CN107164237A - Selenium-enriched cordceps militaris and its cultural method and application - Google Patents
Selenium-enriched cordceps militaris and its cultural method and application Download PDFInfo
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- CN107164237A CN107164237A CN201710433450.3A CN201710433450A CN107164237A CN 107164237 A CN107164237 A CN 107164237A CN 201710433450 A CN201710433450 A CN 201710433450A CN 107164237 A CN107164237 A CN 107164237A
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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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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The present invention provides a kind of selenium-enriched cordceps militaris and its cultural method and application.The present invention prepares nanometer selenium by the method for chemical synthesis and biofermentation, and the selenium-enriching cultivation of Cordyceps militaris is carried out using the nanometer selenium of preparation.The present invention obtains selenium-enriched cordceps militaris using nanometer selenium in the course of cultivation first, and obtained selenium-enriched cordceps militaris is applied in selenium-enriched functional food, health products and feed product.The nanometer selenium that the present invention is used, with environment-friendly, the features such as safe and efficient.The nanometer selenium of low toxicity, high bioactivity, which is applied in the cultivation of selenium-enriched cordceps militaris, makes Selenium-riched Cordyceps Militaris effect significantly, and Organic Selenium form can be completely converted into by bioconversion be present in selenium-enriched cordceps militaris fructification.The cultural method of the present invention, which is resulted in, to be grown, easily survival, and yield is high, the high selenium-enriched cordceps militaris of nutritional quality, is conducive to carrying out popularizing planting, so as to improve planting household income.
Description
Technical field
The present invention relates to cordyceps cultivation technology, specifically, it is related to a kind of selenium-enriched cordceps militaris and its cultural method with answering
With.
Background technology
Selenium (Selenium, Se) element is one of indispensable trace element in humans and animals vital movement, is biological
The required component of a variety of selenoenzyme albumen such as internal sulphur hydrogen reduction protease, de- iodine enzyme, glutathione peroxidase, and participate in
Human body multiple metabolic pathways.Research is found, when body lacks selenium element, can be weakened immunologic mechanism, be reduced the resistance to disease
, there are various corresponding shortage symptoms in power.China has abundant selenium resource, but selenium is extremely uneven in distributed in nature, causes me
The regional selenium deficiency of state more than 2/3rds.Chinese Soclety of Nutrition and FAO/WHO/IAEA joint specialist committee members can determine whether human intaking amount
Optimum range is 60-250 μ g/d, and safe dose is 400 μ g/d, and toxic dose is 800 μ g/d.Selenium excessively will also result in human body danger
Evil, therefore urgently develop safe and efficient selenium source for selenium deficiency crowd.Selenium is main in two kinds of forms of Organic Selenium and inorganic selenium in nature
Deposit, confirmed that the absorption of selenium is relevant with its chemical state in meals, the bioactivity of usual inorganic states selenium is less than Organic Selenium
Bioactivity.Research shows that inorganic selenium (selenate, selenite) toxicity is larger, potentially dangerous as direct selenium-supply source；
And Organic Selenium (selenomethionine, selenocystine, selenium methyl selenocystine, etc.) toxicity is relatively low, and lived with a variety of biologies
Property, including antitumor, anti-oxidant, raising immunity etc..Therefore bioconversion is utilized, inorganic selenium is converted into high activity
Organic Selenium is most safely and effectively one of means of current selenium-rich product.External source addition tetravalence selenium, hexavalent selenium have higher ring
Border risk, while tetravalence selenium, hexavalent selenium are larger to the toxic action of organism, organism is difficult that it efficiently is utilized into not shadow
Ring its own quality.And the toxicity of nanometer selenium is relatively low, organism absorbs to it and tolerable concentration is significantly higher than tetravalence selenium and sexavalence
Selenium, therefore nanometer selenium is safer in biotransformation, more efficient.
Cordyceps militaris (Cordyceps militaris) also known as northern Chinese caterpillar Fungus, it is that sac fungus is sub- to belong on biology point is learned
Door, ergot Zoopagales, Clavicipitaceae, Cordyceps.Cordyceps militaris is a kind of medicine-food two-purpose fungi, containing abundant cordycepic acid, cordycepin,
Adenosine, amino acid isoreactivity composition, and with significant pharmacological action, such as antifatigue, antitumor, resist oxygen lack, strengthen immunity,
Enhancing sexual function etc..Selenium passes through bioconversion in fungi, most of to be present in organic Se form in fructification, thus easily by
Absorption of human body is utilized, but Se content is relatively low in common fruiting bodies of cordyceps militaris, while tolerance and biology of the common Cordyceps militaris to selenium turn
Change ability is relatively low, selenium concentration reach its tolerance limit occur as soon as mycelia and fructification growth it is heavily suppressed, or even extremely
Die.It is a kind of more easy, economy, environmental protection acquisition to prepare nanometer selenium by the method for chemical synthesis and microbial fermentation
The method of elemental selenium.By screening the cordyceps militaris link bacterial strain of resistance to selenium, while carrying out the cultivation of selenium-enriched cordceps militaris using nanometer selenium, effectively drop
The low environmental risk of external source selenium, while higher degree improves organic selenium content in fruiting bodies of cordyceps militaris.People is studied both at home and abroad
The formation of conversion and Organic Selenium each valence state for the selenium that member participates in microorganism has clearer and more definite understanding.Utilize selenium-enriched cordceps militaris
Can completely by inorganic selenium be converted into low toxicity, high activity Organic Selenium form, can more safely and efficiently carry out selenium-supply.
The content of the invention
It is an object of the invention to provide a kind of selenium-enriched cordceps militaris and its cultural method.
It is a further object of the present invention to provide the selenium-enriched cordceps militaris in selenium-enriched functional food, health products and feed product
In application.
It is yet another object of the invention to provide the productions such as the culture medium exploitation selenium-rich chicken feed after being cultivated using selenium-enriched cordceps militaris
Application in terms of product.
In order to realize the object of the invention, the cultural method for the selenium-enriched cordceps militaris that the present invention is provided comprises the following steps：
S1, the high cordyceps militaris link bacterial strain of resistance to selenium of screening；
S2, the high cordyceps militaris link bacterial strain of resistance to selenium for obtaining step S1 screenings are trained in the culture medium added with nanometer selenium
Support, produce selenium-enriched cordceps militaris.
The nanometer selenium that heretofore described nanometer selenium synthesizes for the nanometer selenium and/or biofermentation of chemical synthesis.Wherein,
The nanometer selenium of chemical synthesis, it, which is prepared, can use method disclosed in ZL201410520106.4, the nanometer selenium particle size range of synthesis
For 30-300nm.Biofermentation synthesis nanometer selenium, its prepare can using CN201510047365.4,
Side disclosed in CN201610566900.1, CN201610952997.x, CN201610946230.6 or CN 201610946282.3
Method.
The concentration of nanometer selenium is 2-100mg/l in foregoing method, culture medium described in step S2.
Foregoing method, the method that step S1 screens the high cordyceps militaris link bacterial strain of resistance to selenium is as follows：
S11, collection wild cordyceps militaris, Cordyceps militaris is cleaned, sterilized with 70-75% alcohol (preferably 75% alcohol), will divide
Raw sporophore is cut into small pieces；
S12, the sodium selenite solution of filtration sterilization is added in PDA culture medium, makes the end of sodium selenite in culture medium
Concentration reaches 40-60mg/l (preferably 60mg/l), is down flat after plate, and conidium stalk is inoculated on the flat board containing sodium selenite,
In 18-22 DEG C of lucifuge culture 7-10 days, strain growth situation is observed, the selection mycelial growth preferably high bacterial strain of resistance to selenium is used as richness
The starting strain of selenium Cordyceps militaris；
Foregoing method, step S2 is specific as follows：
S21, the high starting strain of resistance to selenium is inoculated on PDA slant mediums, 7- is cultivated in 18-22 DEG C of constant incubator
10 days, then conidium is forwarded in the conical flask equipped with PD fluid nutrient mediums, liquid amount is 150-200ml/500ml
(preferably 200ml/500ml), 18-22 DEG C in isothermal vibration incubator, 150rpm is cultivated 3-5 days；
Bacterium solution obtained by S22, step S21 takes 5ml to dilute bacterium solution and is inoculated in equipped with richness after 5-10 times of dilution (preferably 10 times)
In the culture bottle of the seleno culture medium containing nanometer, in lucifuge, stuffy, 15-18 DEG C of temperature, relative air humidity 60-65% condition
Lower culture 4-7 days, culture medium can be covered with to mycelia；
S23, mycelia give illumination after covering with, it is ensured that daily light irradiation time 12h, and after growing 3-5 days, mycelia is changed by white
Yellowly, after mycelia is completely reformed into crocus, daily continuous light 24h continues to cultivate 40-50 days at 18-22 DEG C；Group
Entity grows into 5-7cm, and top is expanded, and shows that fruiting bodies of cordyceps militaris is ripe, you can harvesting, obtains selenium-enriched cordceps militaris.
Wherein, described in step S22 rich in nanometer seleno culture medium prepare it is as follows：10- is added into 300ml culture bottles
40g rice, 10-40ml PD fluid nutrient mediums, is then added nanometer selenium, is sealed with sealed membrane, and in high-pressure sterilizing pot, 121 DEG C go out
Bacterium 30min；Sterilizing is cooled to room temperature after terminating, and culture bottle is placed on super-clean bench, opens uviol lamp sterilizing 30min.
The present invention also provides the selenium-enriched cordceps militaris obtained according to above-mentioned cultural method, and it contains 2.8-362 μ g/g Organic Seleniums,
Cordycepin and adenosine content are respectively 6.8-10.3 μ g/g, 2.2-4.1 μ g/g, and total amino acid content is 1.8-2.4 μ g/kg.Together
When selenium-enriched cordceps militaris in contain higher Cordyceps sinensis polysaccharide, Cordyceps sinensis polysaccharide has unnecessary oxygen in antioxidation activity, removing machine body free
Base, improves the abilities such as immunity.
The present invention also provides the selenium-enriched cordceps militaris answering in selenium-enriched functional food, health products and feed product is prepared
With.
The present invention also provides the selenium-enriched functional food prepared by the selenium-enriched cordceps militaris, health products and feed product.Open
The product sent includes but is not limited to selenium-enriched cordceps militaris candy tabletting, selenium-enriched cordceps militaris capsule, the instant corn flour of selenium-rich, and selenium-rich is
Eat millet powder, the instant bean powder of selenium-rich, selenium-enriched cordceps militaris health liquor, selenium-enriched cordceps militaris health protection tea and selenium-rich animal feed etc..
The present invention further provides a kind of selenium-rich chicken feed, preparation method is：Selenium-enriched cordceps militaris is prepared using the above method,
After selenium-enriched cordceps militaris harvest, selenium-enriched cordceps militaris bacteria cake is made with the culture medium of residual, then by selenium-enriched cordceps militaris bacteria cake and dregs of beans,
The raw materials such as corn, stone flour, vitamin, trace element, salt, additive carry out mixing compounding, produce selenium-rich chicken feed.
Preferably, Se content is 300-500 μ g/kg in selenium-rich chicken feed.
In the specific embodiment of the present invention, the main formula of selenium-rich chicken feed is：Selenium-enriched cordceps militaris bacteria cake
600-800g, dregs of beans 100-150g, corn flour 200-300g, stone flour 40-50g, vitamin 2-3g, micro- 2-4g, salt
0.5-1g, additive 1-2g.Chicken farm is daily can to make Se content in egg reach 250-600 μ to chicken feeding selenium-rich chicken feed
g/kg。
The present invention prepares nanometer selenium by the method for chemical synthesis and biofermentation, and pupa is carried out using the nanometer selenium of preparation
The selenium-enriching cultivation of cordyceps sinensis.The present invention obtains selenium-enriched cordceps militaris using nanometer selenium in the course of cultivation first, and by obtained selenium-rich
Cordyceps militaris is applied in selenium-enriched functional food, health products and feed product.The nanometer selenium that the present invention is used, with environment friend
It is good, the features such as safe and efficient.The nanometer selenium of low toxicity, high bioactivity, which is applied in the cultivation of selenium-enriched cordceps militaris, makes pupa worm
Careless selenium-rich effect significantly, and can be completely converted into Organic Selenium form by bioconversion and be present in selenium-enriched cordceps militaris fructification
In.The cultural method of the present invention, which is resulted in, to be grown, easily survival, and yield is high, and the high selenium-enriched cordceps militaris of nutritional quality is conducive to
Popularizing planting is carried out, so as to improve planting household income.
Brief description of the drawings
Fig. 1 is the growing state of the high cordyceps militaris link bacterial strain of resistance to selenium in the embodiment of the present invention 1.
Fig. 2 is selenium-enriched cordceps militaris fructification in the embodiment of the present invention 1.
Fig. 3 is Determination of Selenium result in selenium-enriched cordceps militaris in the embodiment of the present invention 1.
Fig. 4 is Se form in selenium-enriched cordceps militaris in the embodiment of the present invention 1, mainly with selenocystine and selenomethionine two
Organic Selenium form is planted to exist.
Fig. 5 is cordycepin, adenosine HPLC collection of illustrative plates in selenium-enriched cordceps militaris in the embodiment of the present invention 1.
Fig. 6 is amino acid collection of illustrative plates in selenium-enriched cordceps militaris in the embodiment of the present invention 1.
Fig. 7 is raised for selenium-rich chicken feed in the embodiment of the present invention 2 and is obtained Se content in egg.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.Unless otherwise specified, embodiment
In the conventional meanses that are well known to those skilled in the art of used technological means, raw materials used is commercial goods.
The culture medium being related in the present invention is as follows：
PDA culture medium, preparation method is：200g potatoes are cleaned into peeling stripping and slicing, 1000ml water is added, boils to spoon and gently press
It is just broken, multilayer filtered through gauze is used, 20g glucose is added into filtrate, 20g agar powders are settled to 1000ml with 1 water, adjust pH
7.0.After packing, with 121 DEG C of high-pressure sterilizing pot, 30min sterilizings are standby after cooling.
PD fluid nutrient mediums：Potato 200g, glucose 20g, water 1000ml, pH 7.0.
Unless otherwise noted, heretofore described " part " refers to " parts by weight ".
The selenium-enriched cordceps militaris of embodiment 1 and its cultural method
1st, the high cordyceps militaris link bacterial strain of resistance to selenium screening
(1) sampling instrument is got out, wild cordyceps militaris are gathered in suitable when and where.
(2) wild cordyceps militaris of collection are cleaned up, and uses 75% alcohol disinfecting, conidiophore is cut into small pieces.
(3) sodium selenite solution of filtration sterilization is added in PDA culture medium, final concentration reaches 60mg/l, is down flat plate
Afterwards, conidium stalk is inoculated on the flat board containing sodium selenite.It is placed in 18-22 DEG C of lucifuge culture 7-10 days, observation bacterial strain life
Long situation, the selection mycelial growth preferably high bacterial strain of resistance to selenium (Fig. 1).
2nd, the cultivation of selenium-enriched cordceps militaris
(1) PDA slant mediums are prepared, sterilizes and the high strain of resistance to selenium is accessed after cooling down, be placed in 18-22 DEG C of constant incubator
Middle culture 7-10 days.
(2) take 500ml conical flasks to add 200ml PD fluid nutrient mediums, access the high strain of resistance to selenium after sterilizing cooling, be placed in
18-22 DEG C in isothermal vibration incubator, 150rpm is cultivated 3-5 days.
(3) 10-40g rice is added in 300ml culture bottles, 10-40ml PD fluid nutrient mediums add nanometer selenium and caused
Selenium concentration is 2-100mg/l, in 121 DEG C of sterilizing 30min of high-pressure sterilizing pot after being sealed with sealed membrane.
(4) room temperature is cooled to after end subject to sterilization, is put in super-clean bench, open uviol lamp sterilizing 30min.Liquid spawn is diluted
Respectively toward inoculation 5ml strains in every bottle after 10 times.
(5) culture bottle for having connect bacterium is placed in lucifuge, stuffy, temperature control in 15-18 DEG C, relative air humidity control
In 60-65%, cultivate 4-7 days, mycelia can cover with culture medium.
(6) illumination is opened after mycelia is covered with, it is ensured that daily light irradiation time 12h, after growing 3-5 days, white hypha can change
Yellowly.
(7) after mycelia is completely reformed into crocus, daily continuous light 24h continues in 18-22 DEG C of culture to 40-50
My god.
(8) when fructification grows into 5-7cm or so, top is expanded, and shows that fruiting bodies of cordyceps militaris is ripe, you can harvesting (figure
2).Fruiting bodies of cordyceps militaris is scooped up from root using the small iron spoon of elbow, dries or dries.
3rd, selenium-enriched cordceps militaris attributional analysis
(1) total selenium analysis in selenium-enriched cordceps militaris
The selenium-enriched cordceps militaris for cultivating acquisition is crushed, 100 mesh sieves are crossed.Weigh (triplicate) the addition 10ml of 0.2g dense
Nitric acid, cold soaking 24h clears up 1h step by step with micro-wave digestion, and cooling is settled in 100ml volumetric flasks.Use inductively coupled plasma matter
Compose (ICP-MS) (U.S. Agilent company 7500a) and determine total Se content, while selenium standard serial solution 0,1,5,25 is prepared,
50,100 μ g/l, draw standard curve (selenium standard liquid is purchased from Chinese Industrial Standards (CIS) material center), quantification of external standard method.Measurement result
It was found that wherein Se content reaches 2.8-362 μ g/g (Fig. 3) after selenium-enriched cordceps militaris drying prepared by nanometer selenium.
(2) Se form is analyzed in selenium-enriched cordceps militaris
The selenium-enriched cordceps militaris for cultivating acquisition is crushed, 100 mesh sieves are crossed.Weigh (triplicate) the addition 8mg/ml of 0.2g
The 10ml of protease 14.Repeat after mixing, stayed overnight in 37 DEG C of constant-temperature table concussions.After reaction, sample centrifugation obtains supernatant,
0.22 μm of filter membrane is crossed, with high performance liquid chromatography inductive coupling plasma mass spectrometry (HPLC-ICP-MS) analysis wherein Se form.Standard
Product selenocystine (SeCys2), selenomethionine (SeMet) is purchased from Chinese Industrial Standards (CIS) material center, quantified by external standard method.Liquid phase color
Compose condition of work：Anion-exchange column (Hamilton PRP-X100), mobile phase 40mM diammonium hydrogen phosphates, pH 6.Measurement result
It was found that selenium is main in selenium-enriched cordceps militaris has (Fig. 4) in two kinds of Organic Selenium forms of selenocystine and selenomethionine.
(3) cordycepin, adenosine analysis in selenium-enriched cordceps militaris
The selenium-enriched cordceps militaris for cultivating acquisition is crushed, 100 mesh sieves are crossed.Weigh (triplicate) the addition 8ml of 0.5g high
Pure water, ultrasonic 5h (P>450W, 50Hz-100Hz) 60 DEG C, 10ml is settled to water after taking-up, is shaken up.1ml sample liquids are taken,
After 4000rpm centrifugations, rear 0.22 μm of filter membrane.
Cordycepin, adenosine standard items respectively take 10mg (being converted by actual content), are settled to 100ml with mobile phase, shake up.Should
Mixed standard solution cordycepin and adenosine concentration are 100 μ g/ml, and 4 DEG C of refrigerators are preserved, the term of validity 1 month.0.1ml is taken respectively
0.2ml 0.5ml 1.0ml 2.0ml5.0ml mother liquors (100 μ g/ml) are settled to the μ g/l of 10ml standard curves concentration 1.0,2.0 μ
g/l、5.0μg/l、10.0μg/l、20.0μg/l、50.0μg/l.The use condition of high performance liquid chromatography, C18,250mm ×
4.6mm(i.d)5μm；Mobile phase：A acetonitriles, the acetic acid A of B 1.0%:B=1：99；Flow velocity 1.0ml/min；35 DEG C of column temperature；Detect ripple
Long 260nm UV-detectors are opened；The μ l of sample size 10.Cordycepin and adenosine content difference in selenium-enriched cordceps militaris prepared by nanometer selenium
6.8-10.3 μ g/g, 2.2-4.1 μ g/g (Fig. 5) can be reached.It is significantly higher than cordycepin and adenosine content in common Cordyceps militaris.
(4) amino acid analysis in selenium-enriched cordceps militaris
The selenium-enriched cordceps militaris for cultivating acquisition is crushed, 100 mesh sieves are crossed.300mg or so samples are weighed, 10ml water is placed in
Solve in glass tube；Add 7ml-8ml or so 6mol/L hydrochloric acid；Hydrolysis lid is tightened, sample is shaken up, is placed in baking oven and heats
Hydrolysis, hydrolysis temperature is 110 DEG C, is hydrolyzed 22 hours；Hydrolysis pipe is placed to room temperature, and the sample taken out in hydrolysis pipe is added to
In 100ml volumetric flasks, with distilled water diluting to scale.Volumetric flask is shaken up, using 0.2 μm of membrane filtration of syringe-type, takes 2ml to filter
Liquid is placed in eggplant shape rotary evaporation bottle, is removed acid using vacuum rotary evaporator, is cleaned, be evaporated with distilled water, extremely close
It is neutral；Add the dissolving of 2ml distilled water, upper machine analysis.Use INSTRUMENT MODEL：L-8900 Hitachis amino-acid analyzer (L-
8900Amino Acid Analyzer), standard items are：Japan and the hybrid standard product of Guang Chun medicines Co., Ltd., the standard name of an article
Claim：Amino Acids Mixture Standard Solution, Type H, quantification of external standard method.Amino acid analysis result table
It is bright, the amino acid classes and the content no significance difference compared with common Cordyceps militaris that contain of Cordyceps militaris of selenium-rich prepared by nanometer selenium
Different, total amino acid content is in 1.8-2.4 μ g/kg (Fig. 6).
Application of the selenium-enriched cordceps militaris of embodiment 2 in selenium-enriched functional food, health products and feed product
1st, the preparation of selenium-enriched cordceps militaris candy tabletting
Selenium-enriched cordceps militaris prepared by embodiment 1 contains 2.8-362 μ g/g Organic Seleniums, and cordycepin and adenosine content are respectively
6.8-10.3 μ g/g, 2.2-4.1 μ g/g, total amino acid content is in 1.8-2.4 μ g/kg.Simultaneously containing higher in selenium-enriched cordceps militaris
Cordyceps sinensis polysaccharide, Cordyceps sinensis polysaccharide has a unnecessary oxygen radical in antioxidation activity, removing machine body, improves the abilities such as immunity.Will
Selenium-enriched cordceps militaris lies fallow and health-care effect well as raw material, exploitation selenium-enriched cordceps militaris pressed candy tool.
Selenium-enriched cordceps militaris pressed candy is main by 30-40% selenium-enriched cordceps militaris fructifications, 20-40% cornstarch, 10-
20% ferment powder, 15-25% maltodextrins, 0.3-0.7% magnesium stearates composition, by percentage to the quality.
For example, 40% selenium-enriched cordceps militaris is taken, 30% cornstarch, 10% ferment powder；10% maltodextrin, 9.5% sucrose,
0.5% magnesium stearate, in irradiation sterilization 1h after being sufficiently mixed, tabletting produces selenium-enriched cordceps militaris candy tabletting.
Every selenium-enriched cordceps militaris candy tabletting contains 20-60 μ g Organic Seleniums, and the daily ingestion of 1-2 pieces of normal person can be met often
Day to the demand of selenium, daily ingestion of 4-6 of the special population such as illness, old man.Selenium-enriched cordceps militaris pressed candy has plurality kinds of health care
Function, is acted on body immunity, antitumor, anti-oxidant, anti-inflammatory etc. is improved.Pressed candy carries easy simultaneously, and taste is sweet
Sweet tea, instant.
2nd, the preparation of selenium-enriched cordceps militaris capsule
(1) sturdy, golden yellow color the selenium-enriched cordceps militaris fructification of growth is picked out, cleans, dry, be freeze-dried
24h, keeps water content in selenium-enriched cordceps militaris to be less than 10%.
(2) the selenium-enriched cordceps militaris fructification after will be lyophilized is crushed with high speed disintegrator, and 300-400 mesh sieves are crossed after crushing
Obtain the selenium-enriched cordceps militaris powder of exquisiteness.
(3) by selenium-enriched cordceps militaris powder 0.4g every part be sub-packed in the natural capsule of food-grade, after the completion of packing carry out irradiation go out
Bacterium.
(4) every selenium-enriched cordceps militaris capsule contains 20-60 μ g Organic Seleniums, and daily ingestion of 1-2 of normal person can meet often
Day to the demand of selenium, daily ingestion of 4-6 of the special population such as illness, old man.Selenium-enriched cordceps militaris capsule manufacturing process is convenient, without outer
Source material addition, Cordyceps militaris composition is pure, is preferable selenium-supply good merchantable brand.It is quantitative edible, good health-care effect can be played.
3rd, the preparation of selenium-enriched cordceps militaris function nutrition powder
(1) selenium-enriched cordceps militaris corn flour
The instant corn flour of selenium-rich, its composition mainly includes：Cordyceps militaris powder, corn flour, rice flour.
Exquisiteness is crushed to after selenium-enriched cordceps militaris is dried, 20 parts are weighed；Select clean without going mouldy, water content is less than 10%
High-grade maize grain, cleaned, peeling, break, proposes embryo, crushing, weighs 100 parts, add 4% water mix thoroughly after carry out it is expanded, it is swollen
100 mesh sieves are crossed after change；Fresh clean is selected without going mouldy, water content is less than 10% 40 parts of rice, and rice adds 3% after crushing
Water is mixed thoroughly, after the swelling, crosses 100 mesh sieves.
By 20-60 parts of selenium-enriched cordceps militaris powder, 100-300 parts of expanded corn powders, 40-120 parts of puffed rice powder, 20-60 parts
It is the instant corn flour of selenium-rich that DE20 maltodextrin carries out parcel packing after being sufficiently mixed after irradiation sterilization.Often wrap instant jade
Ground rice content is 10-60g, wherein containing 10-100 μ g Organic Seleniums.The daily ingestion of 1-4 bags of normal person can meet daily to selenium
Demand, the daily ingestion of 2-8 bags of special population such as illness, old man.
(2) selenium-enriched cordceps militaris millet powder
The instant millet powder of selenium-rich, main composition has：Selenium-enriched cordceps militaris powder, millet powder, walnut meat powder, white sesameseed powder.
Exquisiteness is crushed to after selenium-enriched cordceps militaris is dried, 10-40 parts are weighed；The millet of high-quality is selected, dust, rice is cleaned
Chaff, controls water content to be less than 10% after drying, then through break, propose embryo, crushing, weigh 100 parts, add 4% water mix thoroughly it is laggard
Row is expanded, expanded to cross 100 mesh sieves afterwards；The walnut kernel (5-20 parts) of high-quality, white sesameseed (5-20 parts) are selected by cleaning, is dried,
Stir-fry.Milling is standby.
By 20-60 parts of selenium-enriched cordceps militaris powder, 100-300 parts of expanded millet powders, 10-40 portions of walnut kernel, 10-40 portions of white sesameseeds
It is the instant millet powder of selenium-rich to carry out parcel packing after irradiation sterilization after being sufficiently mixed.Instant millet powder content is often wrapped for 10-
60g, wherein containing 10-100 μ g Organic Seleniums.The daily ingestion of 1-4 bags of normal person can meet the daily demand to selenium, ill, old
The daily ingestion of 2-8 bags of the special populations such as people.
(3) selenium-enriched cordceps militaris bean powder
The instant bean powder of selenium-rich, main composition has：Selenium-enriched cordceps militaris powder, analysis for soybean powder.
Exquisiteness is crushed to after selenium-enriched cordceps militaris is dried, 20-60 parts are weighed；Good quality soybeans are selected, are cleaned, after drying,
Toast 1h at 130 DEG C, dipping by lye, peeling, defibrination, by the spray drying of soya bean slurries, obtain 100-300 parts of analysis for soybean powder.
By 20-60 parts of selenium-enriched cordceps militaris powder, 100-300 parts of analysis for soybean powder carry out parcel point after being sufficiently mixed after irradiation sterilization
Dress is the instant bean powder of selenium-rich.Instant bean powder content is often wrapped for 10-60g, wherein containing 10-100 μ g Organic Seleniums.Normal person is daily
Intake 1-4 bags can be met daily to the demand of selenium, the daily ingestion of 2-8 bags of special population such as illness, old man.
4th, the preparation of selenium-enriched cordceps militaris health liquor
Selenium-enriched cordceps militaris is compounded with a variety of nourishing based foods, selenium-enriched cordceps militaris health liquor is prepared.
Selenium-enriched cordceps militaris health liquor using selenium-enriched cordceps militaris as main material, be aided with the bark of eucommia, the root of straight ladybell, radix polygonati officinalis, matrimony vine, Poria cocos,
Soaked in jujube kernel, Radix Angelicae Sinensis immersion wine.Every 2 jin of Chinese herbal medicine consumption proportions are：Cordyceps militaris 40-200g；Matrimony vine 30-40g；Bark of eucommia 3-
6g；The root of straight ladybell, jujube kernel 10-20g；Radix polygonati officinalis, Poria cocos 5-10g.
Organic selenium content is in 0.5-15 μ g/ml in selenium-enriched cordceps militaris health liquor, the daily ingestion of 10-20ml selenium-rich pupa of adult
Chinese caterpillar fungus health can improve immunity of organisms, improve sanguimotor effect, while the effect of having tonifying kidney and strengthening yang.
5th, the preparation of selenium-enriched cordceps militaris health protection tea
Using selenium-enriched cordceps militaris as primary raw material, it is aided with STEVIA REBAUDIANA, Jasmine, selenium-enriched cordceps militaris health protection tea is made in rose.
Selenium-enriched cordceps militaris health-care tea formula：Selenium-enriched cordceps militaris 30-40%, STEVIA REBAUDIANA 5-10%, Jasmine 5-10%, rose
Flower 5-10%.
By selenium-enriched cordceps militaris, STEVIA REBAUDIANA, Jasmine, rose drying control moisture content below 10%.By being formulated compounding
Packing, is sterilized later.
Selenium-enriched cordceps militaris health protection tea color is limpid, contains the various active material such as cordycepin, adenosine, selenium, flavones, Neng Gouzeng
Strong body immunity is antifatigue.
Organic selenium content is in 40-100 μ g/g in selenium-enriched cordceps militaris health protection tea, the daily ingestion of 2-4g selenium-enriched cordceps militaris of adult
Health care can improve immunity of organisms, improve sanguimotor effect, while the effect of having tonifying kidney and strengthening yang.
The application of addition product in the selenium-enriched cordceps militaris cultivation of embodiment 3
The preparation of selenium-rich chicken feed：
(1) nanometer selenium has a large amount of selenium-rich culturing bases during selenium-enriched cordceps militaris is prepared after selenium-enriched cordceps militaris harvest
Retain.
(2) the remaining culture medium obtected pupa Chinese caterpillar fungus hypha of selenium-enriched cordceps militaris is had thorough grasp, and Se content reaches 1-200 μ g/g in culture medium.
Culture medium contains the nutriments such as substantial amounts of cordycepin, adenosine, amino acid simultaneously.
(3) the selenium-enriched cordceps militaris bacteria cake that selenium-rich chicken feed is produced during mainly preparing selenium-enriched cordceps militaris using nanometer selenium is original
Material, coordinates dregs of beans, corn, stone flour, vitamin, trace element, salt, additive.
(4) main formula of selenium-rich chicken feed：The main formula of selenium-rich chicken feed is：Selenium-enriched cordceps militaris bacteria cake 600-
800g, dregs of beans 100-150g, corn flour 200-300g, stone flour 40-50g, vitamin 2-3g, micro- 2-4g, salt 0.5-
1g, additive 1-2g.
(5) Se content is in 300-500 μ g/kg in selenium-rich chicken feed, and chicken farm feeds selenium-rich chicken feed, feeding to chicken daily
After 20 days, Se content in egg can be made to reach 250-600 μ g/kg, chicken Se content reaches 160-280 μ g/kg (Fig. 7).
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be modified or improved, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. a kind of cultural method of selenium-enriched cordceps militaris, it is characterised in that comprise the following steps：
S1, the high cordyceps militaris link bacterial strain of resistance to selenium of screening；
S2, the high cordyceps militaris link bacterial strain of resistance to selenium for obtaining step S1 screenings are cultivated in the culture medium added with nanometer selenium, i.e.,
Obtain selenium-enriched cordceps militaris.
2. according to the method described in claim 1, it is characterised in that the nanometer selenium described in step S2 is the nanometer of chemical synthesis
Selenium and/or the nanometer selenium of biofermentation synthesis；Wherein, the nanometer selenium of chemical synthesis, its preparation uses ZL201410520106.4
Disclosed method, the nanometer selenium particle size range of synthesis is 30-300nm；The nanometer selenium of biofermentation synthesis, it, which is prepared, uses
CN201510047365.4, CN201610566900.1, CN201610952997.x, CN201610946230.6 or CN
201610946282.3 disclosed method.
3. method according to claim 2, it is characterised in that the concentration of nanometer selenium is 2- in culture medium described in step S2
100mg/l。
4. according to the method described in claim 1, it is characterised in that step S1 screens the method for the high cordyceps militaris link bacterial strain of resistance to selenium such as
Under：
S11, collection wild cordyceps militaris, Cordyceps militaris is cleaned, uses 70-75% alcohol disinfectings, conidiophore is cut into small pieces；
S12, the sodium selenite solution of filtration sterilization is added in PDA culture medium, makes the final concentration of sodium selenite in culture medium
40-60mg/l is reached, is down flat after plate, conidium stalk is inoculated on the flat board containing sodium selenite, in 18-22 DEG C of lucifuge culture
7-10 days, observe strain growth situation, the selection mycelial growth preferably high bacterial strain of resistance to selenium.
5. the method according to claim any one of 1-4, it is characterised in that step S2 is specific as follows：
S21, by high resistance to selenium inoculation on PDA slant mediums, cultivate 7-10 days in 18-22 DEG C of constant incubator, so
Conidium is forwarded in the conical flask equipped with PD fluid nutrient mediums afterwards, liquid amount is 150-200ml/500ml, in constant temperature shake
18-22 DEG C is swung in incubator, and 150rpm is cultivated 3-5 days；
Bacterium solution obtained by S22, step S21 takes 5ml to dilute bacterium solution and is inoculated in equipped with rich in nanometer seleno culture medium after 5-10 times of dilution
Culture bottle in, under conditions of lucifuge, stuffy, 15-18 DEG C of temperature, relative air humidity 60-65% cultivate 4-7 days, extremely
Mycelia can cover with culture medium；
S23, mycelia give illumination after covering with, it is ensured that daily light irradiation time 12h, after growing 3-5 days, mycelia is transformed into Huang by white
Color, after mycelia is completely reformed into crocus, daily continuous light 24h continues to cultivate 40-50 days at 18-22 DEG C；Work as fructification
5-7cm is grown into, top is expanded, show that fruiting bodies of cordyceps militaris is ripe, you can harvesting, obtain selenium-enriched cordceps militaris；
Wherein, described in step S22 rich in nanometer seleno culture medium prepare it is as follows：10-40g is added into 300ml culture bottles big
Then rice, 10-40ml PD fluid nutrient mediums are added nanometer selenium, are sealed with sealed membrane, in 121 DEG C of sterilizings of high-pressure sterilizing pot
30min；Sterilizing is cooled to room temperature after terminating, and culture bottle is placed on super-clean bench, opens uviol lamp sterilizing 30min.
6. the selenium-enriched cordceps militaris obtained according to any one of the claim 1-5 cultural methods, it is organic that it contains 2.8-362 μ g/g
Selenium, cordycepin and adenosine content are respectively 6.8-10.3 μ g/g, 2.2-4.1 μ g/g, and total amino acid content is 1.8-2.4 μ g/kg.
7. application of the selenium-enriched cordceps militaris described in claim 6 in selenium-enriched functional food, health products and feed product is prepared.
8. the selenium-enriched functional food, health products and the feed product that are prepared as selenium-enriched cordceps militaris described in claim 6.
9. a kind of selenium-rich chicken feed, it is characterised in that its preparation method is：Utilize any one of claim 1-5 methods described system
After standby selenium-enriched cordceps militaris, selenium-enriched cordceps militaris harvest, selenium-enriched cordceps militaris bacteria cake is made with the culture medium of residual, then by selenium-rich pupa worm
Careless bacteria cake and dregs of beans, corn, stone flour, vitamin, trace element, salt, additive carry out mixing compounding, produce selenium enriched chicken feeding
Material.
10. selenium-rich chicken feed according to claim 9, it is characterised in that Se content is 300-500 μ in selenium-rich chicken feed
g/kg。
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| CN111887333A (en) * | 2020-07-31 | 2020-11-06 | 广东粤微生物科技有限公司 | Selenium-rich cordyceps militaris tablet candy and preparation method thereof |
| CN112899169A (en) * | 2021-01-26 | 2021-06-04 | 苏州市十颐健康科技股份有限公司 | Fermentation method of mulberry leaf cordyceps militaris nano-selenium thallus |
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