CN111165273B - Culture method and culture medium for high-yield selenium-rich cordyceps militaris - Google Patents
Culture method and culture medium for high-yield selenium-rich cordyceps militaris Download PDFInfo
- Publication number
- CN111165273B CN111165273B CN202010118155.0A CN202010118155A CN111165273B CN 111165273 B CN111165273 B CN 111165273B CN 202010118155 A CN202010118155 A CN 202010118155A CN 111165273 B CN111165273 B CN 111165273B
- Authority
- CN
- China
- Prior art keywords
- culture
- parts
- cordyceps militaris
- culture medium
- culturing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 116
- 239000001963 growth medium Substances 0.000 title claims abstract description 106
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 102
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 102
- 239000011669 selenium Substances 0.000 title claims abstract description 102
- 238000012136 culture method Methods 0.000 title description 5
- 239000007788 liquid Substances 0.000 claims abstract description 98
- 238000012258 culturing Methods 0.000 claims abstract description 81
- 238000000855 fermentation Methods 0.000 claims abstract description 62
- 230000004151 fermentation Effects 0.000 claims abstract description 62
- 239000007787 solid Substances 0.000 claims abstract description 61
- 238000000034 method Methods 0.000 claims abstract description 22
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 229940091258 selenium supplement Drugs 0.000 claims description 98
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 84
- 238000005286 illumination Methods 0.000 claims description 58
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 52
- 239000002994 raw material Substances 0.000 claims description 46
- 239000001888 Peptone Substances 0.000 claims description 43
- 108010080698 Peptones Proteins 0.000 claims description 43
- 235000019319 peptone Nutrition 0.000 claims description 43
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 42
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 42
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 42
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 42
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 40
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 38
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 38
- 239000000843 powder Substances 0.000 claims description 28
- 235000007164 Oryza sativa Nutrition 0.000 claims description 26
- 240000006394 Sorghum bicolor Species 0.000 claims description 26
- 235000011684 Sorghum saccharatum Nutrition 0.000 claims description 26
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 26
- 229930006000 Sucrose Natural products 0.000 claims description 26
- 235000009566 rice Nutrition 0.000 claims description 26
- 229960004793 sucrose Drugs 0.000 claims description 26
- 239000002609 medium Substances 0.000 claims description 25
- 238000009423 ventilation Methods 0.000 claims description 25
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 21
- 229960001471 sodium selenite Drugs 0.000 claims description 21
- 235000015921 sodium selenite Nutrition 0.000 claims description 21
- 239000011781 sodium selenite Substances 0.000 claims description 21
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 19
- 229910052760 oxygen Inorganic materials 0.000 claims description 19
- 239000001301 oxygen Substances 0.000 claims description 19
- 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 claims description 17
- 239000008103 glucose Substances 0.000 claims description 17
- 235000019156 vitamin B Nutrition 0.000 claims description 17
- 239000011720 vitamin B Substances 0.000 claims description 17
- 229930003270 Vitamin B Natural products 0.000 claims description 16
- 229940088594 vitamin Drugs 0.000 claims description 16
- 229930003231 vitamin Natural products 0.000 claims description 16
- 235000013343 vitamin Nutrition 0.000 claims description 16
- 239000011782 vitamin Substances 0.000 claims description 16
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 16
- 229940041514 candida albicans extract Drugs 0.000 claims description 15
- 239000012138 yeast extract Substances 0.000 claims description 15
- 239000009429 Ginkgo biloba extract Substances 0.000 claims description 13
- 238000012546 transfer Methods 0.000 claims description 5
- 240000007594 Oryza sativa Species 0.000 claims 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 abstract description 24
- 239000002126 C01EB10 - Adenosine Substances 0.000 abstract description 12
- 229960005305 adenosine Drugs 0.000 abstract description 12
- 241000209094 Oryza Species 0.000 description 24
- 238000011218 seed culture Methods 0.000 description 21
- 238000001816 cooling Methods 0.000 description 20
- 230000001954 sterilising effect Effects 0.000 description 20
- 238000002156 mixing Methods 0.000 description 19
- 230000000052 comparative effect Effects 0.000 description 15
- 239000005720 sucrose Substances 0.000 description 12
- 229930003451 Vitamin B1 Natural products 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 238000002791 soaking Methods 0.000 description 10
- 229960003495 thiamine Drugs 0.000 description 10
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 10
- 239000011691 vitamin B1 Substances 0.000 description 10
- 235000010374 vitamin B1 Nutrition 0.000 description 10
- 241000830535 Ligustrum lucidum Species 0.000 description 9
- 235000013399 edible fruits Nutrition 0.000 description 9
- 235000012239 silicon dioxide Nutrition 0.000 description 5
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 4
- 241000190633 Cordyceps Species 0.000 description 3
- 229940068052 ginkgo biloba extract Drugs 0.000 description 3
- 235000020686 ginkgo biloba extract Nutrition 0.000 description 3
- 238000004806 packaging method and process Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000255777 Lepidoptera Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000013475 authorization Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001317465 Bambusa textilis Species 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
- 206010006458 Bronchitis chronic Diseases 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- 206010010970 Cor pulmonale chronic Diseases 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
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 241000255925 Diptera Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000008620 Fagopyrum esculentum Species 0.000 description 1
- 235000009419 Fagopyrum esculentum Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000221775 Hypocreales Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000207834 Oleaceae Species 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000001391 atomic fluorescence spectroscopy Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 208000007451 chronic bronchitis Diseases 0.000 description 1
- 208000026636 chronic pulmonary heart disease Diseases 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
- 230000007812 deficiency Effects 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229940106580 ginkgo biloba leaf extract Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 150000004678 hydrides Chemical class 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229940046001 vitamin b complex Drugs 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/50—Inoculation of spawn
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/60—Cultivation rooms; Equipment therefor
- A01G18/69—Arrangements for managing the environment, e.g. sprinklers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to a method for culturing high-yield selenium-rich cordyceps militaris, which comprises the following steps: s1, plate culture; s2, liquid fermentation culture; s3, hypha culture; s4, color conversion culture; s5, culturing stroma; s6, culturing sporocarp. The invention also provides a liquid fermentation culture medium and a solid culture medium for high-yield selenium-rich cordyceps militaris. The invention improves the output, selenium content and adenosine content of the cordyceps militaris.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a culture method and a culture medium for high-yield selenium-rich cordyceps militaris.
Background
Cordyceps militaris (Cordyceps militaris) also called Cordyceps militaris and Cordyceps militaris, belong to the phylum Eumycota, Ascomycotina, Pyrenomycetes, Clavicipitales, Clavipitaceae, genus Cordyceps, are heterogeneous with Cordyceps sinensis, and are the model species of Cordyceps genus. A large number of researches show that the pharmacological action of cordyceps militaris is very similar to that of cordyceps sinensis, the content and SOD activity of medicinal components of cordycepic acid, cordyceps polysaccharide, cordycepin and the like are higher than those of the cordyceps sinensis, and the cordyceps militaris and cordyceps militaris and cordyceps militaris extract has the effects of resisting tumors, resisting fatigue, reducing blood sugar, resisting bacteria, diminishing inflammation, improving immunity, tonifying lung yin, kidney yang, treating kidney deficiency and the like, and has obvious curative effects on chronic bronchitis and pulmonary heart disease of the old.
The Cordyceps militaris has a relatively wide host range, but has low specificity, is mainly parasitic on larvae, imagoes and pupae of insects such as Lepidoptera, Diptera and Coleoptera, mainly Lepidoptera, and is divided into 11 families of 3 meshes and 19 families. Cordyceps militaris is mainly distributed in tropical and subtropical areas and cold and warm climatic zones. The distribution is more in france, germany, the united kingdom in europe and the united states and canada in north america; the natural resources in China are rich and are found in provinces such as Liaoning, Shaanxi, Shanxi, Anhui, Sichuan, Guizhou, Yunnan, Hubei, Hunan, Jilin, Henan, Guangdong, Guangxi, Shandong, Jiangsu, Gansu and Fujian. Cordyceps militaris grows mainly in the middle and shallow mountains with sufficient sunlight and high humidity, and the stroma is usually differentiated in late summer and early autumn every year.
Selenium is an indispensable trace element in human body, can remove excessive oxygen free radicals in human body, effectively prevents the selenium element from being destroyed by lipid peroxidation of cell membranes, is a trace element necessary for human body, and has physiological functions of oxidation resistance and aging resistance. Based on the medicinal value of cordyceps militaris and the importance of selenium to human bodies, the artificial culture of selenium-rich cordyceps militaris is of great significance.
The application publication date is 2018, 7 and 20, and the application publication number is CN108300667A, which discloses a method for fermentation culture of selenium-rich cordyceps militaris, and the method comprises the steps of adding inorganic selenium and phosphate into a culture medium when the pH of the culture medium of selenium-rich fermented cordyceps militaris hyphae is 8; provided that the inorganic selenium and the phosphate are not added simultaneously. The invention avoids the adverse effect in the fermentation culture of the cordyceps militaris by effectively applying phosphate and sulfate, improves the enrichment efficiency and ensures the safety of the product.
The invention discloses a Chinese patent with an authorization notice date of 2019, 7 and 26 and an authorization notice number of CN106544277B, which discloses a method for preparing cordyceps militaris sporocarp products, and comprises the following steps: (a1) culturing Cordyceps militaris strains by using a seed strengthening culture medium to obtain a seed strengthening culture; (a2) inoculating the seed enhanced culture obtained in the step (a1) into a liquid seed culture medium for culture to obtain liquid seeds; (a3) inoculating the liquid seeds obtained in the step (a2) to a selenium-rich culture medium of the sporocarp for culture to obtain the cordyceps militaris sporocarp. The invention utilizes the selenium-rich yeast powder as an organic nitrogen source for culturing the cordyceps militaris strain sporocarp and provides a selenium source, thereby improving the utilization rate and the absorption rate of selenium and greatly enhancing the selenium content in the sporocarp.
The application publication date is 2018, 12 and 7, and the application publication number is CN108934781A, which discloses a high-yield culture medium for selenium-rich cordyceps militaris, and the culture medium comprises: silkworm chrysalis, glucose, buckwheat bran, peptone, phosphate, inorganic selenium, selenium-rich vinasse, selenium-rich yeast, vitamin B complex and water.
The technical schemes disclosed above all have a certain effect of increasing the selenium content in cordyceps militaris, but the selenium content and the yield are not ideal.
Disclosure of Invention
The invention provides a culture method of high-yield selenium-rich cordyceps militaris, which is used for improving the yield, selenium content and adenosine content of the cordyceps militaris.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for culturing high-yield selenium-rich cordyceps militaris is characterized by comprising the following steps:
s1, plate culture: inoculating the cordyceps militaris strain to a PDA culture medium, and culturing for 12-18 days at 18-25 ℃;
s2, liquid fermentation culture: inoculating the strains with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture for 7-15 days at the temperature of 20-28 ℃ and at the speed of 120-180 r/min to obtain liquid strains;
s3, hypha culture: inoculating the liquid strain to a solid culture medium according to the weight ratio of 8-14%, and culturing for 4-12 days at the temperature of 15-25 ℃ under the conditions of light protection, no oxygen and air relative humidity of 50-75% to obtain hyphae;
s4, color conversion culture: culturing the hyphae for 7-12 days under the conditions that the temperature is 16-25 ℃, the illumination intensity is 150-300 Lx, the illumination time is 10-16 h/d, and the relative humidity of air is 70-90%;
s5, stroma culture: culturing the color-transfer cultured hyphae for 8-18 days under the conditions that the temperature is 18-25 ℃, the illumination intensity is 200-400 Lx, the illumination time is 6-12 h/d, ventilation is carried out for 20-30 min every 4-6 h, and the relative air humidity is 75-95% to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing the fruiting body for 30-40 days under the conditions that the temperature is 19-28 ℃, the illumination intensity is 200-400 Lx, the illumination time is 6-12 h/d, ventilation is kept, and the relative humidity of air is 75-90%.
In the method for culturing high-yield selenium-rich cordyceps militaris provided by the invention, preferably, the liquid fermentation medium is mainly prepared from the following raw materials in percentage by weight: 2-8% of glucose, 0.1-0.6% of monopotassium phosphate, 0.03-0.3% of magnesium sulfate, 0.6-1.9% of peptone, 2-7% of yeast extract, 0-0.009% of vitamin B10.001 and the balance of water.
In the method for culturing high-yield selenium-rich cordyceps militaris provided by the invention, preferably, the solid culture medium is prepared from the following raw materials in parts by weight: 30-80 parts of rice, 5-15 parts of sorghum, 3-14 parts of bran, 0.1-0.6 part of monopotassium phosphate, 0.03-0.25 part of magnesium sulfate, 1-5 parts of cane sugar, 0.1-0.8 part of peptone, 0.2-2.0 parts of yeast powder, 0. 10.001-0.008 part of vitamin B and 80-170 parts of water, and meanwhile, the solid culture medium further comprises 0.1-2.5 parts of selenium yeast or 0.1-2.5 parts of sodium selenite.
In the method for culturing high-yield selenium-rich cordyceps militaris provided by the invention, preferably, the liquid fermentation medium further comprises the following raw materials in percentage by weight: 0.05-0.2% of ginkgo leaf extract.
In the culture method of high-yield selenium-rich cordyceps militaris provided by the invention, more preferably, the solid culture medium further comprises the following raw materials in parts by weight: 0.01 to 0.15 portion of tabasheer.
On the other hand, the technical problem to be solved by the invention is to provide a liquid fermentation medium and a solid culture medium for culturing cordyceps militaris, which are used for improving the yield and the selenium content of the cordyceps militaris.
The liquid fermentation medium is mainly prepared from the following raw materials in percentage by weight: 2-8% of glucose, 0.1-0.6% of monopotassium phosphate, 0.03-0.3% of magnesium sulfate, 0.6-1.9% of peptone, 2-7% of yeast extract, 0-0.009% of vitamin B10.001 and the balance of water. Preferably, the liquid fermentation medium is mainly prepared from the following raw materials in percentage by weight: 2-4% of glucose, 0.15-0.3% of monopotassium phosphate, 0.05-0.1% of magnesium sulfate, 0.6-1.2% of peptone, 2-4% of yeast extract, 10.002-0.004% of vitamin B and the balance of water. Further preferably, the liquid fermentation medium is mainly prepared from the following raw materials in percentage by weight: 3% of glucose, 0.3% of potassium dihydrogen phosphate, 0.08% of magnesium sulfate, 1% of peptone, 2.5% of yeast, vitamin B10.003% and the balance of water.
In the liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris provided by the invention, the liquid fermentation culture medium preferably further comprises the following raw materials in percentage by weight: 0.05-0.2% of ginkgo leaf extract.
The solid culture medium is mainly prepared from the following raw materials in parts by weight: 30-80 parts of rice, 5-15 parts of sorghum, 3-14 parts of bran, 0.1-0.6 part of monopotassium phosphate, 0.03-0.25 part of magnesium sulfate, 1-5 parts of cane sugar, 0.1-0.8 part of peptone, 0.2-2.0 parts of yeast powder, 0. 10.001-0.008 part of vitamin B and 80-170 parts of water, and meanwhile, the solid culture medium further comprises 0.1-2.5 parts of selenium yeast or 0.1-2.5 parts of sodium selenite. Preferably, the solid culture medium is mainly prepared from the following raw materials in parts by weight: 40-60 parts of rice, 5-8 parts of sorghum, 5-8 parts of bran, 0.1-0.2 part of monopotassium phosphate, 0.05-0.1 part of magnesium sulfate, 1.25-2.5 parts of cane sugar, 0.2-0.4 part of peptone, 0.5-1.0 part of yeast powder and 10.002-0.004 part of vitamin B, and meanwhile, the solid culture medium further comprises 0.25-1.5 parts of selenium yeast or 0.25-1.5 parts of sodium selenite. Further preferably, the solid medium is mainly prepared from the following raw materials in parts by weight: 52 parts of rice, 7 parts of sorghum, 6 parts of bran, 0.1 part of monopotassium phosphate, 0.07 part of magnesium sulfate, 1.5 parts of cane sugar, 0.35 part of peptone, 0.7 part of yeast powder and 0.78 part of vitamin B10.003, and meanwhile, the solid culture medium further comprises 0.6 part of selenium yeast or 0.6 part of sodium selenite.
In the solid culture medium for high-yield selenium-rich cordyceps militaris provided by the invention, the solid culture medium preferably further comprises the following raw materials in parts by weight: 0.01 to 0.15 portion of tabasheer.
Concretio silicea Bambusae seu Schizostachyi is dried block of secretion in stem of Gramineae plant such as Bambusa textilis McClure or Schiostachiyum chinensis Rendle. In the present invention, the concretio silicea bambusae is ground into powder for use.
Compared with the prior art, the invention has the following beneficial technical effects:
the temperature adopted in the hypha culture stage is 15-25 ℃, the light is protected, the oxygen is not contained, the relative humidity of air is 50-75%, the temperature adopted in the color transfer culture stage is 16-25 ℃, the illumination intensity is 150-300 Lx, the illumination time is 10-16 h/d, the oxygen is not contained, the relative humidity of air is 70-90%, the temperature adopted in the stroma culture stage is 16-25 ℃, the illumination intensity is 150-300 Lx, the illumination time is 10-16 h/d, the oxygen is not contained, the relative humidity of air is 70-90%, the temperature adopted in the sporophore culture stage is 19-28 ℃, the illumination intensity is 200-400 Lx, the illumination time is 6-12 h/d, the ventilation is kept, and the relative humidity of air is 75-90%, so that the cordyceps militaris has good growth vigor.
The wheat bran added into the solid culture medium has the effects of increasing the yield of cordyceps militaris, enabling fruiting bodies to grow sturdy and enabling color and luster to be better, and the ginkgo leaf extract added into the liquid fermentation culture medium has the effect of further increasing the yield of cordyceps militaris. The yield of the cordyceps militaris reaches more than 60.6 g/bottle.
The concretio silicea bambusae is added into the solid culture medium to promote the enrichment of selenium element in cordyceps militaris, and the glossy privet fruit is added into the solid culture medium to further promote the enrichment of selenium element in cordyceps militaris, wherein the selenium content of cordyceps militaris reaches over 19.319mg/kg and is up to 29.553 mg/kg.
The adenosine content of the cordyceps militaris reaches 2.15 mg/g. The ginkgo leaf extract is added into the liquid fermentation culture medium and the tabasheer is added into the solid culture medium, so that the synergistic effect is realized on the improvement of the adenosine content of the cordyceps militaris, and the adenosine content is up to 3.19 mg/g.
Drawings
FIG. 1 is a bar graph of the average output, average selenium content and average adenosine content of each group of Cordyceps militaris in the present invention;
FIG. 2 is a bar graph of the average output of each group of Cordyceps militaris in the present invention;
FIG. 3 is a bar graph of the average selenium content of each group of Cordyceps militaris in the present invention;
FIG. 4 is a bar graph of the average adenosine content of each group of Cordyceps militaris in the present invention.
Detailed Description
The following further describes embodiments of the present invention with reference to the drawings. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.2kg of glucose, 0.06kg of monopotassium phosphate, 0.003kg of magnesium sulfate, 0.06kg of peptone, 0.7kg of yeast extract and 10.0001kg of vitamin B, and water is added to make up 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 30 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 8kg of rice, 0.5kg of sorghum, 1.4kg of bran, 0.01kg of monopotassium phosphate, 0.003kg of magnesium sulfate, 0.5kg of sucrose, 0.01kg of peptone, 0.02kg of yeast powder, 10kg of vitamin B10.0008 kg of water and 0.25kg of selenium yeast. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1 and selenium yeast, adding water, soaking for 20 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 18 deg.C for 18 days;
s2, liquid fermentation culture: inoculating the strains with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and carrying out constant-temperature shaking culture for 15 days at the temperature of 20 ℃ and at the speed of 180r/min to obtain liquid strains; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated onto the solid medium at a weight ratio of 8, that is, 16g of liquid seed culture was inoculated into each flask. Culturing at 15 deg.C under dark conditions without oxygen and with relative air humidity of 75% for 12 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 25 deg.C under illumination intensity of 300Lx for 10h/d for 7 days in the absence of oxygen and relative air humidity of 70%;
s5, stroma culture: culturing the color-changed mycelium for 8 days under the conditions of 25 ℃ of temperature, 200Lx of illumination intensity, 12h/d of illumination time, ventilation for 30min every 6h and 95% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 19 deg.C under illumination intensity of 400Lx for 12h/d for 300 days under ventilation and relative air humidity of 90% to obtain selenium-rich Cordyceps militaris.
Example 2
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.8kg of glucose, 0.01kg of monopotassium phosphate, 0.03kg of magnesium sulfate, 0.19kg of peptone, 0.2kg of yeast extract and 10.0009kg of vitamin B, and water is added to make up for 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 3kg of rice, 1.5kg of sorghum, 0.3kg of bran, 0.06kg of monopotassium phosphate, 0.025kg of magnesium sulfate, 0.1kg of sucrose, 0.08kg of peptone, 0.2kg of yeast powder, 10.0001kg of vitamin B, 8kg of water and 0.01kg of sodium selenite. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1 and sodium selenite, adding water, soaking for 10 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 25 deg.C for 12 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and carrying out constant-temperature shaking culture for 7 days at the temperature of 28 ℃ and at the speed of 120r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated onto the solid medium at a ratio of 14% by weight, that is, 28g of the liquid seed culture was inoculated into each flask. Culturing at 25 deg.C under dark condition and without oxygen and with relative air humidity of 50% for 4 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 16 deg.C under illumination intensity of 150Lx for 16h/d for 12 days in the absence of oxygen and relative air humidity of 90%;
s5, stroma culture: culturing the color-transfer cultured mycelia for 18 days under the conditions of 18 ℃ of temperature, 400Lx of illumination intensity, 6h/d of illumination time, ventilation for 20min every 4h and 75% of relative air humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 200Lx for 6h/d under ventilation and air relative humidity of 75% for 40 days to obtain selenium-rich Cordyceps militaris.
Example 3
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract and 10.0003kg of vitamin B, and water is added to make up 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 13kg of vitamin B10.0003 kg of water and 0.125kg of sodium selenite. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1 and sodium selenite, adding water, soaking for 12 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Example 4
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract, 0.0003kg of vitamin B and 0.005kg of ginkgo leaf extract, and adding water to make up for 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 13kg of vitamin B10.0003 kg of water and 0.125kg of sodium selenite. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1 and sodium selenite, adding water, soaking for 12 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Example 5
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract and 10.0003kg of vitamin B, and water is added to make up 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 0.0003kg of vitamin B10, 0.001kg of tabasheer, 13kg of water and 0.125kg of sodium selenite. Mixing rice, sorghum, potassium dihydrogen phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1, tabasheer and sodium selenite uniformly, adding water, soaking for 12 hr, adding bran, mixing uniformly, and packaging into 1L culture bottles, wherein each culture bottle contains 200 g. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Example 6
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract, 0.0003kg of vitamin B and 0.005kg of ginkgo leaf extract, and adding water to make up for 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 0.0003kg of vitamin B10, 0.001kg of tabasheer, 13kg of water and 0.125kg of sodium selenite. Mixing rice, sorghum, potassium dihydrogen phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1, tabasheer and sodium selenite uniformly, adding water, soaking for 12 hr, adding bran, mixing uniformly, and packaging into 1L culture bottles, wherein each culture bottle contains 200 g. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Example 7
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract, 0.0003kg of vitamin B and 0.005kg of ginkgo leaf extract, and adding water to make up for 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 0.0003kg of vitamin B10, 0.001kg of tabasheer, 0.05kg of glossy privet fruit, 13kg of water and 0.125kg of sodium selenite. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1, tabasheer, glossy privet fruit and sodium selenite, adding water, soaking for 12 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Example 8
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract, 0.0003kg of vitamin B and 0.02kg of ginkgo biloba extract, and adding water to make up for 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 0.0003kg of vitamin B10, 0.015kg of tabasheer, 0.08kg of glossy privet fruit, 13kg of water and 0.25kg of selenium yeast. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1, tabasheer, glossy privet fruit and selenium yeast, adding water, soaking for 12 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g of each culture bottle is filled. Then sterilizing the subpackaged solid culture medium for 20min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 19 deg.C for 17 days;
s2, liquid fermentation culture: inoculating the strains with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and carrying out constant-temperature shaking culture for 14 days at the temperature of 23 ℃ at 140r/min to obtain liquid strains; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 12% by weight, that is, 24g of the liquid seed culture was inoculated into each flask. Culturing at 24 deg.C under dark conditions without oxygen and with relative air humidity of 65% for 10 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 24 deg.C under illumination intensity of 270Lx for 14h/d in the absence of oxygen and relative air humidity of 80% for 11 days;
s5, stroma culture: culturing the color-transfer cultured mycelia for 14 days under the conditions of 20 ℃ of temperature, 370Lx of illumination intensity, 10h/d of illumination time, ventilation for 25min every 5h and 80% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 25 deg.C under illumination intensity of 390Lx for 10h/d, ventilation and air relative humidity of 80% for 37 days to obtain selenium-rich Cordyceps militaris.
Comparative example 1
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract and 10.0003kg of vitamin B, and water is added to make up 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.6kg of bran, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 0.0003kg of vitamin B10 and 13kg of water. Uniformly mixing rice, sorghum, monopotassium phosphate, magnesium sulfate, sucrose, peptone, yeast powder and vitamin B1, adding water, soaking for 12 hours, adding bran, uniformly mixing, and subpackaging into 1L culture bottles, wherein 200g is filled in each culture bottle. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
Comparative example 2
A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 0.3kg of glucose, 0.03kg of monopotassium phosphate, 0.008kg of magnesium sulfate, 0.1kg of peptone, 0.25kg of yeast extract and 10.0003kg of vitamin B, and water is added to make up 10 kg. The raw materials are fully dissolved in water and then are subpackaged into 1L triangular bottles, and each triangular bottle is subpackaged with 300 mL. Sterilizing the liquid fermentation culture medium at 121 deg.C under 0.105MPa for 25 min. And cooling for later use.
A solid culture medium for high-yield selenium-rich cordyceps militaris is prepared from the following raw materials: 5.2kg of rice, 0.7kg of sorghum, 0.01kg of monopotassium phosphate, 0.007kg of magnesium sulfate, 0.15kg of cane sugar, 0.035kg of peptone, 0.07kg of yeast powder, 13kg of vitamin B10.0003 kg of water and 0.125kg of sodium selenite. Mixing rice, sorghum, potassium dihydrogen phosphate, magnesium sulfate, sucrose, peptone, yeast powder, vitamin B1 and sodium selenite uniformly, adding water, soaking for 12 hr, and packaging into 1L culture bottles, wherein 200g is filled in each culture bottle. Then sterilizing the subpackaged solid culture medium for 30min under the conditions of pressure of 0.105MPa and temperature of 121 ℃. And cooling for later use.
A method for culturing high-yield selenium-rich Cordyceps militaris comprises the following steps:
s1, plate culture: inoculating Cordyceps militaris strain to PDA culture medium, and culturing at 23 deg.C for 15 days;
s2, liquid fermentation culture: inoculating the strain with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture at 25 ℃ for 10 days at 170r/min to obtain a liquid strain; the clumps were broken up using a magnetic stirrer to distribute the strains evenly.
S3, hypha culture: the liquid seed culture was inoculated to the solid medium at a ratio of 10% by weight, that is, 20g of the liquid seed culture was inoculated into each flask. Culturing at 20 deg.C under dark and oxygen-free conditions with relative air humidity of 70% for 8 days to obtain mycelium;
s4, color conversion culture: culturing the mycelium at 23 deg.C under illumination intensity of 280Lx for 12h/d for 10 days in the absence of oxygen and relative air humidity of 80%;
s5, stroma culture: culturing the color-changed mycelium for 14 days under the conditions of 22 ℃ of temperature, 360Lx of illumination intensity, 8h/d of illumination time, ventilation for 25min every 5h and 85% of air relative humidity to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing fruiting body at 28 deg.C under illumination intensity of 380Lx for 10h/d, ventilation and relative air humidity of 85% for 35 days to obtain selenium-rich Cordyceps militaris.
In the present invention, the glossy privet fruit is a dried ripe fruit of Ligustrum lucidum Ait (Ligustrum lucidum Ait.) belonging to Oleaceae. Harvesting in winter, removing branches and leaves, and pulverizing.
Test examples
In order to verify the beneficial technical effects of the invention, the following tests are specially carried out:
cordyceps militaris cultures were performed according to the methods of examples 1 to 8, comparative example 1 and comparative example 2, and the yield, selenium content, adenosine content, shape and color of Cordyceps militaris were examined (100 bottles of samples were randomly drawn from each test group), and the results are shown in Table 1.
1. The selenium content was determined by the first method (hydride atomic fluorescence spectrometry) of GB 5009.93-2017.
2. The adenosine content was measured by the method of "content measurement" of Cordyceps sinensis in the first part of the "Chinese pharmacopoeia" 2015 edition.
Table 1. output, selenium content, adenosine content, shape and color of each group of Cordyceps militaris
As can be seen from table 1, fig. 2, fig. 3 and fig. 4:
1) the yield of comparative example 2 was significantly lower than the yields of examples 1 to 8 and comparative example 1, and the yields of examples 4, 6, 7 and 8 were significantly higher than those of examples 1, 2, 3, 5 and comparative example 1. The comparative example 2 reduces the bran, and because the bran is added into the solid culture medium, a hard layer is not formed on the surface of the solid culture medium, and the bran has good water absorption, certain gaps are formed among matrixes in the culture medium after the bran is added, so that the liquid strain inflow and the mycelium propagation are facilitated, the fruiting body weed production is facilitated, and the yield of the cordyceps militaris is improved. The ginkgo biloba extract was added to the liquid fermentation medium of examples 4, 6, 7 and 8, and thus the ginkgo biloba extract had the purpose of increasing the yield of cordyceps militaris.
2) The selenium content of the cordyceps militaris obtained in the examples 5 to 8 is obviously higher than that of the cordyceps militaris obtained in the examples 1 to 4 and the comparative examples 1 and 2. The selenium content of examples 7 and 8 was again higher than that of examples 5 and 6, and the selenium content of comparative example 1 was the lowest. In examples 5 and 6, the solid medium was enriched with concretio silicea Bambusae; in examples 7 and 8, the solid medium was supplemented with concretio silicea Bambusae and fructus Ligustri Lucidi; in comparative example 1, selenium yeast or sodium selenite was not added to the solid medium. Therefore, the tabasheer has the function of promoting the cordyceps militaris to absorb the enriched selenium element, and the glossy privet fruit has the function of further promoting the cordyceps militaris to absorb the enriched selenium element.
3) The adenosine content of examples 6 to 8 was significantly higher than that of examples 1 to 5, comparative examples 1 and 2. In examples 6 to 8, ginkgo biloba leaf extract was added to the liquid fermentation medium, and tabasheer was added to the solid medium. Therefore, the ginkgo leaf extract and the tabasheer have a synergistic effect on increasing the adenosine content.
4) In examples 1 to 8 and comparative example 1, the shape and color of the fruiting body were robust and golden yellow; in comparative example 2, the shape and color of the neutron entity were easily aged, thin, and pale yellow. In comparative example 2, bran was reduced in the solid medium. Therefore, the bran added into the solid culture medium has the effects of enabling the fruit bodies to grow robustly and have better color.
The embodiments of the present invention have been described in detail with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (3)
1. A method for culturing high-yield selenium-rich cordyceps militaris is characterized by comprising the following steps:
s1, plate culture: inoculating the cordyceps militaris strain to a PDA culture medium, and culturing for 12-18 days at 18-25 ℃;
s2, liquid fermentation culture: inoculating the strains with good growth vigor in the step S1 into a liquid fermentation culture medium by adopting a puncher with the diameter of 1cm, and performing constant-temperature shaking culture for 7-15 days at the temperature of 20-28 ℃ and at the speed of 120-180 r/min to obtain liquid strains;
s3, hypha culture: inoculating the liquid strain to a solid culture medium according to the weight ratio of 8-14%, and culturing for 4-12 days at the temperature of 15-25 ℃ under the conditions of light protection, no oxygen and air relative humidity of 50-75% to obtain hyphae;
s4, color conversion culture: culturing the hyphae for 7-12 days under the conditions that the temperature is 16-25 ℃, the illumination intensity is 150-300 Lx, the illumination time is 10-16 h/d, and the relative humidity of air is 70-90%;
s5, stroma culture: culturing the color-transfer cultured hyphae for 8-18 days under the conditions that the temperature is 18-25 ℃, the illumination intensity is 200-400 Lx, the illumination time is 6-12 h/d, ventilation is carried out for 20-40 min every 4-6 h, and the relative air humidity is 75-95% to obtain fruiting bodies with the length of 4-6 cm;
s6, fruiting body culture: culturing the sporocarps for 30-40 days under the conditions that the temperature is 19-28 ℃, the illumination intensity is 200-400 Lx, the illumination time is 6-12 h/d, ventilation is kept, and the relative humidity of air is 75-90%;
the solid culture medium is mainly prepared from the following raw materials in parts by weight: 30-80 parts of rice, 5-15 parts of sorghum, 3-14 parts of bran, 0.1-0.6 part of monopotassium phosphate, 0.03-0.25 part of magnesium sulfate, 1-5 parts of cane sugar, 0.1-0.8 part of peptone, 0.2-2.0 parts of yeast powder, 0-0.008 part of vitamin B10.001, 0.01-0.15 part of tabasheer and 80-170 parts of water, and meanwhile, the solid culture medium further comprises 0.1-2.5 parts of selenium yeast or 0.1-2.5 parts of sodium selenite;
the liquid fermentation medium is mainly prepared from the following raw materials in percentage by weight: 2-8% of glucose, 0.1-0.6% of monopotassium phosphate, 0.03-0.3% of magnesium sulfate, 0.6-1.9% of peptone, 2-7% of yeast extract, 0.05-0.009% of vitamin B10.001, 0.05-0.2% of ginkgo leaf extract and the balance of water.
2. A liquid fermentation culture medium for high-yield selenium-rich cordyceps militaris is characterized by being mainly prepared from the following raw materials in percentage by weight: 2-8% of glucose, 0.1-0.6% of monopotassium phosphate, 0.03-0.3% of magnesium sulfate, 0.6-1.9% of peptone, 2-7% of yeast extract, 0.05-0.009% of vitamin B10.001, 0.05-0.2% of ginkgo leaf extract and the balance of water.
3. A solid culture medium for high-yield selenium-rich cordyceps militaris is characterized by being mainly prepared from the following raw materials in parts by weight: 30-80 parts of rice, 5-15 parts of sorghum, 3-14 parts of bran, 0.1-0.6 part of monopotassium phosphate, 0.03-0.25 part of magnesium sulfate, 1-5 parts of cane sugar, 0.1-0.8 part of peptone, 0.2-2.0 parts of yeast powder, 0. 10.001-0.008 part of vitamin B, 0.01-0.15 part of tabasheer and 80-170 parts of water, and meanwhile, the solid culture medium further comprises 0.1-2.5 parts of selenium yeast or 0.1-2.5 parts of sodium selenite.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010118155.0A CN111165273B (en) | 2020-02-26 | 2020-02-26 | Culture method and culture medium for high-yield selenium-rich cordyceps militaris |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010118155.0A CN111165273B (en) | 2020-02-26 | 2020-02-26 | Culture method and culture medium for high-yield selenium-rich cordyceps militaris |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111165273A CN111165273A (en) | 2020-05-19 |
| CN111165273B true CN111165273B (en) | 2022-04-15 |
Family
ID=70621040
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010118155.0A Active CN111165273B (en) | 2020-02-26 | 2020-02-26 | Culture method and culture medium for high-yield selenium-rich cordyceps militaris |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111165273B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113711839B (en) * | 2020-05-26 | 2023-05-30 | 中国科学院大连化学物理研究所 | Artificial cultivation management method for increasing thickness of cordyceps militaris fruiting body |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850964A (en) * | 2006-06-02 | 2006-10-25 | 广东省微生物研究所 | Highactive wild Chinese caterpillar fungus strain, and its fruiting body and artificial culture method |
| CN102972204A (en) * | 2012-12-03 | 2013-03-20 | 无限极（中国）有限公司 | Artificial cultivating method for edible mushrooms |
| CN105009931A (en) * | 2015-04-13 | 2015-11-04 | 鲁东大学 | Preparation of liquid strain for pleurotus eryngii and research method of culture technique of high-quality high-yield pleurotus eryngii through liquid strain |
| CN106350458A (en) * | 2016-08-29 | 2017-01-25 | 杭州雪域生物技术有限公司 | Method for improving yield and activity of fermentation products of agaricus blazei murill liquid |
| CN107333564A (en) * | 2017-07-19 | 2017-11-10 | 安康市农业科学研究所 | A kind of high yield, the production method of high selenium Cordyceps militaris |
| CN107699501A (en) * | 2017-11-24 | 2018-02-16 | 湖南新汇制药股份有限公司 | A kind of culture medium of Hericium erinaceus, bioconversion mycelium, the mycelial extract of bioconversion and application thereof |
| CN108834764A (en) * | 2018-06-19 | 2018-11-20 | 贵州省贵福菌业发展有限公司 | A kind of preparation method of oil tea mushroom activated liquid strain |
| CN109182450A (en) * | 2018-09-14 | 2019-01-11 | 江苏品品鲜生物科技有限公司 | A kind of selection of true pleurotus cornucopiae strain |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897275B (en) * | 2010-08-13 | 2011-08-10 | 义乌市丹溪药用生物开发研究所 | Selenium-rich cordyceps culturing method |
| CN106473099A (en) * | 2016-10-12 | 2017-03-08 | 钟术光 | Functional food or alternative tea that effect improves |
| CN108450236A (en) * | 2018-01-30 | 2018-08-28 | 新平源健农业开发有限公司 | A kind of culture medium and preparation method thereof for flat mushroom strain culture |
-
2020
- 2020-02-26 CN CN202010118155.0A patent/CN111165273B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850964A (en) * | 2006-06-02 | 2006-10-25 | 广东省微生物研究所 | Highactive wild Chinese caterpillar fungus strain, and its fruiting body and artificial culture method |
| CN102972204A (en) * | 2012-12-03 | 2013-03-20 | 无限极（中国）有限公司 | Artificial cultivating method for edible mushrooms |
| CN105009931A (en) * | 2015-04-13 | 2015-11-04 | 鲁东大学 | Preparation of liquid strain for pleurotus eryngii and research method of culture technique of high-quality high-yield pleurotus eryngii through liquid strain |
| CN106350458A (en) * | 2016-08-29 | 2017-01-25 | 杭州雪域生物技术有限公司 | Method for improving yield and activity of fermentation products of agaricus blazei murill liquid |
| CN107333564A (en) * | 2017-07-19 | 2017-11-10 | 安康市农业科学研究所 | A kind of high yield, the production method of high selenium Cordyceps militaris |
| CN107699501A (en) * | 2017-11-24 | 2018-02-16 | 湖南新汇制药股份有限公司 | A kind of culture medium of Hericium erinaceus, bioconversion mycelium, the mycelial extract of bioconversion and application thereof |
| CN108834764A (en) * | 2018-06-19 | 2018-11-20 | 贵州省贵福菌业发展有限公司 | A kind of preparation method of oil tea mushroom activated liquid strain |
| CN109182450A (en) * | 2018-09-14 | 2019-01-11 | 江苏品品鲜生物科技有限公司 | A kind of selection of true pleurotus cornucopiae strain |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111165273A (en) | 2020-05-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104030845B (en) | A kind of glossy ganoderma substituting stuff cultivation substratum that with the addition of Grosvenor Momordica | |
| CN104030844B (en) | A kind of glossy ganoderma substituting stuff cultivation substratum that with the addition of Herba Andrographis | |
| CN106222098B (en) | One plant of monascus strain and its application | |
| US20210079339A1 (en) | Fermentation production process of paecilomyces hepialis cs-4 | |
| CN104030848B (en) | A kind of glossy ganoderma substituting stuff cultivation substratum that with the addition of Flower of Chinese Globeflower | |
| CN101988036A (en) | Aureobasidium pullulans strain as well as preparation method and application thereof in producing pigment-free pullulan | |
| CN111165273B (en) | Culture method and culture medium for high-yield selenium-rich cordyceps militaris | |
| CN111500472B (en) | Cordyceps gunnii mycelium rich in flavone and polyphenol and production method thereof | |
| CN109566269A (en) | A kind of culture medium improving cordycepin content in Cordyceps militaris | |
| CN102286324B (en) | Method for preparing gingko golden fungus fruit wine | |
| CN101638669A (en) | Method for producing bulbus fritillariae cirrhosae total alkaloid by adopting cell mass suspension culture | |
| CN101113430A (en) | Method for gaining panax japonicus secondary metabolite by using panax japonicus cell culture and biological transformation technique | |
| CN106748240A (en) | It is the method that culture medium cultivates Chinese caterpillar fungus to solidify nutrient solution with water-loss reducer | |
| CN104087632B (en) | A kind of deep layer liquid fermentation produces the method for Phellinus igniarius (L. ex Fr.) Quel. extracellular polysaccharide | |
| CN104372034B (en) | A kind of method that hairy roots of giant knotweed produces resveratrol and expansion culture | |
| CN102792849A (en) | Preparation method and application of solid ganoderma lucidum mycelia | |
| CN108812193A (en) | A kind of gastrodin cultivation matrix and preparation method thereof | |
| CN112493260B (en) | Pleione bulbocodioides extract and application thereof in inhibiting plant pathogenic fungi | |
| CN100594238C (en) | Production process for improving suspension cell saponin content of American ginseng | |
| CN104030847B (en) | A kind of glossy ganoderma substituting stuff cultivation substratum that with the addition of hops | |
| CN113141969A (en) | Method for cultivating ganoderma lucidum | |
| CN113583880A (en) | Culture medium suitable for preparing generalized cordyceps sinensis liquid fermentation seed liquid and preparation method and culture method thereof | |
| CN111534548A (en) | Method for segmented fermentation of flowers and fruits based on saccharomycetes and lactobacillus plantarum | |
| CN110583371A (en) | Fermentation method of mushroom liquid strain with high activity and high stability | |
| CN1546644A (en) | Production method of Chinese caterpillar fungus powder and active Chinese caterpillar fungus polysaccharide freeze-drying powder for immunity, and product thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240315 Address after: 230000 Room 203, building 2, phase I, e-commerce Park, Jinggang Road, Shushan Economic Development Zone, Hefei City, Anhui Province Patentee after: Hefei Jiuzhou Longteng scientific and technological achievement transformation Co.,Ltd. Country or region after: China Address before: 467036 Henan province Pingdingshan city new city Longxiang Road Patentee before: Henan University of Urban Construction Country or region before: China |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240329 Address after: No. 11, Shezai Bei Er Xiang, Yuanzhou District, Yichun City, Jiangxi Province, 336000 Patentee after: Chen Dong Country or region after: China Patentee after: Chen Xiaoyun Patentee after: Chen Zhifeng Patentee after: Ma Wenjun Address before: 230000 Room 203, building 2, phase I, e-commerce Park, Jinggang Road, Shushan Economic Development Zone, Hefei City, Anhui Province Patentee before: Hefei Jiuzhou Longteng scientific and technological achievement transformation Co.,Ltd. Country or region before: China |
|
| TR01 | Transfer of patent right |