CN1149277C - Spherical thallus of cordyceps - Google Patents
Spherical thallus of cordyceps Download PDFInfo
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- CN1149277C CN1149277C CNB991178939A CN99117893A CN1149277C CN 1149277 C CN1149277 C CN 1149277C CN B991178939 A CNB991178939 A CN B991178939A CN 99117893 A CN99117893 A CN 99117893A CN 1149277 C CN1149277 C CN 1149277C
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- cordyceps
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Abstract
The present invention provides a pure spherical thallus of cordyceps. The bacterial strains of cordyceps can be cultivated on a flat plate, eluted with aseptic water and stirred to be prepared into bacterial liquid, the bacterial liquid is cultivated by liquid culture of a plurality of grades, and then, the pure spherical thallus of cordyceps is prepared. After the liquid culture process of each grade is finished, the bacterial liquid is sieved by a grading sieve. The spherical thallus of cordyceps is characterized in that the spherical thallus is hollow, and protrusions or hairy objects exist on the surface of the spherical thallus. The pure spherical thallus of cordyceps can be directly used for the technical fields of medicines, beverages, food, cosmetics, etc.
Description
(1) technical field: the present invention relates to a kind ofly utilize microorganism culturing and prepare the method for new organism, specifically be utilize Cordyceps strain through flat board cultivate, some grades liquid culture and make spherical thallus, this product can directly or be used for fields such as medicine, drink, food and makeup after the drying.
(2) background technology: in recent years, it is comparatively extensive as the application of health care aspect that Cordyceps sinensis is made various mixed preparations.Existing Cordyceps sinensis deep layer fermenting process exists a difficult point, i.e. the isolating problem of Cordyceps mycelium and substratum.Because the mycelium and the spore of Cordyceps sinensis are very trickle, the substratum particle is bigger than mycelium mostly, and by the physical method press filtration, if the aperture is excessive, then thalline runs off; The aperture is little, and then impurity can't be removed.Adopt centrifugal separation method invalid too.Because of the high compositions close such as protein, amino acid that contain in the impurity, adopt chemical process not only Impurity removal can not have been increased pollution on the contrary with thalline.Owing to be mixed with a large amount of substratum (peanut powder, yeast powder etc.) in the mycelium, seriously restricted mycelial purity.The present inventor utilizes Cordyceps strain, makes pure spherical thallus of cordyceps, can be widely used in fields such as medicine, healthcare products, food and makeup.
(3) summary of the invention: task of the present invention provides a kind of Cordyceps strain that utilizes, and cultivates, after some grades of liquid culture, makes the method for pure spherical thallus of cordyceps through flat board.The spherical thallus of method for preparing can directly apply to fields such as medicine, drink, food and makeup, also can be used for above-mentioned field after the drying.
Cordyceps fungus [Cordyceps sinensis (Berk.) Sacc.], another name: Chinese caterpillar fungus, Cordyceps, Cordyceps sinensis (referring to " Chinese medicinal fungi " the 15th page).The Cordyceps strain that the present invention uses is bought (sealing 4 advertisements referring to the 5th phase of " edible mushrooms " nineteen ninety-five) from academy of agricultural sciences, Shanghai edible mushrooms institute strain plant.
The morphological specificity of Cordyceps strain:
25 ℃～28 ℃ cultivations on nutrient agar, the mycelium shape that obtains, or the shape that presents of spore: bacterium colony circle, neat in edge, smooth, tight, white, fine and soft shape, growth retardation, colony diameter 15mm, bacterium colony is not wet, and rope-like constructed is arranged, no fan-shaped sudden change angle, the greenish orange look of reverse side does not have the pigment that is spread in substratum.
Mycelia is transparent, has every, branch, and diameter 1.5 μ m～2 μ m, the content homogeneous of hyphal cell is the ghost shape after a while, no chlamydospore.Mycelia can be tied when condition is suitable and be gathered into rope form, but in most cases is independent mycelia, does not form rope bundle structure.Conidiophore is transparent, directly or curved slightly, single giving birth to or 2～3 consor, but be not colyliform, end or cardinal extremity do not have obviously and expand (15～40 * 1.5～2) μ m.Conidial head is false head, and diameter 20 μ m * 35 μ m are formed by about 20 conidium adhesive aggregations, and conidium is transparent, and is unicellular, ellipse, and quarter butt shape, lanceolar, straight, wall is thin, the blunt circle in two ends, (2.5～6 * 1.5～1.8) μ m.
The cultivation of Cordyceps strain does not have special regulation to the nutrition source in the substratum, can make to contain carbon source, nitrogenous source and other nutrition source that is usually used in microorganism culturing in the substratum.Wherein nitrogenous source is peptone, urea etc., and carbon source can be than good utilisation glucose, sucrose, wood sugar etc.
For culture condition such as culture temperature, times, to be suitable for the using growth of bacterium to be as the criterion, the pH value scope of substratum is 4.5～6.5, and culture temperature should be at 25 ℃～28 ℃.
The method for preparing spherical thallus of cordyceps of the present invention, step is as follows: 1. Cordyceps strain is cultivated at the nutrient agar upper flat plate, 25 ℃～28 ℃ of culture temperature after waiting to cover with mycelia, are used the sterilized water wash-out, stir then, make suspension; 2. above-mentioned bacterium liquid adding is filled shaking in the bottle of liquid nutrient medium, the rotary cultivation of constant temperature 3 days～4 days, form tiny spherical thallus, culture condition is 25 ℃～28 ℃ of temperature, 50 rev/mins～250 rev/mins of rotating speeds, the liquid nutrient medium that uses is formed and weight percent is: peanut 2%～8%, corn 2%～8%, peptone 0.1%～0.4%, sugar 1.5%～4%, potassium primary phosphate 0.25%～0.75%, sal epsom 0.05%～0.25%, and all the other are water; 3. with the classified screen sizing of above-mentioned coccoid thalline, discard unsetting thing, the fresh liquid nutrient medium that adds by above-mentioned formulated in the container of bigger rotatable cultivation carries out the secondary liquid culture then, the rotary cultivation of constant temperature 1 day～3 days, culture condition is 25 ℃～27 ℃ of temperature, 50 rev/mins～250 rev/mins of rotating speeds, when treating that coccoid thalline diameter increases to 2 times～20 times, change over to again in the bigger container that fills by the rotatable cultivation of the fresh liquid nutrient medium of above-mentioned formulated, press said temperature, rotating speed continues to cultivate, and reaches requirement up to the spherical thallus diameter; 4. the spherical thallus that will 3. go on foot acquisition obtains the spherical thallus of certain specification by the sizing screen sub-sieve, uses sterilized water wash-out substratum again.
Peanut, corn during liquid nutrient medium of the present invention is formed directly decoct nutritive substance and leach but the particle unshaping after cleaning, filter then.
The present invention compared with prior art has substantive distinguishing features and obvious improvement:
That adopts spherical thallus of cordyceps that the inventive method obtains is characterized as spheroid hollow, and spherome surface has projection or hair, and its form is as pearl spherical thallus of a size; Adopt rotary liquid shake cultivation, substratum adopts the full water-soluble material that does not contain vegetable particle, and the spherical thallus of cordyceps diameter of classified cultivation is not less than 2mm, and is bigger more than 1000 times than the mycelium diameter of producing with deep layer fermenting process; Through sterilized water wash-out substratum, thalline is pure, do not have any impurity, solved the Cordyceps powder decon problem that perplexs enterprise for a long time; The Chinese caterpillar fungus spherical thallus that product of the present invention is pure can be widely used in industrial circles such as pharmacy, makeup, food.
(4) embodiment:
The invention will be further described below by specific embodiment:
Embodiment 1 prepares the method for spherical thallus of cordyceps
With potato, glucose, agar, water in 200: 20: 20: 1000 ratio is made dull and stereotyped nutrient agar, and the pH value is 6.5, and is standby after sterilizing.
Add water with the composition of peanut 6%, corn 2%, peptone 0.1%, sugar 1.5%, potassium primary phosphate 0.25%, sal epsom 0.05% and ratio and make liquid nutrient medium, the pH value is 5.5, be sub-packed in respectively in some 250ml triangular flasks, the rotatable container of 1000ml and the rotatable container of 5000ml, standby after sterilizing.
Cordyceps strain is inoculated in this nutrient agar cultivates, 28 ℃ of culture temperature, treat that mycelia is covered with after, use the sterilized water wash-out, use magnetic stirrer, make bacterium evenly be suspended in the liquid; Cultivated 3 days carrying out the rotary liquid one-level of constant temperature in the above-mentioned bacterium liquid adding 250ml triangular flask, 28 ℃ of culture temperature, the bacterium liquid in 100 rev/mins of the rotating speeds, 250ml triangular flask forms tiny spherical thallus; The sizing screen of tiny spherical thallus by certain pore size carried out classification, discard amorphous thalline;
Cultivate carrying out the liquid secondary in the coccoid thalline adding 1000ml container after the above-mentioned classification, the rotary cultivation of constant temperature 3 days, 27 ℃ of temperature, 150 rev/mins of rotating speeds, coccoid thalline diameter increases to 20 times; Sizing screen with certain pore size carries out classification;
The spherical thallus that above-mentioned classification is qualified adds in the container of 5000ml, presses said temperature, rotating speed continuation cultivation; Reach requirement up to the spherical thallus diameter, use sterilized water wash-out spherical thallus substratum on every side then, promptly obtain pure spherical thallus of cordyceps, it is characterized by spheroid hollow, spherome surface has projection or hair.
Each level will contain the sizing screen of the cultivation liquid of spherical thallus by certain pore size after cultivating and finishing, and with large, medium and small spherical thallus separately, split cultivation, until reaching the required diameter specification.
By the requirement of product spherical thallus diameter, decision splits and changes the number of times of culture medium culturing.
Present embodiment obtains the spherical thallus of diameter 200mm specification, and through three grades of liquid culture, incubation time 11 days can obtain spherical thallus dry weight 10g.
Embodiment 2 is applied to food with spherical thallus
With the above-mentioned spherical thallus that after flat board cultivation, liquid culture, makes, directly make the high-grade nourishing drink that suspends.Ratio in 2.5%～5.0% adds in the beverage, adjusts the concentration 4.0%～8.0% of sugar, makes the proportion of beverage that spherical thallus is suspended among the liquid, through pasteurization.
Embodiment 3 is applied to makeup with spherical thallus
Utilize the characteristics of spherical thallus hollow, the ratio in 1%～1.5% adds in the face nursing liquid.The effective ingredient of thalline beauty treatment is infiltrated in the face nursing liquid.Directly smearing face with face nursing liquid or spheroid during use gets final product.
Embodiment 4 is applied to medicine with spherical thallus
Because spherical thallus is extremely pure, the content of its effective constituent Chinese caterpillar fungus gland glucoside is 4 times of conventional zymophyte powder, so Cordyceps fungus powder capsule volume reduces to the 0.25g/ grain by the 0.5g/ grain, and dose reduces to 1/time by 2/time simultaneously.
Claims (2)
1, a kind of method for preparing spherical thallus of cordyceps, step is as follows:
1. Cordyceps strain is cultivated at the nutrient agar upper flat plate, 25 ℃～28 ℃ of culture temperature use the sterilized water wash-out after covering with mycelia, and suspension is made in stirring;
2. above-mentioned bacterium liquid adding is filled shaking in the bottle of liquid nutrient medium, the rotary cultivation of constant temperature 3 days～4 days forms tiny spherical thallus, 25 ℃～28 ℃ of culture temperature, 50 rev/mins～250 rev/mins of rotating speeds; The liquid nutrient medium that uses is formed and weight percent is: peanut 2%～8%, corn 2%～8%, peptone 0.1%～0.4%, sugar 1.5%～4%, potassium primary phosphate 0.25%～0.75%, sal epsom 0.05%～0.25%, and all the other are water;
3. above-mentioned coccoid thalline is carried out sub-sieve by sizing screen, coccoid thalline behind the sub-sieve carries out secondary and cultivates in the fresh liquid nutrient medium by above-mentioned formulated, the rotary cultivation of constant temperature 1 day～3 days, 25 ℃～27 ℃ of culture temperature, 50 rev/mins～250 rev/mins of rotating speeds; When treating that coccoid thalline diameter increases to 2 times～20 times, change in the bigger container that fills by the fresh liquid nutrient medium of above-mentioned formulated, press said temperature, rotating speed continues to cultivate;
4. the spherical thallus that will 3. go on foot acquisition obtains the spherical thallus of certain specification by the sizing screen sub-sieve, with sterilized water wash-out substratum.
2, the method for preparing spherical thallus of cordyceps according to claim 1 is characterized in that peanut, the corn in the described liquid nutrient medium composition do not pulverized, and directly decocts nutritive substance and leach but the particle unshaping after cleaning, and filters then.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991178939A CN1149277C (en) | 1999-10-15 | 1999-10-15 | Spherical thallus of cordyceps |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991178939A CN1149277C (en) | 1999-10-15 | 1999-10-15 | Spherical thallus of cordyceps |
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| Publication Number | Publication Date |
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| CN1293244A CN1293244A (en) | 2001-05-02 |
| CN1149277C true CN1149277C (en) | 2004-05-12 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB991178939A Expired - Fee Related CN1149277C (en) | 1999-10-15 | 1999-10-15 | Spherical thallus of cordyceps |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1323157C (en) * | 2005-09-23 | 2007-06-27 | 田向荣 | Chinese caterpillar fungus bionic culturing method |
| CN103229666B (en) * | 2013-04-27 | 2014-09-10 | 青岛农业大学 | Cordyceps militaris peanut and preparing method thereof |
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1999
- 1999-10-15 CN CNB991178939A patent/CN1149277C/en not_active Expired - Fee Related
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