CN112899169A - Fermentation method of mulberry leaf cordyceps militaris nano-selenium thallus - Google Patents
Fermentation method of mulberry leaf cordyceps militaris nano-selenium thallus Download PDFInfo
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- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 65
- 239000011669 selenium Substances 0.000 title claims abstract description 65
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 60
- 238000000855 fermentation Methods 0.000 title claims abstract description 57
- 230000004151 fermentation Effects 0.000 title claims abstract description 57
- 240000000249 Morus alba Species 0.000 title claims abstract description 55
- 235000008708 Morus alba Nutrition 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 24
- 229940091258 selenium supplement Drugs 0.000 claims abstract description 64
- 239000000284 extract Substances 0.000 claims abstract description 34
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 30
- 241001052560 Thallis Species 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 13
- 239000011781 sodium selenite Substances 0.000 claims abstract description 11
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims abstract description 9
- 229960001471 sodium selenite Drugs 0.000 claims abstract description 9
- 235000015921 sodium selenite Nutrition 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims abstract description 7
- 238000000605 extraction Methods 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 40
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 18
- 239000011347 resin Substances 0.000 claims description 18
- 229920005989 resin Polymers 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 12
- 229910021529 ammonia Inorganic materials 0.000 claims description 9
- 229930013930 alkaloid Natural products 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 239000000287 crude extract Substances 0.000 claims description 6
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 6
- 229930003935 flavonoid Natural products 0.000 claims description 5
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- 239000002609 medium Substances 0.000 claims description 3
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- 239000005017 polysaccharide Substances 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 150000003342 selenium Chemical class 0.000 claims description 3
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- 239000000306 component Substances 0.000 claims 2
- 239000002075 main ingredient Substances 0.000 claims 1
- 239000012533 medium component Substances 0.000 claims 1
- 238000000643 oven drying Methods 0.000 claims 1
- 241000233866 Fungi Species 0.000 abstract description 7
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- 230000000052 comparative effect Effects 0.000 description 6
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- 229910018143 SeO3 Inorganic materials 0.000 description 4
- 229930003944 flavone Natural products 0.000 description 4
- 235000011949 flavones Nutrition 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 3
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- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 description 1
- 241000235349 Ascomycota Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 description 1
- LXBIFEVIBLOUGU-UHFFFAOYSA-N Deoxymannojirimycin Natural products OCC1NCC(O)C(O)C1O LXBIFEVIBLOUGU-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000221775 Hypocreales Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000218231 Moraceae Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 206010039921 Selenium deficiency Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000002479 acid--base titration Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 231100000693 bioaccumulation Toxicity 0.000 description 1
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- 230000036983 biotransformation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- OFEZSBMBBKLLBJ-BAJZRUMYSA-N cordycepin Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)C[C@H]1O OFEZSBMBBKLLBJ-BAJZRUMYSA-N 0.000 description 1
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- LXBIFEVIBLOUGU-JGWLITMVSA-N duvoglustat Chemical compound OC[C@H]1NC[C@H](O)[C@@H](O)[C@@H]1O LXBIFEVIBLOUGU-JGWLITMVSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
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- 150000002213 flavones Chemical class 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- -1 immunity regulation Chemical compound 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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- 230000000050 nutritive effect Effects 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
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Classifications
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- 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
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
The invention discloses a fermentation method of mulberry leaf cordyceps militaris nano-selenium thalli, and belongs to the field of fermentation of edible fungi. Under the condition of obtaining three different mulberry leaf extract components by extraction and separation, the cordyceps militaris extract components are added into a cordyceps militaris liquid fermentation culture medium, sodium selenite with different concentrations is added into the culture medium, fermentation culture is carried out, and finally cordyceps militaris thalli rich in nano selenium particles and mulberry leaf active ingredients are obtained. The invention adopts the liquid state fermentation of the cordyceps militaris, can effectively convert inorganic selenium element into nano-selenium, and combines the nano-selenium with active ingredients in mulberry leaves, and the prepared cordyceps militaris nano-selenium thalli can be applied to food.
Description
Technical Field
The invention relates to the technical field of fermentation of edible fungi, in particular to a fermentation method of nano-selenium thalli of mulberry leaves and cordyceps militaris.
Background
Folium mori, a traditional Chinese medicine, is the dry leaf of mulberry of the family Moraceae, is produced in most areas throughout the country, and deoxynojirimycin extracted from folium mori is combined with flavonoid extracted from radix puerariae to form a new substance, namely, insulin-washing glucose-clearing agent, which is suitable for treating blood sugar and blood fat reduction, inflammation resistance and the like. In 2014, the mulberry leaves have dietary therapy value because the mulberry leaves have rich nutritive value, and are called mulberry sprouts by modern people.
Selenium is a micronutrient necessary for human body, and has multiple functions of delaying senility, enhancing immunity and the like. Selenium deficiency is a ubiquitous problem, and selenium-rich products such as selenium-rich yeast, selenium-rich edible fungus powder and the like play an important role in food nutrition enhancers and medical products. The brick red nano-selenium prepared by the nano technology can be absorbed and utilized by human bodies despite being essentially zero-valent element selenium in chemical nature, can also play the biological and health-care functions of selenium such as immunity regulation, oxidation resistance and the like, and has lower toxicity than other selenium compounds.
The synthesis of nano-selenium is usually carried out by reducing glucose, Vc and other edible reducing agents with strong reducing power under certain conditions by adopting a chemical reduction method. Plants and microorganisms have good effects on bioaccumulation and selenium conversion, but the nano-selenium conversion effect is not very remarkable. The edible fungi has great potential in the aspects of absorption, transformation and enrichment of nano selenium because of good biological enrichment and transformation effects.
Cordyceps militaris (Cordyceps militaris), also called as scarlet caterpiller fungus, belongs to fungi, Ascomycota, Pyrenomycetes, Clavipitales, Clavipitaceae, Cordyceps, contains various bioactive components such as cordycepin, Cordyceps polysaccharide, etc., and has various physiological functions of resisting oxidation, fatigue, tumor, and regulating immunity. The new food raw material of cordyceps militaris is approved by the national ministry of health council in 2014, the limitation on the usage amount, the usage range and the like of cordyceps militaris is eliminated, and the popularization and the eating of cordyceps militaris as a health food are promoted. Related scholars and persons in the industry also use cordyceps militaris to carry out partial research on the biotransformation of selenium, and cordyceps militaris with a certain selenium-rich amount is cultured, but selenium in the product mainly is selenium compounds such as organic selenium and the like, the content is low, and the synthesis of nano selenium is low. Therefore, how to efficiently produce the cordyceps militaris rich in the nano-selenium and the active ingredients of the mulberry leaves according to the actual demand becomes a new research topic.
Disclosure of Invention
The invention aims to provide a fermentation method for producing cordyceps militaris mycelia rich in mulberry leaf extract and nano-selenium.
In order to achieve the purpose, the invention provides the following technical scheme:
in one embodiment, the mulberry leaves are dry mulberry leaves, and are pulverized, extracted with hot water, and passed through D72 resin to obtain different mulberry leaf extracts.
In one embodiment, the tetravalent inorganic selenium salt is sodium selenite, which is added to the fermentation broth 2-4 days after inoculation to make the final concentration of selenium 0.05-18 mM, and the fermentation is continued for 5-9 days.
In one embodiment, the method comprises the steps of:
(1) weighing 50g of dry mulberry leaves, crushing, sieving with a 50-mesh sieve, extracting the mulberry leaf powder and hot water in a material-water ratio of 1: 10-1: 30 in a boiling water bath for 30-90 min, repeating the extraction for three times, and combining the extracting solutions to obtain a mulberry leaf crude extract.
(2) Allowing the crude extract obtained in step (1) to pass through D72 resin to obtain resin passing liquid, and precipitating to obtain folium Mori extract S1;
(3) performing secondary elution on the D72 resin in the step (2), and concentrating the eluent to obtain a mulberry leaf extract S2 and a mulberry leaf extract S3;
(4) preparing a cordyceps militaris liquid fermentation culture medium, adding the obtained mulberry leaf extract, and sterilizing;
(5) transferring Cordyceps militaris strains to a seed culture solution from a flat plate, fermenting for 48h, and centrifuging to obtain Cordyceps militaris hypha seeds;
(6) the cordyceps militaris hypha seeds obtained in the step (5) are prepared according to the proportion of 1: 50-1: mixing the fermentation broth with a mass (wet weight) volume ratio of 100g/mL, and fermenting for 3-6 d at the temperature of 18-25 ℃ at 100-180 r/min;
(7) and (4) adding a sodium selenite solution into the fermentation liquor obtained in the step (6), continuing fermentation until the total fermentation time reaches 7-9 d, and collecting the fermentation product after the fermentation is finished.
In one embodiment, the resin obtained in step (2) is concentrated by liquid and then precipitated by ethanol with 4 times volume, and the obtained mulberry leaf extract S1 mainly contains macromolecular components such as polysaccharide, protein and the like.
In one embodiment, the secondary elution in step (3) is to wash the D72 resin in step (2) with water and then elute with ethanol to obtain ethanol eluate, and the ethanol eluate is concentrated to obtain folium mori extract S2, mainly containing flavonoids; and eluting the D72 resin with ethanol aqueous solution containing 1% ammonia, wherein the volume content of ethanol in the ethanol aqueous solution is 70%, to obtain ammonia water-ethanol eluate, and concentrating the ammonia water-ethanol eluate to obtain folium Mori extract S3, mainly containing folium Mori alkaloids.
In one embodiment, the mulberry leaf extract added to the fermentation medium in the step (4) is S1 and/or S2 and/or S3.
In one embodiment, centrifugation at 3000-5000 r/min and/or suction filtration is carried out after fermentation to obtain cordyceps militaris thallus rich in mulberry leaf extract and nano-selenium, and drying or freeze drying is carried out.
The invention also claims the cordyceps militaris mycelium which is rich in active ingredients of mulberry leaves and nano selenium and is obtained by fermentation by applying the method.
The invention has the technical effects and advantages that:
according to the invention, the cordyceps militaris liquid state fermentation is adopted, inorganic selenium element can be effectively converted into nano-selenium, and the nano-selenium is combined with active ingredients in mulberry leaves, the yield of nano-selenium in the prepared cordyceps militaris thallus rich in mulberry leaf active ingredients and nano-selenium reaches 4.86% (namely 4.86g/100g dry weight of the thallus), and the conversion rate can reach 99.64% at most. The method adopts an edible fungus liquid state fermentation process, has the characteristics of simple and convenient operation, mild conditions, environmental friendliness and the like, and the prepared cordyceps militaris thallus has potential industrial application prospects in the fields of functional foods, selenium-rich food nutrition enhancers, selenium-rich cosmetic raw materials and medical products.
Detailed Description
The method for measuring the contents of total sugar, total protein, total flavone and alkaloid in the mulberry leaf extract and the cordyceps militaris thallus respectively adopts a phenol-sulfuric acid method, a BCA method, an aluminum nitrate method and an acid-base titration method.
The content of selenium in the thalli is measured by adopting a microwave digestion-ICP-AES method.
The method for calculating the yield and the nano-selenium conversion rate comprises the following steps:
the yield is the total mass (g) of the nano selenium in the thallus/fermentation, and the dry weight (g) of the thallus is multiplied by 100 percent;
the conversion rate of nano selenium is equal to the total mole number (mol) of nano selenium in the thallus/mole number (mol) of selenium with valence of 4 and Na2SeO3Mole) x 100%.
Example 1:
weighing 50g of dry folium mori, crushing, sieving by a 50-mesh sieve, extracting the folium mori powder and hot water in a material-water ratio of 1:30 in a boiling water bath for 60min, repeating the steps for three times, and combining the extracting solutions to obtain a folium mori crude extract. Passing the crude extract through D72 resin to obtain resin passing solution, and precipitating to obtain folium Mori extract S1; washing D72 resin with water, eluting with ethanol to obtain ethanol eluate, and concentrating the ethanol eluate to obtain folium Mori extract S2 mainly containing flavonoids; and eluting the D72 resin with ethanol aqueous solution containing 1% ammonia, wherein the volume content of ethanol in the ethanol aqueous solution is 70%, to obtain ammonia water-ethanol eluate, and concentrating the ammonia water-ethanol eluate to obtain folium Mori extract S3, mainly containing folium Mori alkaloids.
Preparing Cordyceps militaris liquid fermentation culture medium, adding the obtained folium Mori extract S1, and sterilizing. Transferring Cordyceps militaris strains to a seed culture solution from a flat plate, fermenting for 48h, centrifuging to obtain Cordyceps militaris hypha seeds, and mixing the Cordyceps militaris hypha seeds with the seed culture solution according to the proportion of 1: mixing the seeds with the fermentation liquid at a mass (wet weight) volume ratio of 50g/mL, fermenting at 20 deg.C and 150r/min for 4d, adding sodium selenite solution and Na2SeO3Making Na in fermentation liquor2SeO3The concentration is 4.0mM (namely the concentration of inorganic selenium in the fermentation liquor is 4.0mM), the fermentation is continued until the total time is 8d, brick red cordyceps militaris mycelium and orange red fermentation liquor are obtained, the yield of the mycelium is 7.23g/L, and the conversion rate of nano-selenium is 86.07%.
Example 2:
on the basis of the embodiment 1, mulberry leaf extracts S1 and S2 are added into the cordyceps militaris culture medium according to the mass ratio of 1:1, the cordyceps militaris thallus yield is 7.58g/L after fermentation, the nano-selenium conversion rate is increased to 89.76%, and the content of total flavonoids is obviously increased.
Example 3:
on the basis of the embodiment 1, mulberry leaf extracts S1, S2 and S3 are added into a cordyceps militaris culture medium according to the mass ratio of 1:1:1, the yield of cordyceps militaris thalli is 7.48g/L after fermentation, the conversion rate of nano-selenium is 92.60%, and the content of total flavone and total alkaloid is obviously improved.
Example 4:
on the basis of example 3, Na in the fermentation liquor2SeO3The concentration is adjusted to 1.0mM, the output of the cordyceps militaris thallus is 8.53g/L after fermentation, and the nano-selenium conversion rate reaches 99.39％。
Example 5:
on the basis of example 4, Na in the fermentation liquor2SeO3The adding time is adjusted to be 3d, and the table fermentation is continued for 5d after the adding, and the result shows that the cordyceps militaris thallus yield is 7.88g/L and the nano-selenium conversion rate is 96.80%.
Comparative example 1:
the specific implementation manner is the same as that of example 1, except that no mulberry leaf extract is added into the cordyceps militaris fermentation medium, and the results show that the cordyceps militaris bacterium amount and the nano-selenium conversion rate are both low, and are respectively 6.31g/L and 70.72%.
Comparative example 2:
the specific embodiment is the same as example 4, except that Na is added immediately when preparing the fermentation solution2SeO3The result shows that the growth of the cordyceps militaris mycelium is obviously inhibited, the growth is slow, the yield of the mycelium is only 4.27g/L at most, and therefore, the conversion rate of the nano-selenium is low and is 53.54%.
The yield of the cordyceps militaris mycelium, the yield and conversion rate of nano-selenium in the mycelium, and the contents of total flavone and biomass in the mycelium are shown in table 1.
In summary, according to the comparative data in examples 1-3 and comparative example 1, it can be seen that the addition of different mulberry leaf extracts can improve the output of cordyceps militaris thallus, while the addition of mulberry leaf extracts S2 and S3 is helpful to improve the content of total flavones and total alkaloids in cordyceps militaris. According to the data of the examples 1 and 4, the addition of sodium selenite in the fermentation liquor has a certain inhibition effect on the growth of the thalli, and the inhibition effect on the growth of the thalli is small at low concentration of sodium selenite, so that the conversion rate is higher. According to the comparative data of example 5 and comparative example 2, the adding time of sodium selenite has important influence on the growth of the thalli and the biological conversion of selenium, the addition is too early, the growth of the thalli is severely inhibited, and the conversion rate of nano-selenium is also severely reduced.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that are within the spirit and principle of the present invention are intended to be included in the scope of the present invention.
Claims (9)
1. A fermentation method of mulberry leaf cordyceps militaris nano-selenium thalli is characterized by comprising the following steps: the mulberry leaf extract is used as the liquid state fermentation medium component of the cordyceps militaris and is fermented in the environment containing tetravalent inorganic selenium salt to produce cordyceps militaris thallus rich in mulberry leaf active components and nano selenium.
2. The method for fermenting the mulberry leaf and cordyceps militaris nano-selenium thalli as claimed in claim 1, wherein the mulberry leaves are dry mulberry leaves, ground, extracted by hot water, and passed through D72 resin to obtain different mulberry leaf extracts.
3. The method for fermenting the nano-selenium thalli of the mulberry leaves and the cordyceps militaris, as claimed in claim 1, wherein the adopted quadrivalent inorganic selenium salt is sodium selenite, and is added into the fermentation liquid respectively 2-4 days after inoculation, so that the final concentration of selenium is 0.05-18 mM, and the fermentation is continued for 5-9 days.
4. The method for fermenting the nano-selenium thalli of the mulberry leaves and the cordyceps militaris is characterized by comprising the following steps of:
(1) weighing 50g of dry mulberry leaves, crushing, sieving with a 50-mesh sieve, extracting the mulberry leaf powder and hot water in a material-water ratio of 1: 10-1: 30 in a boiling water bath for 30-90 min, repeating the extraction for three times, and combining the extracting solutions to obtain a mulberry leaf crude extract.
(2) Allowing the crude extract obtained in step (1) to pass through D72 resin to obtain resin passing liquid, and precipitating to obtain folium Mori extract S1;
(3) performing secondary elution on the D72 resin in the step (2), and concentrating the eluent to obtain a mulberry leaf extract S2 and a mulberry leaf extract S3;
(4) preparing a cordyceps militaris liquid fermentation culture medium, adding the obtained mulberry leaf extract, and sterilizing;
(5) transferring Cordyceps militaris strains to a seed culture solution from a flat plate, fermenting for 48h, and centrifuging to obtain Cordyceps militaris hypha seeds;
(6) the cordyceps militaris hypha seeds obtained in the step (5) are prepared according to the proportion of 1: 50-1: mixing the fermentation broth with a mass (wet weight) volume ratio of 100g/mL, and fermenting for 3-6 d at the temperature of 18-25 ℃ at 100-180 r/min;
(7) and (4) adding a sodium selenite solution into the fermentation liquor obtained in the step (6), continuing fermentation until the total fermentation time reaches 7-9 d, and collecting the fermentation product after the fermentation is finished.
5. The fermentation method of the mulberry leaf cordyceps militaris nano-selenium thalli, which is characterized by comprising the following steps of: concentrating the resin obtained in the step (2), and precipitating with 4 times volume of ethanol to obtain folium Mori extract S1 containing macromolecular components such as polysaccharide and protein as main ingredients.
6. The fermentation method of the mulberry leaf cordyceps militaris nano-selenium thalli, which is characterized by comprising the following steps of: the secondary elution in the step (3) is to wash the D72 resin in the step (2) with water and then elute with ethanol to obtain ethanol eluent, and the ethanol eluent is concentrated into extractum to obtain mulberry leaf extract S2 which is mainly flavonoid; and eluting the D72 resin with ethanol aqueous solution containing 1% ammonia, wherein the volume content of ethanol in the ethanol aqueous solution is 70%, to obtain ammonia water-ethanol eluate, and concentrating the ammonia water-ethanol eluate to obtain folium Mori extract S3, mainly containing folium Mori alkaloids.
7. The fermentation method of the mulberry leaf cordyceps militaris nano-selenium thalli, which is characterized by comprising the following steps of: the mulberry leaf extract added in the fermentation medium in the step (4) is S1 and/or S2 and/or S3.
8. The fermentation method of the mulberry leaf cordyceps militaris nano-selenium thalli as claimed in claims 1 to 4, which is characterized in that: centrifuging at 3000-5000 r/min after fermentation, and/or performing suction filtration to obtain Cordyceps militaris thallus rich in folium Mori extract and nanometer selenium, and drying by oven drying or freeze drying.
9. A product prepared by the method of any one of claims 1 to 8.
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