d, the shaking bed speed was 175r / min, finally transferred to the A component medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultured at 28℃ for 7d, the shaking bed speed was 165r / min, to obtain fermentation product A; (2) The Inonotus obliquus strain was transferred to the B component medium for primary fermentation culture (primary shake flask), cultured at 23℃ for 14d, the shaking bed speed was 185r / min, then transferred to the B component medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultured at 28℃ for 14d, the shaking bed speed was 175r / min, finally transferred to the B component medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultured at 28℃ for 7d, the shaking bed speed was 165r / min, to obtain fermentation product B; (Three) the fermentation A and fermentation B mixed, placed in a petri dish, 28 ℃ environment, culture 14d, get mixed fermentation; (Four) the mixed fermentation was filtered, separation of mycelium and broth, mycelium dried and crushed; broth on the macroporous resin column, respectively, with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol gradient elution, respectively, get water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol five kinds of flow fraction; the five kinds of flow fraction were mixed, get inonotus obliquus fermentation product.

[0017] Comparative example 1 The plant flour is mixed by wheat flour, green bean powder and sorghum powder; the mass ratio of the wheat flour, green bean powder and sorghum powder is 1:1:0.5; The plant starch is mixed by potato starch, corn starch and cassava starch; the mass ratio of the potato starch, corn starch and cassava starch is 0.2:1:0.6; The preparation process of A component medium: (one) 70 parts of purified water was divided into two equal parts, for standby; (two) one of the purified water was heated to 35 ℃, 12 parts of plant flour was added and stirred at 25 r / min for 8 min to obtain pre-prepared solution A; (three) the pre-prepared solution A was cooled to 25 ℃, 4 parts of glucose, 1 part of dipotassium hydrogen phosphate and magnesium sulfate were added at the same time, and stirred at 30 r / min for 23 min to obtain solution A; (four) the other part of purified water was heated to 65 ℃, 6 parts of agar was added and stirred at 65 r / min for 35 min to obtain pre-prepared solution B; (five) the pre-prepared solution B was cooled to 25 ℃, 6 parts of protein peptide was added and stirred at 30 r / min for 22 min to obtain solution B; (six) the solution A and solution B were mixed, sterilized at 120 ℃ after uniform, to obtain A component medium; The preparation process of B component medium: (one) 70 parts of purified water was divided into two equal parts, for standby; (two) one of the purified water was heated to 35 ℃, 13 parts of plant starch was added and stirred at 25 r / min for 7 min to obtain pre-prepared solution C; (three) the pre-prepared solution C was cooled to 25 ℃, 3 parts of glucose and 2 parts of terramycin were added at the same time, and stirred at 30 r / min for 23 min to obtain solution C; (four) the other part of purified water was heated to 65 ℃, 12 parts of sodium hyaluronate was added and stirred at 65 r / min for 35 min to obtain pre-prepared solution D; (five) the pre-prepared solution D was cooled to 25 ℃, 2 parts of chloramphenicol was added and stirred at 30 r / min for 22 min to obtain solution D; (six) the solution C and solution D were mixed, sterilized at 120 ℃ after uniform, to obtain B component medium; The inonotus obliquus fermentation product was prepared by the following method: (I) Inoculate the Inonotus obliquus strain into the A component culture medium for primary fermentation culture (primary flask), and cultivate at 23°C for 14 days under the condition of a shaking speed of 185 r / min. Then inoculate it into the A component culture medium at a 15% inoculation amount for secondary fermentation culture (secondary flask), and cultivate at 28°C for 14 days under the condition of a shaking speed of 175 r / min. Finally, inoculate it into the A component culture medium at a 10% inoculation amount for tertiary fermentation culture (tertiary flask), and cultivate at 28°C for 7 days under the condition of a shaking speed of 165 r / min, to obtain the fermentation product A; (I) Inoculate the Inonotus obliquus strain into the A component culture medium for primary fermentation culture (primary flask), and cultivate at 23°C for 14 days under the condition of a shaking speed of 185 r / min. Then inoculate it into the A component culture medium at a 15% inoculation amount for secondary fermentation culture (secondary flask), and cultivate at 28°C for 14 days under the condition of a shaking speed of 175 r / min. Finally, inoculate it into the A component culture medium at a 10% inoculation amount for tertiary fermentation culture (tertiary flask), and cultivate at 28°C for 7 days under the condition of a shaking speed of 165 r / min, to obtain the fermentation product A; (III) Mix the fermentation product A and the fermentation product B, and place them in a culture dish, and cultivate at 28°C for 14 days, to obtain the Inonotus obliquus fermentation product.

[0018] Comparative Example 2 The preparation process of the A component culture medium is as follows: (I) Divide 70 parts of purified water into two equal parts, and reserve; (II) Heat one part of the purified water to 35°C, add 12 parts of potato starch, and stir at 25 r / min for 8 min to obtain a pre-preparation A; (III) Cool the pre-preparation A to 25°C, and simultaneously add 4 parts of glucose, 1 part of hydrogen phosphate dipotassium, and 1 part of magnesium sulfate, and stir at 30 r / min for 23 min to obtain a solution A; (IV) Heat the other part of the purified water to 65°C, add 6 parts of agar, and stir at 65 r / min for 35 min to obtain a pre-preparation B; (V) Cool the pre-preparation B to 25°C, add 6 parts of protein peptide, and stir at 30 r / min for 22 min to obtain a solution B; (VI) Mix the solution A and the solution B, sterilize at 120°C, and obtain the A component culture medium; Preparation process of the B component culture medium: (1) divide 70 parts of purified water into two equal parts, and reserve; (2) heat one part of the purified water to 35℃, add 13 parts of potato starch, and stir at 25 r / min for 7 min to obtain a pre-prepared solution C; (3) cool the pre-prepared solution C to 25℃, and simultaneously add 3 parts of glucose and 2 parts of terramycin, and stir at 30 r / min for 23 min to obtain a solution C; (4) heat another part of the purified water to 65℃, add 12 parts of sodium hyaluronate, and stir at 65 r / min for 35 min to obtain a pre-prepared solution D; (5) cool the pre-prepared solution D to 25℃, add 2 parts of chloramphenicol, and stir at 30 r / min for 22 min to obtain a solution D; (6) mix the solution C and the solution D, and sterilize at 120℃ after being uniform to obtain the B component culture medium; The Inonotus obliquus fermentation product is prepared by the following method: (1) inoculate the Inonotus obliquus strain into the A component culture medium for primary fermentation culture (primary shake flask), cultivate at 23℃ for 14 days, the rotation speed of the shaking table is 185 r / min, then inoculate into the A component culture medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultivate at 28℃ for 14 days, the rotation speed of the shaking table is 175 r / min, finally inoculate into the A component culture medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultivate at 28℃ for 7 days, the rotation speed of the shaking table is 165 r / min, to obtain the fermentation product A; (2) inoculate the Inonotus obliquus strain into the B component culture medium for primary fermentation culture (primary shake flask), cultivate at 23℃ for 14 days, the rotation speed of the shaking table is 185 r / min, then inoculate into the B component culture medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultivate at 28℃ for 14 days, the rotation speed of the shaking table is 175 r / min, finally inoculate into the B component culture medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultivate at 28℃ for 7 days, the rotation speed of the shaking table is 165 r / min, to obtain the fermentation product B; (3) mix the fermentation product A and the fermentation product B, place in a culture dish, cultivate at 28℃ for 14 days, to obtain a mixed fermentation product; (4) filter the mixed fermentation product, separate the mycelium and the broth, dry the mycelium and crush it; load the broth on a macroporous resin column, and elute with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e. a water fraction, a 20% ethanol fraction, a 40% ethanol fraction, a 60% ethanol fraction and an 80% ethanol fraction; mix the five fractions to obtain the Inonotus obliquus fermentation product.

[0019] Comparative Example 3 The plant flour is a mixture of wheat flour, mung bean flour and sorghum flour; the mass ratio of the wheat flour, the mung bean flour and the sorghum flour is 1:1:0.5; The preparation process of the culture medium is as follows: (1) 70 parts of purified water are divided into two equal parts, and are used as needed; (2) one part of the purified water is heated to 35℃, 12 parts of plant flour are added, and stirred at 25 r / min for 8 min to obtain a pre-prepared solution A; (3) the pre-prepared solution A is cooled to 25℃, 4 parts of glucose, 1 part of dipotassium hydrogen phosphate and magnesium sulfate are added at the same time, and stirred at 30 r / min for 23 min to obtain a solution A; (4) the other part of the purified water is heated to 65℃, 6 parts of agar are added, and stirred at 65 r / min for 35 min to obtain a pre-prepared solution B; (5) the pre-prepared solution B is cooled to 25℃, 6 parts of protein peptides are added, and stirred at 30 r / min for 22 min to obtain a solution B; (6) the solution A and the solution B are mixed, sterilized at 120℃ after being uniformly mixed, and a culture medium is obtained. The fermentation product of Inonotus obliquus is prepared by the following method: (1) the Inonotus obliquus strain is transferred to the culture medium for primary fermentation culture (primary shake flask), cultured at 23℃ for 14 days, the shaking speed is 185 r / min, then transferred to the culture medium for secondary fermentation culture (secondary shake flask) at a 15% inoculation amount, cultured at 28℃ for 14 days, the shaking speed is 175 r / min, finally transferred to the culture medium for tertiary fermentation culture (tertiary shake flask) at a 10% inoculation amount, cultured at 28℃ for 7 days, the shaking speed is 165 r / min, and a fermentation product is obtained; (2) the fermentation product is filtered, the mycelium and the broth are separated, the mycelium is dried and crushed, the broth is added to a macroporous resin column, and then eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e. a water fraction, a 20% ethanol fraction, a 40% ethanol fraction, a 60% ethanol fraction and an 80% ethanol fraction; and the five fractions are mixed to obtain the fermentation product of Inonotus obliquus.

[0020] Comparative Example 4 The plant starch is a mixture of potato starch, corn starch and cassava starch; the mass ratio of the potato starch, the corn starch and the cassava starch is 0.2:1:0.6; The preparation process of the culture medium is as follows: (1) 70 parts of purified water are divided into two equal parts, and are used as needed; (2) one part of the purified water is heated to 35℃, 13 parts of plant starch are added, and stirred at 25 r / min for 7 min to obtain a pre-prepared solution A; (3) the pre-prepared solution A is cooled to 25℃, 3 parts of glucose and 2 parts of terramycin are added at the same time, and stirred at 30 r / min for 23 min to obtain a solution A; (4) the other part of the purified water is heated to 65℃, 12 parts of sodium hyaluronate are added, and stirred at 65 r / min for 35 min to obtain a pre-prepared solution B; (5) the pre-prepared solution B is cooled to 25℃, 2 parts of chloramphenicol are added, and stirred at 30 r / min for 22 min to obtain a solution B; (6) the solution A and the solution B are mixed, sterilized at 120℃ after being uniformly mixed, and a culture medium is obtained. The Inonotus obliquus fermentation product is prepared by the following method: (I) The Inonotus obliquus strain is transferred to a subculture medium for primary fermentation culture (primary shake flask), cultured at 23°C for 14 days, the shaking speed is 185 r / min, then transferred to a subculture medium at a 15% inoculation amount for secondary fermentation culture (secondary shake flask), cultured at 28°C for 14 days, the shaking speed is 175 r / min, finally transferred to a culture medium at a 10% inoculation amount for tertiary fermentation culture (tertiary shake flask), cultured at 28°C for 7 days, the shaking speed is 165 r / min, to obtain a fermentation product; (II) The fermentation product is filtered, the mycelium and the broth are separated, the mycelium is dried and crushed; the broth is subjected to macroporous resin column, and then eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol respectively to obtain five fractions, i.e. water fraction, 20% ethanol fraction, 40% ethanol fraction, 60% ethanol fraction and 80% ethanol fraction; the five fractions are mixed to obtain the Inonotus obliquus fermentation product.

[0021] Performance detection ① The total sugar content of the Inonotus obliquus fermentation products prepared in Examples 1-3 and Comparative Examples 1-4 is determined by the phenol-sulfuric acid method, and the determination results are shown in Table 1: Table 1 .

[0022] ② The α-glucosidase inhibitory activity of the Inonotus obliquus fermentation products prepared in Examples 1-3, Comparative Examples 1-4 and the control group with acarbose as the positive sample is determined by the glucose oxidase method, the test concentration is 100 mg / ml, and the results are shown in Table 2: Table 2 .

[0023] ③ The α-glucosidase inhibitory activity of the Inonotus obliquus fermentation products prepared in Examples 1-3, Comparative Examples 1-4 and the control group with acarbose as the positive sample is determined by the glucose oxidase method, the test concentration is 25 mg / ml, and the results are shown in Table 3: Table 3 .

[0024] It can be seen from the comparison of the determination results of each group in Table 1, Table 2 and Table 3 that the hypoglycemic effect of the Inonotus obliquus fermentation product prepared in Example 2 is the best, and is comparable to that of acarbose.

[0025] The above embodiments only express several implementation manners of the present application, and the description is more specific and detailed, but it should not be understood as a limitation on the patent scope of the present application. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are within the protection scope of the present application.

Claims

1. A fermentation product of Inonotus obliquus, characterized in that, The birchwood fermentation product is obtained by fermentation of birchwood strains, and is prepared by the following method: (a) The Inonotus obliquus strain was transferred to component A medium for primary fermentation culture (primary shake flask) and cultured at 23°C for 14 days with a shaker speed of 185 r / min. Then, 15% of the inoculum was transferred to component A medium for secondary fermentation culture (secondary shake flask) and cultured at 28°C for 14 days with a shaker speed of 175 r / min. Finally, 10% of the inoculum was transferred to component A medium for tertiary fermentation culture (tertiary shake flask) and cultured at 28°C for 7 days with a shaker speed of 165 r / min to obtain fermentation product A. (ii) The Inonotus obliquus strain was transferred to component B medium for primary fermentation culture (primary shake flask) and cultured at 23℃ for 14 days with a shaker speed of 185 r / min. Then, 15% of the inoculum was transferred to component B medium for secondary fermentation culture (secondary shake flask) and cultured at 28℃ for 14 days with a shaker speed of 175 r / min. Finally, 10% of the inoculum was transferred to component B medium for tertiary fermentation culture (tertiary shake flask) and cultured at 28℃ for 7 days with a shaker speed of 165 r / min to obtain fermentation product B. (iii) Mix ferment A and ferment B, place them in a petri dish, and incubate at 28°C for 14 days to obtain a mixed ferment; (iv) Filter the mixed fermentation material to separate the mycelium and the bacterial solution. After drying, the mycelium is pulverized. After the bacterial solution is loaded onto a macroporous resin column, it is eluted with water, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol in a gradient to obtain five fractions: water fraction, 20% ethanol fraction, 40% ethanol fraction, 60% ethanol fraction and 80% ethanol fraction. The five fractions are mixed to obtain the fermentation product of Inonotus obliquus.

2. The *Inonotus obliquus* fermentation product according to claim 1, characterized in that, The raw materials for preparing component A of the culture medium, by weight, include: 10-15 parts plant flour, 3-6 parts glucose, 0.8-1.2 parts dipotassium hydrogen phosphate, 1-2 parts magnesium sulfate, 5-8 parts protein peptides, 4-8 parts agar, and 60-80 parts purified water; the raw materials for preparing component B of the culture medium include: 12-15 parts plant starch, 2-4 parts glucose, 1-3 parts oxytetracycline, 1-3 parts chloramphenicol, 8-16 parts sodium hyaluronate, and 60-80 parts purified water.

3. The *Inonotus obliquus* fermentation product according to claim 1, characterized in that, The preparation process of the A component culture medium is as follows: (i) Divide purified water into two equal parts by weight and set aside; (ii) Heat one part of the purified water to 35°C, add plant flour and stir at 25 r / min for 5-10 min to obtain pre-prepared solution A; (iii) Cool the pre-prepared solution A to 25°C, add glucose, dipotassium hydrogen phosphate and magnesium sulfate at the same time, and stir at 30 r / min for 20-25 min to obtain solution A; (iv) Heat the other part of the purified water to 65°C, add agar and stir at 65 r / min for 30-40 min to obtain pre-prepared solution B; (v) Cool the pre-prepared solution B to 25°C, add protein peptides and stir at 30 r / min for 20-25 min to obtain solution B; (vi) Mix solution A and solution B, and sterilize at 120°C to obtain the A component culture medium.

4. The *Inonotus obliquus* fermentation product according to claim 1, characterized in that, The preparation process of the B component culture medium is as follows: (i) Divide purified water into two equal parts by weight and set aside; (ii) Heat one part of the purified water to 35°C, add plant starch and stir at 25 r / min for 5-10 min to obtain pre-prepared solution C; (iii) Cool the pre-prepared solution C to 25°C, add glucose and oxytetracycline at the same time, stir at 30 r / min for 20-25 min to obtain solution C; (iv) Heat the other part of the purified water to 65°C, add sodium hyaluronate and stir at 65 r / min for 30-40 min to obtain pre-prepared solution D; (v) Cool the pre-prepared solution D to 25°C, add chloramphenicol and stir at 30 r / min for 20-25 min to obtain solution D; (vi) Mix solution C and solution D, and sterilize at 120°C to obtain the B component culture medium.

5. The fermentation product of *Inonotus obliquus* according to claim 3, characterized in that, The plant flour is made by mixing wheat flour, mung bean flour and sorghum flour; the mass ratio of wheat flour, mung bean flour and sorghum flour is 1:1:0.

5.

6. The *Inonotus obliquus* fermentation product according to claim 4, characterized in that, The plant starch is a mixture of potato starch, corn starch and cassava starch; the mass ratio of potato starch, corn starch and cassava starch is 0.2:1:0.

6.

7. The application of a fermentation product of Inonotus obliquus in hypoglycemic drugs.