CN105543111B - Aweto mycelium plate method and solid medium used based on food materials - Google Patents
Aweto mycelium plate method and solid medium used based on food materials Download PDFInfo
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- CN105543111B CN105543111B CN201610082382.6A CN201610082382A CN105543111B CN 105543111 B CN105543111 B CN 105543111B CN 201610082382 A CN201610082382 A CN 201610082382A CN 105543111 B CN105543111 B CN 105543111B
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- aweto
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- aweto mycelium
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- 239000007787 solid Substances 0.000 title claims abstract description 69
- 239000000463 material Substances 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 39
- 235000013305 food Nutrition 0.000 title claims abstract description 26
- 239000004615 ingredient Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 244000144977 poultry Species 0.000 claims abstract description 8
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 7
- 150000002016 disaccharides Chemical class 0.000 claims abstract description 5
- 150000002772 monosaccharides Chemical group 0.000 claims abstract description 4
- 241001248610 Ophiocordyceps sinensis Species 0.000 claims description 37
- 241000894006 Bacteria Species 0.000 claims description 24
- 239000002054 inoculum Substances 0.000 claims description 20
- 235000012054 meals Nutrition 0.000 claims description 15
- 230000001954 sterilising effect Effects 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 13
- 238000004659 sterilization and disinfection Methods 0.000 claims description 12
- 244000062793 Sorghum vulgare Species 0.000 claims description 11
- 235000019713 millet Nutrition 0.000 claims description 11
- 244000061456 Solanum tuberosum Species 0.000 claims description 10
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
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- 235000013594 poultry meat Nutrition 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 7
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- 239000000203 mixture Substances 0.000 claims description 5
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims description 3
- 235000015895 biscuits Nutrition 0.000 claims description 3
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- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 2
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- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
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- 239000002609 medium Substances 0.000 abstract description 51
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- 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 description 8
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- 235000015579 Alternanthera sessilis Nutrition 0.000 description 4
- 241000255789 Bombyx mori Species 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 206010062717 Increased upper airway secretion Diseases 0.000 description 2
- 241001416980 Paecilomyces hepiali Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
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- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 208000026435 phlegm Diseases 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
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- 235000015099 wheat brans Nutrition 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
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- 240000008025 Alternanthera ficoidea Species 0.000 description 1
- 241000272165 Charadriidae Species 0.000 description 1
- 241001480006 Clavicipitaceae Species 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
- 241000190633 Cordyceps Species 0.000 description 1
- 241001640558 Cotoneaster horizontalis Species 0.000 description 1
- 240000003176 Digitaria ciliaris Species 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 208000000616 Hemoptysis Diseases 0.000 description 1
- 241000130660 Hepialidae Species 0.000 description 1
- 241000330899 Hepialus Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 240000001307 Myosotis scorpioides Species 0.000 description 1
- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 1
- 240000004713 Pisum sativum Species 0.000 description 1
- 235000010582 Pisum sativum Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000004922 Vigna radiata Species 0.000 description 1
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 1
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 1
- 240000001417 Vigna umbellata Species 0.000 description 1
- 235000011453 Vigna umbellata Nutrition 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000003288 anthiarrhythmic effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
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- 238000006243 chemical reaction Methods 0.000 description 1
- 208000013116 chronic cough Diseases 0.000 description 1
- 238000010276 construction Methods 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
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 201000001881 impotence Diseases 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000000629 knee joint Anatomy 0.000 description 1
- 239000010808 liquid waste Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
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- 238000011069 regeneration method Methods 0.000 description 1
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- 238000012502 risk assessment Methods 0.000 description 1
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- 238000000926 separation method Methods 0.000 description 1
- 238000010563 solid-state fermentation Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001256 tonic effect Effects 0.000 description 1
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Classifications
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- 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)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention discloses a kind of aweto mycelium solid medium, by following weight content at being grouped as: partial size≤2mm major ingredient 30~50%, fowl poultry kind 0.1~15%, greengrocery 0.1~15%, carbohydrate 0.1~15%, surplus are water；The major ingredient is non-waxy or beans, and the fowl poultry kind and greengrocery constitute auxiliary material, and the carbohydrate is monosaccharide or disaccharide.The present invention further simultaneously discloses the preparation method of above-mentioned aweto mycelium solid medium, and the present invention further simultaneously discloses the aweto mycelium plate method based on food materials carried out using above-mentioned solid medium.Culture medium of the invention makes culture medium have biggish specific surface area and good loose gas permeability completely using food materials as major ingredient and auxiliary material, can grow for aweto mycelium and provide good nutrition condition and physical space.
Description
Technical field
The invention belongs to biomaterial breeding method field, in particular to a kind of solid for aweto mycelium is trained
Support base and cultural method.
Background technique
Cordyceps sinensis (Ophiocordyceps sinensis) is Lepidoptera Hepialidae Genus Hepialuss larva by Chinese coat
Spore (Hirsutella sinensis) parasitism is formed by entomogenous fungi complex, and in China, folklore application is more than 2,000 years existing
History, the record of related document can be traced to before thousand.It is recorded according to edition Pharmacopoeia of the People's Republic of China is gone through, primary efficacy
It is kidney tonifying benefit lung, hemostasis and phlegm；For treating deficiency of kidney essence, impotence and seminal emission, soreness of waist and knee joint, chronic cough and dyspnea of deficiency type, overstrain cough hemoptysis.It is modern
Medicine and pharmaceutical research prove that cordyceps sinensis has the function of a variety of treatments and tonic: having expansion branch to respiratory system
Tracheae, relieving cough and asthma, eliminating the phlegm anti-inflammatory effect；Have the effects that anti-arrhythmia, anti-myocardial ischemina to cardiovascular system；It is right
Immune function is in dual regulation, also has anticancer activity and sedative-hypnotic effect and antifatigue effect.Cordyceps sinensis again because
Its mild-natured power is slow, and the flat tonifying yin sun of energy adverse reaction will not occur even if many people take for a long time throughout the year, thus be also
The nourishing diet good food of person worn with age, postoperative physically weak person and sub-health population.Aweto parasitics is harsh, host bat
The habitat of moth category insect is unique, is mainly distributed on the alpine meadow ecosystem of China Qinghai-Tibet Platean height above sea level 3500m~5000m,
Bat moth completes a life cycle and usually requires 3~4 years, so the natural resources of cordyceps sinensis are very rare.In huge city
Under the driving of field demand and economic interests, the excavation for robbing formula, which brings the regeneration of Cordyceps Resources, to be seriously threatened, and is also given
Local fragile ecological environment brings heavy pressure.Exploitation cordyceps sinensis artificial substituting product be solve wild resource scarcity with it is huge
Big medicinal and health care demand contradiction protects the effective way of China's sources of three rivers ecological safety.Though the artificial culture of cordyceps sinensis
It has succeeded but has not yet accomplished scale production, applied most common cordyceps sinensis artificial substituting product at present, come from and utilize the winter
Worm summer grass phorozoon, that is, Clavicipitaceae fungi Hirsutella sinensis (Hirsutella sinensis) or concomitance bacterium Paecilomyces hepiali chen
The mycelium that (Paecilomyces hepiali) etc. passes through deep fermentation culture.The liquid fermentation of aweto mycelium
Production has numerous patents report, such as application number 03135623.0,85107991,2011101568723,
200410025517.2,200410101728.X, 200710142960.1 etc., their common feature (1) needs fermentor, equipment
Investment is big；(2) production process needs continuous force feed filtrated air, and energy consumption is high；(3) complex process need to produce bacterium by being separated by solid-liquid separation
Filament, thus generates a large amount of organic liquid wastes, must governance for reaching standard discharge, not only waste of resource but also increased environmental protection pressure.So liquid is deep
Layer fermented cordyceps sinensis mycelia product, price is still expensive, fails edible as the daily invigorant of general public.
Application No. is 03135622.2 patents of invention, disclose a kind of solid fermentation working system of aweto, should
Patent replaces liquid fermentation tank with solid-state fermentation tank, allows Hirsutella sinensis containing solid-baseds such as sucrose, yeast and dried silkworm chrysalis meals
Grown in matter, finished product is solid culture, and also without discharging of waste liquid in production, but the method is still needed to fermentor, and need to
Force feed filtrated air in tank, energy consumption is high.Application No. is 200510014467.2 patents of invention, disclose a kind of cordyceps sinensis
The preparation method of thallus oatmeal, the method is using wheat or oatmeal as the solid medium of aweto, 2 weeks or so mycelia after inoculation
Body wraps up wheat, terminates culture.Since the main component of wheat is starch, the composition of nutrition composition and the place of aweto
Main bat moth polypide differs greatly, it is difficult to sufficiently meet the nutritional need of aweto growth.There are more patents of invention, such as Shen
Please number be 200910197957.9,201110228542.0,2104164.4,200810147785.X, 99117265.5,
Solid medium in the solid fermentation method of 201310398242.6,201210509164.8 equal reported awetos exists
On the basis of major ingredient cereal, dried silkworm chrysalis meal, wheat bran, peptone, yeast extract, beef extract, potassium dihydrogen phosphate are added, magnesium sulfate etc.
One or more are auxiliary material, to supplement the nutrition that cereal is lacked.However, these auxiliary materials and improper food materials, silkworm chrysalis therein
It will affect the palatability of product with special odor, some will lead to chemical residues.Further, since aweto is aerobic
Bacterium, mycelium are grown on the surface of the gap of solid medium attachment cereal-granules etc., and the general solid culture time is relatively
Long, the solid culture time as disclosed in the patent of invention application No. is 200910197957.9 is up to 8~12 weeks.
Summary of the invention
The aweto mycelium plate method that the technical problem to be solved in the present invention is to provide a kind of based on food materials and
Solid medium used, the culture medium make culture medium have biggish specific surface completely using food materials as major ingredient and auxiliary material
Long-pending and good loose gas permeability can grow for aweto mycelium and provide good nutrition condition and physical space.
In order to solve the above technical problem, the present invention provides a kind of aweto mycelium solid mediums, by following
Weight content at being grouped as:
Surplus is water；
The major ingredient is non-waxy or beans, and the fowl poultry kind and greengrocery constitute auxiliary material, and the carbohydrate is single
Sugar or disaccharide.
Improvement as aweto mycelium solid medium of the invention:
The non-waxy is millet, corn, long-grained nonglutinous rice, polished rice, Semen Coicis, wheat；
The beans is soybean, kidney bean, pea, red bean, semen sojae atricolor, mung bean, semen viciae fabae；
The fowl poultry kind is poultry meat, poultry or the flesh of fish, and the greengrocery is potato, stem tuber, root tuber, leaf vegetables, melon and fruit or mushroom
(that is, being its edible portion)；
The monosaccharide is glucose, fructose；
The disaccharide is sucrose, maltose.
Remarks explanation:, can be directly as major ingredient when non-waxy is millet；When non-waxy be corn, long-grained nonglutinous rice,
When polished rice, Semen Coicis, wheat, it need to be crushed to partial size≤2mm in advance；Similarly, above-mentioned all beans need to be crushed in advance partial size≤
2mm。
Above-mentioned greengrocery is its edible portion, and therefore, potato, melon and fruit etc. are required to carry out peeling processing.
The present invention goes back while providing the preparation method of above-mentioned aweto mycelium solid medium, including walks as follows
It is rapid:
1., major ingredient cleaned up；
2., auxiliary material is cleaned after crush, homogenate, obtain slurry；
3., will clean after major ingredient, slurry, carbohydrate and water mix, steamed or boiled mature meal；
4., ripe meal handled through high-temperature sterilization, after being cooled to room temperature aweto mycelium solid medium is (referred to as solid
Body culture medium)；It is to be seeded.
Remarks explanation:
Steam or boil mature meal refer to it is cooked or meal braizing machine cooks with electric cooker, pressure cooker etc..How to judge whether rice is ripe,
This is routine techniques.
High-temperature sterilization processing refers to the 20min that sterilizes at 121 DEG C, this is also routine techniques.
The present invention goes back while providing to be consolidated using the aweto mycelium based on food materials that above-mentioned solid medium carries out
Body cultivation is selected aweto phorozoon Hirsutella sinensis (Cordyceps sinensis), is included the following steps:
1), aweto phorozoon Hirsutella sinensis (Cordyceps sinensis) is prepared into second class inoculum；
In the second class inoculum, the content of Hirsutella sinensis (Cordyceps sinensis) is >=108cfu/ml；
2) sterilization processing first, is carried out to incubator；
Then second class inoculum is inoculated into solid medium by 3~20% (V/W) inoculum concentrations (for example, 5%), then set
It is cultivated in 18~25 DEG C 6~8 days (mycelia is full of culture medium gap and surface at this time) in incubator after sterilizing, it is fresh to obtain mycelium
Cake；
Above-mentioned 3~20% (V/W) inoculum concentration refers to the second level bacterium that 3~20ml is inoculated on the solid medium of every 100g
Kind.
As the improvement of aweto mycelium plate method of the invention, the cultivation is further comprising the steps of:
3), the fresh biscuit of mycelium is dry:
Prior to 85~90 DEG C of the fresh cake of mycelium (for example, 85 DEG C) is dried into 30min, moisture content is dried to then at 55 DEG C and is lower than
8%；
Remarks illustrate: above-mentioned 85 DEG C are the microorganism in killing baking oven the purpose of drying 30min；55 DEG C are dried to moisture content and are lower than
8% is the volatilization loss for preventing functional mass such as cordycepin therein under high temperature, can smell its fragrance during the drying process；
4) it, crushes:
The resulting drying mycelia cake of step 3) be crushed into 80 mesh, obtain aweto mycelium dry powder.Vacuum packaging.
Further improvement as aweto mycelium plate method of the invention:
The step 1) are as follows:
Hirsutella sinensis (Cordyceps sinensis) slant strains are inoculated into PDA plate, obtain activated spawn；Specifically
Are as follows: under aseptic technique, picking aweto inclined-plane lawn is inoculated in PDA plate, cultivates 5d at 15~25 DEG C, obtain
Aweto activates lawn；
Activated spawn is inoculated into PDA liquid medium, in 18~22 DEG C of oscillation (100~200r/min) cultures 3~5
It, obtains first order seed bacterium solution；
By the inoculum concentration of 0.5~15% (v/v), first order seed bacterium solution is inoculated into PDA liquid medium, in 18~22
DEG C oscillation (100~200r/min) cultivate 3~5 days second class inoculum (aweto secondary seed solution), (be placed in 4 DEG C for use
Under it is spare).
Further improvement as aweto mycelium plate method of the invention:
Aweto phorozoon Hirsutella sinensis (Cordyceps sinensis) is purchased from Chinese agriculture microorganism fungus kind
Preservation administrative center, deposit number: ACCC50542.
Further improvement as aweto mycelium plate method of the invention:
It is described that sterilization processing is carried out to incubator are as follows: 75% (volume %) alcohol is sprayed to incubator and opens ozone hair
Raw device 30min is to carry out sterilization.
In the present invention, PDA plate, PDA liquid medium belong to well-known technique.
The formula of PDA solid medium and the preparation method are as follows: potato 200g, and 0.5h is boiled in peeling stripping and slicing, filtered through gauze,
Add sucrose 20g, adds distilled water to 1000ml, agar 18g.In pressure 1.05kg/cm2, sterilize at 121.3 DEG C of temperature 20min.
The formula of PDA liquid medium and the preparation method are as follows: potato 200g, and 0.5h is boiled in peeling stripping and slicing, filtered through gauze,
Add sucrose 20g, adds distilled water to 1000ml.In pressure 1.05kg/cm2, sterilize at 121.3 DEG C of temperature 20min.
In solid medium of the invention,
The effect of major ingredient is: the main donor of carbon source, stilt hypha body attachment；
The effect of auxiliary material and carbohydrate is: the main donor of nitrogen source, and replenishing vitamins, minerals and soluble carbon source；
The effect of water is: maintaining solid medium appropriated moisture.
In aweto mycelium plate method of the invention,
The resulting aweto mycelium dry powder of the present invention in actual use, takes dry powder 3~10g/ times, is brewed i.e. with boiling water
It is drinkable.It takes within one day 1~2 time.
The present invention has following significant advantage:
(1) the host's bat moth polypide nutrition for selecting conventional food materials to simulate aweto by compatibility is constituted, to keep away
Exempt from the prior art because using dried silkworm chrysalis meal, wheat bran, peptone, yeast extract, beef extract, potassium dihydrogen phosphate, magnesium sulfate etc. very
Rule food materials cause the edible taste of aweto mycelium product bad or there are chemical residues for auxiliary material；
(2) it selects non-waxy as solid medium major ingredient, can prevent culture medium from excessively bonding and influence gas permeability,
To guarantee the good loose gas permeability of culture medium, and then aerobic environment is built for aweto mycelium growth；
(3) for example non-Semen setariae of small particle cereal is selected, or big grain cereal is crushed, non-waxy corn is such as broken for small particle
Corn is broken or small particle cereal and greater particle size cereal such as long-grained nonglutinous rice, polished rice etc. are mixed, to improve the specific surface of solid medium
Product, to increase attachment space of the aweto mycelium in solid medium.
That is, the present invention, which reduces its partial size by broken grains, improves its specific surface area, and select non-waxy kind with
It reduces the caking property of solid medium and improves its loose gas permeability, be beneficial to promote growth, in terms of existing technologies,
The time required to culture can be substantially reduced.
(4) homogenate and major ingredient boiling maturation meal is made in culture auxiliary material, has both made starch gelatinization in cereal, easily by aweto
Filament utilizes；Can be sufficiently mixed again with the nutritional ingredient in auxiliary material, with improve solid medium nutrition at uniformity, favorably
It is absorbed in aweto mycelium；The appropriate aqueous amount that can control ripe meal by the adjusting of excess water simultaneously, makes solid medium
Dry and wet is appropriate, to maintain aerobic condition.
(5) available raw material type multiplicity, formula flexibly, can satisfy different taste preferences and more specifically seek
The demand of supporting has high commodity value and market potential.
(6) production cost and equipment requirement are lower, compare existing the relevant technologies, and hardly possible is accepted in the achievements conversion for reducing enterprise
Degree, market-oriented potentiality further strengthen.
(7) production procedure is clear, succinct, efficient, the raw material being related to without known and need the harmful components that control, convenient for into
Key point control and risk analysis in row production, thereby guarantee that product in process of production and the safety of finished product, with this into
One step improves Related product in the reliability of food safety level.
Specific embodiment
The preparation method of aweto secondary seed solution (second class inoculum), successively follows the steps below:
Under aseptic technique, picking aweto inclined-plane lawn ----Hirsutella sinensis (Cordyceps
Sinensis) slant strains are inoculated in PDA plate, in 15~25 DEG C of culture 5d, obtain aweto activation lawn；Picking winter worm
Summer grass bacterium activation lawn is inoculated in PDA liquid medium, and (100~200r/min) is vibrated at 18~22 DEG C and cultivates 5d, obtains the winter
Worm summer grass bacterium primary seed solution (first order seed bacterium solution)；First order seed bacterium solution is inoculated in PDA liquid by 10% (v/v) inoculum concentration
Culture solution is cultivated 5 days in 18~22 DEG C of oscillations (100~200r/min), obtains second class inoculum (aweto secondary seed solution),
It (is placed in spare at 4 DEG C) for use.
In the second class inoculum, the content of Hirsutella sinensis (Cordyceps sinensis) is >=108cfu/ml。
Embodiment 1,
One, aweto mycelium solid medium and preparation method, the formula of the culture medium is by following weight percent
Food materials composition:
The preparation method of the culture medium is implemented as follows:
1., non-glutinous yellow millet rinsed well with tap water, as the culture medium major ingredient；
2., clean peeled potatoes and rabbit meat crush and be homogenized, obtain slurry；
3., will clean after the slurry that is prepared of major ingredient, auxiliary material mixed with sucrose and water, be cooked into auto-electric cooker using pottery ripe
Meal；
4., ripe meal is sub-packed in 15cm culture dish, sterilize 20min at 121 DEG C, it is cooling after solid medium, for use.
Two, aweto mycelium plate method successively follows the steps below:
1., first to culture chamber interior wall spray 75% (volume %) alcohol, restart ozone generator 30min, to incubator
Carry out disinfection sterilizing；
2., under aseptic technique, be inoculated with second class inoculum (aweto two by 5% (V/W) of solid medium weight
Grade seed liquor), inoculation is placed in the incubator of sterilized sterilization treatment, is cultivated 7 days in 20 DEG C；Aweto mycelium at this time
Surface and gap full of solid medium terminate culture, obtain the fresh bacteria cake of aweto mycelium.
Above-mentioned 5% (V/W) inoculum concentration refers to the second class inoculum that 5ml is inoculated on the solid medium of every 100g.
3., the fresh biscuit of mycelium it is dry:
Fresh bacteria cake is placed in baking oven, prior to 85 DEG C baking 30min are to inactivate the microorganism in baking oven；It is dried at 55 DEG C
It dry (moisture content is lower than 8%), obtains aweto mycelium and does bacteria cake；
4., aweto mycelium do bacteria cake crushed 80 mesh, obtain aweto mycelium dry powder；Vacuum packaging.
Aweto mycelium dry powder drinking method are as follows: take dry powder 3~10g/ times, brewed with 100ml or so boiling water
It drinks.
Embodiment 2,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1 (that is, the preparation method of solid medium, aweto mycelium plate method
Equivalent integers 1.)
Embodiment 3,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 4,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 5,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 6,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 7,
Aweto mycelium solid culture based formulas is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 8,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 9,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 10,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Embodiment 11,
The formula of aweto mycelium solid medium is made of the food materials of following weight percent:
Remaining content is equal to embodiment 1.
Experiment 1 measures 1~embodiment of above-described embodiment, the 11 fresh bacterium of resulting aweto mycelium using colony counting method
The content of aweto in cake, obtains result and is listed in the table below 1.
Table 1
| Embodiment | Aweto mycelium content (the fresh bacteria cake of cfu/g) |
| 1 | 5.1×106 |
| 2 | 4.3×106 |
| 3 | 3.8×106 |
| 4 | 3.2×106 |
| 5 | 4.7×106 |
| 6 | 5.3×106 |
| 7 | 4.9×106 |
| 8 | 3.9×106 |
| 9 | 5.4×106 |
| 10 | 5.7×106 |
| 11 | 4.6×106 |
Comparative example 1-1,
Peeled potatoes, rabbit meat, sucrose in the solid culture based formulas of embodiment 1 is changed to non-glutinous yellow millet,
That is, formula is as follows:
| Food materials | Percentage (%) |
| Major ingredient: non-glutinous yellow millet | 46 |
| Water | 44.0 |
Remaining content is equal to embodiment 1.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, solid culture primary surface and gap not
See that aweto mycelium covers.The aweto of fresh bacteria cake (the fresh bacteria cake of aweto mycelium) is measured using plate count
Filament content is only 2.4 × 103cfu/g。
Comparative example 1-2,
Rabbit meat in the solid culture based formulas of embodiment 1 is changed to non-glutinous yellow millet, that is, formula is as follows:
| Food materials | Percentage (%) |
| Major ingredient: non-glutinous yellow millet | 49.2 |
| Auxiliary material: peeled potatoes | 4.5 |
| Sucrose | 2.3 |
| Water | 44.0 |
Remaining content is equal to embodiment 1.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, solid culture primary surface and gap not
See that aweto mycelium covers.Using the aweto mycelium content that plate count measures fresh bacteria cake be only 8.7 ×
103cfu/g。
Comparative example 1-3,
Peeled potatoes in the solid culture based formulas of embodiment 1 are changed to non-glutinous yellow millet, that is, formula is as follows:
Remaining content is equal to embodiment 1.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, on the surface and gap of solid medium
There is a little aweto mycelium to cover but not as good as 1 densification of embodiment.The Chinese caterpillar fungus bacterial filament of fresh cake is measured using plate count
Body content is 9.3 × 104The fresh cake of cfu/g.
Comparative example 1-4,
Sucrose in the solid culture based formulas of embodiment 1 is changed to non-glutinous yellow millet, that is, formula is as follows:
Remaining content is equal to embodiment 1.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, on the surface and gap of solid medium
There is aweto mycelium to cover but not as good as 1 densification of embodiment.Contained using the aweto mycelium that plate count measures fresh cake
Amount is 3.8 × 105The fresh bacteria cake of cfu/g.
Comparative example 2-1,
The preparation method of the solid medium of embodiment 1 is modified to successively follow the steps below:
1., non-glutinous yellow millet rinsed well with tap water, as the culture medium major ingredient；
2., clean peeled potatoes, rabbit meat be cut into the block that size is about 10 × 10 × 10mm；
3., will clean after major ingredient, the resulting block of step 2) mixed with sucrose and water, be cooked into auto-electric cooker using pottery ripe
Meal；
4., ripe meal is sub-packed in 15cm culture dish, sterilize 20min at 121 DEG C, it is cooling after solid medium, for use.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, on the surface and gap of solid medium
There is aweto mycelium to cover but not as good as 1 densification of embodiment.The Chinese coat spore mycelium of fresh cake is measured using plate count
Amount is 6.1 × 105The fresh bacteria cake of cfu/g.
Comparative example 2-2,
Corn in the formula of 2 solid medium of embodiment broken (partial size≤2mm) is changed to whole kernel corn, remaining is equal to
Embodiment 2.
In aweto mycelium plate method, when being cultivated 7 days at 20 DEG C, on the surface and gap of solid medium
There is a small amount of aweto mycelium to cover.Use plate count measure the Chinese coat spore mycelia scale of construction of fresh cake for 8.4 ×
104The fresh bacteria cake of cfu/g.
Comparative example 2-3,
Step in the aweto mycelium plate method of cancellation embodiment 1 is 1., that is, does not carry out in advance to incubator
Sterilization；Remaining is equal to embodiment 1.When cultivating 7 days at 20 DEG C, aweto mycelium is full of solid medium at this time
Surface and gap, but there is mould spot, scrap of the product in solid culture primary surface.
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair
Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (7)
1. aweto mycelium solid medium, it is characterized in that by following weight content at being grouped as:
The major ingredient is non-waxy, and the fowl poultry kind and greengrocery constitute auxiliary material, and the carbohydrate is monosaccharide or disaccharide；
The non-waxy is millet, corn, long-grained nonglutinous rice, polished rice, Semen Coicis or wheat；
The fowl poultry kind is poultry meat, poultry or the flesh of fish, and the greengrocery is potato, leaf vegetables, melon and fruit or mushroom；
The monosaccharide is glucose or fructose；
The disaccharide is sucrose or maltose.
2. the preparation method of aweto mycelium solid medium as described in claim 1, it is characterized in that including following step
It is rapid:
1., major ingredient cleaned up；
2., auxiliary material is cleaned after crush, homogenate, obtain slurry；
3., will clean after major ingredient, slurry, carbohydrate and water mix, steamed or boiled mature meal；
4., ripe meal handled through high-temperature sterilization, after being cooled to room temperature aweto mycelium solid medium.
3. preparing the aweto mycelium solid culture based on food materials that resulting solid medium carries out using claim 2
Method is selected aweto phorozoon Hirsutella sinensis (Cordyceps sinensis), it is characterized in that including the following steps:
1), aweto phorozoon Hirsutella sinensis (Cordyceps sinensis) is prepared into second class inoculum；
In the second class inoculum, the content of Hirsutella sinensis (Cordyceps sinensis) is >=108cfu/ml；
2) sterilization processing first, is carried out to incubator；
Then second class inoculum is inoculated into solid medium by 3~20% (V/W) inoculum concentrations, be subsequently placed in after sterilizing in incubator
It is cultivated 6~8 days in 18~25 DEG C, obtains the fresh cake of mycelium.
4. aweto mycelium plate method according to claim 3, it is characterized in that the cultivation further includes following
Step:
3), the fresh biscuit of mycelium is dry:
By prior to 85~90 DEG C baking 30min of the fresh cake of mycelium, moisture content is dried to then at 55 DEG C lower than 8%；
4) it, crushes:
The resulting drying mycelia cake of step 3) be crushed into 80 mesh, obtain aweto mycelium dry powder.
5. aweto mycelium plate method according to claim 3 or 4, it is characterized in that:
The step 1) are as follows:
Hirsutella sinensis (Cordyceps sinensis) slant strains are inoculated into PDA plate, obtain activated spawn；
Activated spawn is inoculated into PDA liquid medium, in 18~22 DEG C shaken cultivation 3~5 days, obtain first order seed bacterium solution；
By the inoculum concentration of 0.5~15% (v/v), first order seed bacterium solution is inoculated into PDA liquid medium, is vibrated in 18~22 DEG C
Culture 3~5 days, obtains second class inoculum.
6. aweto mycelium plate method according to claim 3 or 4, it is characterized in that:
Aweto phorozoon Hirsutella sinensis (Cordyceps sinensis) is purchased from Chinese agriculture Microbiological Culture Collection
Administrative center, deposit number: ACCC50542.
7. aweto mycelium plate method according to claim 3 or 4, it is characterized in that:
It is described that sterilization processing is carried out to incubator are as follows: to open ozone generator after spraying 75% alcohol of volumetric concentration to incubator
20~30min is to carry out sterilization.
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| US6558943B1 (en) * | 2000-09-05 | 2003-05-06 | Sun Ten Pharmaceutical Co., Ltd. | Method for propagating fungi using solid state fermentation |
| CN101695257A (en) * | 2009-10-30 | 2010-04-21 | 上海芝草生物技术有限公司 | Solid fermentation method for cordyceps sinensis |
| CN103937690A (en) * | 2014-04-29 | 2014-07-23 | 高锦乐 | Solid state fermentation method for cordyceps sinensis |
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|---|---|---|---|---|
| US6558943B1 (en) * | 2000-09-05 | 2003-05-06 | Sun Ten Pharmaceutical Co., Ltd. | Method for propagating fungi using solid state fermentation |
| CN101695257A (en) * | 2009-10-30 | 2010-04-21 | 上海芝草生物技术有限公司 | Solid fermentation method for cordyceps sinensis |
| CN103937690A (en) * | 2014-04-29 | 2014-07-23 | 高锦乐 | Solid state fermentation method for cordyceps sinensis |
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