CN102612985A - Production technology for cordyceps militaris mycelium - Google Patents
Production technology for cordyceps militaris mycelium Download PDFInfo
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Abstract
The invention discloses a production technology for cordyceps militaris mycelium. The production technology mainly comprises the procedures of preparation of a test tube stock culture, culture medium preparation and culturing mode of a liquid strain, culture medium preparation and culturing mode of mycelium, and the like. The defects of the materials adopted by a culture medium are complex, the manufacturing cost is high and the liquid strain culturing condition is high in the prior art are changed by adjusting a formula and a technology according to the invention; after the mycelium is cultured and subjected to color transfer, the main components and pharmacology of the mycelium are more approached to those of the natural cordyceps militaris; and the production technology has the advantages of simple method, low comprehensive production cost, capability of performing large-scale production, and higher developing value.
Description
Technical field
The invention belongs to the biology techniques field, be specifically related to the mycelial production technology of a kind of Cordyceps militaris.
Background technology
Northern Chinese caterpillar Fungus; Claim Cordyceps militaris again; It is that section ergot fungus cordyceps sinensis Cordycrps sinensis (Berk.) Sacc colonizes in stroma and the complex of larva corpse on the larvae section insect larvae; Be a kind of traditional famous and precious tonic Chinese herbal medicine material, be listed as three big invigorants with natural ginseng, pilose antler.Its property of medicine is gentle, equal edible throughout the year, and the old or young, sick, weak, virtual person is all suitable, than the invigorant of other kinds medical value is widely arranged.Since ancient times, people all it as invigorant and pharmaceutical products.
Natural wild cordyceps mainly is grown in high height above sea level, high and cold area, and the formation of growing environment is very harsh and complicated, and people constantly increase and irrational excavating the demand of Chinese caterpillar fungus in addition, cause natural Cordyceps sinensis natural resources to gradually reduce.Therefore, in order to solve the medicine source, scientific and technical personnel have to meet the need of market through the artificial cultivation Cordyceps militaris, but for these years, tame Cordyceps militaris complex manufacturing, production cycle are grown, yield poorly etc. and perplexing people always.
Through the effort of researcher, a kind of brand-new Cordyceps militaris product---the cordyceps filament has developed, and this mycelium just is to use biological submerged fermentation technology to obtain.
Agricultural University Of Anhui had once carried out the analysis of active ingredient to the cordyceps filament; Its result shows: the amino acid kind is consistent with the Cordyceps sinensis crude drug in the mycelium; But total amino acid content and TEAA are apparently higher than the Cordyceps sinensis crude drug; The ratio that essential amino acid accounts for total amino acid also is higher than the latter; The content of threonine, valine, isoleucine, leucine, lysine is farther far above the latter in the essential amino acid, and the content of crude protein, total reducing sugar and mannitol all is higher than the Cordyceps sinensis crude drug in the mycelium; During mycelial fatty acid is formed; Unsaturated fatty acid accounts for 85.38% of fatty acid composition; Wherein polyvalent unsaturated fatty acid such as linoleic acid, linolenic acid accounts for 58.87% of fatty acid composition, and linoleic content accounts for (51.04%) over half that whole fatty acid is formed; Fermentation mycelium also contains abundant to metallic elements such as the very important Zn of health, Fe, Ca, and the content with Ca is height especially.
Produce the cordyceps filament with biological submerged fermentation technology, characteristics such as it is with short production cycle, technology is simple, production efficiency is high, stable are the main developing direction of artificial culture Cordyceps militaris product.
In the existing technology; Number of patent application is that 02134573.2 Deel-layer fermentation process for culturing Brazilian cordyceps mycelia and number of patent application are 93104709.9 producing process of Chinese caterpillar fungus hypha fermentation; All be to use expensive fermentation tank to cultivate; Fermentation condition is high, and raw material that medium uses is complicated, cost is high.What number of patent application was that 01141350.6 a kind of CORDYCEPS bacterium powder class green feed additive adopts is solid fermentation, and production technology is simple, but needs general 2 months time to cultural hypha well, and the production cycle is long.The mycelium physiological property that above-mentioned technology is produced is all not mature enough, does not contain high activity materials such as the distinctive cordycepin of Chinese caterpillar fungus, SOD, Cordyceps sinensis polysaccharide.
Summary of the invention
The objective of the invention is to overcome existing technical deficiency and use expensive fermentation tank to cause fermentation condition high cost height, strain cultivation condition height and medium to use shortcomings such as raw material complicacy in the prior art through adjusting production technology and filling a prescription to change; The technology that is provided is the fermentation tank that a kind of liquid spawn need not involve great expense when cultivating, and is to use rotary shaking table to cultivate; The raw material that uses is cast aside expensive mineral element, and most use is easy to get, the crops leftovers of cheapness; Whole technology pattern condition of culture is simple, the cycle is short, efficient is high, stable etc.Importantly mycelium makes its physiological property ripe after being cultured to annesl, contains high activity materials such as the distinctive cordycepin of Chinese caterpillar fungus, SOD, Cordyceps sinensis polysaccharide.
The present invention realizes through following technical scheme:
1, the female compound method of planting of test tube:
(1) medium proportioning: potato 100 grams, corn flour 15 grams, analysis for soybean powder 12 grams, white sugar 20 grams, dried silkworm chrysalis meal 15 grams, agar 18 grams, 1000 milliliters in water;
(2) preparing process: after potato dug skin 1000 ml waters are broken into slurry, be added into corn flour, analysis for soybean powder, white sugar, dried silkworm chrysalis meal, agar more successively according to quantity, then solution is heated to the boiling back and divides and install in the test tube, the branch loading amount is 1/4 of a test tube; The test tube that branch is installed is positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 40 minutes under 121 ℃ the high pressure, hot conditions; Put, be cooled to slant tube to the test tube of the poison that disappeared;
(3) tube: screening meets suitability for industrialized production, mycelial growth is vigorous, color and luster normal and FFI one-level Cordyceps militaris inclined-plane is female plants, the slant tube that switching goes into to disinfect under gnotobasis, and inserting kind of a block size is 0.3～0.5cm
2
(4) cultivate: being placed on temperature to the test tube of good kind of switching is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) annesl: when the medium face length in the test tube expires mycelia, promptly under the luminosity of 100～200 luxs, made public 3～4 days, mycelia transfers salmon pink to.
2, the compound method of liquid spawn culture medium:
(1) proportioning of medium: potato 1000 grams, corn flour 50 grams, analysis for soybean powder 120 grams, flour 100 grams, white sugar 200 grams, dusty yeast (cream) 80 grams, dried silkworm chrysalis meal 150 grams, 10000 milliliters in water;
(2) preparing process: after potato dug skin 10000 ml waters are broken into slurry; Be added into corn flour, analysis for soybean powder, flour, white sugar, dusty yeast (cream), dried silkworm chrysalis meal more successively according to quantity; Then solution being heated to boiling back branch installs in the Clear glass bottles and jars of high temperature high voltage resistant; Charge weight is for holding the half the of tool actual capacity, then with cotton with the bottleneck jam-pack; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 60 minutes under 121 ℃ the high pressure, hot conditions.
3, the cultural method of liquid spawn:
(1) inoculation: under gnotobasis, inserts the test tube mother kind that turns over look to the bottling liquid medium of the poison that disappeared, every bottle graft is gone into 4～6 ferfas pieces, and static cultivation 3～5 days;
(2) cultivate: bacterial classification sprout, free of contamination bottling liquid medium moves on to rotary shaking table and dynamically cultivates; The shaking table rotary speed is 150～300 rev/mins; The rotation of shaking table is also rotated liquid nutrient medium thereupon; The clinker collision of the rotation of bacterial classification liquid medium within and medium the inside constantly point is torn into several bacterium balls (sheet, silk) down, and acquisition has the liquid spawn of enriching bacterium ball (sheet, silk) after 3 days.
4, the mycelial compound method of solid:
(1) proportioning of medium: 30 jin in rice, 30 jin in rice bran, 30 jin in wheat bran, 3 jin of corn flour, 3 jin of analysis for soybean powder, 4 jin of sugar, 130 jin in water;
(2) the even back of above-mentioned various mixing of materials branch is installed in the plastic sack, the branch loading amount is the half the of plastic sack actual capacity, tightens sack then; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 90 minutes under 121 ℃ the high pressure, hot conditions;
(3) under gnotobasis, insert cultured liquid spawn to the packed medium of the poison that disappeared, the access amount of liquid spawn is 3～5% of a solid material;
(4) being placed on temperature to the packed medium that connects kind is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) after the medium in the bag is had thorough grasp by mycelia, promptly under the luminosity of 100～200 luxs, to make public 3～4 days, mycelia transfers salmon pink to.
5, mycelial processing method:
The mycelium that is cultured to annesl nature or mechanical drying to solids water content under 60 ℃ of environment are promptly obtained the cordyceps filament of high-quality below 10%.
Compared with prior art, the existing following advantage of the present invention:
1, the raw material that uses of the present invention simple, be prone to purchase, cost is low.
2, the appearance tool that uses of the present invention also simple, be prone to purchase, cost is low.
3, the present invention adopts rotary shaking table to carry out strain cultivation, and liquid spawn does not need multistage cultivation in incubation simultaneously, not only shortens the production cycle, has also reduced the risk because of the easy infection of multistage switching spawn culture simultaneously.
What 4, the present invention cultivated is that mycelium is after being cultured to annesl, to make its physiological property ripe, contains high activity materials such as the distinctive cordycepin of Chinese caterpillar fungus, SOD, Cordyceps sinensis polysaccharide.
Embodiment
Below in conjunction with embodiment method of the present invention is further specified.
The mycelial production technology of a kind of Cordyceps militaris, embodiment is following:
1, the female compound method of planting of test tube:
(1) medium proportioning: potato 100 grams, corn flour 15 grams, analysis for soybean powder 12 grams, white sugar 20 grams, dried silkworm chrysalis meal 15 grams, agar 18 grams, 1000 milliliters in water;
(2) preparing process: after potato dug skin 1000 ml waters are broken into slurry, be added into corn flour, analysis for soybean powder, white sugar, dried silkworm chrysalis meal, agar more successively according to quantity, then solution is heated to the boiling back and divides and install in the test tube, the branch loading amount is 1/4 of a test tube; The test tube that branch is installed is positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 40 minutes under 121 ℃ the high pressure, hot conditions; Put, be cooled to slant tube to the test tube of the poison that disappeared;
(3) tube: screening meets suitability for industrialized production, mycelial growth is vigorous, color and luster normal and FFI one-level Cordyceps militaris inclined-plane is female plants, the slant tube that switching goes into to disinfect under gnotobasis, and inserting kind of a block size is 0.3～0.5cm
2
(4) cultivate: being placed on temperature to the test tube of good kind of switching is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) annesl: when the medium face length in the test tube expires mycelia, promptly under the luminosity of 100～200 luxs, made public 3～4 days, mycelia transfers salmon pink to.
2, the compound method of liquid spawn culture medium:
(1) proportioning of medium: potato 1000 grams, corn flour 50 grams, analysis for soybean powder 120 grams, flour 100 grams, white sugar 200 grams, dusty yeast (cream) 80 grams, dried silkworm chrysalis meal 150 grams, 10000 milliliters in water;
(2) preparing process: after potato dug skin 10000 ml waters are broken into slurry; Be added into corn flour, analysis for soybean powder, flour, white sugar, dusty yeast (cream), dried silkworm chrysalis meal more successively according to quantity; Then solution being heated to boiling back branch installs in the Clear glass bottles and jars of high temperature high voltage resistant; Charge weight is for holding the half the of tool actual capacity, then with cotton with the bottleneck jam-pack; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 60 minutes under 121 ℃ the high pressure, hot conditions.
3, the cultural method of liquid spawn:
(1) inoculation: under gnotobasis, inserts the test tube mother kind that turns over look to the bottling liquid medium of the poison that disappeared, every bottle graft is gone into 4～6 ferfas pieces, and static cultivation 3～5 days;
(2) cultivate: bacterial classification sprout, free of contamination bottling liquid medium moves on to rotary shaking table and dynamically cultivates; The shaking table rotary speed is 150～300 rev/mins; The rotation of shaking table is also rotated liquid nutrient medium thereupon; The clinker collision of the rotation of bacterial classification liquid medium within and medium the inside constantly point is torn into several bacterium balls (sheet, silk) down, and acquisition has the liquid spawn of enriching bacterium ball (sheet, silk) after 3 days.
4, the mycelial compound method of solid:
(1) proportioning of medium: 30 jin in rice, 30 jin in rice bran, 30 jin in wheat bran, 3 jin of corn flour, 3 jin of analysis for soybean powder, 4 jin of sugar, 130 jin in water;
(2) the even back of above-mentioned various mixing of materials branch is installed in the plastic sack, the branch loading amount is the half the of plastic sack actual capacity, tightens sack then; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 90 minutes under 121 ℃ the high pressure, hot conditions;
(3) under gnotobasis, insert cultured liquid spawn to the packed medium of the poison that disappeared, the access amount of liquid spawn is 3～5% of a solid material;
(4) being placed on temperature to the packed medium that connects kind is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) after the medium in the bag is had thorough grasp by mycelia, promptly under the luminosity of 100～200 luxs, to make public 3～4 days, mycelia transfers salmon pink to.
5, mycelial processing method:
The mycelium that is cultured to annesl nature or mechanical drying to solids water content under 60 ℃ of environment are promptly obtained the cordyceps filament of high-quality below 10%.
Claims (5)
1. mycelial production technology of Cordyceps militaris is characterized in that the female compound method of planting of test tube is following:
(1) medium proportioning: potato 100 grams, corn flour 15 grams, analysis for soybean powder 12 grams, white sugar 20 grams, dried silkworm chrysalis meal 15 grams, agar 18 grams, 1000 milliliters in water;
(2) preparing process: after potato dug skin 1000 ml waters are broken into slurry, be added into corn flour, analysis for soybean powder, white sugar, dried silkworm chrysalis meal, agar more successively according to quantity, then solution is heated to the boiling back and divides and install in the test tube, the branch loading amount is 1/4 of a test tube; The test tube that branch is installed is positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 40 minutes under 121 ℃ the high pressure, hot conditions; Put, be cooled to slant tube to the test tube of the poison that disappeared;
(3) tube: screening meets suitability for industrialized production, mycelial growth is vigorous, color and luster normal and FFI one-level Cordyceps militaris inclined-plane is female plants, the slant tube that switching goes into to disinfect under gnotobasis, and inserting kind of a block size is 0.3～0.5cm
2:
(4) cultivate: being placed on temperature to the test tube of good kind of switching is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) annesl: when the medium face length in the test tube expires mycelia, promptly under the luminosity of 100～200 luxs, made public 3～4 days, mycelia transfers salmon pink to.
2. the mycelial production technology of a kind of Cordyceps militaris according to claim 1 is characterized in that the compound method of liquid spawn culture medium is following:
(1) proportioning of medium: potato 1000 grams, corn flour 50 grams, analysis for soybean powder 120 grams, flour 100 grams, white sugar 200 grams, dusty yeast (cream) 80 grams, dried silkworm chrysalis meal 150 grams, 10000 milliliters in water;
(2) preparing process: after potato dug skin 10000 ml waters are broken into slurry; Be added into corn flour, analysis for soybean powder, flour, white sugar, dusty yeast (cream), dried silkworm chrysalis meal more successively according to quantity; Then solution being heated to boiling back branch installs in the Clear glass bottles and jars of high temperature high voltage resistant; Charge weight is for holding the half the of tool actual capacity, then with cotton with the bottleneck jam-pack; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 60 minutes under 121 ℃ the high pressure, hot conditions.
3. the mycelial production technology of a kind of Cordyceps militaris according to claim 1 is characterized in that the cultural method of liquid spawn is following:
(1) inoculation: under gnotobasis, inserts the test tube mother kind that turns over look to the bottling liquid medium of the poison that disappeared, every bottle graft is gone into 4～6 ferfas pieces, and static cultivation 3～5 days;
(2) cultivate: bacterial classification sprout, free of contamination bottling liquid medium moves on to rotary shaking table and dynamically cultivates; The shaking table rotary speed is 150～300 rev/mins; The rotation of shaking table is also rotated liquid nutrient medium thereupon; The clinker collision of the rotation of bacterial classification liquid medium within and medium the inside constantly point is torn into several bacterium balls (sheet, silk) down, and acquisition has the liquid spawn of enriching bacterium ball (sheet, silk) after 3 days.
4. the mycelial production technology of a kind of Cordyceps militaris according to claim 1 is characterized in that the mycelial compound method of solid is following:
(1) proportioning of medium: 30 jin in rice, 30 jin in rice bran, 30 jin in wheat bran, 3 jin of corn flour, 3 jin of analysis for soybean powder, 4 jin of sugar, 130 jin in water;
(2) the even back of above-mentioned various mixing of materials branch is installed in the plastic sack, the branch loading amount is the half the of plastic sack actual capacity, tightens sack then; Next be positioned in the high-pressure bottle at 1.5 kilograms/cm
2Pressure, temperature are to keep sterilization 90 minutes under 121 ℃ the high pressure, hot conditions;
(3) under gnotobasis, insert cultured liquid spawn to the packed medium of the poison that disappeared, the access amount of liquid spawn is 3～5% of a solid material;
(4) being placed on temperature to the packed medium that connects kind is that 18～22 ℃, relative moisture are that 75～85% half-light environment was cultivated 4～5 days down;
(5) after the medium in the bag is had thorough grasp by mycelia, promptly under the luminosity of 100～200 luxs, to make public 3～4 days, mycelia transfers salmon pink to.
5. the mycelial production technology of a kind of Cordyceps militaris according to claim 1 is characterized in that mycelial processing method is following:
The mycelium that is cultured to annesl nature or mechanical drying to solids water content under 60 ℃ of environment are promptly obtained the cordyceps filament of high-quality below 10%.
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| CN103030452A (en) * | 2013-01-17 | 2013-04-10 | 李卫旗 | Method for preparing liquid mycelium fermentation culture medium formula of selenium-rich cordyceps militaris |
| CN103460981A (en) * | 2013-07-24 | 2013-12-25 | 山西省医药与生命科学研究院 | Novel cordyceps militaris culturing method |
| CN103548571A (en) * | 2013-10-29 | 2014-02-05 | 中国海洋大学 | Culture method of cordyceps mycelia with high-yield acidic polysaccharides and acidic polysaccharides |
| CN103621315A (en) * | 2013-12-12 | 2014-03-12 | 福建农林大学 | Method for cultivating cordyceps militaris by using Chinese herbal medicine nutrient solution |
| CN103858673A (en) * | 2014-03-18 | 2014-06-18 | 鲁东大学 | Method for planting cordyceps militaris by beer residue |
| CN104004660A (en) * | 2014-05-05 | 2014-08-27 | 安徽金豪生态农业科技有限公司 | Shake-flask spawn culture composition of cordyceps militaris |
| CN104170650A (en) * | 2014-08-14 | 2014-12-03 | 安徽亿源中药饮片科技有限公司 | Cordyceps militaris industrialized artificial cultivation method |
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| CN103460981A (en) * | 2013-07-24 | 2013-12-25 | 山西省医药与生命科学研究院 | Novel cordyceps militaris culturing method |
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