CN104686204A - Cordyceps sinensis naked barley - Google Patents

Cordyceps sinensis naked barley Download PDF

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Publication number
CN104686204A
CN104686204A CN201510124844.1A CN201510124844A CN104686204A CN 104686204 A CN104686204 A CN 104686204A CN 201510124844 A CN201510124844 A CN 201510124844A CN 104686204 A CN104686204 A CN 104686204A
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China
Prior art keywords
naked barley
cordyceps
cordyceps sinensis
cordyceps militaris
barley
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CN201510124844.1A
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Chinese (zh)
Inventor
蔡宇杰
杨帆
白玉
管政兵
廖祥儒
张大兵
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Jiangnan University
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Jiangnan University
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Priority to CN201510124844.1A priority Critical patent/CN104686204A/en
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms

Abstract

Cordyceps sinensis naked barley takes naked barley as a culture medium; cordyceps sinensis and cordyceps militaris are subjected to double-bacteria mixed solid-state fermentation, and the cordyceps sinensis naked barley belongs to the technical field of biological engineering. The naked barley obtained by double-bacteria bioconversion has life cultivation and health preservation effects and has a good application prospect.

Description

Chinese caterpillar fungus naked barley
Technical field
Adopting Cordyceps sinensis (Ophiocordyceps sinensis) and Cordyceps militaris (Cordyceps militaris), take naked barley as medium, carries out two bacterium mixed solid fermentation, belongs to technical field of bioengineering.Utilizing bacterial strain Cordyceps militaris preserving number to be CICC 14013, bacterial strain Ophiocordyceps sinensis preserving number is CICC 14088, is produced obtain nutrition and health care naked barley by two bacterium mixed solid fermentation.
Background technology
Cordyceps (Cordyceps) is under the jurisdiction of mycota (Fungi), Ascomycota (Ascomycota), Ascomycetes (Ascomycetes), excrement shell bacterium subclass (Sordariomycetidae), Hypocreales (Hypocreales), Clavicipitaceae (Clavicipitaceae).Cordyceps sinensis fungus great majority are medicinal fungis, wherein Cordyceps sinensis (Ophiocordyceps sinensis) is generally well known, applicating history is also remote, and the application study of Cordyceps militaris (Cordyceps militaris) is the most deep.
Cordyceps sinensis is recorded in 710 years Christian eras " moon king medicine is examined " the earliest according to textual research, records first appeared in Qing Dynasty Wu Yi Lip river " new compilation of materia medica " (1757 Christian eras) with medicine, thinks its " sweet flat; to protect lung kidney-nourishing; hemostasis and phlegm, phthisical cough, control diaphragm disease all good ".Cordyceps sinensis is China's tradition rare Chinese medicine, and it and ginseng, pilose antler are described as Chinese medicine Triratna together.It has mend and not high, temperature and not fiery, grow and oiliness pharmacodynamic profile, have more negative and positive with the unique effects of mending, make it become nourishing panacea, be described as " kings of hundred medicines " by physician.
Cordyceps militaris is one of Chinese medicinal material of being of great rarity of China, and its medical value is on the books very early, and Li Shizhen (1518-1593 A.D.) (1518) is pointed out in Compendium of Material Medica, and cicada fungus can cure mainly " children's hangs in sky frightened epilepsy, the morbid night crying of babies, palpitaition ".Cordyceps militaris is easy to large-scale artificial and cultivates, and its active component and pharmacological action are similar to Cordyceps sinensis again.
The current grain raw material that Cordyceps sinensis or Cordyceps militaris is applied to ferments by wide coverage.Such as: patent 200610117997.4 adopts Cordyceps militaris industrialization to cultivate discarded object and makes rice flour product, Chinese caterpillar fungus is pulverized rear mixing with the careless highland barley fried and makes health products by patent 200710050135.9, patent 200710166278.6 adopts wheat to cultivate fruiting bodies of cordyceps militaris, patent 200810068734.8 adopts adopting red yeast rice synergistic to ferment thus improves the output of fruiting bodies of cordyceps militaris and cordycepin, patent 200910218363.1 discloses a kind of method being improved primary medicinal component by Poria cocos and Cordyceps militaris solid co-fermentation, patent 201110055425.9 utilizes solid state fermentation to produce Cordyceps militaris fermentation rice, patent 201110348239.4 take lotus seeds as matrix, carries out the solid state fermentation of Cordyceps militaris or winter grass summer grass, patent 201110064459.4 discloses a kind of preparation method of Cordyceps militaris paddy food, patent 201310150267.4 and 201310151228.6 respectively with soya bean and peanut for matrix, carry out solid state fermentation after inoculation Cordyceps militaris, but need in process to add insect enzymolysis liquid, cause cost and difficult quality to control, patent 201210064068.7 discloses a kind of with glossy ganoderma, Phellinus etc. can the mushroom entity of efficient cellulase-producing as medium major ingredient, grain raw material is auxiliary material, inoculate into Cordyceps militaris, Cordyceps sinensis, mushroom, Hericium erinaceus, the bacterial classifications such as acupuncture needle aunt carry out Mixed culture, make health products or additive, but this complex process, edible and medical fungi fruit body is expensive, and after autoclaving, destroy the active ingredient of edible and medical fungi fruit body, complicated component and uncontrollable after other strain fermentation again, the strain combination of various complexity also will cause the instability of end product quality.
In sum, the solid ferment process of current Cordyceps militaris or Cordyceps sinensis all can add mineral salt, insect hydrolyzate or other medicinal ingredient, to improving Chinese caterpillar fungus thalline or cordycepin output and drug effect.Different additives can cause complicated component and import external pollution source, and is not easy to set up quality index system.In addition, the solid ferment process of Cordyceps militaris often using cordycepin (Cordycepin) as important quality index.Cordycepin, is also referred to as 3'-Deoxyadenosine (3-deoxyadenosine), and since self-discovery, its pharmacological action is constantly made known.Personnel study discovery after deliberation, and cordycepin has immunological regulation, antitumor, anti-leucocyte, the multiple pharmacological effect such as antimycotic, antiviral.What is interesting is, cordycepin is mainly stored in Cordyceps militaris, and content in Cordyceps sinensis is atomic, but the disappearance of cordycepin does not affect the remarkable efficacy of Cordyceps sinensis.Therefore, these two kinds of effects of Cordyceps sinensis and Cordyceps militaris are similar, but the incomplete same bacterial classification of composition has complementarity.
Summary of the invention
Based on current situation, the present invention take naked barley as medium, when not adding other additive any, only naked barley is carried out expanding treatment, after sterilizing, Simultaneous vaccination Cordyceps sinensis and Cordyceps militaris carry out Mixed culture, finally obtain the health care naked barley of high cordycepin content and high bioactivity.Naked barley (formal name used at school: Secalecereale), also known as rye, is a kind of cereal very wide in Temperate Region in China distribution.The naked barley raw material that the present invention adopts is easy to get, and method is implemented simple, quality controllable, has wide application and development prospect.
The present invention's Cordyceps strain Ophiocordyceps sinensis preserving number used is CICC14088, and cordyceps militaris link bacterial strain Cordyceps militari preserving number is CICC 14013.
Technical scheme of the present invention is as follows:
(1) part naked barley adopts the twin-screw extruder containing three sections to process.1st section is preheating zone, and serviceability temperature is 40-60 DEG C, and the 2nd district is traffic zone, and temperature is 120-160 DEG C, and the 3rd district is slaking district, and temperature is 140-180 DEG C.The water content of expanded front naked barley is 10%-20%.
(2) part naked barley mechanical crushing is 0.1-2mm to particle diameter.
(3) mixed with certain proportion with the naked barley of mechanical crushing by the naked barley of extrusion, the naked barley proportion of extrusion is 1-100%.Keep the skin wet after mixing and make water content reach 20%-70%, 121 DEG C of insulations sterilizing in 20 minutes.After cooling, access Cordyceps sinensis, inoculum concentration is 1-10%.Meanwhile, access Cordyceps militaris, inoculum concentration is 1-10%.Initial pH and fermentation process pH is nature, and fermentation temperature is 15-28 DEG C, and envionmental humidity is 60%-100%, and fermentation period is 5-25 days.The content of final cordycepin reaches as high as 7.9%.
The Cordyceps sinensis that the present invention is used and Cordyceps militaris spawn preservation slant medium formula are: glucose 20g/l, potato juice 200g/l, agar 20g/l, pH nature.
The seed culture based formulas of Cordyceps sinensis of the present invention and Cordyceps militaris is: glucose 20g/l, potato juice 200g/l, pH nature.After slant strains is inoculated into the triangular flask that seed culture medium is housed, the incubation time of seed is 96 hours, and the cultivation temperature of seed is 25 DEG C.
Cordycepin content adopts liquid chromatography for measuring.Assay method refers to: Wei Huiping, Ye little Li, Zhang Huaying, Li Xuegang, Zhong Yunjun, with the research of Cordyceps militaris solid fermentation method High-efficient Production cordycepin, and CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1998,33 (19), 2159-2162.
Beneficial effect of the present invention: by expanded, makes naked barley starch gelatinization and naked barley protein denaturation at short notice.Expanded naked barley is fermented after mixing with mechanical crushing naked barley, the nutriment making thalline to utilize puffing process to produce fast on the one hand, on the other hand mechanical crushing naked barley is the equal of that solid state fermentation carrier makes fermentation substrate keep loose favourable thalli growth, and when thalli growth is vigorous, thalline produces a large amount of extracellular enzyme and can decompose further again and utilize mechanical crushing naked barley.This process, without the need to adding other material, ensures that the quality height of fermented product is controlled.The common fermentation of Cordyceps sinensis and Cordyceps militaris two kinds of similar bacterial classifications of character, on the one hand the tunning of similar quality bacterial classification can not change overall pharmacology pharmacy proterties, can be in harmonious proportion two kinds of strain fermentation products defect separately on the other hand.The method simple possible, is easy to accomplish scale production, is with a wide range of applications.
Embodiment
Embodiment 1
Adopt the DS56-III type twin-screw extruder process naked barley that Jinan Ju Xian plant equipment Co., Ltd produces, naked barley used is commercially available, and measuring its moisture content through moisture teller is 14.5%.Setting charging rate is 120kg/h.Table 1 lists temperature combination and the puffed degree of three sections.When the temperature in preheating zone, traffic zone, slaking district is respectively 60,160,190 DEG C, the puffed degree of naked barley is the highest as can be seen from Table 1, under this temperature condition, in naked barley, supplement a small amount of sterile water further or naked barley is dried further, investigating puffed degree during different moisture content.As can be seen from Table 2, moisture content is that expansion effect is best when about 14-18%, therefore can directly carry out expanded with commercially available naked barley.
Table 1: the different each section different temperatures combination of extruder obtains the puffed degree of expanded naked barley
Table 2: the puffed degree after different water cut naked barley is expanded
Moisture content % 10.1 12.1 14.5 16.2 17.2 18.3 19.1
Puffed degree 1.5 1.8 2.4 2.4 2.3 2.3 2.1
Embodiment 2
Be inoculated into by aweto strain and be equipped with in the 250ml triangular flask of 50ml medium, medium consists of: glucose 20g/l, and potato juice 200g/l, pH nature, 25 DEG C, shaking speed 150 turns/min, cultivates 96 hours, as inoculation seed.
Be inoculated into by Cordyceps militaris spawn and be equipped with in the 250ml triangular flask of 50ml medium, medium consists of: glucose 20g/l, and potato juice 200g/l, pH nature, 25 DEG C, shaking speed 150 turns/min, cultivates 96 hours, as inoculation seed.
Get puffed degree be 2.4 expanded naked barley and mechanical crushing degree be 1mm naked barley mixes, amount to 50g, adding distil water 40g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate after cooling into Cordyceps sinensis and each 5ml of Cordyceps militaris seed liquor, control environment humidity 90% in constant temperature and humidity incubator, temperature is 25 DEG C, ferments and measures cordycepin content after 15 days.Cordycepin output under different condition combination is as shown in table 3.When expanded naked barley and mechanical crushing naked barley ratio are 3:2, cordycepin produces the highest as can be seen from Table 3, can reach 6.5%.
Table 3: solid state fermentation cordycepin content under expanded naked barley and mechanical crushing degree different proportion
Expanded naked barley weight (g) Mechanical crushing naked barley weight (g) Cordycepin content (%)
50 0 3.0
1 49 3.3
25 25 4.7
10 40 3.5
40 10 3.2
20 30 5.4
30 20 6.5
Embodiment 3
Get puffed degree be 2.4 expanded naked barley 30g and mechanical crushing degree be 2mm naked barley 20g mixes, adding distil water 40g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate into cultured Cordyceps sinensis and each 5ml of Cordyceps militaris seed liquor after cooling, control environment humidity 80% in constant temperature and humidity incubator, and temperature is 15 DEG C, ferment 20 days afterwards measure cordycepin content be 5.3%.
Embodiment 4
Get puffed degree be 2.4 expanded naked barley 30g and mechanical crushing degree be 0.5mm naked barley 20g mixes, adding distil water 50g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate into cultured Cordyceps sinensis and each 6ml of Cordyceps militaris seed liquor after cooling, control environment humidity 90% in constant temperature and humidity incubator, and temperature is 20 DEG C, ferment 5 days afterwards measure cordycepin content be 1.9%.
Embodiment 5
Get puffed degree be 2.4 expanded naked barley 30g and mechanical crushing degree be 1mm naked barley 20g mixes, adding distil water 45g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate into cultured Cordyceps sinensis seed liquor 5ml after cooling, control environment humidity 90% in constant temperature and humidity incubator, and temperature is 20 DEG C, ferment 25 days afterwards measure cordycepin content be 0.03%.The fermentation product that this process obtains is designated as sample 1.
Get puffed degree be 2.4 expanded naked barley 30g and mechanical crushing degree be 1mm naked barley 20g mixes, adding distil water 45g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate into cultured Cordyceps militaris seed liquor 5ml after cooling, control environment humidity 90% in constant temperature and humidity incubator, and temperature is 20 DEG C, ferment 25 days afterwards measure cordycepin content be 7.8%.The fermentation product that this process obtains is designated as sample 2.
Get puffed degree be 2.4 expanded naked barley 30g and mechanical crushing degree be 1mm naked barley 20g mixes, adding distil water 40g again, put into 500ml triangular flask, 121 DEG C of sterilizing 20min, inoculate into cultured Cordyceps sinensis and Cordyceps militaris seed liquor 5ml after cooling, control environment humidity 90% in constant temperature and humidity incubator, and temperature is 20 DEG C, ferment 25 days afterwards measure cordycepin content be 7.9%.The fermentation product that this process obtains is designated as sample 3.
Get the Kunming mouse 40 purchased from Shanghai Slac Experimental Animal Co., Ltd., Individual Quality 18-22g, male and female half and half.Be divided into 4 groups at random, often organize 10, be respectively sample 1,2,3 administration group and blank group.In first 7 days of experiment, every day, gavage gave different solid state fermentation samples, and the sample size of every mouse gavage is 0.5g.Blank group gavage to be unfermentable puffed degree be 2.4 0.5g naked barley.7th day, every mouse peritoneal injected 2% starch 2ml, after 24h, every mouse peritoneal injects 2% chicken red blood cell 1ml, after 30min, broken end, Intraperitoneal injection 0.2ml heparin, get peritoneal fluid smear, Rui Shi-Ji's nurse Sa dyeing 1min, salt solution gradient washes, oily sem observation phagocytic percentage, phagocytic index, get spleen, thymus gland is weighed, calculate spleen index, thymus index.
Spleen index=spleen average quality (mg)/average body quality (g).
Thymus index=thymus gland average quality (mg)/average body quality (g).
The cell number of the quantity of phagocytic index=engulf/engulf.
The phagocyte number of the cell number of phagocytic rate=engulf/total.
Spleen index is listed, thymus index, phagocytic index and phagocytic rate in table 4.
Table 4: different fermentations sample is on the impact of mouse spleen, thymic factor D injection, Peritoneal Macrophage Phagocytosis
Group Thymus index Spleen index Phagocytic rate (%) Phagocytic index
Blank group 1.81±0.29 3.11±0.25 47.2±7.83 1.8±0.2
1 group, sample 2.33±0.42 4.21±0.31 58.4±10.2 2.1±0.3
2 groups, sample 2.24±0.34 3.83±0.26 65.3±10.2 2.2±0.3
3 groups, sample 3.31±0.35 6.08±0.45 86.5±11.1 3.2±0.2
Index and spleen index and thymus index are the important parameters directly reflecting Immunologic States, and sample 1-3 group all significantly can promote the growth of mouse spleen and thymus gland relative to blank group as can be seen from Table 4, the immune level effect of being significantly improved.The Be very effective that worm winter, worm summer and Cordyceps militaris carry out 3 groups, the sample of mixed culture fermentation is better than 1 group, the sample of Cordyceps sinensis list strain fermentation and 2 groups, the sample of the single strain fermentation of Cordyceps militaris.
1 group, sample fermenting compared to single bacterium of worm winter, worm summer and Cordyceps militaris 3 groups, sample carrying out mixed culture fermentation and 2 groups, sample significantly can promote the phagocytic function of Turnover of Mouse Peritoneal Macrophages as can be seen from Table 4, the anti-infective of body and anti-tumor capacity can be improved, strengthen the immunologic function of body, stable state of keeping fit.

Claims (2)

1. the present invention adopts naked barley to be matrix, carries out Cordyceps sinensis and Cordyceps militaris mixed solid fermentation.
2. mixed solid fermentation according to claim 1, is characterized in that part naked barley need carry out expanding treatment.
CN201510124844.1A 2015-03-20 2015-03-20 Cordyceps sinensis naked barley Pending CN104686204A (en)

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Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5526808A (en) * 1978-08-12 1980-02-26 Hokushin Kousan Yuugen Artifical cultivation of cordyceps
JPH0962A (en) * 1995-06-22 1997-01-07 Hokuto Sangyo Kk Artificial culture of plant worm
CN1243678A (en) * 1998-08-01 2000-02-09 刘瑞林 Technology for producing instant food by fungus cultivation
CN1342425A (en) * 2000-09-11 2002-04-03 张国防 Nutritive food and its production method
CN101066090A (en) * 2007-05-28 2007-11-07 李治德 Aweto activated reinforced immunological feed additive and its production process
CN102349649A (en) * 2011-10-08 2012-02-15 杨毅 Production method of novel edible and medicinal fungi solid fermented powder
CN102578559A (en) * 2012-03-07 2012-07-18 云南省农业科学院生物技术与种质资源研究所 Medical fungus health care product fermented by taking soybean curb residue as matrix and preparation method of medical fungus health care product
CN103271281A (en) * 2013-06-09 2013-09-04 食品行业生产力促进中心 Method for producing cordyceps militaris fermentation coarse cereals through solid-state fermentation
CN103751479A (en) * 2014-01-28 2014-04-30 福建华尔康生物科技有限公司 Chinese caterpillar fungus combination containing plants and fungal polysaccharide and preparation method thereof
CN104137975A (en) * 2014-06-16 2014-11-12 浙江百兴生物科技有限公司 Health mushroom fruit and vegetable enzyme and preparation method thereof

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5526808A (en) * 1978-08-12 1980-02-26 Hokushin Kousan Yuugen Artifical cultivation of cordyceps
JPH0962A (en) * 1995-06-22 1997-01-07 Hokuto Sangyo Kk Artificial culture of plant worm
CN1243678A (en) * 1998-08-01 2000-02-09 刘瑞林 Technology for producing instant food by fungus cultivation
CN1342425A (en) * 2000-09-11 2002-04-03 张国防 Nutritive food and its production method
CN101066090A (en) * 2007-05-28 2007-11-07 李治德 Aweto activated reinforced immunological feed additive and its production process
CN102349649A (en) * 2011-10-08 2012-02-15 杨毅 Production method of novel edible and medicinal fungi solid fermented powder
CN102578559A (en) * 2012-03-07 2012-07-18 云南省农业科学院生物技术与种质资源研究所 Medical fungus health care product fermented by taking soybean curb residue as matrix and preparation method of medical fungus health care product
CN103271281A (en) * 2013-06-09 2013-09-04 食品行业生产力促进中心 Method for producing cordyceps militaris fermentation coarse cereals through solid-state fermentation
CN103751479A (en) * 2014-01-28 2014-04-30 福建华尔康生物科技有限公司 Chinese caterpillar fungus combination containing plants and fungal polysaccharide and preparation method thereof
CN104137975A (en) * 2014-06-16 2014-11-12 浙江百兴生物科技有限公司 Health mushroom fruit and vegetable enzyme and preparation method thereof

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吴振强: "《固态发酵技术与应用》", 31 March 2006, 北京:化学工业出版社 *

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Application publication date: 20150610