CN110387334A - A kind of method of fast culture antrodia mycelia - Google Patents
A kind of method of fast culture antrodia mycelia Download PDFInfo
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- CN110387334A CN110387334A CN201810357719.9A CN201810357719A CN110387334A CN 110387334 A CN110387334 A CN 110387334A CN 201810357719 A CN201810357719 A CN 201810357719A CN 110387334 A CN110387334 A CN 110387334A
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- 238000000034 method Methods 0.000 title claims abstract description 17
- 241000123370 Antrodia Species 0.000 title claims abstract description 12
- 241001486992 Taiwanofungus camphoratus Species 0.000 claims abstract description 27
- 239000000843 powder Substances 0.000 claims abstract description 24
- 241000894006 Bacteria Species 0.000 claims abstract description 21
- 238000000855 fermentation Methods 0.000 claims abstract description 18
- 230000004151 fermentation Effects 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 11
- 239000007787 solid Substances 0.000 claims abstract description 6
- 238000011081 inoculation Methods 0.000 claims abstract description 4
- 230000004913 activation Effects 0.000 claims abstract description 3
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- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 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
- 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
- 238000002386 leaching Methods 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 241001416980 Paecilomyces hepiali Species 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 235000013312 flour Nutrition 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 2
- 239000008272 agar Substances 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 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
- 238000001914 filtration Methods 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract description 2
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- 244000005700 microbiome Species 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 14
- 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 description 10
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 5
- 229960005305 adenosine Drugs 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 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 4
- 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 4
- 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 4
- 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 4
- 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 4
- 239000002028 Biomass Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 241000386927 Cinnamomum micranthum f. kanehirae Species 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
- 241000233866 Fungi Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000190633 Cordyceps Species 0.000 description 1
- 241000219071 Malvaceae Species 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 240000007313 Tilia cordata Species 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000002456 anti-arthritic effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 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
- 238000009395 breeding Methods 0.000 description 1
- 239000002537 cosmetic Substances 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
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 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
- 230000037111 immune power Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 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
- 230000001151 other effect Effects 0.000 description 1
- 230000000144 pharmacologic 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
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000033 toxigenic Toxicity 0.000 description 1
- 230000001551 toxigenic effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 150000003648 triterpenes Chemical class 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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- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
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- Botany (AREA)
- Mycology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract
The invention belongs to microorganisms technical fields, and in particular to a method of with peacilomyce hepiahi bacterium powder fast culture antrodia mycelia.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 peacilomyce hepiahi bacterium powder 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 quickly and stably cultivate the higher antrodia mycelia of medical value using this method.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of to use peacilomyce hepiahi bacterium powder fast culture Antrodia camphorata
Mycelial method.
Background technique
Antrodia camphorata (Antrodia cinnamomea) it is a kind of distinctive rare medicinal fungi in Taiwan Province, China, mainly contain
There are the chemical components such as triterpene, polysaccharide, adenosine, has antifatigue, antiallergy, lowering blood pressure and blood fat, liver protecting, enhancing immune
Power, prevention cardiovascular disease and other effects have high research and proper value in industries such as medicine, food, cosmetics.At present
The artificial cultivation method of Antrodia camphorata includes cinnamomum kanehirai cultivation basswood method, plate method and solution fermentation.But cinnamomum kanehirai linden
The Antrodia camphorata low output that cultivation incubation time length (needing 2-3), toxigenic capacity are high, obtain, plate method and liquid hair
The antrodia mycelia that ferment method obtains, medicinal component are low.Therefore, a set of new breeding method suitable for Antrodia camphorata, In are established
Fast culture, which goes out the higher Antrodia camphorata of medical value, in short period becomes the emphasis of vast researcher research.
Paecilomyces hepiali chen (Paecilomyces hepiali) it is the asexual generation being separated to from fresh cordyceps sinensis
Bacterial strain and state approval can be used for the fungi strain of health food, have Antiarthritic, strengthen immunity, it is antibacterial, resist it is tired
The pharmacological actions such as labor, antitumor, anti-oxidant.By liquid deep layer fermenting culture, peacilomyce hepiahi bacterium filament obtained can
As the artificial substituent of wild cordyceps, develop into drug or health food.The present invention is by liquid deep layer fermenting culture
Peacilomyce hepiahi bacterium powder is as additive, for the liquid fermentation of Antrodia camphorata, then by stringent fermentation, quickly
Obtain the higher antrodia mycelia of medical value.
Summary of the invention
The purpose of the present invention is to provide a kind of methods with peacilomyce hepiahi bacterium powder fast culture Antrodia camphorata.
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.Adenosine and the higher peacilomyce hepiahi bacterium powder of Quantitative Determination of Ergosterol are chosen as fermentation training
Support the raw material of base.By peacilomyce hepiahi bacterium powder 5-15g, peptone 10-20g, potato leaching powder 20-30g, glucose 20-
30g, water 1000mL prepare fermentation medium, 121 DEG C of sterilizings natural cooling, spare after twenty minutes.It is inoculated with the inoculum concentration of 5-10%
Antrodia camphorata seed liquor carries out shaking table culture under conditions of temperature is 25-30 DEG C, revolving speed is 100-150r/min.To 20-30 days
It grows filtering fermentation liquor after a large amount of 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. peacilomyce hepiahi bacterium powder selects: using potato leaching powder, corn flour and dried silkworm chrysalis meal as raw material, by 8:1:1,
The proportion of 8:2:1 and 9:1:1 prepares fluid nutrient medium, passes through liquid deep layer fermenting culture peacilomyce hepiahi bacterium filament.Then
It by mycelium freeze-drying, crushes, peacilomyce hepiahi bacterium powder is made.Choose adenosine and the quasi- blueness of the higher bat moth of Quantitative Determination of Ergosterol
Mould powder is used for the liquid fermentation of Antrodia camphorata.Peacilomyce hepiahi bacterium strain for liquid deep layer fermenting is preserved in the micro- life in Yunnan Province
Object research institute Culture Collection Center, deposit number are YIM Yu2009661.
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: peacilomyce hepiahi bacterium powder 5g, peptone
10g, potato leaching powder 20g, glucose 25g, water 1000mL, pH are natural.
6. inoculation and culture: with 8% inoculum concentration inoculation Antrodia camphorata seed liquor in fermentation medium, temperature be 29 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.Every bottle can receive Antrodia camphorata dry product weight about 4g.
8. the Antrodia camphorata analysis of effective component of peacilomyce hepiahi bacterium powder culture
The antrodia mycelia biomass of peacilomyce hepiahi bacterium powder culture is much higher than control group (common Antrodia camphorata), illustrates bat
Moth paecilomycerol powder has facilitation to the culture of Antrodia camphorata.Through detecting, above-mentioned drying, crushing antrodia mycelia in, often
1g is respectively 9.18mg, 46.73mg, 1.72mg, 5.72 μ g containing polysaccharide, PEARLITOL 25C, adenosine, ergosterol, hence it is evident that is better than bat
Bat moth paecilomycerol powder 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 1 peacilomyce hepiahi bacterium powder culture of table compares (n=3)
| Sample | Mycelial biomass g/200mL | Polysaccharide mg/g | PEARLITOL 25C mg/g | Adenosine mg/g | Ergosterol μ g/g |
| The Antrodia camphorata of peacilomyce hepiahi bacterium powder culture | 3.93±0.21 | 9.18±0.60 | 46.73±2.72 | 1.72±0.02 | 5.72±0.02 |
| Peacilomyce hepiahi bacterium powder | - | 7.79±0.41 | 31.89±3.33 | 1.35±0.02 | 3.51±0.02 |
| Common Antrodia camphorata | 2.01±0.12 | 3.36±0.27 | 32.96±3.29 | 1.49±0.10 | 1.06±0.02 |
Note: mycelial biomass refers to the weight of mycelium dry matter in unit volume fermentation liquid；Common Antrodia camphorata refers to and does not add
The antrodia mycelia of the fermentation medium culture of peacilomyce hepiahi bacterium powder.
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 (4)
1. a kind of method of fast culture antrodia mycelia, it is characterised in that:
A, Antrodia camphorata strain is inoculated in the PDA solid medium activated spawn of improvement；
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: Paecilomyces hepiali chen
Bacterium powder 5-15g, peptone 10-20g, potato leaching powder 20-30g, glucose 20-30g, water 1000mL, pH are natural.
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| CN110387333A (en) * | 2018-04-20 | 2019-10-29 | 云南云百草实验室有限公司 | A method of with blue level ground cordyceps culture Antrodia camphorata |
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| 刘燕隔: "牛樟芝菌株培养条件优化、抗疲劳及保肝作用研究", 中国优秀硕士学位论文全文数据库（电子期刊）医药卫生科技辑, no. 9, pages 057 - 27 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110387333A (en) * | 2018-04-20 | 2019-10-29 | 云南云百草实验室有限公司 | A method of with blue level ground cordyceps culture Antrodia camphorata |
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