CN1372002A

CN1372002A - Method for producing cordycepin powder and active immne cordyceps sinensis polysaccharide frozen dried powder and products thereof

Info

Publication number: CN1372002A
Application number: CN 01106767
Authority: CN
Inventors: 孙悦迎; 陈瑞真
Original assignee: Individual
Current assignee: Individual
Priority date: 2001-02-26
Filing date: 2001-02-26
Publication date: 2002-10-02

Abstract

The present invention adopts modern tissue cell culture technology to separate and obtain agamotype cell of cordyceps sinensis for sexual carposporophyte of natural cordyceps sinensis, makes purification and naturalization, and utilizes the processes of autoamtic control fermentation, spray-drying and cell-breaking, etc. to produce cordyceps sinensis fungus pwoder; then uses the cordyceps sinensis fungus powder as raw material and adopts composite enzyme extraction method and adsorption method to prepare active immune cordyceps sinenssis polysaccharide freeze-dried powder.

Description

The production method and the product of Cordyceps fungus powder and active immne Cordyceps polysaccharide lyophilized powder

Content of the present invention belongs to biochemical article preparing technical field, relates to a kind of method and product that utilizes method that Cordyceps strain prepares Cordyceps powder and product and the above bacterium powder raw material of a kind of basis to extract Chinese caterpillar fungus immunocompetence polysaccharide.

Cordyceps sinensis (Cordyceps sinensis) is a kind of famous and precious edible and medicinal fungi that is distributed in ground high mountain such as China Sichuan, Tibet, Qinghai, Yunnan, Gansu and subalpine meadow area, belongs to Ascomycetes, Clavicipitaceae.Because compositions such as contained cordycepic acid and cordycepin have expansion tracheae, calmness, anti-various bacteria and effect such as hypotensive in the Cordyceps sinensis, be used as medicine with exsiccant stroma and polypide on the traditional Chinese medicine, can be used for treatment and consider various disease conditions such as impairment and cough phlegm blood, asthma, pain in the back, pulmonary tuberculosis, chronic nephritis, diabetes, tumour, seminal emission impotence, spontaneous sweating, insomnia forgetfulness, Chinese caterpillar fungus itself also contains multiple human body essential amino acid and the necessary vitamins B of human body hematopoiesis in addition ₁₂, thereby it has splendid medical treatment and nourishing function, and in recent years, society's being in great demand to it.In view of the residing growing environment of wild cordyceps is high and cold particular surroundings; its host bat moth needs just can finish epoch more than 3 years; cause the worm grass resources deficiency of natural propagation; produce for contradiction originally very outstanding; artificial in recent years in addition excessively digging; ecotope is seriously damaged, and some geographic Cordyceps sinensis have been on the verge of exhaustion, and setting up effective sfgd. also has been very urgent.Therefore; this area demands seeking out the anamorph cell paste and products thereof of the Cordyceps sinensis of novel Tong Guo suitability for industrialized production urgently; make it become the surrogate of natural cordyceps sporophore; to alleviate the imbalance between supply and demand of Chinese medicinal materials; preserve the ecological environment, also enter the world market simultaneously and produce road of high value added product wound for China's herbal medicine.

One of purpose of the present invention is at the prior art present situation, and a kind of Cordyceps fungus powder and production method thereof of alternative natural cordyceps is provided; It is that raw material extracts kind of the production method and the product of active immne Cordyceps polysaccharide lyophilized powder that another object of the present invention is to provide more than one to state the Cordyceps fungus powder product.

Be used to realize that the whole design idea of foregoing invention purpose is such: the feature of dividing teleomorph and anamorph according to the life history of Cordyceps sinensis, adopt modern histocyte culture technique, separate the anamorph cell that obtains Cordyceps sinensis from the sexual sporophore of natural cordyceps, after it is carried out purifying and instructionization, utilize the automatic operations such as fermentation method and spraying drying, cytoclasis of controlling to produce Cordyceps fungus powder; Then be that raw material is prepared active immne Cordyceps polysaccharide lyophilized powder by operations such as prozyme extraction method and absorption methods with the Cordyceps fungus powder.

Production method of the present invention and product comprise according to natural cordyceps sporophore pure strain and prepare the production method of Cordyceps fungus powder and product and be two kinds of the production method of feedstock production active immne Cordyceps polysaccharide lyophilized powder and products with the Cordyceps fungus powder.

The method of production Cordyceps fungus powder of the present invention is made of following production process:

(1), the acquisition of good pure bacterial strain bacterial classification: the fresh individuality that selects the natural cordyceps sporophore, from bacterial strain, filter out the bacterial strain of 3～8% relatively stable and better performances with the primary dcreening operation of routine and multiple sieve method, obtain Chinese synnema spore (synnematum sinense Yines sp new) bacterial strain bacterial classification by isolating methods of conventional microflora such as separation, mutagenesis, rejuvenation and contrasts successively then;

(2), use the fermentative Production winter worm summer herb thallus: comprise two kinds of solid fermentation method and solution fermentations, wherein:

A, solid fermentation working system---making the bacterial classification that obtains through operation () is to inoculate on 6.8～7.2 the comprehensive ADP substratum at pH value, and makes it to make solid fermentation with substratum and cultivated 15～20 days, and culture temperature is 16～25 ℃; Substratum is separated with the growth thalline, and isolated thalline is carried out centrifugal cryodrying, obtain dry thalline;

B, liquid fermenting working system---will insert pH value through the bacterial classification that operation () obtains is to inoculate in 6.8～7.2 the comprehensive ADP liquid medium jar fermenter, and makes it to do submerged fermentation cultivated 15～20 days in liquid medium, and culture temperature is 16～25 ℃; Thereafter the method by evaporation concentration and Plate Filtration is separated the thalline after liquid medium and the growth, and isolated thalline is carried out spraying drying, obtains dry thalline;

(3), ultrafine powder is produced in pulverizing: will make the ultrafine powder of granularity for the Cordyceps fungus powder of-300 orders (particle diameter is 4～10 microns) through the dry thalline that operation (two) obtains with vibromill;

(4), end properties detects, packs, puts in storage.

The product of Cordyceps fungus powder of the present invention is the product of cell wall breaking rate＞95% that makes through above-mentioned operation ()～(four).Because cell and micro mist are the bases that pharmaceuticals improve drug effect, in, the main pharmacodynamics composition of Western medicine is unobstructed all is distributed in the cell and in the intercellular substance, and in cell, behind cell wall breaking, effective constituent comes out in its cell, the cell wall breaking rate is high more, and the drug effect of product is just high more, and onset speed is also fast more.The determination and analysis result who bacterium powder finished product is done through units such as Shaanxi Province's examination and test of products institute, Shaanxi Province health and epidemic prevention station, the Shaanxi Province academy of traditional Chinese medicine, the attached medical colleges in Shanghai Communications University, Xi'an shows: the content of this product inner nucleotide class material, N.F,USP MANNITOL, active immne Cordyceps polysaccharide etc. all is greater than or equal to the content of natural cordyceps, it also contains each seed amino acid, trace element and organoselenium etc. in addition, its sanitary index meets national standard, and antitumous effect is obvious.Thisly not only reach the performance that can substitute natural cordyceps from chemical constitution, nutrition, pharmacological action by the Cordyceps fungus powder that manually makes, and the tissue morphology of its product also reached the peak demand of international pharmaceutical industry to Chinese medicine, but the product one-time-reach-place is directly integrated with the world market.

The finished product of the present invention are to utilize Cordyceps fungus powder to extract immunocompetence polysaccharide and other product.In recent years, a lot of in this area about fungi immunocompetence STUDY ON POLYSACHAROSE report, but product seldom, and its major cause is exactly to extract the cost height of polysaccharide production technique (as hot water extraction method, organic solvent extraction method, conventional enzyme extraction method etc.), active low at present.The present invention is directed to the defective of prior art, adopt the extracting method of combined-enzyme method as Cordyceps polysaccharide, this method has overcome prior art and has destroyed the three-dimensional arrangement of bioactive molecules and reduce shortcoming such as biological activity ability in leaching process, improved extraction yield, obtain the high Cordyceps polysaccharide of immunocompetence, also reduced production cost simultaneously.Operational path by Cordyceps fungus powder production immunocompetence Cordyceps polysaccharide is: be lower than utilize under 40 ℃ of conditions proteolytic ferment, amylolytic enzyme and cellulolytic enzyme respectively with the Cordyceps fungus powder effect, extract polysaccharide component wherein; Obtaining content by the frozen drying method again is 75～85% polysaccharide raw product; Then utilize proteolytic ferment, amylolytic enzyme and cellulolytic enzyme to extract again with the raw product effect respectively again, and utilize absorption method that extract is again decoloured and handle the back and obtain content at the refinishing polyose product more than 95%, again with after the freeze-drying of refinishing polyose product, obtain active immne Cordyceps polysaccharide lyophilized powder at last.

Compare with traditional extraction polysaccharide method, utilize reaction conditions gentleness, the enzyme reaction character of combined-enzyme method extraction polysaccharide single-minded, under general condition, can not destroy or influence 1 of the three-dimensional arrangement of polysaccharide and macromolecular main chain structure, the poly-hydroxy group that anti-tumor activity is played an important role on the 3-glycosidic link skeleton, thus reach the impregnable purpose of protection active group physiological activity; In addition, the reaction times of prozyme extraction method weak point, enzyme source can utilize saccharifying enzyme production, and it is simple for process, cost is low, can produce at any time as required, have overcome difficult extraction in general enzyme source and the costly problem in enzyme source.The Cordyceps polysaccharide that obtains according to aforesaid method has the particularly immune antitumor action of pharmacological action widely, its anti-tumor activity mainly relies on host's reaction, that is to say that it produces antibody by activation phagocytic cell and stimulation and reaches the physiological function that improves the human immunological competence, after antibody produces, just show as intensive anti-tumor activity and the restraint very strong to cancer cells, add the effect of cordycepin and organoselenium, its drug effect is relatively extensively also practical, thereby we can say that it is prevention and the functional matrix for the treatment of all diseases.

The production technique of product of the present invention is carried out biotechnology production fully; do not adopt any chemical substance; the starting material of using also all derive from agricultural byproducts; do not produce any source of pollution; the product accessory that obtains has the rare Chinese medicine material of high added value and advanced rare biochemical special effect medicine raw material both at home and abroad; with high content of technology, competitive power is strong, market outlook good; the practicing of its technical scheme can effectively be alleviated the contradiction of protection environment and human's demand, and realizes that manually instruction and batch production production surrogate have been founded precedent for rare, natural crude drugs for China.

Claims

1, a kind of method that is used to produce Cordyceps fungus powder is characterized in that comprising following production process:

1.1, the acquisition of good pure bacterial strain bacterial classification: the fresh individuality that selects the natural cordyceps sporophore, from bacterial strain, filter out the bacterial strain of 3～8% relatively stable and better performances with the primary dcreening operation of routine and multiple sieve method, obtain Chinese synnema spore bacterial strain bacterial classification by isolating methods of conventional microflora such as separation, mutagenesis, rejuvenation and contrasts successively then;

1.2, use the fermentative Production winter worm summer herb thallus: comprise two kinds of solid fermentation method and solution fermentations:

1.2a, the solid fermentation working system---making the bacterial classification that obtains through 1.1 operations is to inoculate on 6.8～7.2 the comprehensive ADP substratum at pH value, and makes it to make solid fermentation with substratum and cultivates 15～20 days, culture temperature is 16～25 ℃; Substratum is separated with the growth thalline, and isolated thalline is carried out centrifugal cryodrying, obtain dry thalline;

1.2b, the liquid fermenting working system---will insert pH value through the bacterial classification that 1.1 operations obtain is to inoculate in 6.8～7.2 the comprehensive ADP liquid medium jar fermenter, and makes it to do submerged fermentation cultivated 15～20 days in liquid medium, culture temperature is 16～25 ℃; Thereafter the method by evaporation concentration and Plate Filtration is separated the thalline after liquid medium and the growth, and isolated thalline is carried out spraying drying, obtains dry thalline;

1.3, pulverize and to produce ultrafine powder: will make granularity through the dry thalline that 1.2 operations obtain with vibromill is-300 purpose ultrafine powders.

2, a kind of Cordyceps fungus powder is characterized in that the product of said bacterium powder for cell wall breaking rate＞95% that makes according to the described production method of claim 1.

3, a kind of method of extracting active immne Cordyceps polysaccharide lyophilized powder according to the described Cordyceps fungus powder of claim 2, it is characterized in that be lower than utilize under 40 ℃ of conditions proteolytic ferment, amylolytic enzyme and cellulolytic enzyme respectively with the Cordyceps fungus powder effect, extract polysaccharide component wherein; Obtaining content by the frozen drying method again is 75～85% polysaccharide raw product; Then utilize proteolytic ferment, amylolytic enzyme and cellulolytic enzyme to extract again with the raw product effect respectively again, and utilize absorption method that extract is again decoloured and handle the back and obtain content at the refinishing polyose product more than 95%, again with after the freeze-drying of refinishing polyose product, obtain active immne Cordyceps polysaccharide lyophilized powder at last.

4, a kind of active immne Cordyceps polysaccharide lyophilized powder is characterized in that the product of said lyophilized powder for making according to claim 1 and 2 described production methods.