CN112410228A - Culture of inonotus obliquus biotransformation mycelium and application in reducing blood sugar - Google Patents
Culture of inonotus obliquus biotransformation mycelium and application in reducing blood sugar Download PDFInfo
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- CN112410228A CN112410228A CN202011289873.0A CN202011289873A CN112410228A CN 112410228 A CN112410228 A CN 112410228A CN 202011289873 A CN202011289873 A CN 202011289873A CN 112410228 A CN112410228 A CN 112410228A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The culture of the biotransformation mycelium of the inonotus obliquus and the application of the biotransformation mycelium of the inonotus obliquus in reducing blood sugar. The preparation method of the mycelium composition comprises the following steps: mixing a crop mixture of corn, especially black corn, containing corn kernels, corncobs and corn stigma with rice bran, especially millet rice bran, according to a specific ratio, and adding water to form a culture medium; inoculating the inonotus obliquus strain to a packaging medium such as a bottled medium for culturing; and taking out the whole culture and drying to obtain the culture. The invention efficiently obtains the inonotus obliquus biotransformation mycelium product with hypoglycemic bioactivity by a unique culture method.
Description
Technical Field
The present invention relates generally to the culture of Inonotus obliquus mycelium having a hypoglycemic effect.
Background
Inonotus obliquus is a fungus which parasitizes under the skin of living standing trees such as white birch, silver birch, elm, alder, etc. or on the withered trunk of felled trees, and belongs to the genus Fuscoporia of the order Nostocales, the family Polyporaceae, the class Basidiomycotina. The product contains Fuscoporia obliqua polysaccharide, lanosterol and oxidized triterpenes, Fuscoporia obliqua alcohol and Fuscoporiarin, melanin, tannin compounds, lignin, etc., and has immunity enhancing effect.
The wild resources of the inonotus obliquus are rare, and the problems of sclerotium growth, active ingredients and the like need to be overcome by artificial domestication and cultivation.
Disclosure of Invention
The invention aims to provide a inonotus obliquus biotransformation mycelium product with high hypoglycemic activity.
According to a first aspect of the present invention, there is provided a method for preparing a composition of biotransformed mycelium of Inonotus obliquus, comprising:
providing a corn mixture consisting of corn kernels, corn cobs and corn silk, wherein the corn mixture consists of the following components in parts by weight: 65-75% of corn kernels, 20-30% of corncobs and 3-7% of corn stigma;
providing rice bran;
the corn mixture and the rice bran form uniformly mixed culture medium powder, and the weight of the culture medium powder comprises the following components: 75-95% of corn mixture and 5-25% of rice bran;
adding water into the culture medium powder to form a packaged culture medium, wherein the weight of the added water is 50-150% of the weight of the culture medium powder;
inoculating the inonotus obliquus strain to the packaging culture medium for packaging culture, wherein the culture temperature is 18-30 ℃, and the culture time is 40-60 days;
and taking out all the encapsulated cultures and drying to obtain the biotransformation mycelium composition.
According to a preferred embodiment of the present invention, the corn mixture is composed of corn kernels, corn cobs and corn stigma of black corn crops, and the rice bran is millet rice bran. Compared with common light-colored corn, the black corn has advantages in the aspects of crude fiber, mineral substances (the content of iron is up to 20 percent), vitamin (such as VB1, VB2 and the like) component content and the like, which can cause that the black corn can generate strong synergistic promotion effect on the growth of inonotus obliquus mycelium under the condition of being matched with millet rice bran which has high fiber content, high porosity and difficult caking (the strong mycelium obtained by culturing can see a spot).
In addition, through years of research, the inventor believes that a proper amount of corn stigma, particularly black corn stigma, has the effect of drawing pindianjing on the efficient accumulation of active ingredients such as triterpenes in hyphae.
According to the present invention, the corn blend preferably comprises 70% corn kernels, 25% corn cobs and 5% corn silks by weight.
According to the invention, the composition of the culture medium powder is preferably 80-90% by weight of the corn mixture and 10-20% by weight of the rice bran.
According to the invention, the granularity of the culture medium powder is preferably 10-20 meshes.
In accordance with the present invention, the Inonotus obliquus strain is preferably an Inonotus obliquus (Fr) Pilat strain of the genus Fuscoporia of the family Polyporaceae.
According to the invention, the water and the medium powder are preferably of equal weight in the encapsulated medium.
According to a second aspect of the present invention, there is also provided an inonotus obliquus bioconversion product comprising the inonotus obliquus bioconversion mycelium composition prepared according to the above-described method.
According to a third aspect of the present invention, there is also provided a method for extracting an active material from a composition of biotransformed mycelium of Inonotus obliquus, comprising:
and (3) carrying out water extraction on the inonotus obliquus biotransformation mycelium composition at 55-65 ℃ for 3-6 hours to obtain a corresponding extracting solution, wherein the weight of the extracting water is 8-12 times of that of the mycelium composition.
The extraction method according to the present invention further preferably includes:
and (3) concentrating and drying the obtained extracting solution at 55-65 ℃ under the vacuum condition of 0.04-0.06 MP until a solid extract is obtained.
The inventor finds that the extraction process is also very important, and the extraction method (especially by controlling the water extraction temperature and the subsequent drying temperature to be about 60 ℃) is simple and easy to implement and can carry out high-efficiency extraction without damaging the effective components.
According to a fourth aspect of the present invention, there is also provided an extract of an active substance of Inonotus obliquus obtained by the above extraction method.
According to other aspects of the present invention, the above-mentioned Fuscoporia obliqua bioconversion product or Fuscoporia obliqua active substance extract can be used for effectively reducing blood glucose.
In conclusion, according to the unique culture method of the present invention, the biologically transformed mycelium product of inonotus obliquus with hypoglycemic bioactivity is obtained with high efficiency.
Detailed Description
The present invention is further illustrated by the following specific examples and comparative examples.
Culture substrate raw Material preparation
The black corn seeds are purchased from Chinese agricultural academy of sciences for field planting, and after the seeds are mature, the corn kernels, the corn cobs and the corn silk are respectively mixed according to the weight ratio of 70%, 25% and 5% and then crushed to form a black corn mixture for later use.
The common yellow corn seeds purchased in the market are planted in the field in the same way, and after the corn seeds, the corn cobs and the corn stigma are ripened, the corn grains, the corn cobs and the corn stigma are respectively mixed according to the weight ratio of 70 percent, 25 percent and 5 percent and then crushed to form a yellow corn mixture for later use.
Millet bran and rice bran are purchased on the market for later use.
Inonotus obliquus (Fr) Pilat strain of Inonotus obliquus (Fuscoporia obliqua) of Fuscoporia of Polyporaceae is purchased (from "northeast edible fungi research institute") and pre-cultured into mother strain for use.
Example 1
Mixing the black corn mixture and millet bran at a weight ratio of 80% to 20%, and adding purified water to obtain culture medium.
The resulting culture medium was bottled (500ml) and sterilized before inoculation with the strain stock.
Culturing at 25 deg.C for 55 days, taking out the dense mycelium block material in the bottle, and oven drying to obtain the biotransformation mycelium composition.
Mixing the mycelium composition with purified water at a weight ratio of 1:10, extracting with water at 60.2 deg.C for 4 hr, and drying under 0.05MPa at 60 deg.C to obtain brown solid extract IN-1.
Example 2
Mixing the black corn mixture and millet bran at a weight ratio of 90% to 10%, and adding purified water to obtain culture medium.
The obtained culture medium is bottled and sterilized, and then the strain mother strain is inoculated.
Culturing at 28 deg.C for 52 days, taking out the dense mycelium block material from the bottle, and oven drying to obtain the biotransformation mycelium composition.
Mixing the mycelium composition with purified water at a weight ratio of 1:9, extracting with water at 60 deg.C for 4.2 hr, and drying at 60 deg.C under 0.05MPa to obtain brown solid extract IN-2.
Comparative example 1
The same procedure as in example 1 was repeated except that the black corn mixture in example 1 was replaced with the yellow corn mixture. The final solid extract was labeled IN-3.
Comparative example 2
The same procedure as in example 2 was repeated except that only the millet bran in example 2 was replaced with rice bran. The final solid extract was labeled IN-4.
Extract activity assay
The following animal experiments were performed on the Inonotus obliquus extract of the present invention to test its related activity.
1. Test materials
The animals are healthy clean male mice with the weight of 18-22 g. The test instrument includes a blood glucose meter and a syringe. The test reagent is alloxan. The extracts were IN-1, IN-2, IN-3 and IN-4 obtained according to the above examples and comparative examples.
2. Test method
2.1 establishment of Tetraoxypyrimidine diabetic mouse model
After the mice are adaptively raised for 7 days, the mice are fasted (without water supply) for 12 hours, and venous blood is taken to detect the blood sugar concentration. Injecting alloxan with concentration of 1% twice to establish hyperglycemia model, wherein the dosage of the first injection is higher than that of the second injection, and supplementing 5% of glucose after 2h of injection.
2.2 grouping
The test mice were randomly divided into 5 groups of 10 mice each, namely a normal group, an IN-1 group, an IN-2 group, an IN-3 group and an IN-4 group.
2.3 treatment method
Infusing equal volume of corresponding extract into stomach of IN-1 group, IN-2 group, IN-3 group and IN-4 group, and infusing equal volume of normal saline into stomach of normal group. After 4 weeks of observation, fasting plasma glucose was periodically measured.
2.4 blood glucose assay in Normal mice
50 mice were divided into 5 groups at random. Respectively comprises normal group, IN-1 group, IN-2 group, IN-3 group and IN-4 group. The equivalent volume of the corresponding extract of each group was 400mg/kg by intragastric administration, and the equivalent volume of normal saline was intragastric administration for 28 days, once a day. Periodically taking venous blood to measure blood sugar.
2.5 hyperglycemic white mouse blood glucose assay
50 mice were randomly divided into 5 groups, namely a normal group, an IN-1 group, an IN-2 group, an IN-3 group and an IN-4 group. Alloxan (280mg/kg) was injected intraperitoneally, resulting in a diabetic model. Animals not contributing to the model (fasting for 12h, blood glucose less than 11.1mmol/L) were removed by fasting for 12h before injection and venous blood for 72h after injection for blood glucose determination. The intragastric administration mode of each group is the same as 2.4, 1 time per day, and for 28 consecutive days, venous blood is periodically taken to measure blood sugar.
3. Test results
3.1 Effect of extracts on blood glucose in Normal mice
As shown in table 1, the blood glucose values of the mice in each group after administration were not significantly different from those of the normal group; the blood sugar value of the mice after the administration of each group has no significant difference with the blood sugar value before the administration.
TABLE 1 Effect of extracts on blood glucose in Normal mice
Note: blood glucose values are expressed as mean ± standard deviation. Indicates p <0.05, with statistically significant differences.
3.2 Effect of extracts on blood glucose in diabetic mice
As shown in table 2, there was no significant difference in blood glucose values between groups before treatment; on day 0, fasting blood glucose was high in each diabetic group; on day 7, a significant drop IN blood glucose occurred IN IN-1 and IN-2 groups (p < 0.05); on day 14, the blood glucose in both groups decreased by an average of 25%; on day 21, the blood glucose in these two groups decreased by an average of 40%; on day 28, the blood glucose levels in both groups were close to normal levels. The blood glucose decreased relatively slowly IN IN-3 and IN-4 groups compared to both groups, and also decreased significantly by 28 days.
TABLE 2 Effect of extracts on blood glucose in diabetic mice
Note: blood glucose values are expressed as mean ± standard deviation. Indicates that p <0.05 was statistically significantly different compared to day 0.
The test results show that the IN extracts can reduce the blood sugar of mice with hyperglycemia induced by alloxan to a certain extent, thereby improving the symptoms of mice with diabetes induced by alloxan. However, the hypoglycemic effects of the IN-1 and IN-2 extracts are clearly better than those of the IN-3 and IN-4 extracts.
Claims (10)
1. A method for preparing a composition of biotransformed mycelium of Inonotus obliquus comprises:
providing a corn mixture consisting of corn kernels, corn cobs and corn silk, wherein the corn mixture consists of the following components in parts by weight: 65-75% of corn kernels, 20-30% of corncobs and 3-7% of corn stigma;
providing rice bran;
the corn mixture and the rice bran form uniformly mixed culture medium powder, and the weight of the culture medium powder comprises the following components: 75-95% of corn mixture and 5-25% of rice bran;
adding water into the culture medium powder to form a packaged culture medium, wherein the weight of the added water is 50-150% of the weight of the culture medium powder;
inoculating the inonotus obliquus strain to the packaging culture medium for packaging culture, wherein the culture temperature is 18-30 ℃, and the culture time is 40-60 days;
and taking out the whole packaged culture and drying to obtain the biotransformation mycelium composition.
2. The method of claim 1, wherein the corn mixture is composed of corn kernels, corn cobs and corn silk of black corn, and the rice bran is millet rice bran.
3. The method according to claim 1, wherein the powder of the culture medium has a particle size of 10 to 20 mesh.
4. The process according to claim 1, wherein the strain of Inonotus obliquus (Fr) Pilat is Inonotus obliquus (Inonotus obliquus) of Fuscoporia of Polyporaceae.
5. A Inonotus obliquus bioconversion product comprising an Inonotus obliquus bioconversion mycelium composition prepared according to the method of any one of claims 1-4.
6. A method for extracting active substances from a composition of biotransformed mycelium of Inonotus obliquus prepared according to one of the methods of claims 1 to 4, comprising:
and (3) carrying out water extraction on the inonotus obliquus biotransformation mycelium composition at 55-65 ℃ for 3-6 hours to obtain a corresponding extracting solution, wherein the weight of the extracting water is 8-12 times of that of the mycelium composition.
7. The method of claim 6, further comprising:
and (3) concentrating and drying the obtained extracting solution at 55-65 ℃ under the vacuum condition of 0.04-0.06 MP until a solid extract is obtained.
8. An extract of Inonotus obliquus active substance obtained by the method according to claim 6.
9. Use of a biotransformation product of Inonotus obliquus according to claim 5 for lowering blood glucose.
10. Use of an extract of inonotus obliquus active substance according to claim 8 for lowering blood glucose.
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| CN115181680A (en) * | 2020-11-18 | 2022-10-14 | 山西运奕道生物科技有限公司 | Active substance extraction and blood sugar reducing application in inonotus obliquus culture |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115181680A (en) * | 2020-11-18 | 2022-10-14 | 山西运奕道生物科技有限公司 | Active substance extraction and blood sugar reducing application in inonotus obliquus culture |
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| CN112956683A (en) * | 2021-02-23 | 2021-06-15 | 山西运奕道生物科技有限公司 | Chaba calcium tablet and preparation method thereof |
| CN113278662A (en) * | 2021-05-27 | 2021-08-20 | 山西运奕道生物科技有限公司 | Method for extracting black corn anthocyanin by using inonotus obliquus |
| CN113278662B (en) * | 2021-05-27 | 2022-07-01 | 山西运奕道生物科技有限公司 | Method for extracting black corn anthocyanin by using inonotus obliquus |
Also Published As
| Publication number | Publication date |
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| CN115181680A (en) | 2022-10-14 |
| CN115181680B (en) | 2023-09-05 |
| CN112410228B (en) | 2023-04-07 |
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