CN104073531B - A kind of deep layer liquid fermentation produces the method for Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide - Google Patents
A kind of deep layer liquid fermentation produces the method for Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide Download PDFInfo
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Abstract
The invention belongs to microbial technology field, be specifically related to a kind of method that deep layer liquid fermentation produces Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide.The specifically production method of the Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide of a kind of industrially scalable, the method is that the Marasmius androsaceus (L.ex Fr.) Fr. strain activated is carried out expansion joined by being linked into seed tank, again the liquid seeds that expansion prepares is linked in the special liquid culture medium producing polysaccharide and ferments, then fermentation liquid is separated, extract extracellular polysaccharide.The present invention is directed to above-mentioned the deficiencies in the prior art study, on the basis of lab scale and pilot scale continue to optimize fermentation medium (especially carbon source), promote Biomass and extracellular polysaccharide, solve be amplified to industrialized production runs into by lab scale Fermentation Process of Parameter control, extraction equipment type selecting, the difficult problem such as optimization, finally being successfully grafted onto 10 20t fermentation tanks carries out industrialization production, and produce the Marasmius polysaccharide of high-quality smoothly, be truly realized continuously, automatization, the production of industrialization.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of deep layer liquid fermentation and produce Marasmius androsaceus (L.ex Fr.) Fr. born of the same parents
The method of exo polysaccharides.
Background technology
Marasmius androsaceus (L.ex Fr.) Fr., formal name used at school is Marasmius androsaceus (L.ex Fr.) Fr. another name Marasmius equicrinis Muell., dark brown
Marasmiuses etc., belong to Basidiomycetes, Agaricales (Agaricales), Bai Mo section (ThichoLomataceae), cuticle
Umbrella belongs to (Marasmius).It is born on the deadwood above remote, thickly forested mountains, cool, damp, acid ground, is mainly distributed
Save in China Jilin, Hunan, Hubei, Guangdong etc..China's morning among the people useful wild Marasmius androsaceus (L.ex Fr.) Fr. root shape bacterium
Rope treats fracture pain, traumatic injury etc..According to Guangdong institute of microbiology (1975), this institute is with wide
State the first pharmaceutical factory etc. uses artificial fermentation culture method, it is thus achieved that Marasmius androsaceus (L.ex Fr.) Fr. fermentation culture medium, then is concentrated
" the marasmius androsaceus capsule " made and medicated wine, 600 example according to clinical verification, it was demonstrated that to trigeminal neuralgia, ischium
Neuralgia, migraine, supraorbital neuralgia, facial nerve paralysis, facial spasm and rheumatic arthritis etc. all have
Certain curative effect.Centering, light leprosy neuralgia, lumbar muscle strain etc. also have certain mitigation, are to pacify human body
Entirely, the analgesia having no side effect and antirheumatic.The medicine made with it at present, except " marasmius androsaceus capsule "
Outward, also have " marasmius androsaceus tincture ", " marasmius androsaceus injection " and " CNNS's gardan " etc..Clinic and market effect
Very well.Recent studies indicate that the healthy nutritive value of Marasmius androsaceus (L.ex Fr.) Fr. is the highest, can promote nervous tissue and
The inflammation of fibrous connective tissue disappears, and improves blood circulation and the histotrophic nutrition situation of local, recovers function of nervous system.
Human body can be exempted from by the compositions such as aminoacid that bacterium is contained within, saccharide as food and the adding ingredient of health product
Epidemic disease power improves and defying age has certain effect.The function of other component of Marasmius androsaceus (L.ex Fr.) Fr. is also in studying
Stage.
Increasing scholar begins one's study the active ingredient of Marasmius androsaceus (L.ex Fr.) Fr. and pharmacological action in recent years, but mesh
Before domestic wild Marasmius androsaceus (L.ex Fr.) Fr. resource scarcity, the artificial culture success rate of sporophore is low and poor repeatability, Gu
Body cultivation period length, pollution rate are high, are easily limited by surrounding, it is impossible to meet the growing market demand.
And deep layer liquid fermentation technology has the advantages such as fermentation period is short, high yield, pollution rate are low, not by surrounding
Limit, resource apparatus utilization rate advantages of higher, thus favored by increasing researchers.Both at home and abroad
Also the researcher having a small amount of universities and colleges carried out research to the submerged fermentation of Marasmius androsaceus (L.ex Fr.) Fr., but often it is main
Emphasis is how to obtain higher Biomass, and only small part researcher pays close attention to the functional component of Marasmius androsaceus (L.ex Fr.) Fr.
The research of polysaccharide, but mainly extracted by mycelium and obtain polysaccharide, the byproduct fermentation filtrate as fermentation is big
All directly drain as waste liquid, waste substantial amounts of precious resources, and add environmental protection pressure, the row of improving
Dirty cost.
Summary of the invention
The invention aims to solve the technical bottleneck of industrial-scale production Marasmius polysaccharide, use
Liquid fermentation produce Marasmius polysaccharide overcome solid fermentation cycle length, investment is big, be subject to seasonal restrictions, difficult
With drawbacks such as industrialization, use this invention can continuously, automatization, the CNNS producing high-quality of industrialization
Marasmius polysaccharide.
To achieve these goals, the present invention is carried out essentially according to following step:
The industrialization deep layer liquid fermentation of the present invention produces the method for Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide and includes following step
Rapid:
A kind of method preparing Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid, after accessing sterilizing by Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid
Fermentation medium, inoculum concentration is 10～20%, and ventilating ratio is 1:0.5 1.5vvm, tank pressure 0.02～0.05Mpa,
Cultivate 46 days at 22～30 DEG C of bottom fermentations, obtain Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid；Wherein said fermentation medium
Formula is: glucose 1-3%, starch 1-3%, yeast powder 0.2-1%, Semen Maydis pulp 0.5-2%, potassium dihydrogen phosphate
0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, 20000u/ml high temperature resistant α-
Amylase 0.001-0.007% (v/v), regulates pH value 6.5-7.5.Except amylase in above-mentioned culture medium composition
" % " in addition, all represents g/100ml.
Described defoamer can be vegetable oil (Oleum Glycines, the Oleum Brassicae campestris) material such as class, organic ethers.
Wherein, described Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid is preferably prepared by the following method and obtains:
1) first order seed is cultivated: activated shaking flask strain accesses the liquid seed culture medium after sterilizing, inoculation
Amount is 0.1～2%, and ventilating ratio is 1:0.5 1.5vvm, tank pressure 0.02～0.05Mpa, cultivates at 22～30 DEG C
5～6 days, obtain Marasmius androsaceus (L.ex Fr.) Fr. first order seed culture fluid；
2) secondary seed is cultivated: by step 1) the first order seed culture fluid of gained accesses the liquid strain after sterilizing
Sub-culture medium, inoculum concentration is 10～20%, and ventilating ratio is 1:0.5 1.5vvm, tank pressure 0.02～0.05Mpa,
Cultivate 3～4 days at 22～30 DEG C, obtain described Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid.
Described liquid seed culture medium formula is preferably: glucose 2-4%, dried silkworm chrysalis meal 0.1-0.5%, Semen Glycines
Cake powder 1-3%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%,
Regulation pH value 6.5-7.5." % " in above-mentioned culture medium composition all represents g/100ml.
Described fermentation culture terminal is: fungus ball becomes broken, filtrate is muddy；Reducing sugar is down to minimum and slightly fluctuates,
Reducing sugar controls 0.3% (g/100g) below；Microscopy mycelia is in small, broken bits, without living contaminants.
A kind of method that Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide extracts, the method comprises the steps:
1) separate: by separated for the Marasmius androsaceus (L.ex Fr.) Fr. fermentation culture of claim 1 gained, obtain CNNS's cuticle
Umbrella supernatant；
2) concentrating: by step 1) filtrate of gained carries out using two grade low-temps to concentrate, and obtain concentrating proportion and be
The concentrated pulp of 1.25～1.40；
3) precipitate with ethanol, drying: add 95% ethanol of concentrated pulp weight 3-5 times amount, with alcohol meter detection mixing
Liquid alcoholic strength about 65-80% (v/v), stands 8-16h, precipitate carries out vacuum drying, temperature 50-75 DEG C,
Vacuum is not less than 0.085MPa；Drying to dried object moisture is 3～8% (g/100g), obtains outside born of the same parents
Sugar.
Wherein, described step 1) described in separation use horizontal screw centrifuge to carry out, rotating speed controls
2500‐3500r/min。
Described step 2) employed in secondary concentration be: one-level use 1 3t double effect evaporator concentrate, will send out
It is 1.15 1.20 (50 DEG C of heat are surveyed) that ferment liquid is concentrated to proportion, and two grades use 1 3t haplo-effect concentrator to concentrate, will
Proportion is that the concentrated solution of 1.1 1.20 (50 DEG C of heat are surveyed) continues to be concentrated to proportion to be 1.25～1.40 (50 DEG C of heat are surveyed)
Concentrated pulp；Concentration process is required to control temperature and is not higher than 75 DEG C, and vacuum is not less than 0.085MPa.
In the present invention, precipitate with ethanol is to use 1 3t Alcohol-settling tank, and precipitate with ethanol number of times is the most, and the content of the outer sugar of born of the same parents is the highest.Very
The empty vacuum drying oven used of drying, born of the same parents' outer sugar content of present invention extraction is 20 50%.
The present invention has a following prominent advantage:
The present invention is directed to above-mentioned the deficiencies in the prior art study, continue to optimize fermentation culture at lab scale and pilot scale
On the basis of base (especially carbon source), lifting Biomass and extracellular polysaccharide, solve and be amplified to industry by lab scale
Fermentation Process of Parameter controls that metaplasia runs in producing, extraction equipment type selecting, the difficult problem such as optimization, the most successful final grafting
Carry out industrialization production to 10 20t fermentation tanks, and produce the Marasmius polysaccharide of high-quality smoothly, very
Just achieving continuously, automatization, the production of industrialization.
In order to improve the yield of purpose product extracellular polysaccharide, the present invention has carried out excellent in the carbon source of fermentation medium
Change and combination: use the compound prescription of fugitive glucose and long-acting starch.Earlier fermentation, to grow mycelium, carries
High biological content is main, therefore with the addition of the glucose carbon source that thalline is preferentially selected in culture medium, breeds rapidly；
The content of fermentation later stage purpose to be improved product, therefore carbon source provides thalline relatively to utilize slower starch, and
Adding appropriate amount of starch enzyme in culture medium, control thalline quantity is at proper size, and the raw metabolite born of the same parents of pregnancy ceased are outer sugared.
In order to provide born of the same parents the yield of outer sugar, the present invention optimizes the formula of fermentation medium, uses for carbon source part
Compound prescription, both ensure that the Biomass of early stage thalline, had ensured again the yield of later stage metabolite.
The employing present invention according to the market demand and product standard, can prepare the polysaccharide of the high-quality of different content, many
Sugar content can be 20 50%.
Detailed description of the invention
The present invention is further illustrated below by way of specific embodiment.But the detail of embodiment is only used for explaining
The present invention, should not be construed as limited overall technical solution.
The Marasmius androsaceus (L.ex Fr.) Fr. that following example use is bought in Institute of Microorganism, Academia Sinica's common micro-organisms
Center, strain coding CGMCC NO5.0152.As a example by this strain bacterium, describe the concrete side of the present invention in detail
Case.But the inventive method is common to all of Marasmius androsaceus (L.ex Fr.) Fr., it is not limited in this bacterial strain.
Embodiment 1
Industrialization deep layer liquid fermentation of the present invention produces the method for Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide and enters in the steps below
OK:
1, the preparation of Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid: by the Marasmius androsaceus (L.ex Fr.) Fr. shaking flask bacterial strain that activated by inoculum concentration 1%
Being inoculated in seed culture medium (300L), wherein seed culture medium consists of glucose 3%, dried silkworm chrysalis meal 0.3%, Huang
Soybean cake powder 1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value
7.0, ventilating ratio is 1:1.0vvm, tank pressure 0.03Mpa, 26～28 DEG C of shaken cultivation 5 Tian Ji get CNNS cuticles
Umbrella primary seed solution；Primary seed solution is accessed secondary seed medium (3000L) by inoculum concentration 15%, wherein two
Level seed culture medium consist of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium dihydrogen phosphate 0.3%,
Magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, and ventilating ratio is 1:1.0vvm, tank pressure 0.03Mpa,
26～28 DEG C of cultivations i.e. obtain Marasmius androsaceus (L.ex Fr.) Fr. secondary seed solution in 3 days；Secondary seed solution is accessed by inoculum concentration 18%
Fermentation medium (7500L), wherein fermentation medium consist of glucose 2%, starch 2%, yeast powder 0.5%,
Semen Maydis pulp 1.5%, potassium dihydrogen phosphate 0.2%, Magnesium sulfate heptahydrate 0.2%, defoamer Oleum Glycines 0.03%, high temperature resistant
Alpha amylase (Zaozhuang outstanding person promise enzyme biology company limited, 20000u/ml) 0.002% (v/v), regulates pH value
7.0；Ventilating ratio is 1:0.5vvm, tank pressure 0.03Mpa, 26～28 DEG C cultivate 4 days, ferment to fungus ball become broken,
Filtrate is muddy；Reducing sugar is down to minimum 0.2% and slightly fluctuates；Microscopy mycelia is in small, broken bits, without living contaminants.I.e.
Obtain Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid 8850Kg.
2, Marasmius androsaceus (L.ex Fr.) Fr. separation of fermentative broth: above-mentioned Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid 8850Kg is centrifuged by sleeping spiral shell
Machine separates, and rotating speed controls at 3000r/min, and disengaging time, at 2h, obtains Marasmius androsaceus (L.ex Fr.) Fr. filtrate 7524Kg
With the wet mycelium (water content is 80%) of 1105Kg, wet mycelium directly cold drying can pulverize prepared fermentation
Marasmius androsaceus (L.ex Fr.) Fr. mycopowder.
4, cryoconcentration: the Marasmius androsaceus (L.ex Fr.) Fr. filtrate 7524Kg vacuum of step 2 gained is transferred to 3t economic benefits and social benefits
Vaporizer, is 0.085MPa in vacuum, and thickening temperature is concentrated into proportion under the conditions of being 65 DEG C be 1.20 (50
DEG C heat is surveyed) concentrated solution, then concentrated solution vacuum is transferred to 1.5t single-action ball-type concentrator, in vacuum is
0.09MPa, thickening temperature continues under the conditions of being 60 DEG C to be concentrated into the concentration that proportion is 1.30 (50 DEG C of heat are surveyed)
Slurry 280kg.
5, precipitate with ethanol, drying: add 95% wine of weight 1100kg in the Alcohol-settling tank filling 280kg concentrated pulp
Essence, with alcohol meter detection mixed liquor alcoholic strength 72% (v/v), stands 12h, separates bottom sediment with true
Empty drying baker is dried, temperature 60 C, vacuum 0.09MPa；Dry 12h and obtain born of the same parents outer sugar 132.8kg.
Detection moisture is 4%, and polyoses content is 30%.
Embodiment 2
1, the preparation of Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid: by the Marasmius androsaceus (L.ex Fr.) Fr. shaking flask bacterial strain that activated by inoculum concentration
1.5% is inoculated in seed culture medium (300L), wherein seed culture medium consist of glucose 3%, dried silkworm chrysalis meal 0.3%,
Soybean cake powder 1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH
Value 7.0, ventilating ratio is 1:1.0vvm, tank pressure 0.03Mpa, and 25～27 DEG C of shaken cultivation 6 Tian Ji get CNNSs are little
Skin umbrella primary seed solution；Primary seed solution is accessed secondary seed medium (3000L) by inoculum concentration 15%, wherein
Secondary seed medium consist of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium dihydrogen phosphate 0.3%,
Magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, and ventilating ratio is 1:1.0vvm, tank pressure
0.03Mpa, 25～27 DEG C of cultivations i.e. obtain Marasmius androsaceus (L.ex Fr.) Fr. secondary seed solution in 3 days；By secondary seed solution by inoculation
Amount 18% access fermentation medium (15000L), wherein fermentation medium consists of glucose 2%, starch 2%, ferment
Female powder 0.7%, Semen Maydis pulp 1.5%, potassium dihydrogen phosphate 0.2%, Magnesium sulfate heptahydrate 0.2%, defoamer Oleum Brassicae campestris
0.03%, thermostable α-amylase (Zaozhuang outstanding person promise enzyme biology company limited, 20000u/ml) 0.002% (v/v),
Regulation pH value 7.0；Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, cultivates 4 days for 25～27 DEG C, fermentation
To fungus ball change is broken, filtrate is muddy；Reducing sugar is down to minimum 0.2% and slightly fluctuates；Microscopy mycelia is in small, broken bits, nothing
Living contaminants, had both obtained Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid 17700Kg.
2, the separation of Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid: by above-mentioned Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid 17700Kg by sleeping spiral shell
Centrifuge separates, and obtains the wet mycelium (water content of Marasmius androsaceus (L.ex Fr.) Fr. filtrate 15540Kg and 1800Kg
It is 75%), wet mycelium directly cold drying can pulverize prepared fermentation Marasmius androsaceus (L.ex Fr.) Fr. mycopowder.Concrete technology bar
Part is: rotating speed controls at 3200r/min, and disengaging time is at 4.5h.
4, cryoconcentration: the Marasmius androsaceus (L.ex Fr.) Fr. filtrate 15540Kg vacuum of step 2 gained is transferred to 3t economic benefits and social benefits
Vaporizer, is 0.09MPa in vacuum, and thickening temperature is concentrated into proportion under the conditions of being 63 DEG C be 1.18 (50
DEG C heat is surveyed) concentrated solution, then concentrated solution vacuum is transferred to 1.5t single-action ball-type concentrator, in vacuum is
0.09MPa, thickening temperature is concentrated into the concentrated pulp that proportion is 1.35 (50 DEG C of heat are surveyed) under the conditions of being 65 DEG C
570kg。
5, precipitate with ethanol, drying: add 95% wine of weight 2300kg in the Alcohol-settling tank filling 570kg concentrated pulp
Essence, with alcohol meter detection mixed liquor alcoholic strength 71% (v/v), stands 10h, separates bottom sediment with true
Empty drying baker is dried, temperature 65 DEG C, vacuum 0.09MPa；Dry 18h and obtain born of the same parents outer sugar 260kg.Inspection
Surveying moisture is 4.5%, and polyoses content is 32%.
Embodiment 3
CNNS is produced in industrialization for the compound prescription of fugitive glucose and long-acting starch in the more preferably checking present invention
The superiority of Marasmius polysaccharide, has done again several contrast test.Test only have adjusted in liquid fermentation medium
Carbon source glucose or (with) consumption of starch, comparative example 1 does not contains starch, concentration of glucose is 1.5%；
Not containing glucose in comparative example 2, starch concentration is 2.5%, and in comparative example 3, glucose content is 2.5%,
Content of starch is 0.5%；Remaining condition is all with embodiment 1, and result of the test is shown in Table 1:
Table 1
Claims (4)
1. the method preparing Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid, it is characterized in that the fermentation medium after Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid is accessed sterilizing, inoculum concentration is 10 ~ 20%, ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, cultivate 3 ~ 5 days at 22 ~ 30 DEG C of bottom fermentations, obtain Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid；Wherein said fermentative medium formula is: glucose 1-3%, starch 1-3%, yeast powder 0.2-1%, Semen Maydis pulp 0.5-2%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, the Thermostable α-Amylase 0.001-0.007%v/v of defoamer 0.01-0.05%, 20000u/ml, regulates pH value 6.5-7.5；
Wherein, described Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid is prepared by the following method and obtains:
1) first order seed is cultivated: activated shaking flask strain accesses the liquid seed culture medium after sterilizing, inoculum concentration is 0.1 ~ 2%, and ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, cultivate 5 ~ 6 days at 22 ~ 30 DEG C, obtain Marasmius androsaceus (L.ex Fr.) Fr. first order seed culture fluid；
2) secondary seed is cultivated: the first order seed culture fluid of step 1) gained accesses the liquid seed culture medium after sterilizing, inoculum concentration is 10 ~ 20%, and ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, cultivate 3 ~ 4 days at 22 ~ 30 DEG C, obtain described Marasmius androsaceus (L.ex Fr.) Fr. seed culture fluid；
Described liquid seed culture medium formula is: glucose 2-4%, dried silkworm chrysalis meal 0.1-0.5%, soybean cake powder 1-3%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, regulates pH value 6.5-7.5.
2. the extracting method of a Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide, it is characterised in that the method comprises the steps:
1) Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid is prepared: prepare Marasmius androsaceus (L.ex Fr.) Fr. fermentation liquid in accordance with the method for claim 1；
2) separate: by separated for the Marasmius androsaceus (L.ex Fr.) Fr. fermentation culture of gained, obtain Marasmius androsaceus (L.ex Fr.) Fr. supernatant；
3) concentrating: by step 2) filtrate of gained carries out using two grade low-temps to concentrate, obtains concentrating the concentrated pulp that proportion is 1.25～1.40；
4) precipitate with ethanol, drying: add 95% ethanol of concentrated pulp weight 3-5 times amount, is 65-80% with alcohol meter detection mixed liquor alcoholic strength, stands 8-16h, precipitate carries out vacuum drying, temperature 50-75 DEG C, vacuum is not less than 0.085MPa；Drying to dried object moisture is 3～8%, and wherein moisture unit is g/100g, obtains the outer sugar of born of the same parents.
The extracting method of Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide the most according to claim 2, it is characterised in that described step 2) described in separation use horizontal screw centrifuge carry out, rotating speed controls at 2500-3500r/min.
The extracting method of Marasmius androsaceus (L.ex Fr.) Fr. extracellular polysaccharide the most according to claim 2, it is characterized in that the secondary concentration employed in described step 3) is: one-level uses double effect evaporator to concentrate, it is 1.15-1.20 that fermentation liquid is concentrated to proportion, two grades use haplo-effect concentrators to concentrate, and will proportion be that the concentrated solution of 1.15-1.20 continues to be concentrated to the concentrated pulp that proportion is 1.25～1.40；Concentration process is required to control temperature and is not higher than 75 DEG C, and vacuum is not less than 0.085MPa.
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