CN102174414A - Application of new morchella costata M8-13 liquid fermentation substance to development of health-care products and medicaments - Google Patents

Abstract

The invention relates to newly separated morchella M8-13 and application thereof to development of health-care products and medicaments.

Description

A kind of new application of costae morel M8-13 liquid fermentate in healthcare products and pharmaceutical developments

Technical field

The present invention relates to a kind of new isolating morel M8-13 and the application in medicine and healthcare products exploitation thereof.

Background technology

Morel (Morchella sp.) is world-renowned (medicine) edible mushrooms, is called as in the U.S. " land fish ", and more American-European developed countries think that it is the senior tonic of human nutrition.Distribution is all arranged all over the world.Morchella esculenta (L.) Pers sporophore contains the number of chemical composition, measure according to one's analysis, contain moisture 12.869g in every 100g dry product morel, fat 3.829g, protein 22.069g, wherein protein content is 2 times of auricularia auriculajudae, 1.5 times of (Long Zhenghai etc. of mushroom, 1997), and very easily be absorbed by the body, this point is that other high-protein foods hardly match.It is abundant that morel contains the necessary aminoacids content of 10 seed amino acids, particularly human body, exceeds 25% one 40% of general edible mushrooms.VITAMIN and mineral element A wide selection of colours and designs in the morel, the content height contains vitamins B and nicotinic acid, vitamin H, adjoins the alcohol of trembling, folic acid, multiple amino acids etc.Per 100 gram morels contain mineral element potassium 2.89g, phosphorus 1.59g, and be respectively 7 times and 4 times of Cordyceps sinensis: the content of zinc is 4.3 times of mushroom, 4 times of Hericium erinaceus (Bull. Ex Fr.) Pers.; The content of iron is 31 times of mushroom, 12 times (Chen XiangDong etc., 2002) of Hericium erinaceus (Bull. Ex Fr.) Pers..The element that also contains needed by human body such as calcium, magnesium, sodium, chromium in the morel is especially with the organic germanium content height.The existence of several mineral materials makes morel seem that more the personal value is out of the ordinary.Domestic and international research mainly concentrates on polysaccharide, amino acid and fats compound, about enzyme, Pyronone antibiotic, inorganic elements, pigment, alcohol, sterol and saponins report [1-3] is arranged also in addition.Because the mycelium of morel not only contains the various nutritive substances and the activeconstituents of sporophore, and can also keep the local flavor of sporophore uniqueness preferably, and be suitable for industrial fermentation production, so Morchella esculenta (L.) Pers mycelium submerged fermentation research has obtained development rapidly in recent years, the report of development and use mycelium, fermented liquid is also a lot.Advantages such as that the submerged fermentation technology has is easy to operate, mycelium propagation is fast, with short production cycle, output is big are that main raw material extracts various nutritive ingredients of morel and activeconstituents with submerged fermentation mycelium and fermented liquid, can be significantly reduced to this, enhance productivity.But, the present domestic launch that does not also have aspects such as ripe Morchella esculenta (L.) Pers mycelium and polysaccharide.

Morel is nutritious to have higher pharmaceutical use, develops series health-care products and will have vast market prospect.The wild toadstool stock number is limited, artificial culture is unsuccessful, wild toadstool world market factors such as supply falls short of demand, makes morel price go up year by year (reaching 2000 yuan/kilogram).Therefore, will be by toadstool ferment exploitation healthcare products for solving many-sided difficult medical problem such as human, resistance low at immunity function.

Macro fungi polysaccharide immunostimulant mechanism mainly is to stimulate scavenger cell, lymphocyte, T lymphocyte, natural killer cell (NK cell), killer cell, B cell from each level of immune organ, immunocyte and immune molecule, increase their quantity, strengthen their activity, and then impel immunity system to play a role.Polysaccharide increases their weight from the growth and the differentiation of immune organ level promotion thymus gland and spleen, and then promotes the quantity of T cell, B cell and scavenger cell in the immune organ to increase.Polysaccharide promotes maturation, differentiation and the secretion of T cell, B cell, monocyte, scavenger cell, mastocyte, killer cell, natural killer cell from the immunocyte level.Morchellaceae (Morchellaceae), not only nutritious, unique flavor, morel also has higher drug effect, and its thalline contains a large amount of compositions such as polysaccharide, has than the powerful antitumor effect, regulate immunologic function, antifatigue, and can alleviate the toxic side effect that cancer patients's chemicotherapy causes.This shows that morel is a kind of rare edible mushrooms with DEVELOPMENT PROSPECT, and can export goods and earn foreign currency, it has higher using value in fields such as medicine, food, strong product, makeup.Morel contains higher nutritive ingredients such as protein carbohydrate multiple amino acids multivitamin; Identical with the Cordyceps sinensis function, be a kind of any hormone natural tonic without any side effects that do not contain, often eat not only and can improve immunizing power, also can eliminate facial color spot macula lutea freckle and make whiteness of skin tenderize [2].But at present, still not deep enough to the extraction and purification research of morel effective constituent, about morel the restraining effect of bacterium etc. is not found relevant report substantially.

The problems of the prior art

Morel is as a kind of rare edible and medicinal fungi, and its research still rests on morphology and ecological level, to the research of its nutrition biochemical character seldom.Domesticly Morchella esculenta (L.) Pers polysaccharide was not carried out than systematic study so far.In recent years, domestic scholars had obtained certain progress to the research of Morchella esculenta (L.) Pers polysaccharide.Jia Jianhui etc. ^[11]Morchella esculenta (L.) Pers mycelium and fermented liquid with submerged fermentation are that main raw material extracts Morchella esculenta (L.) Pers polysaccharide, the Several Factors that influences polysaccharide extract rate has been carried out the contrast experiment respectively, and polysaccharide has been carried out purifying and component evaluation.Wei Yun etc. ^[12]From the fermented morel liquid of liquid submerged fermentation gained, extract 3 kinds of morel holosaccharides, analyzed its component and structure.Lie group ^[13]Carried out the Morchella esculenta (L.) Pers polysaccharide Determination on content with the anthrone colorimetry, for the further investigation of Morchella esculenta (L.) Pers polysaccharide is laid a good foundation.

And Morchella esculenta (L.) Pers mycelium is suitable with sporophore on nutritive ingredient ^[15]Because morel can not carry out large-scale artificial culture, therefore in edible Morchella esculenta (L.) Pers sporophore, people more lay particular emphasis on the fermentation research of Morchella esculenta (L.) Pers mycelium.

J.Sguest cultivated the morel mycelium pellet first in fermentor tank in 1958, and carried out 25000 gallons production test, since the sixties in last century, states such as U.S., method begin its mycelium of scale operation, in order to make seasonings, its product is subjected to human consumer's welcome deeply.At present ^[14]Can carry out the fermentative production of the following scale of 500L abroad, and the equipment of toadstool ferment occur being suitable for specially.Though China's research comparatively lags behind, in the lab scale stage in still being at present, Recent study has obtained bigger progress.People have carried out extensive studies: Liu Shiwang etc. to morel liquid spawn, liquid nutrient medium ^[16](1998) explored Submerged Culture of Morchella conica condition and screened substratum, Zhao Liangqi etc. ^[17](1999) by shaking the conditions suitable that bottle orthogonal test has been studied the Morchellaconica liquid fermenting.Now, people are devoted to seek the optimum fermentation condition of morel mycelia, reach higher in the hope of the morel mycelial yield, make benefit higher.

Summary of the invention

Verified in practice, have only to obtain higher vigor, bacterial strain that production performance is good, just might realize successfully fermenting and producing, reduce production costs.By natural seed selection eliminate that vigor is poor, bacterial strain a little less than the fermentation capacity, for further strain improvement and fermentative production are beaten next solid starting point and basis.Therefore, the present invention sets about from strain improvement, mainly the preliminary screening that the existing 24 strain morels in this laboratory are produced polysaccharide and mycelium dry weight.From the prerequisite that the advantage of bacterial classification aspect is produced as later large scale fermentation, biomass also reaches higher level when being intended to improve the exocellular polysaccharide amount.

In one embodiment, the invention provides a kind of costae morel (MorchellaCostata) M8-13, its preserving number is CGMCC3899.

In another embodiment, the invention provides the purposes that described costae morel M8-13 is used for fermentative production costae Morchella esculenta (L.) Pers mycelium.

In another embodiment, the invention provides described costae morel M8-13 or its mycelium purposes in the preparation healthcare products.Preferably, wherein said healthcare products are used for: reducing blood-fat, press down oxidation, radioprotective, antitumor, raise immunity, antifatigue, and alleviate the toxic side effect that cancer patients's chemicotherapy causes.

In another embodiment, the invention provides described costae morel M8-13 or its mycelium and be used for the treatment of purposes in the disease medicament in preparation.Preferably, wherein said disease comprises: tumour, hyperlipidemia, immune dysfunction.

In another embodiment, the invention provides the method that a kind of usefulness described costae morel M8-13 or its mycelium fermentation are produced fungus polysaccharide, comprising:

(1) spore of the described costae morel of fermentation culture M8-13;

Polysaccharide in the fermented liquid of (2) centrifugation step (1);

(3) sedimentary polysaccharide in extraction and the purification step (2).

Preferably, wherein the method for purification of step (3) is a water extraction and alcohol precipitation method.

In another embodiment, the invention provides described costae morel M8-13 or its mycelium purposes in the preparation that is used for the bacteriostatic medicine, preferably, wherein said bacterium is a streptococcus aureus.

Description of drawings

The mycelium (right side) that Fig. 1 shows morel M8-13 sporophore (left side) and cultivates.

Fig. 2 show morel M8-13 solid culture (left side) and M8-13 intracellular organic matter to the inhibition of streptococcus aureus (in), control strain (right side).

Fig. 3 shows the comparison of M8 series morel biomass, and lowercase is represented 5% significance level of difference, and capitalization is represented 1% significance level of difference; Same letter represents that difference does not show.

Fig. 4 shows the comparison that M8 series morel produced exocellular polysaccharide on the 5th day, and lowercase is represented 5% significance level of difference, and capitalization is represented 1% significance level of difference; Same letter represents that difference is not remarkable, different letter representation significant differences.

Fig. 5 shows the comparison of M8 and the 5th day product of M9 series morel intracellular polyse, and lowercase is represented 5% significance level of difference, and capitalization is represented 1% significance level of difference; Same letter represents that difference is not remarkable, different letter representation significant differences.

Embodiment

The mensuration screening of the screening of bacterial strain of the present invention, evaluation, polysaccharide, the screening of anti-microbial activity etc. all are in China, and carry out in the Xie Zhanling of Qinghai University professor's the laboratory Xining, Qinghai.Morel is that world-renowned wild food (medicine) is used bacterium, and the tender and crisp deliciousness of meat, nutrition and pharmaceutical use are high, so demonstrate great potentiality to be exploited in fields such as food, medicine, health care, makeup.In recent years, because artificial deforestation, soil erosion, the plague of rats, human over-activity and economic interests are chased etc., Qinghai-Tibet morel resource sharply reduces.

The present inventor collects and successful separation and purification Qinghai-Tibet Platean morchella 24 strain strain isolated pure growths (belong to yellow morel and black morel respectively and contained 5-6 kind), and test strain: M8 and the serial bacterium of M9 are Microbiological Lab in May, 2008 and adopt morel in May, 2009 from the Qinghai-Tibet Platean and separate and obtain by Qinghai University's bio-science.The contriver also selects for use morel 51009 (Morchella esculenta 51009, the Chinese Academy of Agricultural Sciences) bacterial strain to do contrast.

The collection of embodiment 1 morel

The morel surface treatment of gathering from the Qinghai-Tibet Platean is clean, after 95% ethanol wiping, be put in the clean plate or wide-necked bottle, place under 40 ℃ in the baking oven, after treating a large amount of ejections of spore, its spore of picking, line on PDA (potato 20%, sucrose 2%, agar 1.5-2%, pH nature) flat board.Cultivate after one day for 25 ℃ and use microscopic examination, wait to have observed single spore germination after, again the substratum cutting-out of sprouting place is shifted on another PDA flat board.Be kept in the PDA slant medium.

The contriver carries out liquid culture to these bacterial strains, and all bacterial strains all earlier activate liquid fermentation medium (KH on the PDA solid medium ₂PO ₄0.1%, MgSO ₄0.1%, sucrose 2%, peptone 0.5%), it forwards in the liquid fermentation medium of the bottled liquid 60mL of 150mL triangle again, under 25 ℃, cultivate under the condition of 100r/min.

In the 24 strain morels of being cultivated, the restraining effect of its biomass, exocellular polysaccharide and corresponding staphylococcus aureus thereof is studied.By analysis (morphological specificity of sporophore, for example mycelial width and habit) and its ITS sequence, identify bacterial strain costae morel (Morchella Costata) M8-13 to its morphological feature.Having carried out in the born of the same parents and the extracellular antiseptic experiment screening to 23 strain morels, is lactic acid intestinal bacteria, subtilis, streptococcus aureus for the examination bacterial classification; And measured polysaccharide content in the mycelium cells of 23 strain morels with the anthrone sulfuric acid process.

Embodiment 2 screening biomass and the high bacterial strains of exopolysaccharides:

The bacterial classification that the inclined-plane whiteruss is sealed up for safekeeping is forwarded on the PDA solid medium, 25 ℃ of following constant temperature illumination cultivation, and wherein M8 series and M51009 cultivated 5 days.Carry out actication of culture in the PDA substratum after, it is forwarded in the liquid fermentation medium of the bottled liquid 60mL of 150mL triangle, every kind of bacterial strain is done 3 parallel controls again, cultivates under the condition of 25 ℃ and 100r/min.

(1) precipitation of exocellular polysaccharide:

To cultivate the 3rd day, the 4th day, the 5th day fermented morel liquid sucking-off 4mL respectively, with the centrifugal 15min of 3500r/min, each centrifuge tube is inhaled the 3mL supernatant liquor, adds the 9mL dehydrated alcohol, and-10 ℃ precipitate 24 hours.

(2) mensuration of exocellular polysaccharide:

Sedimentary polysaccharide with the centrifugal 15min of 3500r/min, is abandoned supernatant, add 3mL distilled water and in 60 ℃ of water-baths, dissolve, dilute 30 times, be made into polysaccharide soln, measure polysaccharide with the anthrone sulfuric acid process, get polysaccharide soln 2mL, place ice-water bath 5min after, in ice-water bath, add anthrone developer (anthrone: H ₂SO ₄=1g: 500mL treats that anthrone dissolves afterwards) 6mL, mixing, boiling water bath 15min, and then place ice-water bath 5min, balance 10min under the room temperature, use UV-9200 ultraviolet-visible pectrophotometer (Beijing Rayleigh Analytical Instrument Co.,Ltd) in 620nm place colorimetric, the result as shown in Figure 4.Glucose typical curve equation is: y=9.5256x-0.0027 (R ²=0.9993)

The exocellular polysaccharide content of table 1 bacterial strain

(3) mensuration of mycelium dry weight:

With 8 days fermented liquid of fermentation with filtered through gauze to obtain mycelium, wash repeatedly 3 times with distilled water, put in the baking oven and dry to constant weight under 55 ℃, weigh, the result as shown in Figure 3.

The mycelium dry weight of table 2 bacterial strain

Embodiment 3 intracellular polyses are measured:

Take by weighing 0.020g left and right sides exsiccant Morchella esculenta (L.) Pers mycelium, add 1mL water grinding altogether at twice, boil and extract 30min, the centrifugal 20min of cooling back 2500r/min.Get supernatant liquor and place a centrifuge tube, in residue, add 1mL water again, boil and extract 30min, collect supernatant liquor and place above-mentioned same centrifuge tube.In supernatant liquor, add the 7mL dehydrated alcohol polysaccharide separated out, behind the precipitation 30min with the centrifugal 20min of 2500r/min, abandoning supernatant, residue adds same centrifugal abandoning supernatant after the 3.5mL ethanol stirring and washing again.With resolution of precipitate, be settled to 2mL with about 80 ℃ of hot water after the cooling, the anthrone sulfuric acid process is measured polysaccharide, and is the same, referring to Fig. 5.

The purifying of embodiment 4 isolated strains M8-13 bacterial strains:

The sample of isolated strains M8-13 comes from morchella sporophore spore separation thing.Be to support and purifying by a series of inferior being commissioned to train on screening culture medium.Screening culture medium comprises 0.6% peptone, 0.1%K ₂HPO ₄, 0.05%MgSO ₄7H ₂O and 1% glucose, pH 7.0.Cultivated 3-5 days down at 28 ℃.Then with well-grown bacterium colony by inferior be commissioned to train to support be transferred on the fresh PDA substratum.From the bacterium colony of even growth, arrive liquid fermentation medium: KH with one ring transition of transfering loop picking ₂PO4 0.1%, MgSO ₄0.1%, sucrose 2%, peptone 0.5%.Under 25 ℃, in shaking table, cultivate with the rotating speed of 100rmp.For preservation, bacterial strain is inoculated on the PDA substratum and preserves under 4 ℃ of conditions.

Evaluation on embodiment 5 morel M8-13 morphology and the molecules:

The mycelium of morel M8-13 is continued to cultivate 5 days on the PDA substratum, be used for measuring morphological feature, as the mycelia width, the formation of spore and the shape of spore.With CTAB method (vanBurik et al., 1998) from 5 days mycelium of PDA substratum growth, extracting genomic dna, utilization has forward ITS5 primer, and 5 '-GGAAGTAAAAGTCGTAACAAGG-3 ' and ITS4 reverse primer 5 '-TCCTCCGCTTATTGATATGC-3 ' PCR are with Yang et al. (2007) method amplification ITS DNA.The order-checking back is carried out sequential analysis by NCBI BLAST database to the PCR product.By with the comparison of other bacterial strains ITS sequence, combining form is learned feature, determines its classification position according to the sequence similarity degree.

The PCR reaction system of ITS all adopts following system:

PCR reaction system (25 μ L)

ITS gene PCR amplification program is as follows:

1) pre-sex change: 94 ℃ of 1min

2) sex change: 94 ℃ of 15s

Annealing: 58 ℃ of 15s

Extend: 72 ℃ of 1min

30 circulations

3) benefit is flat: 72 ℃ of 5min

Get 5 μ L amplified productions after PCR finishes and carry out electrophoresis on 1% sepharose, ultraviolet lamp detects down and the band of record amplified production on sepharose.Agents useful for same is all given birth to the worker available from Shanghai.

As can be seen from Fig. 1, the mycelium of M8-13 presents the down-like of doing of typical white, spreads all over bacterium colony (Fig.3A) everywhere.Mycelium is formed ((Fig.3B) .) by many intervals.In order to identify morel 8-13, its mycelium is inoculated on the solid medium solid, cultivates 6 days full wares at 25 ℃.

Identify at the genome relevant with 18S ribosomal gene partial sequence, genomic dna among the extraction bacterial strain M8-13 is as the template of pcr amplification, and by round pcr, internal transcription transcribed spacer 1 increases, 5.8S ribosomal RNA gene, internal transcription transcribed spacer 2 and 28S ribosomal RNA gene partial sequence.The sequence information that pcr amplification goes out is used to do BLAST research in ncbi database.BLAST result shows that M8-13 analyzes section, wherein 99% and the costae morel consistent.The analysis of combining form and molecules, we are decided to be the costae morel with M8-13 spare, and be deposited in (No. 3, A, DaTun Road, Chaoyang District, BeiJing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on June 13rd, 2010 according to the rule of culture presevation, Institute of Microorganism, Academia Sinica's postcode: 100101), preserving number: CGMCC3899.

Data processing and errot analysis:

Adopt the DPS data processing software to carry out errot analysis: the new multipole difference of Duncan multiple comparisons method is adopted in test of significance between the different strains.

Embodiment 6 morel M8-13 bacteriostatic tests:

(1) cultivates and preparation for the examination bacterium: the lactic acid intestinal bacteria (Livestock Research and Veterinary Academy of Qinghai Prov. provides), subtilis, the streptococcus aureus that preserve are forwarded on the beef extract-peptone solid medium and activate, place 25 ℃ of intelligent illumination boxs to cultivate 18h.Therefrom the picking bacterium is transferred in the beef extract-peptone liquid nutrient medium then, and 37 ℃ of following 100r/min cultivate 10h, obtain various original bacteria liquids.Draw fermented liquid (fermented liquid is got respectively and cultivated the 3rd, 4,6, the 8 day) mixing of 20 μ L original bacteria liquids (being respectively the original bacteria liquid of lactic acid intestinal bacteria, subtilis, streptococcus aureus) and 180 μ L morels, make for examination bacterium liquid 1.Be used for the outer antibiotic screening of material of born of the same parents.

(2) fermented liquid of morel M8-13 screens antibacterial experiment:

Get 50 μ L and coat on the foster base of beef extract-peptone training solid, cultivate 18h for 25 ℃ for examination bacterium liquid 1.Enumeration.Each is subjected to sample to do 2 parallel laboratory tests, and does contrast (the examination bacterium liquid that supplies of 20 μ L adds the distilled water of 180 μ L).

(3) the bacteriostatic test screening carried out of Morchella esculenta (L.) Pers mycelium:

A. antimicrobial filter paper sheet preparation

Get 0.010g exsiccant Morchella esculenta (L.) Pers mycelium, add 0.3mL water and grind.The circular filter paper sheet (d=9.00mm) of sterilizing and drying is crossed is put into ground mycelium liquid, soak 30min.

B. the mensuration of inhibition zone size

Draw 20 μ L and evenly be applied on the beef extract-peptone flat board, smoothen for examination bacterium liquid 1.Each dull and stereotyped going up is pasted two antimicrobial filter paper sheets, and filter paper is 15.00mm at least from the plate edge.Take out after placing 25 ℃ of intelligent illumination boxs to cultivate 18h, measure antibacterial circle diameter with ruler.Contrast is M51009.

The result:

Judge that according to polysaccharide yield, mycelial biomass, bacteriostatic test etc. are comprehensive filtering out morel M8-13 bacterial strain is optimum strain, can be used for the development research of protective foods and medicine.The cultural characteristic of morel M8-13 bacterial strain sees Table 3, and sporophore and mycelium are seen Fig. 1.Bacteriostatic test the results are shown in Table 4, Fig. 2.

The cultural characteristic of table 3 morel M8-13 bacterial strain

The mycelium pellet number is about 25 in +++" the express liquid fermentation culture

Table 4 morel M8-13 liquid fermenting mycelium is to aureus with inhibition

Morel M8-13 is better to aureus with inhibition, and lactic acid intestinal bacteria and subtilis are not had the obvious suppression effect; Morel M8-13, fermenting can reach 5.867g/L in the mycelia soma on the 8th day, and the 3rd day exocellular polysaccharide that ferment can reach 0.299g/L, and the mycelium intracellular organic matter reaches 20 millimeters to the inhibition zone of streptococcus aureus.Average speed of growth 8.0mm every day, mycelium diameter 6.64-11.96 μ m sees Table 5, Fig. 3, Fig. 4.

In the table 5 morel M8-13 liquid fermenting born of the same parents, exocellular polysaccharide and biomass

This bacterial strain has very strong Medicines and Health Product exploitation potential.

Reference (REFERENCES)

[1] orchid advances, Cao Wenling, Xu Jintang. Chinese morchella fungus resource [J]. and resources science, 1999,21 (2): 56-61.

[2] Li Juan, Wang Zhen, Yao Liangtong, etc. Morchella esculenta Polysaccharides Studies [J]. JOURNAL OF MICROBIOLOGY, 2005,25 (4): 89-91.

[3] Chen XiangDong, Zhu Rong, orchid advances. morel progress [J]. edible mushrooms journal, 2002,9 (2): 56-61.

[4] Liu Minli. wild bacterium inorganic elements such as morel is analyzed [J]. Chinese wild plant resource, 1994, (2): 42-44.

[5] noble scholar. health care treasure---morel [J]. extraordinary economic animals and plants, 2000 (3): 38.

[6] Li Hua. Morchella esculenta (L.) Pers sporophore part chemical constitution study [D]. Jilin: Jilin Agriculture University's master thesis, 2006.05.01.

[7] Bai Lan. the structure of the progress of fungus polysaccharide---fungus polysaccharide, purification and application [J]. forest fruit in Hebei.

[8]Jeng-Leun?Mau，Shih-JengHuang，Chin-Chu?Chen.Antioxidant?properties?of?methanolic?extracts?from?Grifola?frondosa，Morchella?esculenta?and?Termitomyces?albuminosus?mycelia[J].FoodChemistry，2004，111-118.

[9] Sun Xiaoming, Zhang Weiming, Wu Suling, etc. morel antifatigue effect research [J]. Chinese wild plant money grinds 2009,24 (4): 445-448.

[10] Tang Maozhi, Bei Feng. edible fungi food is as the advantage and the characteristics [J] of functional foodstuff. food science and technology, 1999,7 (3): 5-6.

[11] Jia Jianhui, Lv Xiaolian, Fan Liqing. the extraction separation of submerged fermentation Morchella esculenta (L.) Pers polysaccharide and purifying research [J]. Food science, 2002,23 (4): 59-62.

[12] Wei Yun, Zhang Tian helps. the separation and purification of Morchella esculenta (L.) Pers polysaccharide and composition structural analysis [J]. and plant resources and environment journal, 2000,9 (2): 14-17.

[13] Lie group. anthrone colorimetric method for determining Morchella esculenta (L.) Pers polysaccharide and test evaluation [J]. Chinese sanitary inspection magazine, 2000,10 (1): 31-32.

[14] Fu Xiaoyan. Morchella esculenta (L.) Pers mycelium is cultivated and mycelia differentiation research [D]. Beijing: Capital Normal University, 2006.05.20.

[15] Zhao Guiyun, Ma Qingbin. the research of Submerged Culture of Morchella conica based formulas [J]. edible mushrooms, 2002,6:15-16.

[16] Liu Shiwang, Liang Zongqi, Liu Aiying. Study on Submerged Culture of Morchella conica preliminary study [J]. edible mushrooms journal, 1998,5 (3): 31-37.

[17] Zhao Liangqi, Li Zhiqiang, Hou Guixiang. the research of morel liquid state fermentation. fungus system, 1999,18 (1): 94-99.

[18] Ou Chao, Wang Di, Li Zhengpeng. morel liquid submerged fermentation condition [J]. food and biotechnology journal, 2007,26 (2): 26-33.

[19] Wang Ying, Sun Yongmei, Meng Xiangyan. the optimization [J] of thick handle Submerged Culture of Morchella conica condition. edible mushrooms .2007 (6): 6-8.

[20] Xing Zengtao, Sun Fangfang, Liu Jingsheng. Morchellaconica liquid culture Study on Conditions [J]. edible mushrooms journal, 2004,11 (4): 38-43.

[21] Zhang Guanglun, Zhang Weiming. research of morel and utilization [J]. Chinese wild plant resource, 1999,18 (1): 1-4.

[22] Song green pepper, Zou Zuohua, Wang Hongyin waits the experimental study of grinding side and health-care effect thereof [J] of .EF-11 nutritive medium. study carefully Food science, 1996,17 (7): 52-57.

Claims (10)

1. a costae morel (Morchella Costata) M8-13, its preserving number is CGMCC3899.

2. the described costae morel of claim 1 M8-13 is used for the purposes of fermentative production costae Morchella esculenta (L.) Pers mycelium.

3. claim 1 described costae morel M8-13 or its mycelium are in the purposes of preparation in the healthcare products.

4. the described purposes of claim 3, wherein said healthcare products are used for: reducing blood-fat, press down oxidation, radioprotective, antitumor, raise immunity, antifatigue, and alleviate the toxic side effect that cancer patients's chemicotherapy causes.

5. claim 1 described costae morel M8-13 or its mycelium are used for the treatment of purposes in the disease medicament in preparation.

6. the described purposes of claim 5, wherein said disease comprises: tumour, hyperlipidemia, immune dysfunction.

7. method of producing fungus polysaccharide with claim 1 described costae morel M8-13 or its mycelium fermentation comprises:

(1) spore of the described costae morel of fermentation culture claim 1 M8-13;

Polysaccharide in the fermented liquid of (2) centrifugation step (1);

(3) sedimentary polysaccharide in extraction and the purification step (2).

8. the described method of claim 7, wherein the method for purification of step (3) is a water extraction and alcohol precipitation method.

9. claim 1 described costae morel M8-13 or its mycelium are in the purposes of the preparation that is used for the bacteriostatic medicine.

10. the described purposes of claim 9, wherein said bacterium is a streptococcus aureus.

Images