CN110387333A - A method of with blue level ground cordyceps culture Antrodia camphorata - Google Patents
A method of with blue level ground cordyceps culture Antrodia camphorata Download PDFInfo
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- 241001486992 Taiwanofungus camphoratus Species 0.000 title claims abstract description 26
- 241000190633 Cordyceps Species 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000000855 fermentation Methods 0.000 claims abstract description 15
- 230000004151 fermentation Effects 0.000 claims abstract description 15
- 239000007787 solid Substances 0.000 claims abstract description 7
- 238000011081 inoculation Methods 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 230000004913 activation Effects 0.000 claims abstract description 3
- 230000006872 improvement Effects 0.000 claims abstract description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 16
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 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 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 239000000706 filtrate Substances 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 4
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- 241000233866 Fungi Species 0.000 claims description 3
- 238000002386 leaching Methods 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims description 2
- 241000233855 Orchidaceae Species 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 claims description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 2
- 239000008272 agar Substances 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 235000005822 corn Nutrition 0.000 claims description 2
- 235000013312 flour Nutrition 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 240000001307 Myosotis scorpioides Species 0.000 claims 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims 1
- 235000012015 potatoes Nutrition 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 239000002893 slag Substances 0.000 claims 1
- 239000007858 starting material Substances 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 claims 1
- 241000123370 Antrodia Species 0.000 abstract description 9
- 238000001914 filtration Methods 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract description 2
- 230000000996 additive effect Effects 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 13
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 6
- 241001264174 Cordyceps militaris Species 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 2
- RQOCXCFLRBRBCS-UHFFFAOYSA-N (22E)-cholesta-5,7,22-trien-3beta-ol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CCC(C)C)CCC33)C)C3=CC=C21 RQOCXCFLRBRBCS-UHFFFAOYSA-N 0.000 description 2
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 2
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 2
- 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 2
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 2
- DNVPQKQSNYMLRS-NXVQYWJNSA-N Ergosterol Natural products CC(C)[C@@H](C)C=C[C@H](C)[C@H]1CC[C@H]2C3=CC=C4C[C@@H](O)CC[C@]4(C)[C@@H]3CC[C@]12C DNVPQKQSNYMLRS-NXVQYWJNSA-N 0.000 description 2
- 230000000202 analgesic effect Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- DNVPQKQSNYMLRS-SOWFXMKYSA-N ergosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H](CC[C@]3([C@H]([C@H](C)/C=C/[C@@H](C)C(C)C)CC[C@H]33)C)C3=CC=C21 DNVPQKQSNYMLRS-SOWFXMKYSA-N 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 150000003648 triterpenes Chemical class 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000723347 Cinnamomum Species 0.000 description 1
- 241000723346 Cinnamomum camphora Species 0.000 description 1
- 241000386927 Cinnamomum micranthum f. kanehirae Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 240000008397 Ganoderma lucidum Species 0.000 description 1
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 1
- 241000023980 Ophiocordyceps lanpingensis Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- SEGLCEQVOFDUPX-UHFFFAOYSA-N di-(2-ethylhexyl)phosphoric acid Chemical compound CCCCC(CC)COP(O)(=O)OCC(CC)CCCC SEGLCEQVOFDUPX-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical group 0.000 description 1
- 125000001033 ether group Chemical group 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to microorganisms technical fields, and in particular to a method of with blue level ground cordyceps culture Antrodia camphorata.Main operational steps are as follows: the strain of Antrodia camphorata is inoculated in the PDA solid medium of improvement, carries out actication of culture；Then seed liquor will be made on the strain inoculation liquid medium within of activation；Using blue level ground cordyceps as additive, fermentation medium is prepared, is sterilized spare；Antrodia camphorata seed liquor is inoculated in fermentation medium and carries out shaking table culture；After culture, filtering fermentation liquor, filter residue is required antrodia mycelia.The present invention be it is pioneering, can be stablized using this method, continuously, in bulk cultivate the higher antrodia mycelia of medical value.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a method of with blue level ground cordyceps culture Antrodia camphorata.
Background technique
Antrodia camphorata (Antrodia cinnamomea) also known as antrodia, camphor tree mushroom, cinnamomum kanahirai hay mushroom, red antrodia, blood ganoderma lucidum, it is a kind of
The rare medicinal fungi being grown in cinnamomum kanehirai.Modern pharmacological studies have shown that Antrodia camphorata has antitumor, liver protection, blood pressure lowering, drop
Cholesterol inhibits the physiological activity such as platelet aggregation, starts the hot spot gradually researched and developed at medical treatment, health care product since 1990, at present
It has been used for treating a variety of diseases.Although Antrodia camphorata drug effect is significant, growth course is extremely slow, and medicinal ingredient is also unstable,
Want very difficult with the common cultivation method acquisition higher Antrodia camphorata thallus of medical value.
Blue level ground cordyceps sinensis (Ophiocordyceps lanpingensis) it is the line Cordyceps that this laboratory was delivered in 2013
Novel species has closer affiliation with cordyceps sinensis.Studies have shown that the blue level ground cordyceps manually cultivated and Xizang Wild winter worm
Summer 15 kinds of active component contents of grass are very much like, can partially be used as medicine instead of cordyceps sinensis；It is extracted from blue level ground cordyceps
For Thick many candies to drosophila anti-aging significant effect, 0.2% Thick many candies can obviously delay female, male drosophila average life span, most long-lived
Life and half death time；Blue level ground cordyceps sinensis has stronger analgesic activity, petroleum ether moiety, acetic acid second in its extractive from fermentative
Ester moiety, dehydrated alcohol part and water, which mention part, periphery property analgesic effect.The blue level ground Cordyceps Militaris that the present invention will be cultivated manually
Powder is as additive, for the liquid fermentation of Antrodia camphorata, then by stringent fermentation, it is final obtain medical value compared with
High antrodia mycelia.
Summary of the invention
The purpose of the present invention is to provide a kind of method with blue level ground cordyceps culture Antrodia camphorata, this method culture
Antrodia mycelia medical value is higher.
To achieve the above object, the present invention adopts the following technical scheme:
Antrodia camphorata strain inoculated activated in the PDA solid medium of improvement to (bacterial strain is in preservation on March 1 in 2017
In Yunnan Prov. Inst. of Microbiology Culture Collection Center, deposit number is YIM Yu201702).Then the strain of activation is seeded in
Seed liquor is made on fluid nutrient medium.Well-grown blue level ground cordyceps militaris sporocarp is chosen, is lyophilized, pulverization process, orchid is made
Level ground cordyceps.By blue level ground cordyceps 5-15g, peptone 10-20g, potato leaching powder 20-30g, glucose 20-30g, water
1000mL configures fermentation medium, 121 DEG C of sterilizings natural cooling, spare after twenty minutes.Antrodia camphorata is inoculated with the inoculum concentration of 5-10%
Seed liquor carries out shaking table culture under conditions of temperature is 25-30 DEG C, revolving speed is 100-150r/min.It was grown to 20-30 days big
By filtering fermentation liquor after amount mycelium.Mycelium precipitating cleans 3 times, 65 DEG C of drying through distilled water, is sealed.
Specific embodiment
Following embodiment can be with the invention will be further described:
1. PDA solid medium prepare: take do not germinate, the potato of paleness, clean peeling, weigh 200g, be cut into 1 centimetre
The fritter of left and right square.The potato fritter cut is put into about 1000mL water, is kept for 30 minutes after boiling with warm fire.Terminate
Filtered through gauze is used afterwards, and obtaining filtrate is potato juice, and filtrate is complemented to 1000mL.Then glucose 20g, peptone is added
5g, agar 20g, pours into test tube respectively after mixing evenly, sterilizes 20 minutes at 121 DEG C, takes out rear-inclined and places, solidification to be cooled
After can be used for transferred species.
2. fluid nutrient medium prepare: take do not germinate, the potato of paleness, clean peeling, weigh 200g, be cut into 1 centimetre
The fritter of left and right square.The potato fritter cut is put into about 1000mL water, is kept for 30 minutes after boiling with warm fire.Terminate
Filtered through gauze is used afterwards, and obtaining filtrate is potato juice, and filtrate is complemented to after 1000mL and is distributed into 500mL/ bottles (triangular flask is
1000mL specification).Every bottle of addition glucose 10g, corn flour 5g, peptone 5g, yeast extract 3g, magnesium sulfate 1.0g, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.5g, stirs evenly, beyond the Great Wall tampon, in 121 DEG C of sterilizings natural cooling, spare after twenty minutes.
3. the production of hybrid seeds: taking Antrodia camphorata strain, the solid slope culture medium that access step 1 makes under aseptic condition, 28 DEG C are protected from light
Culture.When mycelia covers surface about 70%, mycelia can be accessed the fluid nutrient medium production of hybrid seeds.Production of hybrid seeds process is as follows: from slant medium
On take 1 fritter (size about 3-5mm × 3-5mm) mycelia to be put into the triangular flask equipped with fluid nutrient medium, in 120r/min, 28 DEG C
Lower culture about 10 days, it is long to suitable size to bacterium ball, it can be used to be inoculated with.
4. the culture and processing of blue level ground cordyceps sinensis: using rice and dried silkworm chrysalis meal as culture medium raw material, it is real to cultivate blue level ground cordyceps sinensis
Body.Well-grown blue level ground cordyceps militaris sporocarp was chosen at 40 days or so, is lyophilized, pulverization process, blue level ground cordyceps are made.
Bacterial strain for cultivating blue level ground cordyceps militaris sporocarp is preserved in Yunnan Prov. Inst. of Microbiology Culture Collection Center, deposit number YIM
Yu201301。
5. fermentation medium is prepared: preparing fermentation medium by formula, being distributed into 200mL/ bottles, (triangular flask is 500mL rule
Lattice), 121 DEG C of sterilizings natural cooling, spare after twenty minutes.Fermentative medium formula are as follows: blue level ground cordyceps 5g, peptone 10g,
Potato leaching powder 20g, glucose 25g, water 1000mL, pH are natural.
6. inoculation and culture: with 10% inoculum concentration inoculation Antrodia camphorata seed liquor in fermentation medium, temperature be 28 DEG C,
Revolving speed carries out shaking table culture under conditions of being 120r/min.
7. harvesting: by filtering fermentation liquor after growing a large amount of antrodia mycelias in triangular flask.Mycelium is precipitated through distilling
Water cleans 3 times, 65 DEG C of drying, is sealed.
8. the Antrodia camphorata analysis of effective component of blue level ground cordyceps culture
Through detecting, above-mentioned drying, crushing antrodia mycelia in, every 1g contain polysaccharide, PEARLITOL 25C, total triterpene, ergosterol
Respectively 30.48mg, 70.15mg, 34.63mg, 4.72 μ g, hence it is evident that better than blue level ground cordyceps and common Antrodia camphorata (see Table 1).
Multiple batches of culture is carried out according to the method described above, it is as a result stable.
The Antrodia camphorata effective component of the blue level ground cordyceps culture of table 1 compares (n=3)
| Sample | Polysaccharide mg/g | PEARLITOL 25C mg/g | Total triterpene mg/g | Ergosterol μ g/g |
| The Antrodia camphorata of blue level ground cordyceps culture | 30.48±2.67 | 70.15±2.08 | 34.63±1.49 | 4.72±0.12 |
| Blue level ground cordyceps | 23.79±4.47 | 51.39±3.33 | 28.57±1.53 | 1.51±0.02 |
| Common Antrodia camphorata | 3.36±0.27 | 32.96±3.29 | 16.04±0.32 | 1.06±0.02 |
Note: common Antrodia camphorata refers to the antrodia mycelia for not adding the fermentation medium culture of blue level ground cordyceps.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (5)
1. a kind of method with blue level ground cordyceps culture Antrodia camphorata, it is characterised in that:
A, Antrodia camphorata strain is inoculated in the PDA solid medium of improvement, carries out actication of culture；
B, the preparation of seed liquor will be carried out on the strain inoculation liquid medium within of activation；
C, take 500mL triangular flask to be packed into 160-200mL fermentation medium, moist heat sterilization and after cooling down, by 5-10%(w/w) connect
Antrodia camphorata seed liquor is added in kind amount, and shaking table culture is carried out under conditions of temperature is 25-30 DEG C, revolving speed is 100-150r/min.
2. according to the method described in claim 1, it is characterized in that improveing PDA solid medium raw material proportioning described in a are as follows: Ma Ling
Potato juice 1000mL, glucose 5-25g, peptone 5-15g, agar 15-20g.
Here potato juice is obtained by following step: into the water by the peeled potatoes being cut into small pieces, potato and water
Weight ratio is 1:5-1:6, boils rear warm fire and keeps 25-40min, filters off slag, and filtrate is potato juice, following potato juice with
This is identical.
3. according to the method described in claim 1, it is characterized in that liquid medium starting material described in b matches are as follows: potato juice
1000mL, glucose 5-25g, corn flour 5-25g, peptone 5-15g, yeast extract 5-10g, magnesium sulfate 1.0-3.5g, phosphoric acid
Potassium dihydrogen 1.0-2.0g.
4. according to the method described in claim 1, it is characterized in that the formula of fermentation medium described in c are as follows: blue level ground cordyceps
5-15g, peptone 10-20g, potato leaching powder 20-30g, glucose 20-30g, water 1000mL, pH are natural.
5. orchid level ground cordyceps according to claim 4, it is characterised in that bacterium powder be by blue level ground Chinese caterpillar fungus strain solid fermentation,
Freeze-dried again and pulverization process refines.
Here blue level ground Chinese caterpillar fungus strain is preserved in Yunnan Prov. Inst. of Microbiology Culture Collection Center, deposit number YIM
Yu201301。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| TWI819429B (en) * | 2021-12-10 | 2023-10-21 | 賴孟煜 | Fungal fusant strain, method of manufacturing the same and composition including the same |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110387334A (en) * | 2018-04-20 | 2019-10-29 | 云南云百草实验室有限公司 | A kind of method of fast culture antrodia mycelia |
| TWI819429B (en) * | 2021-12-10 | 2023-10-21 | 賴孟煜 | Fungal fusant strain, method of manufacturing the same and composition including the same |
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