CN105384844A - Method for extracting water soluble beta-glucan from bamboo fungus sporophore - Google Patents
Method for extracting water soluble beta-glucan from bamboo fungus sporophore Download PDFInfo
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- CN105384844A CN105384844A CN201510948161.8A CN201510948161A CN105384844A CN 105384844 A CN105384844 A CN 105384844A CN 201510948161 A CN201510948161 A CN 201510948161A CN 105384844 A CN105384844 A CN 105384844A
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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Abstract
The present invention discloses a method for extracting water soluble beta-glucan from bamboo fungus sporophore, the water soluble beta-glucan is prepared by the steps of bamboo fungus crude polysaccharide preparation, bamboo fungus beta-glucan preparation, water soluble beta-glucan preparation, and the like, bamboo fungus sporophore polysaccharide prepared by the method is beta-glucan with alpha-configuration being removed, and the beta-glucan has good water solubility. Infrared spectroscopy shows that the product is the beta-glucan, phenol-sulfuric method detects that the polysaccharide content is greater than 50%, and the polysaccharide average molecular weight is 2000-200000 Dalton in variety. In the prior art, a particular configuration of bamboo fungus is separated and further purified to obtain high purity polysaccharide, but the separation process is essentially different from the method, according to the method, by acid hydrolysis and water re-dissolution method, the molecular weight of the bamboo fungus polysaccharide is controlled within a certain range for enhancement of the water-solubility of the bamboo fungus polysaccharide, the yield of high molecular weight segment bamboo fungus polysaccharide is improved, and industrial production of bamboo fungus beta-water-soluble polysaccharide is facilitated.
Description
Technical field
the present invention relates to and a kind ofly from dictyophora phalloidea sporophore, extract method of water-soluble beta dextran and products thereof.The activeconstituents belonging to Chinese medicine extracts field.
Background technology
dictyophora phalloidea is one of most precious in the world edible mushrooms, just be listed in one of " careless eight delicacies " from ancient times, it has bottle-green cap, the columned stem of snowy white, peach egg type volva, have on stem top and enclose careful pure white netted skirt and spread out downwards from cap, be called " snow skirt fairy maiden ", " flower of mountain delicacy ", " flower of fungi ", " in bacterium queen " by people.Before the history that China eats dictyophora phalloidea can trace back to more than 1000 year.Chinese's numerous in the Meng is recorded and is said in " Shiliao Bencao ": " the cizu woods moon in summer meets rain, a juice work ground, and raw straw mat is like deer horn, white, edible ", this is the record the earliest about dictyophora phalloidea.Dictyophora phalloidea is cool in nature, and taste is sweet, and micro-hardship, is used as medicine from ancient times, has description in successive dynasties medicine ancient books and records more." Chinese Pharmacopoeia " is recorded, and dictyophora phalloidea has the several functions such as kidney invigorating and YANG supporting, beneficial stomach gut purge, anti-senility, anti-inflammatory analgetic, fat-reducing.The nutritional health function of modern science to dictyophora phalloidea has abundant theoretical investigation and experiment to prove, dictyophora phalloidea is a kind of high protein, low-fat natural health care, its thalline contains abundant nutritive ingredient, amino acid contained nearly 21 kinds, than any one edible mushrooms all at height, also has the different polysaccharide component such as multivitamin, trace element and semi-lactosi, glucose, seminose, wood sugar simultaneously.Edible dictyophora phalloidea can reduce the content of cholesterol in human body, reduces hoarding of Abdominal Wall Fat, and the long-term edible content that can also adjust hematic acid and lipid acid in body is hypotensive and natural non-defective unit that is reducing blood-fat.
along with the application of dictyophora phalloidea is more and more extensive, the activeconstituents of dictyophora phalloidea is extracted in order to a more popular problem.Dictyophora fungus polysaccharide is one of main active ingredient of dictyophora phalloidea, and the extracting solution for dictyophora fungus polysaccharide has had some to report.But these documents are all the extraction to dictyophora phalloidea total polysaccharides, dictyophora fungus polysaccharide coexists with α and β two kinds of configurations in dictyophora phalloidea, the separation of these two kinds of configurations all do not mentioned by the current document for the technical study of dictyophora phalloidea 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 dictyophora fungus polysaccharide is no exception.Dictyophora fungus polysaccharide macromolecule 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 dictyophora fungus polysaccharide.
Summary of the invention
the technical problem that the present invention mainly solves is: the suitability for industrialized production problem 1, extracting beta glucan in dictyophora phalloidea sporophore.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 dictyophora phalloidea 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 dictyophora fungus polysaccharide.Improve the yield of dictyophora fungus polysaccharide, add water-soluble, be more conducive to absorption of human body.
in order to solve the problems of the technologies described above, the object of the present invention is to provide a kind of method extracting water-soluble beta dextran from bamboo shoot sporophore.
from dictyophora phalloidea sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
step 1, preparation dictyophora phalloidea Crude polysaccharides:
get dictyophora phalloidea sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains dictyophora phalloidea Crude polysaccharides;
step 2, preparation dictyophora phalloidea beta glucan:
get dictyophora phalloidea Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain dictyophora phalloidea beta glucan;
step 3, prepare water-soluble beta dextran:
get dictyophora phalloidea beta glucan, add alkali lye and stir extraction, extracting solution acid adjustment basicity 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, obtains water-soluble beta dextran.
in described step 1, cross 50 object sieves after dictyophora phalloidea 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 dictyophora phalloidea 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.
the specific descriptions of extracting method of the present invention:
dictyophora phalloidea sporophore is pulverized and is obtained powder, powder alkali lye lixiviate, and filter cleaner, adds ethanol in filtrate, and leave standstill, filtration is precipitated; This precipitation and dictyophora phalloidea Crude polysaccharides.Dictyophora phalloidea beta glucan is a part for dictyophora phalloidea Crude polysaccharides, its quite a few be positioned on the cell walls of fungi, in order to extract dictyophora phalloidea 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 dictyophora phalloidea raw sugar after 2 hours.
add water after precipitation drying stirring, and then add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate and obtains residue.
the residue of this step and dictyophora phalloidea beta glucan but water-soluble poor.This step adopts the precipitation (dictyophora phalloidea 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.
residue adds alkali lye and stirs 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 bamboo shoot 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 bamboo shoot 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 dictyophora fungus 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 dictyophora fungus polysaccharide, also water-soluble by improve it to the molecular weight control of dictyophora fungus polysaccharide.The present invention is by acid hydrolysis, and the molecular weight that the method that water redissolves controls dictyophora fungus polysaccharide enhances the water-soluble of dictyophora fungus polysaccharide within the specific limits, improves the yield of dictyophora fungus polysaccharide macromolecule section, is conducive to suitability for industrialized production dictyophora phalloidea β water-soluble polysaccharide.
extracting method of the present invention to obtain dictyophora phalloidea 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 50%, and polysaccharide molecular-weight average 2000-200000 dalton is not etc.
Embodiment
below in conjunction with embodiment, the invention will be further described, it should be understood that these embodiments only for the object of illustration, never limit the scope of the invention.
embodiment 1
from dictyophora phalloidea sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
step 1, preparation dictyophora phalloidea Crude polysaccharides:
get dictyophora phalloidea sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains dictyophora phalloidea Crude polysaccharides;
step 2, preparation dictyophora phalloidea beta glucan:
get dictyophora phalloidea Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain dictyophora phalloidea beta glucan;
step 3, prepare water-soluble beta dextran:
get dictyophora phalloidea beta glucan, add alkali lye and stir extraction, extracting solution acid adjustment basicity 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, obtains water-soluble beta dextran.
in described step 1, cross 50 object sieves after dictyophora phalloidea 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 dictyophora phalloidea 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.
embodiment 2
take dictyophora phalloidea 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 4.6 liters.Filtrate adds the dehydrated alcohol of 18.4 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 65% 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 3
take dictyophora phalloidea 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 8.3 liters.Filtrate adds the dehydrated alcohol of 33.2 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 58% 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 4
take dictyophora phalloidea 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 16.8 liters.Filtrate adds the dehydrated alcohol of 67.2 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 60% 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.
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 dictyophora phalloidea sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
Step 1, preparation dictyophora phalloidea Crude polysaccharides:
Get dictyophora phalloidea sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains dictyophora phalloidea Crude polysaccharides;
Step 2, preparation dictyophora phalloidea beta glucan:
Get dictyophora phalloidea Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain dictyophora phalloidea beta glucan;
Step 3, prepare water-soluble beta dextran:
Get dictyophora phalloidea beta glucan, add alkali lye and stir extraction, extracting solution acid adjustment basicity 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, obtains water-soluble beta dextran.
2. a kind of method extracting water-soluble beta dextran from dictyophora phalloidea sporophore according to claim 1, is characterized in that: in described step 1, crosses 50 object sieves, obtain powder after dictyophora phalloidea sporophore being pulverized.
3. a kind of method extracting water-soluble beta dextran from dictyophora phalloidea 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 dictyophora phalloidea 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 dictyophora phalloidea 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 dictyophora phalloidea 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 dictyophora phalloidea Crude polysaccharides stirs, and fully dissolves.
7. a kind of method extracting water-soluble beta dextran from dictyophora phalloidea 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.
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| CN1165148A (en) * | 1996-05-10 | 1997-11-19 | 中国科学院大连化学物理研究所 | Method for preparing dictyophora fungus polysaccharide |
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| CN101503477A (en) * | 2008-09-28 | 2009-08-12 | 武汉理工大学 | Method for extracting triple helix Dictyophora phalloidea polysaccharide |
| 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 |
| CN103044566A (en) * | 2013-01-04 | 2013-04-17 | 合肥工业大学 | Method for preparing antioxidant polysaccharide from dictyophora indusiata aqueous extract residues |
| CN104479040A (en) * | 2014-12-21 | 2015-04-01 | 曾丹 | Process for extracting bamboo fungus polysaccharide |
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2015
- 2015-12-17 CN CN201510948161.8A patent/CN105384844A/en active Pending
Patent Citations (6)
| 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 'beta'-glucan-mannan preparations by autolysis of cells under certain pH, temperature and time conditions |
| CN1165148A (en) * | 1996-05-10 | 1997-11-19 | 中国科学院大连化学物理研究所 | Method for preparing dictyophora fungus polysaccharide |
| CN101503477A (en) * | 2008-09-28 | 2009-08-12 | 武汉理工大学 | Method for extracting triple helix Dictyophora phalloidea polysaccharide |
| 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 |
| CN103044566A (en) * | 2013-01-04 | 2013-04-17 | 合肥工业大学 | Method for preparing antioxidant polysaccharide from dictyophora indusiata aqueous extract residues |
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