CN102242070B

CN102242070B - Method for artificially culturing paecilomyces cicadae and application of culturing product thereof

Info

Publication number: CN102242070B
Application number: CN2011101224684A
Authority: CN
Inventors: 陈祝安; 孙长胜; 赵伟; 李成
Original assignee: Zhejiang Faya Biological Pharmaceutical Co Ltd
Current assignee: Pan Asia Biopharmaceutical Co ltd
Priority date: 2010-05-14
Filing date: 2011-05-12
Publication date: 2013-05-08
Anticipated expiration: 2031-05-12
Also published as: CN102242070A

Abstract

The invention discloses a method for artificially culturing paecilomyces cicadae and application of a culturing product thereof. The method for artificially culturing paecilomyces cicadae in large scales comprises the steps of: preparing strains, dosing and packing into a box, sterilizing, inoculating, solid-fermenting, collecting and the like. The paecilomyces cicadae is cultured by utilizing grains, such as corn flour, bran, wheat, barley, rice, millet, broomcorn and the like and culture mediums of bagasse, cane sugar, shell powder, silkworm chrysalis meal and potassium nitrate. The raw materials are obtained from local resources, a large amount of production cost is saved, the culturing period is shortened, and cordyceps sobolifera obtained by culturing has the advantages of high quality and favorable stability and the like. A culture obtained by the invention can be used for preparing foods, health-care products, drugs and cosmetics with functions of fighting tumor, regulating immunity, reducing blood sugar, blood fat and blood pressure, improving eye sight, resisting radiation, dispelling heat and easing pains, calming and hypnotizing, nourishing and strengthening, improving kidney function and the like.

Description

A kind of method of Paecilomyces cicadae artificial culture and the application of cultured products thereof

Technical field

The invention belongs to the artificial cultivation technique field of medicinal fungi, be specifically related to a kind of application of method and cultured products thereof of Paecilomyces cicadae artificial culture.

Background technology

Cicada fungus, formal name used at school Paecilomyces cicadae (Paecilomyces cicadae) is the product after some cicada nymph is subjected to Paecilomyces cicadae bacterium parasitism, is a kind of bacterium worm complex body.This bacterium was named by Miquel in 1838 and is the mould Isaria cicadae of cicada Isaria.After this multiple synonym appears.As cicada grass Cordyceps cicadae, base is given birth to Isaria Isaria basili, Xin Kelai spherical shell spore Sphaeria sinclairii, the worm shell bacterium Torrubia caespitosa of growing thickly, Xin Kelai Chinese caterpillar fungus Cordyceps sinclairii, Harry's Isaria Isaria hariottii, the fine and soft Cordyceps sobolifera of little cicada, Xin Kelai Isaria Isaria sinclairii, the Munch Isaria Isaria mokanshawii of Soviet Union and racemosus Isaria Isaria arbuscu/a etc.Medical science thinks that cicada fungus is a kind of valuable ingredient of traditional Chinese medicine close with Cordyceps sinensis.

The perfect stage of Paecilomyces cicadae, there is the scholar to think large cicada grass Cordyceps cicadae.Bar-shaped or the horn shape of large cicada grass seed seat, 10-80mm * 3-5mm sends from host's head, and singly give birth to or grow thickly, brown, perithecium pseudovum shape buries, the thecaspore column.What widely distribute at nature is Paecilomyces cicadae, and large cicada grass is rare.

Cicada fungus contains the effective constituents such as glycogen, cordycepic acid, multiple indispensable amino acid, PEARLITOL 25C, multiple alkaloid, ergosterol.17 seed amino acids, polysaccharide, N.F,USP MANNITOL contained in cicada fungus are all close with Cordyceps sinensis, and the content of heavy metal As, Hg, Pb is all low than Cordyceps sinensis, and wherein Hg does not detect cicada fungus, illustrates that cicada fungus has more security than Cordyceps sinensis.By the comparison of ingredients of cicada fungus and Cordyceps sinensis, can find out that Paecilomyces cicadae and Cordyceps sinensis have similar activeconstituents.

The artificial culture Paecilomyces cicadae has the advantages such as the natural condition of not relying on, culture cycle is short, quality product is guaranteed, can be mass-produced, provide new way for solving wild cicada fungus shortage of resources problem, studies show that, tame cicada fungus main active ingredient is not less than wild cicada fungus, particularly polysaccharide content is very high, and very high activeconstituents total amount indicates that its effect may be better than wild cicada fungus.Yet in prior art, the method for artificial culture Paecilomyces cicadae often exists culture cycle longer, so the higher problem of cost, and Paecilomyces cicadae has the parasexuality phenomenon, bacterial strain optimization be produce crucial.

Summary of the invention

The object of the invention is to provides a kind of application of method and cultured products thereof of Paecilomyces cicadae artificial culture by improving the cultural method of prior art.

For achieving the above object, the present invention takes following technical scheme:

A kind of method of Paecilomyces cicadae artificial culture comprises the steps:

1) bacterial classification preparation: comprise that the inclined-plane kind prepares and one-level kind preparation process;

Wherein, the inclined-plane kind prepares and comprises the steps: Paecilomyces cicadae bacterial strain (Paecilomyces cicadae) is transplanted in PSA substratum (potato sucrose nutrient agar) slant tube, cultivated 5～7 days under 22 ℃～25 ℃, good kind saves backup to select growing way;

Described Paecilomyces cicadae bacterial strain can adopt ordinary method to obtain from the nature separation and purification, also can adopt existing preservation or commercially available Paecilomyces cicadae bacterial strain.

The one-level kind prepares and comprises the steps: in inclined-plane kind access potato sucrose nutrient solution, and at 22 ℃～25 ℃, oscillation frequency is to cultivate 3～7 days under the condition of 130～150r/min, occurs to the Erlenmeyer flask after a large amount of mycelium pellets standby as the one-level kind;

2) the one-level kind that makes inoculation: with step 1) is linked in solid medium;

3) fermentation: postvaccinal cultivation box moves in culturing room, at 20 ℃～25 ℃, cultivates 30 days～50 days under the condition of relative humidity 60%～80%;

4) gathering of coremium: when the coremium height of culture reaches the 9cm left and right, can gather when the surface has a small amount of conidium to occur.

Better, step 1) in, contain following component (containing in every 1000ml substratum) in described PSA substratum: potato 200g, sucrose 20g, agar 20g, its preparation method is: will remove the peel potato and dice to be placed in and add water 1000ml in container, boil 20min～30min, filter, add sucrose and agar, supply water to 1000ml, then heating for dissolving divides to be filled in vitro, loading amount is 1/5～1/4 of test tube length, tampon, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave beyond the Great Wall, while hot test tube is placed on skewback cooling.

Better, step 1) in, following component (containing in every 1000ml nutrient solution) contained in described potato sucrose nutrient solution: potato 200g, sucrose 20g; Its preparation method is: will remove the peel potato and dice to be placed in and add water 1000ml in container, boil 15min～30min, filter, add sucrose, supply water to 1000ml, then heating for dissolving divides to be filled in Erlenmeyer flask, and loading amount is 20%～30% of Erlenmeyer flask capacity, tampon beyond the Great Wall, with the kraft paper parcel, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave again, taking-up is cooled to below 20 ℃.

Better, step 2) in, the step of described inoculation is: after the temperature of the solid medium in cultivating box is cooled to below 20 ℃, inoculate under aseptic condition, the inoculum size according to 3～5% is cultivated the access of one-level kind in box.

Better, step 2) in, the component title that contains in described solid medium and the weight part of each component are:

Matrix 42～46 weight parts;

Bagasse 2～10 weight parts;

Oyster shell whiting 0.1～0.5 weight part;

Dried silkworm chrysalis meal 2～5 weight parts;

Saltpetre 0.1～0.5 weight part;

Water 44～56 weight parts;

Wherein, described matrix is selected from a kind of in following combination: the mixture of Semen Maydis powder, wheat bran and sucrose; Wheat; Barley; The mixture of grain and sucrose; The mixture of rice and sucrose; The mixture of Chinese sorghum and sucrose.

Preferably, in the mixture of described Semen Maydis powder, wheat bran and sucrose, in the gross weight of the mixture of described Semen Maydis powder, wheat bran and sucrose, the weight percent of Semen Maydis powder, wheat bran and sucrose is respectively: Semen Maydis powder 43%～55%, wheat bran 44%～55%, sucrose 1%～2%.

Preferably, in the mixture of described grain and sucrose, in the gross weight of the mixture of described grain and sucrose, the weight percent of grain and sucrose is respectively: grain 98%～99%, sucrose 1%～2%.

Preferably, in the mixture of described rice and sucrose, in the gross weight of the mixture of described rice and sucrose, the weight percent of rice and sucrose is respectively: rice 98%～99%, sucrose 1%～2%.

Preferably, in the mixture of described Chinese sorghum and sucrose, in the gross weight of the mixture of described Chinese sorghum and sucrose, the weight percent of Chinese sorghum and sucrose is respectively: Chinese sorghum 98%～99%, sucrose 1%～2%.

Preferably, the preparation method of described solid medium is: first Semen Maydis powder, wheat bran or wheat, barley, grain, rice, Chinese sorghum are boiled, sucrose, saltpetre are dissolved in water, then with bagasse, oyster shell whiting, dried silkworm chrysalis meal proportionally mixes.

Preferably, also contain peptone (fish meal) in described solid medium, and the weight part of described peptone (fish meal) is 0.5 weight part.

Preferably, step 2) in, the carbon-nitrogen ratio of described solid medium is 80～98.

Better, step 3) in, require daylighting at solid fermentation later stage coremium growth phase, and envrionment temperature compares the mycelial growth stage in early stage and need hang down 2～3 ℃, need maintain the circulation of air in culturing process.

Better, step 4) in, described concrete steps of gathering are: remove breathable sealing film, with instrument, culture is taken out and separate coremium, the oven dry classification uses bag film vacuum packaging, mycoplasma to be placed in-20 ℃ of storages after drying.

The product that the present invention cultivates gained is comprised of mixture three parts of coremium, conidium, mycelium and substratum surplus materials, can be used for preparing have antitumor, regulate immune, hypoglycemic, reducing blood-fat, hypotensive, make eye bright, radioprotective, antipyretic-antalgic, tranquilizing soporific, strengthening by means of tonics, improve food or healthcare products or medicine or the makeup of the function such as renal function; Described food comprises the products such as biscuit, nutritive powder, described healthcare products comprise the products such as health promoting wine, oral liquid, described medicine comprises the medicine with effects such as antitumor, strengthening immunity, and described makeup comprise the products such as facial mask, cleansing milk, sunscreen or oil, soap, body wash, shampoo.

The prepared cultured products of method of the present invention can be for the preparation of following material: cicada fungus tea, cicada wine, cicada fungus soy sauce, cicada fungus amino acid seasoning, cicada fungus coffee, cicada fungus milk preparation, cicada fungus candy, cicada fungus honeybee product, cicada fungus puffed food, cicada fungus chewing gum, cicada fungus herbal cuisine, cicada fungus spore oil, cicada fungus capsule, cicada fungus medicine materical crude slice, cicada fungus electuary, cicada fungus cosmetics or cicada fungus feed.

1, cicada fungus tea: the preparation method of cicada fungus tea cuts into sheet with coremium, adds appropriate warm water and can brew into cicada fungus tea.Also coremium can be smashed, be packaged into the tea bag style, add appropriate warm water and brew and form, can within a short period of time the nutritive ingredient in raw material fully be disengaged when making tea.Except above-mentioned preparation method, can also prepare with traditional methods such as parch, but the original effective constituent of this method destructible.

2, cicada wine: the method that tradition prepares wine has percolation process and cold-maceration.Cold-maceration be with the direct soak at room temperature of coremium in the wine base, generally need about 6 months, extract is few, if improving soaking temperature can reduce soak time and improve extract content, its concrete grammar: coremium and mycoplasma are pulverized-〉 are dropped into container-〉 add wine base-〉 certain temperature hot dipping a few hours and constantly stir-〉 change a certain proportion of finings of liquid hot dipping again-〉 merging leach liquor-〉 add-〉 stir-〉 filter-〉 finished product.Can also be on the basis of hot dipping ultrasonic extraction, this method is than the available more extract effective constituent of hot dipping only.Concrete implementation step is as follows: coremium and mycoplasma are pulverized-〉 are soaked in beverage wine-〉 ultrasonic extraction-〉 rectification.The ethanol concn of temperature-time, ultrasonic extraction temperature-time and wine base all need be tested and be determined its top condition.The wine base can be white wine, barley wine, old road wine, yellow rice wine, red wine etc., considers to think that red wine is that the wine base can complement each other with the pharmacological action of cicada fungus itself from nutritive ingredient, pharmaceutical use.Can also add Chinese medicinal materials in the preparation of cicada wine, as Herba Cistanches, the tuber of multiflower knotweed, first meat, ginseng, matrimony vine, the Radix Astragali etc.

3, seasonings:

1) cicada fungus soy sauce: the preparation method of cicada fungus soy sauce mixes Manufactured soy sauce stoste by a certain percentage with the coremium of artificial culture and the extracting solution of mycoplasma, the steps include: that 1. soya bean/dregs of beans, flour/wheat/wheat bran raw material processing-〉 inoculation-〉 fermentation make soy sauce stoste.The coremium of the artificial culture after 2. pulverizing and mycoplasma-〉 ultrasonic hot water extraction-〉 filter to get extracting solution; Residue carries out ultrasonic hot water extraction-〉 filter to get extracting solution again; The soy sauce stoste that 3. united extraction liquid-〉 vacuum concentration makes fermentation with concentrated after extracting solution mix by a certain percentage-〉 caramel colour prepare burden-〉 add batching sweeting agent, freshener-〉 sterilization-〉 filtration-〉 can-〉 finished product.

2) cicada fungus amino acid seasoning: invented in recent years synthetic gourmet powder or purpose compound flavour enhancer with nourishing function, to add trace element and VITAMIN etc. on the basis of Sodium Glutamate monosodium glutamate mostly, thereby change the single seasoning function of monosodium glutamate, but the seasonings nourishing function of most of nourishing functions is single, and the cicada fungus amino acid seasoning can play plurality of health care functions in the situation of adding single component.Technical scheme is for pulverizing coremium and the mycoplasma-〉 decoction filtering in suitable quantity of water of artificial culture, vacuum concentration-〉 after concentrated solution with monosodium glutamate mix and blend or spraying drying-〉 finished product.With the method for mix and blend or spraying, concentrated solution is being permeated attached, white crystal particle monosodium glutamate or powdery monosodium glutamate are developed into the cicada fungus amino acid seasoning, technique is simple, cost is low, has greatly strengthened its nourishing function.

4, protective foods:

1) cicada fungus coffee, milk preparation and candy: the compound method of cicada fungus coffee, milk preparation and candy comprises direct preparing process, refining extraction method.These products have the adjusting body function, build up health, and eliminate the body metabolism waste product, delay senility and the effect of beauty and health care.Compound method is as follows: direct preparing process, and mutually composite with a certain proportion of coffee finished product, milk powder and raw sugar etc. after the coremium of artificial culture is pulverized, in bulk or packed after mixing.Refining extraction method, extract coremium and the mycoplasma of having pulverized artificial culture with ethanol (certain density wine) heat reflow method, when collecting phegma to nothing alcohol flavor, standby after standby or dry transpiring moisture after filtering, residue is collected whole extracting solutions after three alcohol heat reflux extract repeatedly, residue is abandoned, the pulverizing of extract after dry transpiring moisture is composite with coffee finished product, milk preparation and raw sugar, make various packed, bottled, box-packed cicada fungus coffees, milk powder, milk, sour milk, calcium milk, cheese, cold drink milk preparation and candy.

2) cicada fungus honeybee product: honeybee product is that honey is done, the products such as honey, propolis, royal jelly and pollen general designation honeybee product.On market, the kind of honey except pure honey, also has Mel, clover honey, acacia honey etc. now, mostly is greatly plant milk extract and is combined with honey.The present invention is combined traditional Chinese medicine ingredients with honey, have more the medicine edibleness, has to regulate body function, build up health, remove the body metabolism waste product, delay senility and the effect such as beauty and health care.Its compound method is: extract the coremium of artificial culture and the activeconstituents in mycoplasma, it is joined in honey with certain proportion, can obtain finished product.

3) cicada fungus puffed food: fragrant and sweet, multiple tastes that puffed food is crisp is a lot of youngster, especially child's favorite.Its main raw material is the grains such as corn, flour, millet, potato, pea, and the after the swelling surface is sprayed the batchings such as grease, salt and seasonings and makes.If the cicada fungus activeconstituents is added in puffed food, also can accomplish the health care effect when amusement and recreation.Concrete grammar can be sprayed on the extract vacuum concentration that extracts in artificial culture cicada fungus coremium and mycoplasma after the puffed food finished surface dry, also can replace common seasonings with cicada fungus seasonings in foregoing invention, monosodium glutamate, the general soy sauce of cicada fungus soy sauce replacement etc. are with the explained hereafter cicada fungus puffed food of general puffed food as the cicada fungus amino acid seasoning replaces.

4) cicada fungus chewing gum: on sale on the market is fruit flavour type and there is no the chewing gum of traditional Chinese medicine ingredients substantially mostly.Not only can cleaning oral cavity, promoting digestion if add traditional Chinese medicine ingredients in chewing gum, stimulate circulation and take exercise outside facial muscles, can also play the effects such as strengthening by means of tonics, antifatigue anti-stress, antipyretic-antalgic, immunomodulatory.With the coremium of artificial culture and the extracts active ingredients in mycoplasma out, mix with natural gum, add the mediations compactings such as syrup (powder), peppermint, sweeting agent to form.Document record is also arranged, can will make respectively polysaccharide layer, cordycepic acid layer and Nucleotide layer after each effective constituent purifying, the cordycepic acid layer in the centre respectively with the polysaccharide layer and the Nucleotide layer bonding the chewing gum finished product.

5) cicada fungus herbal cuisine: general cicada fungus herbal cuisine is cicada fungus matrimony vine pork liver soup, and the preparation method cleans the medicinal materials such as wild cicada fungus, matrimony vine, puts in proportion pot, fries in shallow oil soup with boiling in pork liver soup, and ripe rotten rear iodized salt gets final product.Rare due to wild cicada fungus resource can even replace higher than wild cicada fungus artificial culture product coremium with nutritive value, except the pork liver generally used, matrimony vine, can also with plant, animal class, dietotherapeutic class and Chinese medicine class compatibility.Plant food comprises: marine products category, grain class work in-process, melon and fruit seed class, medical, edible mushroom, greengrocery, wild vegetable class and beans and goods class, the animal class comprises muroid, marine products category, as goods class, freshwater fish, mountain bird, poultry, livestock products category, wild beast games, insects and batrachia, the dietotherapeutic class has Chinese yam, a species of orchid seed of jog's tears, Semen Sesami Nigrum, Semen Phaseoli, French beans, longan aril, hawthorn, date etc., and the Chinese medicine class comprises ginseng, Radix Panacis Quinquefolii, Stigma Croci, pseudo-ginseng, rhizoma Gastrodiae, the red sage root, the bighead atractylodes rhizome etc.The required condiment class of herbal cuisine also has green onion, ginger, garlic, soy sauce, cooking wine, pepper, capsicum, monosodium glutamate, vinegar and brown sugar etc. except salt compounded of iodine.Cicada fungus herbal cuisine of the present invention is with low cost, be the herbal cuisine series that integrates nourishing, health care, prevention, treatment, beauty treatment, skin care, fat-reducing, establishing-Yang, and have antifatigue, anti-ageing, antiviral, radioprotective, anti-hypoxia, strengthening immunity, memory, improve the quality of living, the effects such as restorative function, two-ways regulation body member function.

6) cicada fungus spore oil: the preparation method of cicada fungus spore oil is cicada fungus artificial culture product spore powder effective constituent after broken wall, in the extraction spore powder, removes impurity and invalid components, namely gets the cicada fungus spore oil.Its effective constituent has polysaccharide, total triterpene and adenosine etc.

5, health care medicine:

The preparation method of cicada fungus capsule, medicine materical crude slice, electuary is as follows: coremium and the mycoplasma of artificial culture are pulverized, got capsule with pharmaceutical excipient mixing packing after lyophilization.The extract of separation and Extraction effective constituent from the coremium of artificial culture and mycoplasma, with after auxiliary material and functional factor mix medicine materical crude slice, auxiliary material is selected: sweeting agent is cane sugar powder, maltose, fructose, maltose, protein sugar, trehalose, lactose and semi-lactosi, raffinose, Xylitol, honey, mannitol, sweet glycan, mill taro powder, Radix Glycyrrhizae, hawthorn, pseudo-ginseng, ginseng, glossy ganoderma; Acidic flavoring agent is citric acid, oxysuccinic acid, tartrate; Other additives are CMC, gelatin, agar, dextrin, molecular distillation mono-stearin, sucrose fatty ester, edible talcum powder; Functional factor is vegetable polysaccharides, nucleic acid, flavones, vitamins C, zinc, calcium, selenium.The present invention has the advantages such as cost is low, instant, Nutrition and health care value is high, mouthfeel is good.Electuary is the extract of separation and Extraction effective constituent from the coremium of artificial culture and mycoplasma, mixes with extract after the residue oven dry after filtering is pulverized, then dries and mix rear packing or can with other auxiliary materials, and this method can be removed composition invalid in raw material.

6, cosmetics:

Add the extracting solution that extracts in cicada fungus artificial culture product spore powder, coremium and mycoplasma in the manufacture craft of washing composition and skin care product, can reduce washing composition and skin care product to the stimulation of skin.Because extracting solution itself is yellow transparent, therefore need to consider the color problem on the techniques such as preparation color make-up base oil, color, pearly-lustre.The activeconstituents of cicada fungus has nourishing, beauty treatment, the anti-ageing effect of waiting for a long time, the effect of having promoted washing composition and skin care product, and the skin quality that has also reduced the generation after personage's life-time service color make-up of liking to be beautiful is coarse, obscure, and the problems such as pigementation are arranged.

7, other products:

1) cicada fungus feed: the preparation method of cicada fungus feed smashs the oven dry of substandard products mycoplasma to pieces to become the cicada fungus feed in feed rear directly adding by a certain percentage as additive, need to do experimentation on animals.This law has reduced invalid components and cost, does not need to extract polysaccharide and concentration.

The doctor thinks that cicada fungus is a kind of valuable ingredient of traditional Chinese medicine close with Cordyceps sinensis, replace the wild cicada fungus of scarcity of resources and develop its deep processed product with cicada fungus artificial culture product (spore powder, coremium and mycoplasma) with low cost, making people can enjoy victory and be worth than the dietotherapeutic that the artificial cicada fungus of wild cicada fungus and relative low price brings.

Compared to the prior art, the present invention has the following advantages: the artificial culture Paecilomyces cicadae method that adopts of the present invention is easy, production cost is low, the cultivation cycle is short, and products obtained therefrom has the advantages such as quality is high, good stability.

Embodiment

Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.

Embodiment 1

1, the preparation of bacterial classification and cultivation:

(1) bacterial classification preparation: preparation potato sucrose nutrient agar or potato sucrose nutrient solution are standby;

1. the inclined-plane kind prepares: the wild cicada fungus that gathers from the Yunnan Head streams obtains by the separate tissue purifying, after tube, cultivates 5 days under 22 ℃, selects good standby of growing way;

2. the one-level kind prepares: the potato sucrose nutrient solution is packed in Erlenmeyer flask, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, under aseptic condition, standby inclined-plane kind is accessed in Erlenmeyer flask after cooling, at 22 ℃, oscillation frequency is to cultivate 5 days under the condition of 130r/min, occurs to the Erlenmeyer flask after a large amount of mycelium pellets standby;

The PSA Medium Proportion that present embodiment adopts is every 1000ml substratum: potato 200g, sucrose 20g, agar 20g, its preparation method is diced to be placed in and is added water 1000ml in container for remove the peel potato, boils 20min, filtration, add sucrose and agar, supply water to 1000ml, then heating for dissolving divides to be filled in vitro, loading amount is 1/4 of test tube length, tampon, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave beyond the Great Wall, while hot test tube is placed on skewback cooling.

The potato sucrose nutrient solution proportioning that present embodiment adopts is every 1000ml nutrient solution: potato 200g, sucrose 20g, and its preparation method is diced to be placed in and is added water 1000ml in container for removing the peel potato, boils 30min, filter, add sucrose, supply water to 1000ml, heating for dissolving, then divide and be filled in Erlenmeyer flask, loading amount is 30% of Erlenmeyer flask capacity, tampon beyond the Great Wall, then wrap up with kraft paper, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, taking-up is cooled to below 20 ℃.

The batching mounted box: the preparation solid medium, the substratum for preparing to be packed into cultivate in box, charge amount is 15%, seals with breathable sealing film; The described solid medium proportioning of present embodiment is matrix 42 weight parts, bagasse 2 weight parts, oyster shell whiting 0.1 weight part, dried silkworm chrysalis meal 2 weight parts, saltpetre 0.1 weight part, peptone (fish meal) is 0.5 weight part, water is 56 weight parts, its mesostroma is comprised of Semen Maydis powder, wheat bran and sucrose, and in the gross weight of matrix, and the weight percent of Semen Maydis powder is 55%, the weight percent of wheat bran is 44%, the weight percent of sucrose is 1%.Its preparation method is: first Semen Maydis powder, wheat bran are boiled, sucrose, saltpetre are dissolved in water, more proportionally mix with bagasse, oyster shell whiting, dried silkworm chrysalis meal.

(2) sterilization: will cultivate box and put into Autoclave, 30min sterilizes under 0.15MPa pressure;

(3) inoculation: after solid medium is cooled to below 20 ℃, inoculate under aseptic condition, every 300ml one-level kind meets 50-60 and only cultivates box;

(4) fermentation: postvaccinal cultivation box moves to culturing room at 20 ℃-25 ℃, cultivate 30d-50d under relative humidity 60%-80%, later stage coremium growth phase requires daylighting, and envrionment temperature is compared the mycelial growth stage in early stage need hang down 2～3 ℃, need ventilation in good time every day, keep cultivating room air fresh;

(5) gather: when the coremium height of culture reaches the 9cm left and right, can gather when the surface has a small amount of conidium to occur; When gathering, remove sealed membrane, with instrument, culture is taken out and separate coremium, the oven dry classification, the cooling rear bag film vacuum packaging of using, mycoplasma is placed in-20 ℃ of storages.

2, output and nature examination:

In the present embodiment, the cultured products of gained is comprised of surplus materials mixture three parts of coremium, conidium, mycelium and substratum, and output is coremium 100g-110g (water ratio is below 8%)/1kg solid medium.Prove through pharmacological experiment, in the present embodiment, the cultured products of Paecilomyces cicadae has the effect that improves renal function.Compare with natural cicada fungus, because natural cicada fungus is had relatively high expectations to growing environment, output is less, and therefore the cultured product relative low price of the Paecilomyces cicadae of the present embodiment under artificial controlled condition can be used for extensive Paecilomyces cicadae artificial culture.

3, the application of cultured products:

The cultured products that makes in the present embodiment is for the preparation of cicada fungus tea, and the preparation method of cicada fungus tea cuts into sheet with coremium, adds appropriate warm water and can brew into cicada fungus tea.

Embodiment 2

1, the preparation of bacterial classification and cultivation:

1. the inclined-plane kind prepares: the wild cicada fungus that gathers from the Yunnan Head streams obtains by the separate tissue purifying, after tube, cultivates 7 days under 25 ℃, selects good standby of growing way;

2. the one-level kind prepares: the potato sucrose nutrient solution is packed in Erlenmeyer flask, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, in cooling rear access Erlenmeyer flask, at 25 ℃, oscillation frequency is to cultivate 3 days under the condition of 150r/min, occurs to the Erlenmeyer flask after a large amount of mycelium pellets standby;

The PSA Medium Proportion that present embodiment adopts is every 1000ml substratum: potato 200g, sucrose 20g, agar 20g, its preparation method is diced to be placed in and is added water 1000ml in container for remove the peel potato, boils 20min-30min, filtration, add sucrose and agar, supply water to 1000ml, then heating for dissolving divides to be filled in vitro, loading amount is the long 1/5-1/4 of test tube, tampon, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave beyond the Great Wall, while hot test tube is placed on skewback cooling.

The potato sucrose nutrient solution proportioning that present embodiment adopts is every 1000ml nutrient solution: potato 200g, sucrose 20g, its preparation method is diced to be placed in and is added water 1000ml in container for removing the peel potato, boil 15min-30min, filter, add sucrose, supply water to 1000ml, heating for dissolving, then divide and be filled in Erlenmeyer flask, loading amount is 20%～30% of Erlenmeyer flask capacity, tampon beyond the Great Wall, then wrap up with kraft paper, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, taking-up is cooled to below 20 ℃.

(2) batching mounted box: the preparation solid medium, the substratum for preparing to be packed into cultivate in box, charge amount is 15%-25%, seals with sealed membrane; The described solid medium proportioning of present embodiment is matrix 46 weight parts, bagasse 5 weight parts, and oyster shell whiting 0.4 weight part, dried silkworm chrysalis meal 2.5 weight parts, saltpetre 0.1 weight part, water are 44 weight parts; Its mesostroma is wheat.

Its preparation method is: first wheat is boiled, sucrose, saltpetre is dissolved in water, then with bagasse, oyster shell whiting, dried silkworm chrysalis meal proportionally mixes.

(3) sterilization: will cultivate box and put into Autoclave, sterilization 30m in-60min under 0.15MPa pressure;

(4) inoculation: after solid medium is cooled to below 20 ℃, inoculate under aseptic condition, every 300ml one-level kind meets 50-60 and only cultivates box;

(5) solid fermentation: postvaccinal cultivation box moves to culturing room at 20 ℃-25 ℃, cultivate 30d-50d under relative humidity 60%-80%, the mycelial growth stage in early stage needs shading, later stage coremium growth phase requires daylighting, and envrionment temperature is compared the mycelial growth stage in early stage need hang down 2～3 ℃, need ventilation in good time every day, keep cultivating room air fresh;

(6) gather: when the coremium height of cicada fungus culture reaches the 9cm left and right, can gather when the surface has a small amount of conidium to occur; When gathering, remove sealed membrane, with instrument, the culture medium culturing thing is taken out and separate coremium, the oven dry classification, the cooling rear bag film vacuum packaging of using, mycoplasma is placed in-20 ℃ of storages.

The artificial culture Paecilomyces cicadae method that adopts of the present invention is easy, production cost is low, the cultivation cycle is short, and products obtained therefrom has the advantages such as quality is high, good stability.

2, output and nature examination:

In the present embodiment, the cultured products of gained is comprised of surplus materials mixture three parts of coremium, conidium, mycelium and substratum, and output is coremium 100g-110g (water ratio is below 8%)/1kg solid medium.Prove through pharmacological experiment, in the present embodiment the cultured products of Paecilomyces cicadae have antitumor, regulate immune, hypoglycemic, reducing blood-fat, hypotensive, make eye bright, radioprotective, antipyretic-antalgic, tranquilizing soporific, strengthening by means of tonics, improve the effect of renal function.Compare with natural cicada fungus, because natural cicada fungus is had relatively high expectations to growing environment, output is less, and therefore the cultured product relative low price of the Paecilomyces cicadae of the present embodiment under artificial controlled condition can be used for extensive Paecilomyces cicadae artificial culture.

3, the application of culture:

The cultured products that makes in the present embodiment is for the preparation of the cicada fungus capsule, and its preparation method is as follows: coremium and the mycoplasma of artificial culture are pulverized, got capsule with pharmaceutical excipient mixing packing after lyophilization.

Embodiment 3

1, the preparation of bacterial classification and cultivation:

2. the one-level kind prepares: the potato sucrose nutrient solution is packed in Erlenmeyer flask, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, in cooling rear access Erlenmeyer flask, at 23 ℃, oscillation frequency is to cultivate 5 days under the condition of 130r/min, occurs to the Erlenmeyer flask after a large amount of mycelium pellets standby;

The potato sucrose nutrient agar proportioning that present embodiment adopts is every 1000ml substratum: potato 200g, sucrose 20g, agar 20g, its preparation method is diced to be placed in and is added water 1000ml in container for removing the peel potato, boil 20min-30min, filter, add sucrose and agar, supply water to 1000ml, heating for dissolving, then divide and be filled in vitro, loading amount is the long 1/5-1/4 of test tube, tampon, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave beyond the Great Wall, while hot test tube is placed on skewback cooling.

The potato sucrose nutrient solution proportioning that present embodiment adopts is every 1000ml nutrient solution: potato 200g, sucrose 20g, and its preparation method is diced to be placed in and is added water 1000ml in container for removing the peel potato, boils 15min-30min, filter, add sucrose, supply water to 1000ml, heating for dissolving, then divide and be filled in Erlenmeyer flask, loading amount is 20% of Erlenmeyer flask capacity, tampon beyond the Great Wall, then wrap up with kraft paper, be placed in the 30min that sterilizes under 0.15MPa pressure in Autoclave, taking-up is cooled to below 20 ℃.

(2) batching mounted box: the preparation solid medium, the substratum for preparing to be packed into cultivate in box, charge amount is 15%, seals with sealed membrane; The described solid medium proportioning of present embodiment is matrix 46 weight parts, bagasse 10 weight parts, and oyster shell whiting 0.5 weight part, dried silkworm chrysalis meal 5 weight parts, saltpetre 0.5 weight part, water are 44 weight parts; Its mesostroma is grain and sucrose (in the gross weight of grain and sucrose, the weight percent of grain is 98%, and the weight percent of sucrose is 2%).Its preparation method is: first grain is boiled, sucrose, saltpetre are dissolved in water, more proportionally mix with bagasse, oyster shell whiting, dried silkworm chrysalis meal.

(3) sterilization: will cultivate box and put into Autoclave, 30min sterilizes under 0.15MPa pressure;

(4) inoculation: after solid medium is cooled to below 20 ℃, inoculate under aseptic condition, every 300ml one-level kind connects 80 and cultivates box;

(5) solid fermentation: postvaccinal cultivation box moves to culturing room at 25 ℃, relative humidity was cultivated 50 days for 80% time, later stage coremium growth phase requires daylighting, and envrionment temperature is compared the mycelial growth stage in early stage need hang down 2～3 ℃, the ventilation of need in good time windowing every day keeps cultivating room air fresh;

2, output and nature examination:

3, the application of culture:

The cultured products that makes in the present embodiment is for the preparation of cicada wine, and its preparation method is as follows: the coremium of artificial culture and mycoplasma are pulverized-〉 are dropped into container-〉 add wine base-〉 certain temperature hot dipping a few hours and constantly stir-〉 change a certain proportion of finings of liquid hot dipping again-〉 merging leach liquor-〉 add-〉 stir-〉 filter-〉 finished product

Measurement of the polysaccharide content in the cultured products that makes in embodiment 4 embodiment 1-3:

1, reagent

A liquid: 0.1% glucose reference liquid

Glucose 0.1g after the accurate weighing oven dry, to 100ml, concentration is 0.1% with the distilled water constant volume.

B liquid: 5% phenol solution.

Essence is got and is taken the 5g re-distilled phenol, uses the distilled water constant volume to 100ml.

2, determination

Accurately take each 0.5g of artificial culture Paecilomyces cicadae coremium that makes in wild cicada fungus, embodiment 1-3, add 10ml distilled water, then ultrasonic extraction 10min puts into 90 ℃ of water-baths, hot water lixiviate 2h, and the centrifugal 10min of 4000r/min obtains supernatant liquor.Repeat above-mentioned steps once, merge supernatant liquor, be used for polysaccharide and detect.

3, sample polysaccharide determination

1) dilution of sample liquid is 12.5 times.

2) rob the rear sample liquid of accurate absorption 1ml dilution with application of sample, add 1ml distilled water, then add 5% phenol solution, shake up.Add the 5ml98% vitriol oil.Shake up.Detect after being cooled to room temperature.

3) adopt ultraviolet-visible pectrophotometer to detect, wavelength 490nm.

4, Specification Curve of Increasing

Accurately draw 0.1% glucose reference liquid (A liquid) 0.0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0ml is in test tube, and test tube is numbered 1,2,3,4,5,6,7,8,9,10,11, and wherein No. 1 test tube is blank liquid.Add water to respectively 2.0ml, accurately add respectively 5% phenol solution 1.0ml, shake up and add rapidly vitriol oil 5.0ml, shake up, be cooled to room temperature, measure absorbancy with ultraviolet-visible spectrophotometry at the wavelength place of 490nm, take absorbancy as ordinate zou, the concentration of glucose is X-coordinate, and the drawing standard curve gets equation of linear regression: y=0.1475 ^*X+0.0226, R2=0.9915.Mass concentration is good relationship in the 0.1-0.6mg/ml scope.

5, calculation formula

Polysaccharide content=aCV/W

A: extension rate, C: sugared concentration (mgmL in the vat liquor that is obtained by regression equation calculation.), V: volume (ml); W: be the weight (mg) of sample.

6, detected result

Table 1 sample test result

Result in table 1 proves: the polysaccharide content in the cicada fungus cultured products that makes in the present embodiment is higher than wild cicada fungus, and product quality is stable, is of very high actual application value.

Embodiment 5, the Paecilomyces cicadae artificial culture culture renal function to 5/6 rat remnant model

The culture of tested material: embodiment 1 (artificial cicada fungus).

Positive control drug: losartan, Hangzhou Mo Shadong pharmaceutical Co. Ltd produces; Natural cicada fungus, the place of production are Zhejiang.

Laboratory animal:

70 of SPF level male SD rats, at 2 monthly ages, body weight 200 ± 10g is provided by Shanghai Univ. of Traditional Chinese Medicine's Experimental Animal Center.

Grouping and modeling:

SD rat adaptability is divided into 2 groups after feeding for 1 week at random: 62 of blank group (sham operated rats) 8 and experimental group (operation group).Adopt the most of cutting method of 5/6 kidney to set up the experimental rat model: the 1st operation, the left kidney of operation group rat always excises renal cortex weight and accounts for 2/3 of left kidney weight; Carry out the 2nd operation after 7d, right kidney is cut entirely.Two operations is for 5/6 of excision two kidney gross weights.Sham operated rats SD rat adopts same step to expose kidney, separates kidney peplos, but refuses the nephrectomy.54 of operation treated animal survivals, sham operated rats is all survived 8.In 2 weeks after the 2nd operation, all animal row inner canthus arteriovenous clump blood samplings are carried out serum creatinine (Scr) and are detected.And with 54 operation group rats, be divided into 5 groups according to the Scr stratified random: 12 of model control group (model group), 10 of losartan groups, 11 of natural cicada fungus groups, the heavy dose of group of artificial cicada fungus (A group) 10,11 of artificial cicada fungus small dose group (B group).The losartan group, natural cicada fungus group, the artificial heavy dose of group of cicada fungus (A group), artificial cicada fungus small dose group (B group) rat is 4 weeks beginning medicine feed after the 2nd operation, and treatment time is 42d.When experiment finishes, survival of rats is 44, each 8 of sham operated rats, the artificial heavy dose of groups of cicada fungus (A group), respectively 7 of model group, losartan group, natural cicada fungus group, artificial cicada fungus small dose group (B group).

Dosage:

Natural cicada fungus group, the heavy dose of group of artificial cicada fungus (A group) rat are pressed 4gkg ^-1D ^-1Consumption, artificial cicada fungus small dose group (B group) is pressed 2gkg ^-1D ^-1Consumption adds administration in feed, and every daily physiological saline gavage 1 time; Losartan group rat is adopted administration by gavage, presses 30mgkg ^-1D ^-1Consumption is dissolved in physiological saline, and every day, gavage was 1 time.All rat grouping sub-cage rearings, every cage 4-5 rat, the whole rats of experimental session freely drink water, the commercially available blocky-shaped particle normal diet of taking food.

Testing tool and method:

Experiment detects Scr, blood urea nitrogen (BUN), the albumin (Alb) of all rats when finishing; Collect the 24h urine, measure quantity of proteinuria.Above index adopts Hitachi's 7170 type automatic biochemistry analyzers to measure.

With HE dyeing and PAS dyeing nephridial tissue pathology sheet inspection glomerular sclerosis, wherein, glomerular sclerosis is defined as that capillary lumen subsides and/or vitreous degeneration.The semi-quantitative method of employing Rajj etc. is only assessed the glomerular sclerosis degree.Under light microscopic (* 200) visual field, 40 renal glomeruluss are observed in every section at least, renal glomerulus ratio shared according to the glomerular sclerosis kitchen range is divided into 0-++++, 0 expression renal glomerulus is without sclerosis, 1 renal glomerulus of+expression 25% impaired, ++ 26%-50% is impaired in expression, ++ 51%-75% is impaired in+expression, ++ ++ 76%-100% is impaired in expression.Then calculate glomerular sclerosis index (GSI).

Adopt SPSS Version 11.0 for Windows to carry out data statistics, with mean ± standard deviation

Expression relatively adopts One-Way ANOVA to analyze between group, the statistical significance level is P＜0.05.

Test-results:

Found that, the Scr of rat, BUN value, each treatment group significantly reduces (P＜0.05) than model group, each group of artificial cicada fungus (heavy dose of group, small dose group) is compared no difference of science of statistics (P＞0.05) with natural cicada fungus, the seralbumin value of rat, sham operated rats, losartan group, natural cicada fungus group, the artificial heavy dose of group of cicada fungus (A group) significantly raise (P＜0.05), and the artificial heavy dose of group of cicada fungus (A group) is compared no difference of science of statistics (P＞0.05) with natural cicada fungus group and sham operated rats; The 24h urine albumen amount of rat, each group is compared with sham operated rats, all obviously increase (P＜0.05), but losartan group, natural cicada fungus group, the artificial heavy dose of group of cicada fungus (A group) compares with model group, and obvious minimizing (P＜0.05) is arranged.See Table 1.

Table 1 rat model hematology and urine detection result are relatively

Annotate: compare with sham operated rats, ^ΔP＜0.05; Compare with model group, ^*P＜0.05; Compare with natural cicada fungus group, ^#P＜0.05 conventional H E and PAS dyeing, observe the nephridial tissue pathology under light microscopic and change: rats in sham-operated group renal glomerulus vessel open, clear in structure, after PAS dyeing, renal glomerulus collagen is thin-line-shaped distribution; In model group Renal Glomeruli In Rats mesangial region matrix, severe increases severe hyperplasia in companion's mesangial cell, and part Capillary loops pressurized is inaccessible, and hyaloid pathology, and matter lymphocytic infiltration between the companion are seen protein cast in the part uriniferous tubules.Each treatment group glomerular sclerosis degree alleviates (P＜0.05) than model group is obvious, be followed successively by B group＞A group＞losartan group＞natural cicada fungus group, and A group, B group and natural cicada fungus group are compared no difference of science of statistics (P＞0.05).

See Table 2

The glomerular sclerosis index respectively organized by table 2 and matrix positive area ratio compares

Annotate: compare with sham operated rats, ^ΔP＜0.05; Compare with model group, ^*P＜0.05; Compare with natural cicada fungus group, ^#P＜0.05

Embodiment 6 acute oral toxicity tests

A. the culture of sample title: embodiment 1

B. sample preparation: take sample 10000mg, adding distil water is to 40ml, fully after mixing as given the test agent.

C. laboratory animal: 20 of Kunming mouses, male and female half and half, body weight 19-22 gram is provided by Shanghai Slac Experimental Animal Co., Ltd..Production licence number: SCXK (Shanghai) 2007-0005.20-25 ℃ of receptacle temperature, relative humidity: 40-70%.

Laboratory animal occupancy permit number: SYXK (Shanghai) 2007-0008.Mouse feed is provided by Suzhou two lion laboratory animal feed technology Services Co., Ltd, registration card number: the E of Soviet Union raises new word (2002) 006.

D. experimental technique:

1. animal fasting (can't help water) was selected each 10 of female, male mouse by the body weight requirement after 16 hours, divided to be put in two mouse cages, with the poor 3g that is no more than of body weight between the sex mouse.

2. given the test agent is adopted the maximum tolerated dose method that laboratory animal is contaminated, mouse is by only weighing, and the gavage capacity is by each 0.4ml/20g batheroom scale, and secondary is to mouse stomach in 24 hours, and gavage interval time is 6 hours.

3. after the contamination, observe general state, body weight change, toxicity symptom and the death condition etc. of animal.Observation period is a week.

4. weigh to animal again in the experiment end.Dead animal and the animal of putting to death that expires are carried out necrotomy, and the visual inspection general pathology changes situation.

5. experiment whole process and observed content are all done detail record, press maximum tolerated dose method test-results, try to achieve chmice acute per os MTD.

E. result:

Table 6 male and female chmice acute Oral toxicity test-results

1. main physical signs performance:

Each treated animal of duration of test is movable normal, and the hair color glossiness is good, has no any poisoning sign and death; Expire and put to death animal, each internal organs situation of gross anatomy visual inspection, no abnormality seen.

2. female mice: MTD＞10000mg/kg

Male mice: MTD＞10000mg/kg

F. conclusion:

Sample to the maximum tolerated dose MTD of male and female chmice acute Oral toxicity test all greater than 10000mg/kg.Belong to actual nontoxic level.

Claims

1. the method for a Paecilomyces cicadae artificial culture, comprise the steps:

2) inoculation: the one-level kind that makes in step 1) is linked in solid medium;

4) gathering of coremium and mycoplasma,

The component title that contains in described solid medium and the weight part of each component are:

Matrix 42～46 weight parts;

Bagasse 2～10 weight parts;

Oyster shell whiting 0.1～0.5 weight part;

Dried silkworm chrysalis meal 2～5 weight parts;

Saltpetre 0.1～0.5 weight part;

Water 44～56 weight parts;

2. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that step 2) in, the step of described inoculation is: after the temperature of the solid medium in cultivating box is cooling, inoculate under aseptic condition, the inoculum size according to 3～5% is with in one-level kind access culture vessel.

3. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that, in the mixture of described Semen Maydis powder, wheat bran and sucrose, in the gross weight of the mixture of described Semen Maydis powder, wheat bran and sucrose, the weight percent of Semen Maydis powder, wheat bran and sucrose is respectively: Semen Maydis powder 43%～55%, wheat bran 44%～55%, sucrose 1%～2%.

4. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that, in the mixture of described grain and sucrose, in the gross weight of the mixture of described grain and sucrose, the weight percent of grain and sucrose is respectively: grain 98%～99%, sucrose 1%～2%.

5. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that, in the mixture of described rice and sucrose, in the gross weight of the mixture of described rice and sucrose, the weight percent of rice and sucrose is respectively: rice 98%～99%, sucrose 1%～2%.

6. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that, in the mixture of described Chinese sorghum and sucrose, in the gross weight of the mixture of described Chinese sorghum and sucrose, the weight percent of Chinese sorghum and sucrose is respectively: Chinese sorghum 98%～99%, sucrose 1%～2%.

7. the method for the Paecilomyces cicadae artificial culture described in claim 1-6, is characterized in that, the carbon-nitrogen ratio of described solid medium is 80～98.

8. the method for the Paecilomyces cicadae artificial culture described in claim 1-6, is characterized in that, also contains peptone in described solid medium, and the weight part of described peptone is 0.5 weight part.

9. the method for the Paecilomyces cicadae artificial culture described in claim 1-6, it is characterized in that, the preparation method of described solid medium is: first the component except sucrose in matrix is boiled, sucrose, saltpetre are dissolved in water, with bagasse, oyster shell whiting, dried silkworm chrysalis meal proportionally mixes again.

10. the method for Paecilomyces cicadae artificial culture as described in claim 1, it is characterized in that, in step 3), require daylighting at solid fermentation later stage coremium growth phase, and envrionment temperature is compared the mycelial growth stage in early stage need hang down 2～3 ℃, need maintain the circulation of air in culturing process.

11. the cultured products of a Paecilomyces cicadae, made by Paecilomyces cicadae cultivation of method described in arbitrary claim in claim 1～10, prepared cultured products is comprised of mixture three parts of coremium, conidium, mycelium and substratum surplus materials.

12. the application of the cultured products of the Paecilomyces cicadae described in claim 11 in preparation food, healthcare products, medicine and makeup.

13. the application of the cultured products of Paecilomyces cicadae described in claim 12, it is characterized in that, described food, healthcare products, medicine and makeup comprise: cicada fungus tea, cicada wine, cicada fungus soy sauce, cicada fungus amino acid seasoning, cicada fungus coffee, cicada fungus milk preparation, cicada fungus candy, cicada fungus honeybee product, cicada fungus puffed food, cicada fungus chewing gum, cicada fungus herbal cuisine, cicada fungus spore oil, cicada fungus capsule, cicada fungus medicine materical crude slice, cicada fungus electuary, cicada fungus cosmetics or cicada fungus feed.

14. the application of the cultured products of Paecilomyces cicadae described in arbitrary claim in claim 12 or 13, it is characterized in that, that described healthcare products and medicine have is antitumor, it is immune, hypoglycemic to regulate, reducing blood-fat, hypotensive, make eye bright, radioprotective, antipyretic-antalgic, tranquilizing soporific, strengthening by means of tonics or improve the function of renal function.