CN103070011A - Preparation method of selenium-rich cordyceps sinensis mycelia - Google Patents
Preparation method of selenium-rich cordyceps sinensis mycelia Download PDFInfo
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- CN103070011A CN103070011A CN2013100057128A CN201310005712A CN103070011A CN 103070011 A CN103070011 A CN 103070011A CN 2013100057128 A CN2013100057128 A CN 2013100057128A CN 201310005712 A CN201310005712 A CN 201310005712A CN 103070011 A CN103070011 A CN 103070011A
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Abstract
The invention provides a preparation method of selenium-rich cordyceps sinensis mycelia. The method comprises the following steps of: inoculating cordyceps sinensis stains in a liquid fermentation culture medium with brewer's grains as a matrix; and culturing for 4-8 days under the conditions of 20-30 DEG C and 120-200 r/min to obtain the selenium-rich cordyceps sinensis mycelia in fermentation liquor, wherein the liquid fermentation culture medium is prepared from the following components: 10-50 g/L of brewer's grains, 10-30 mg/L of Na2SeO3 and water serving as a solvent with pH (Potential of Hydrogen) of 4-8. The obtained fermentation liquor can be further separated by the conventional method to obtain the selenium-rich cordyceps sinensis mycelia. Compared with the prior art, the method has the advantages of mild reaction conditions, low production cost, less equipment investment and easiness in realization of industrialization, and the obtained mycelia have stronger biological activity, so that a breakthrough is expected to be made in prevention and treatment of animal diseases.
Description
(1) technical field
The present invention relates to a kind of preparation method of rich selenium aweto mycelium.
(2) background technology
Chinese caterpillar fungus (Cordyceps) is the distinctive famous and precious medicinal fungi of China.Contain several physiological active substances in the Chinese caterpillar fungus, TCM Literature think Cordyceps sinensis distinguish the flavor of sweet warm in nature, the effect that secret lean gas is arranged, protect lung kidney tonifying, hemostasis and phlegm.The modern medicine clinical research shows, the diseases such as Cordyceps sinensis can treat that phthisical cough phlegm blood, night sweat, impotence, seminal emission, jaundice are sick, pulmonary tuberculosis and old weak, chronic cough and asthma have strong taking in concurrently to hold back calm effect.The contained cordycepin of Cordyceps sinensis can obviously strengthen the ability of red blood cell adhesion tumour cell, suppresses growth and metastasis of tumours, can obviously promote leukocyte and platelet counts, improves rapidly the symptoms such as vomiting behind the chemicotherapy is felt sick, poor appetite, epilation, insomnia.But natural cordyceps yield is very rare, and therefore, people adopt the cultivation aweto mycelium to substitute natural cordyceps.At present, the production of cultural mycelium is main mainly with liquid deep layer fermenting.
Selenium is essential a kind of element in the humans and animals body, and scarce selenium can cause multiple symptom.Lack selenium such as human body and can bring out angiocardiopathy, cancer, Keshan disease and Kaschin-Beck disease etc., and the scarce selenium main manifestations of animal is white muscle disease and poor growth etc.When for example ox lacks selenium, skeletal muscle and muscular degeneration of heart can occur, poor growth is become thin, and calf diarrhea when pig lacks selenium, shows as the liver malnutrition; White muscle disease appears in piglet, Sang Kanxin, and stomach ulcer etc., chicken lacks selenium then the pancreas degeneration necrosis can occur, cerebromalacia, Adult Chicken laying rate and incubation rate all descend degradation.
Periodical " Hubei University's journal (natural science edition) " 03 phase article " the mycelial fermented and cultured of selenium-rich cordyceps and biological function research thereof " in 2006 employing bacterial classification Cordyceps Sinensis, carry out liquid fermentation take sucrose, soybean meal and corn flour as matrix, in fermentation medium, add micro-Na
2SeO
3, the cordyceps mycelia recovery rate reaches 2.52%, and wherein Se content reaches 7.90 mg/kg.In the document, although obtained rich selenium mycelium, the matrix that this method is used is sucrose, soybean meal etc., and cost up does not possess economic advantages.
Brewer's grains (dregs of beans, wheat bran) etc. are because raw material is different, production technology is different, moisture is different, so that nutritive value is variant, but consider from technology, quality, environmental protection, investment, benefit and market several respects, by microorganism with this class grain slag be converted into nutritious, good palatability, added value food additives or microbiological feed high, remarkable in economical benefits are one of most economical effective methods.Contain a large amount of nutriments in the waste residue, behind microbial fermentation, can be transformed into more effectively bioactivator, not only can improve the nutritive value of feed as animal feed, more can improve resistance against diseases and the production capacity of livestock and poultry.If with the further separation and purification of these active components, can be used as food additives or even medicine.By biotechnology brewer's grains are carried out deep processing development, can greatly improve the availability of brewer's grains and the added value of deep processed product, turning waste into wealth for enterprise provides new approaches, promotes the economic benefit of beer producers.
For this reason, the present application people attempts with the medium of agricultural production accessory substance vinasse as fermentation, and Se content carries out rich selenium liquid state fermentation as the fermentation index to Cordyceps sinensis in the cordyceps mycelia, produces feed addictive.
(3) summary of the invention
In order to obtain the higher selenium-rich cordyceps mycelium of nutrition and medical value, overcome fermentation costs too high, cause the shortcoming that is unfavorable for suitability for industrialized production, the invention provides that a kind of new condition of culture is easy to realize, production cost is low, equipment investment is few, is easy to realize the mycelial production technology of industrialized selenium-rich cordyceps.
The technical solution used in the present invention is:
A kind of preparation method of rich selenium aweto mycelium, described method comprises: with aweto strain (conventional bacterial strain, commercial getting) be seeded in the liquid fermentation medium that brewer's grains are matrix, cultivated 4 ~ 8 days under 20 ~ 30 ℃, 120 ~ 200r/min condition, in zymotic fluid, obtain described selenium-rich cordyceps mycelium; Described liquid fermentation medium is prepared by following composition: brewer's grains 10 ~ 50g/L, Na
2SeO
310 ~ 30mg/L, solvent are water, and pH 4 ~ 8.The gained zymotic fluid can further separate the described selenium-rich cordyceps mycelium of acquisition according to a conventional method.The cordyceps mycelia of gained is pulverized, ammonium oxalate solution extracts 1 ~ 4 h, stimulate mouse spleen lymphocyte with extract, observation of cell increment situation, the cell appreciation rate was that 296.32%(is the data of embodiment 8 gained when the result showed 500 μ g/mL), the selenium-rich cordyceps mycelium that visible the inventive method makes has stronger biologically active.
Preferably, described liquid fermentation medium is prepared by following composition: brewer's grains 20 ~ 40g/L, Na
2SeO
315 ~ 25mg/L, solvent are water, and pH 5 ~ 7.
Preferably, described fermentation is carried out under 22 ~ 28 ℃, 120 ~ 180r/min condition.
Thalline inoculum concentration of the present invention is conventional inoculum concentration, generally at 5 ~ 15%(v/v, and seed liquor and fermentation medium volume ratio), being preferably 10%, seed liquor can be carried out according to a conventional method seed culture and be obtained.
Concrete, the inventive method can be carried out as follows:
1) seed liquor is cultivated: first the mother on the slant medium for preparing is planted with the inoculation shovel and get approximately 20 mm
2The lawn access seed culture medium (sucrose 50g/L, peptone 10g/L, yeast extract 10g/L, solvent are water) of size, sample-loading amount is in the triangular flask of 100mL/250mL, 25 ℃, the top fermentation of 160r/min constant temperature oscillator 3 days, forms small bacterium ball;
2) fermentation medium preparation: brewer's grains 10 ~ 50g/L, Na
2SeO
310 ~ 30mg/L, solvent are water, and pH 4 ~ 8(adjusts in order to 0.1N HCl and 0.1N NaOH);
3) sterilization of 120 ℃ of fermentation mediums is 20 minutes, and seed liquor is with 5 ~ 15%(v/v, seed liquor and fermentation medium volume ratio) inoculum concentration is inoculated in fermentation medium, cultivated 4 ~ 8 days under 20 ~ 30 ℃, 120 ~ 200r/min condition.
4) the gained zymotic fluid filters through 60 mesh sieves, and repeatedly washes three times, refilters, and drying namely gets the selenium-rich cordyceps mycelium.
The present invention can by the control condition of culture, obtain the cordyceps mycelia of certain selenium enrichment; And the prepared selenium-rich cordyceps mycelium of the present invention has good biologic activity.
Compared with prior art, the inventive method reaction condition is gentle, production cost is low, equipment investment is few, is easy to realize industrialization, and the mycelium of acquisition has stronger biologic activity, thereby is expected to make a breakthrough in prevention and the treatment of Animal diseases.
(4) embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1:
Starting strain: No. 1, bacterial strain Cordyceps sinensis is available from the honest bacterium industry of Shandong Liangshan County Science and Technology Development Center.
Seed culture: the mother on the slant medium for preparing plants with the inoculation shovel and gets approximately 20 mm
2Lawn access seed culture medium (sucrose 50g/L, peptone 10g/L, the yeast extract 10g/L of size, solvent is water), sample-loading amount is in the triangular flask of 100mL/250mL, 25 ℃, the top fermentation of 160r/min constant temperature oscillator 3 days, forms small bacterium ball in the seed liquor.
Optimize the concentration of brewer's grains in the fermentation medium, select respectively 10g/L, 20g/L, 30g/L, 40g/L and 50g/L add 25mg/L Na
2SeO
3The pH value is adjusted into 6, and the seed liquor inoculum concentration is 10%, is 22 ℃ in temperature, the shaking flask rotating speed is under the 160r/min condition, cultivated 6 days, zymotic fluid filters through 60 mesh sieves, and repeatedly washes three times, refilter, drying namely gets the selenium-rich cordyceps mycelium, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 2:
Take Cordyceps sinensis as starting strain, the concentration of brewer's grains is 30 g/L, selects respectively 10 mg/L, 15 mg/L, 20 mg/L, the Na of 25 mg/L and 30 mg/L
2SeO
3, other conditions namely get the selenium-rich cordyceps mycelium with embodiment 1, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 3:
Take Cordyceps sinensis as starting strain, the Na of 25 mg/L
2SeO
3, the pH value is adjusted into 4,5, and 6,7 and 8, other conditions namely get the selenium-rich cordyceps mycelium with embodiment 2, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 4:
Take Cordyceps sinensis as starting strain, the pH value is adjusted into 6, and inoculum concentration is respectively 5%, 8%, and 10%, 12% and 15%, other conditions namely get the selenium-rich cordyceps mycelium with embodiment 3, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 5:
Take Cordyceps sinensis as starting strain, inoculum concentration is 10%, is respectively 20 ℃ in temperature, and 22 ℃, 25 ℃, 28 ℃ and 30 ℃, other conditions namely get the selenium-rich cordyceps mycelium with embodiment 4, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 6:
Take Cordyceps sinensis as starting strain, temperature is 22 ℃, and the shaking flask rotating speed is respectively 120,140,160,180 and the 200r/min condition under, other conditions namely get the selenium-rich cordyceps mycelium with embodiment 5, the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 7:
Take Cordyceps sinensis as starting strain, the shaking flask rotating speed is under the 160r/min condition, cultivates respectively 4,5,6,7 and 8 days, and other conditions namely get the selenium-rich cordyceps mycelium with embodiment 6, and the Organic Selenium amount the results are shown in following table in the gained mycelia scale of construction and the mycelium.
Embodiment 8:
The cordyceps mycelia of gained is pulverized, is crossed 60 mesh sieves, with 15 times of 100 ℃ of 3%(w/w) ammonium oxalate solution extracts 2 h, and 4000 leave heart 20min, supernatant concentration.Be made into respectively 50,100,200,300, the extract of 400 and 500 μ g/mL stimulates mouse spleen lymphocyte, cell appreciation rate such as following table.
Claims (3)
1. the preparation method of a rich selenium aweto mycelium, described method comprises: aweto strain is seeded in the liquid fermentation medium that brewer's grains are matrix, cultivated 4 ~ 8 days under 20 ~ 30 ℃, 120 ~ 200r/min condition, in zymotic fluid, obtain described selenium-rich cordyceps mycelium; Described liquid fermentation medium is prepared by following composition: brewer's grains 10 ~ 50g/L, Na
2SeO
310 ~ 30mg/L, solvent are water, and pH 4 ~ 8.
2. the method for claim 1 is characterized in that: described liquid fermentation medium is by following composition preparation: brewer's grains 20 ~ 40g/L, Na
2SeO
315 ~ 25mg/L, solvent are water, and pH 5 ~ 8.
3. the method for claim 1 is characterized in that: described fermentation is carried out under 20 ~ 28 ℃, 120 ~ 180 r/min conditions.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263601A (en) * | 2014-10-11 | 2015-01-07 | 哈尔滨艾博雅食品科技开发有限公司 | Manufacturing method of cordyceps militaris juice wine |
| CN104381615A (en) * | 2014-12-12 | 2015-03-04 | 福建农大科技开发总公司 | Method for producing cordyceps sinensis feed additive and product |
| CN105441338A (en) * | 2015-12-31 | 2016-03-30 | 浙江工业大学 | Cordyceps flower production method |
| CN105505795A (en) * | 2015-12-31 | 2016-04-20 | 浙江工业大学 | Production method of iron-rich cordyceps flower |
| CN105524843A (en) * | 2015-12-31 | 2016-04-27 | 浙江工业大学 | Selenium-rich cordyceps sinensis flower production method |
| CN107211725A (en) * | 2017-05-12 | 2017-09-29 | 江苏绿洲园艺绿化有限公司 | A kind of production method of selenium-rich cordyceps flower |
| CN111349648A (en) * | 2020-02-28 | 2020-06-30 | 浙江工业大学 | Method for introducing agrobacterium-mediated exogenous gene into cordyceps militaris cells |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263601A (en) * | 2014-10-11 | 2015-01-07 | 哈尔滨艾博雅食品科技开发有限公司 | Manufacturing method of cordyceps militaris juice wine |
| CN104381615A (en) * | 2014-12-12 | 2015-03-04 | 福建农大科技开发总公司 | Method for producing cordyceps sinensis feed additive and product |
| CN104381615B (en) * | 2014-12-12 | 2018-01-02 | 福建农林大学 | Produce the method and product of aweto feed addictive |
| CN105441338A (en) * | 2015-12-31 | 2016-03-30 | 浙江工业大学 | Cordyceps flower production method |
| CN105505795A (en) * | 2015-12-31 | 2016-04-20 | 浙江工业大学 | Production method of iron-rich cordyceps flower |
| CN105524843A (en) * | 2015-12-31 | 2016-04-27 | 浙江工业大学 | Selenium-rich cordyceps sinensis flower production method |
| CN107211725A (en) * | 2017-05-12 | 2017-09-29 | 江苏绿洲园艺绿化有限公司 | A kind of production method of selenium-rich cordyceps flower |
| CN111349648A (en) * | 2020-02-28 | 2020-06-30 | 浙江工业大学 | Method for introducing agrobacterium-mediated exogenous gene into cordyceps militaris cells |
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Application publication date: 20130501 |