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CN105368899A - Method for extracting water-soluble beta glucan from pleurotus ferulae lenzi sporocarp - Google Patents

Method for extracting water-soluble beta glucan from pleurotus ferulae lenzi sporocarp Download PDF

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Publication number
CN105368899A
CN105368899A CN201510948166.0A CN201510948166A CN105368899A CN 105368899 A CN105368899 A CN 105368899A CN 201510948166 A CN201510948166 A CN 201510948166A CN 105368899 A CN105368899 A CN 105368899A
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Prior art keywords
resina ferulae
ferulae mushroom
water
soluble beta
pleurotus ferulae
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陈喜军
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Heilongjiang Zhongsheng Biological Engineering Co Ltd
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Heilongjiang Zhongsheng Biological Engineering Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/14Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C12P19/08Dextran

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
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  • Life Sciences & Earth Sciences (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
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  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The invention provides a method for extracting water-soluble beta glucan from pleurotus ferulae lenzi sporocarp. The method includes the following steps: 1, preparation of pleurotus ferulae lenzi coarse polysaccharide, wherein pleurotus ferulae lenzi sporocarp is picked and crushed, powder is obtained, the powder is extracted with alkali liquor, filtration is conducted to remove residues, ethyl alcohol is added to the filter liquor, the filter liquor stands still and is filtered, sediment is obtained, and the pleurotus ferulae lenzi coarse polysaccharide is obtained; 2, preparation of pleurotus ferulae lenzi beta sporocarp, wherein the pleurotus ferulae lenzi coarse polysaccharide is picked and dried, then water is added to the dried pleurotus ferulae lenzi coarse polysaccharide, stirring is conducted, alpha amylase is added to the mixture for hydrolysis, the hydrolysis liquid is filtered, filter liquor is removed, residues are obtained, and the pleurotus ferulae lenzi beta sporocarp is obtained; 3, preparation of the water-soluble beta glucan, wherein the pleurotus ferulae lenzi beta sporocarp is picked, alkali liquor is added, stirring and extraction are conducted, the pH value of the extraction liquor is adjusted to 3.5-6.8, heating and hydrolysis are conducted, the hydrolysis liquid is filtered, ethyl alcohol is added to the filter liquor, the liquor stands still, separation, sediment and drying are conducted, and the water-soluble beta glucan is obtained. By means of the method, the problem of water solubility of the beta glucan is solved, and absorption, utilization and product development of the pleurotus ferulae lenzi beta sporocarp are more facilitated.

Description

A kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore
Technical field
the activeconstituents that the present invention relates to Chinese medicine extracts field, particularly a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore.
Background technology
resina Ferulae mushroom has another name called Pleurotus ferulae Lanzi, belong to Basidiomycotina, Hymenomycetes, Agaricales, Pleurotaceae, pleurotus is the edible mushrooms of the medicinal and edible dual-purpose that a kind of aridity grass land grows, be described as good names such as " the white glossy ganoderma in Western Paradise " " boletes on grassland ", be mainly distributed in the ground such as India, France, Xinjiang, China.Nineteen eighty-three China scientific worker carries out artificial training cultivation to wild edible Resina Ferulae mushroom and obtains successfully.At present, started in Barkol county to promote, and obtained good economic and social benefit.
the sporophore of Resina Ferulae mushroom is as white as polished jade, fine and tender taste, plump crisp cunning, people attacked by giving off a strong fragrance, unique flavor, detect through state food quality surveillance inspection center and show: Resina Ferulae mushroom contains the trace elements such as very high fungus polysaccharide, selenium, germanium, chromium, Vc, Ve, Vd, Va unsaturated fatty acids, and 17 seed amino acids of needed by human.Modern pharmacology experiment proves, fungus polysaccharide has enhancing body immunity, physiological equilibrium, anticancer, the effect of anti-cancer, unsaturated fatty acids, SOD has prevention of arterial to harden, lowering blood pressure, antidotal effect, Methionin, arginine has protection liver, strengthen effect of children ' s intelligence development, Vd, Va can promote human calcium, phosphorus absorbs, prevent children rachitis, the choice drug of richets and middle-aged and old osteoporosis, stomach trouble is being treated when the Resina Ferulae mushroom that utilizes among the people on ground, typhoid fever, the various diseases such as reducing blood-fat what, especially the blackbone Chicken cooking is edible, to pregnant woman, postpartum what blood, the large benefit effects such as tonifying Qi is empty.
along with the application of Resina Ferulae mushroom is more and more extensive, the activeconstituents of Resina Ferulae mushroom is extracted in order to a more popular problem.Resina Ferulae mushroom polysaccharide is one of main active ingredient of Resina Ferulae mushroom, and the extracting solution for Resina Ferulae mushroom polysaccharide has had some to report.But these documents are all the extraction to Resina Ferulae mushroom total polysaccharides, Resina Ferulae mushroom polysaccharide coexists with α and β two kinds of configurations in Resina Ferulae mushroom, the separation of these two kinds of configurations all do not mentioned by the current document for the technical study of Resina Ferulae mushroom total polysaccharides, only start with from extraction total polysaccharides itself, the total polysaccharides extracted is further purified, is separated the polysaccharide all one obtaining a certain configuration.
polysaccharide molecular weight from several thousand to millions of not etc., molecular weight is larger, and it is water-soluble poorer, and Resina Ferulae mushroom polysaccharide is no exception.Resina Ferulae mushroom polysaccharide macro-molecular amount part aqueous is poor, makees solvent be difficult to extract completely in separation and Extraction with water, even if use alkali lye to extract, although extraction yield is high, is still not dissolved in water when obtaining polysaccharide product application after drying.Human body absorbs bad for this macromole Resina Ferulae mushroom polysaccharide.
Summary of the invention
the object of the present invention is to provide a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore, solve the water solubility problems of beta glucan, more be conducive to absorbing and product development of Resina Ferulae mushroom beta glucan, improve the yield of Resina Ferulae mushroom polysaccharide, add water-soluble, be more conducive to absorption of human body.
object of the present invention is achieved through the following technical solutions:
from Resina Ferulae mushroom sporophore, extract a method for water-soluble beta dextran, comprise the following steps:
step 1, prepare Resina Ferulae mushroom Crude polysaccharides:
get Resina Ferulae mushroom sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains Resina Ferulae mushroom Crude polysaccharides;
step 2, prepare Resina Ferulae mushroom beta glucan:
get Resina Ferulae mushroom Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain Resina Ferulae mushroom beta glucan;
step 3, prepare water-soluble beta dextran:
get Resina Ferulae mushroom beta glucan, add alkali lye and stir extraction, extracting solution acid adjustment basicity is to pH3.5-6.8, and heating hydrolysis, hydrolyzed solution filters, and adds ethanol in filtrate, leaves standstill, and precipitation separation is also dry, obtains water-soluble beta dextran.
in described step 1, cross 40 object sieves after Resina Ferulae mushroom sporophore being pulverized, obtain powder.
in described step 1,1-10 hour is got in the alkali lye lixiviate adding 5-20 times of weight in powder, and the concentration of alkali lye used is preferably 0.1-1.0 mol/L.
in described step 1, filter and adopt filter cloth to filter, wherein filter cloth aperture is 0.5-5 micron.
in described step 1 or step 3, the dehydrated alcohol adding 1-4 times of volume in filtrate leaves standstill 2 hours.
in described step 2, the water adding 5-10 times of weight after the drying of Resina Ferulae mushroom Crude polysaccharides stirs, and fully dissolves.
in described step 3, concentration of lye is 0.1-0.4 mol/L; Described heating hydrolysis is preferably heated to 80-100 DEG C, and hydrolysis time is 1-4 hour.
beneficial effect of the present invention:
1, the suitability for industrialized production problem extracting beta glucan in Resina Ferulae mushroom sporophore is solved.What prior art extracted extraction usually is total polysaccharides, and method of the present invention extraction obtains beta glucan, and can be applicable to industrial production.
2, solve the water solubility problems of beta glucan, be more conducive to absorbing and product development of Resina Ferulae mushroom beta glucan.The water insoluble the present invention of the dextran that molecular weight is large utilizes acid hydrolysis technology to reduce the molecular weight of macromolecule Resina Ferulae mushroom polysaccharide.Improve the yield of Resina Ferulae mushroom polysaccharide, add water-soluble, be more conducive to absorption of human body.
Embodiment
embodiment 1
take Resina Ferulae mushroom sporophore 1 kilogram pulverizing, cross 50 objects sieves, powder adds sodium hydroxide solution 5 liters of lixiviates 1 hour of 0.1 mol/L, and the plate basket of 0.5 micron pore size filter cloth is crossed and filtered residue, obtains filtrate 4.7 liters.Filtrate adds the dehydrated alcohol of 18.8 liters, leaves standstill after 2 hours and filters to obtain precipitation with the plate basket of 0.5 micron pore size cloth, weigh totally 150 grams after precipitation drying.Precipitation adds 750 ml water stirring at room temperature 10 minutes, adds 1 gram of Fungal Alpha amylase liquid (Shandong Jie Nuo biological enzyme company limited), and with the salt acid for adjusting pH value to 5.5 of 6 mol/L, 50 degrees Celsius are incubated 24 hours.Then filter with the plate basket of 0.5 micron pore size cloth, discard filtrate, residue adds 0.8 liter, 1.0 mol/L sodium hydroxide and stirs extraction 2 hours, extracting solution acid adjustment basicity is to pH2.0, in 90 degrees Centigrade sealing hydrolysis 1 hour, the plate basket of hydrolyzed solution 0.5 micron pore size cloth filters, filtrate adds 3.2 liters of dehydrated alcohols, leave standstill precipitation separation after 2 hours and drying is weighed as 110 grams of infrared spectras shows that this product is beta glucan, it is 45% that Phenol-sulphate acid method surveys polysaccharide content, and it is 2060 dalton that high performance liquid phase gel chromatography records molecular-weight average; After testing, obtain beta glucan and have good water-soluble.
embodiment 2
take Resina Ferulae mushroom sporophore 1 kilogram pulverizing, cross 50 objects sieves, powder adds sodium hydroxide solution 10 liters of lixiviates 5 hours of 1 mol/L, and the plate basket of 0.5 micron pore size filter cloth is crossed and filtered residue, obtains filtrate 7.2 liters.Filtrate adds the dehydrated alcohol of 28.8 liters, leaves standstill after 2 hours and filters to obtain precipitation with the plate basket of 0.5 micron pore size cloth, weigh totally 142 grams after precipitation drying.Precipitation adds 1.4 premium on currency stirring at room temperature 10 minutes, adds 0.6 gram of Fungal Alpha amylase liquid (purchased from Cheng Xing bio tech ltd, Wuxi), and with the salt acid for adjusting pH value to 5.8 of 2 mol/L, 50 degrees Celsius are incubated 24 hours.Then filter with the plate basket of 0.5 micron pore size cloth, discard filtrate, residue adds 0.8 liter, 1.0 mol/L sodium hydroxide and stirs extraction 2 hours, extracting solution acid adjustment basicity is to pH6.8, in 100 degrees Centigrade sealing hydrolysis 4 hours, the plate basket of hydrolyzed solution 0.5 micron pore size cloth filtered, and filtrate adds 3.2 liters of dehydrated alcohols, leave standstill precipitation separation drying is weighed as 102 grams after 2 hours, be thing of the present invention.Infrared spectra shows that this product is beta glucan, and it is 48% that Phenol-sulphate acid method surveys polysaccharide content, and it is 198056 dalton that high performance liquid phase gel chromatography records molecular-weight average; After testing, obtain beta glucan and have good water-soluble.
embodiment 3
take Resina Ferulae mushroom sporophore 1 kilogram pulverizing, cross 50 objects sieves, powder adds sodium hydroxide solution 20 liters of lixiviates 10 hours of 0.5 mol/L, and the plate basket of 2 micron pore size filter clothes is crossed and filtered residue, obtains filtrate 19.5 liters.Filtrate adds the dehydrated alcohol of 78 liters, leaves standstill after 2 hours and filters to obtain precipitation with the plate basket of 2 micron pore size cloth, weigh totally 143 grams after precipitation drying.Precipitation adds 1 premium on currency stirring at room temperature 10 minutes, adds 1 gram of Fungal Alpha amylase liquid (Shandong Jie Nuo biological enzyme company limited), pH value to 5.5, and 50 degrees Celsius are incubated 24 hours.Then filter with the plate basket of 4 micron pore size cloth, discard filtrate, residue adds 0.8 liter, 0.6 mol/L sodium hydroxide and stirs extraction 2 hours, extracting solution acid adjustment basicity is to pH5.8, in 80 degrees Centigrade sealing hydrolysis 2 hours, the plate basket of hydrolyzed solution 2 micron pore size cloth filtered, and filtrate adds 3.2 liters of dehydrated alcohols, leave standstill precipitation separation drying is weighed as 101 grams after 2 hours, be thing of the present invention.Infrared spectra shows that this product is beta glucan, and it is 40% that Phenol-sulphate acid method surveys polysaccharide content, and it is 40460 dalton that high performance liquid phase gel chromatography records molecular-weight average; After testing, obtain beta glucan and have good water-soluble.
embodiment 4
the specific descriptions of extracting method of the present invention:
a, Resina Ferulae mushroom sporophore are pulverized and are obtained powder, powder alkali lye lixiviate, and filter cleaner, adds ethanol in filtrate, and leave standstill, filtration is precipitated; This precipitation and Resina Ferulae mushroom Crude polysaccharides.Resina Ferulae mushroom beta glucan is a part for Resina Ferulae mushroom Crude polysaccharides, its quite a few be positioned on the cell walls of fungi, in order to extract Resina Ferulae mushroom beta glucan to greatest extent, the present invention selects alkali lye to extract, alkali lye dosage is 5-20 times of raw material weight, the concentration of alkali lye chooses arbitrary concentration in 0.1-1.0 mol/L, extraction time 1-10 hour.Alkali lye can use sodium hydroxide, potassium hydroxide, zinc hydroxide, and the preparations such as organic bases, consider Cost Problems, and alkali lye generally chooses sodium hydroxide solution.Extracting liquid filtering removing residue, filtrate adds the dehydrated alcohol of 1-4 times of volume, leaves standstill and filters to obtain precipitation and Resina Ferulae mushroom raw sugar after 2 hours.
add water after b, precipitation drying stirring, and then add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate and obtains residue.
the residue of this step and Resina Ferulae mushroom beta glucan but water-soluble poor.This step adopts the precipitation (Resina Ferulae mushroom Crude polysaccharides) in previous step to be raw material, add α-amylase (because different manufacturers product enzyme activity is different, dosage difference is huge, and those skilled in the art can determine according to the vigor of practical situation and enzyme the consumption adding α-amylase; Consumption is more or a little lessly do not affect effect of the present invention) undissolved for water α configuration polysaccharide hydrolysis is become solvable, discard hydrolyzed solution, simultaneously hydrolyzed solution has also dissolved most of residual monosaccharide in Crude polysaccharides and oligosaccharides, play the effect of purifying, although also a small amount of beta glucan may be there is in hydrolyzed solution, but consider cost recovery, therefore cast out.Only being for further processing to a large amount of beta glucans in residue makes it become solubilized.α-amylase chooses commercially available various models.Hydrolysising condition, enzyme dosage is according to the α-amylase specification sheets of each producer different model.
c, residue add alkali lye and stir extraction, and extracting solution acid adjustment degree is to pH2.0-6.8, and heating hydrolysis, hydrolyzed solution filters, and adds ethanol in filtrate, leaves standstill, and precipitation separation is also dry, to obtain final product.
the residue adopting step b) is raw material, add alkali lye to extract, alkali lye dosage is 5-20 times of raw material weight, extracting solution acid adjustment alkali is to pH2.0-6.8,80-100 DEG C of heating hydrolysis 1-4 hour, hydrolyzed solution filters, and the dehydrated alcohol that filtrate adds 1-4 times of volume precipitates, precipitation separation drying is product of the present invention, namely Resina Ferulae mushroom mushroom water-soluble beta dextran afterwards.
alkali alkali lye can use sodium hydroxide, potassium hydroxide, zinc hydroxide, the preparations such as organic bases, preferred sodium hydroxide, and liquid concentration chooses arbitrary concentration in 0.1-1.0 mol/L.PH can use hydrochloric acid, acetic acid, sulfuric acid, acetic acid etc.Any one value in pH2.0-6.8, just different pH, hydrolysis intensity is different, and after hydrolysis, gained final product weight-average molecular weight is different.
carry out being separated and being further purified obtaining high purity polysaccharide to a certain configuration of Resina Ferulae mushroom mushroom although have in prior art, but its separation method and the present invention have essential distinction, at method many uses column chromatography existing after the water extract-alcohol precipitation of routine, Crude polysaccharides is separated, clip a certain specific part in Crude polysaccharides and obtain highly purified Resina Ferulae mushroom polysaccharide for scientific research, this and industrial method of the present invention have remarkable difference, and the present invention is except carrying out except separation and Extraction to the beta comfiguration of Resina Ferulae mushroom polysaccharide, also water-soluble by improve it to the molecular weight control of Resina Ferulae mushroom polysaccharide.The present invention is by acid hydrolysis, and the molecular weight that the method that water redissolves controls Resina Ferulae mushroom polysaccharide enhances the water-soluble of Resina Ferulae mushroom polysaccharide within the specific limits, improves the yield of Resina Ferulae mushroom polysaccharide macro-molecular amount section, is conducive to suitability for industrialized production Resina Ferulae mushroom β water-soluble polysaccharide.
extracting method of the present invention to obtain Resina Ferulae mushroom fruitbody polysaccharide be the beta glucan eliminating α configuration polysaccharide, and plant beta glucan and have good water-soluble.It is beta glucan that infrared spectra shows product of the present invention, and Phenol-sulphate acid method detects polysaccharide content and is greater than 40%, and polysaccharide molecular-weight average 2000-200000 dalton is not etc.
the foregoing is only preferred embodiment of the present invention, is only illustrative for the present invention, and nonrestrictive.Those skilled in the art understand, and can carry out many changes in the scope of the spirit limited in the claims in the present invention to it, amendment, even equivalence, but all will fall within the scope of protection of the present invention.

Claims (7)

1. from Resina Ferulae mushroom sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
Step 1, prepare Resina Ferulae mushroom Crude polysaccharides:
Get Resina Ferulae mushroom sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains Resina Ferulae mushroom Crude polysaccharides;
Step 2, prepare Resina Ferulae mushroom beta glucan:
Get Resina Ferulae mushroom Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain Resina Ferulae mushroom beta glucan;
Step 3, prepare water-soluble beta dextran:
Get Resina Ferulae mushroom beta glucan, add alkali lye and stir extraction, extracting solution acid adjustment basicity is to pH3.5-6.8, and heating hydrolysis, hydrolyzed solution filters, and adds ethanol in filtrate, leaves standstill, and precipitation separation is also dry, obtains water-soluble beta dextran.
2. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, is characterized in that: in described step 1, crosses 40 object sieves, obtain powder after Resina Ferulae mushroom sporophore being pulverized.
3. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, it is characterized in that: in described step 1,1-10 hour is got in the alkali lye lixiviate adding 5-20 times of weight in powder, and the concentration of alkali lye used is preferably 0.1-1.0 mol/L.
4. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, is characterized in that: in described step 1, and filter and adopt filter cloth to filter, wherein filter cloth aperture is 0.5-5 micron.
5. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, is characterized in that: in described step 1 or step 3, and the dehydrated alcohol adding 1-4 times of volume in filtrate leaves standstill 2 hours.
6. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, is characterized in that: in described step 2, and the water adding 5-10 times of weight after the drying of Resina Ferulae mushroom Crude polysaccharides stirs, and fully dissolves.
7. a kind of method extracting water-soluble beta dextran from Resina Ferulae mushroom sporophore according to claim 1, is characterized in that: in described step 3, and concentration of lye is 0.1-0.4 mol/L; Described heating hydrolysis is preferably heated to 80-100 DEG C, and hydrolysis time is 1-4 hour.
CN201510948166.0A 2015-12-17 2015-12-17 Method for extracting water-soluble beta glucan from pleurotus ferulae lenzi sporocarp Pending CN105368899A (en)

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Citations (4)

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Publication number Priority date Publication date Assignee Title
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CN101870740A (en) * 2010-06-13 2010-10-27 华南师范大学 A kind of extracellular polysaccharide extract of Morchella apex and its preparation method and application
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Publication number Priority date Publication date Assignee Title
CN1192247A (en) * 1995-07-05 1998-09-02 卡尔顿和联合酿酒有限公司 Production of β-glucan-mannan preparations by autolysis of cells under certain pH, temperature and time conditions
CN101597342A (en) * 2009-07-14 2009-12-09 哈尔滨众生北药生物工程有限公司 From Hericium erinaceus, extract method of water-soluble beta dextran and products thereof
CN101870740A (en) * 2010-06-13 2010-10-27 华南师范大学 A kind of extracellular polysaccharide extract of Morchella apex and its preparation method and application
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