CN105481994A - Method for extracting water-soluble beta-glucan from poria cocos fungus entities - Google Patents
Method for extracting water-soluble beta-glucan from poria cocos fungus entities Download PDFInfo
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- CN105481994A CN105481994A CN201510948152.9A CN201510948152A CN105481994A CN 105481994 A CN105481994 A CN 105481994A CN 201510948152 A CN201510948152 A CN 201510948152A CN 105481994 A CN105481994 A CN 105481994A
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- C—CHEMISTRY; METALLURGY
- 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 invention provides a method for extracting water-soluble beta-glucan from poria cocos fungus entities. The method comprises the following steps of 1 poria cocos fungus crude polysaccharide preparing, wherein the poria cocos fungus entities are taken and smashed to obtain powder, the powder is leached with alkali liquor and filtered to remove residues, ethyl alcohol is added in filtrate, standing and filtering are performed to obtain a precipitate, and poria cocos fungus crude polysaccharide is obtained; 2 poria cocos fungus beta-glucan preparing, wherein the poria cocos fungus crude polysaccharide is taken, dried and then stirred by adding water, alpha-amylase is added for hydrolysis, hydrolysate is filtered, filtrate is discarded to obtain residues, and poria cocos fungus beta-glucan is obtained; 3 water-soluble beta-glucan preparing, wherein the poria cocos fungus beta-glucan is taken and stirred and extracted by adding alkali liquor, the pH of an extracting solution is regulated to 2.0-6.8, heating hydrolysis is performed, hydrolysate is filtered, ethyl alcohol is added in filtrate, standing, separating and precipitating and drying are performed, and the water-soluble beta-glucan is obtained. According to the method, the problem of the water solubility of the beta-glucan is solved, absorption and utilization of the poria cocos beta-glucan are better facilitated, and the yield of the poria cocos polysaccharide is increased.
Description
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 Poria fungus sporophore.
Background technology
poria cocos, also known as jade is clever, Fu is clever, Wan Linggui, Fu Tu.The dry sclerotia of Shi Ni shelf fungus section fungi Poria cocos, often colonizes on pine root.Polysaccharide is mainly pachyman (pachyman), content reaches as high as 75%, for one has β (1 → 3) the Glucopyranose glycan of β (1 → 6) Glucopyranose glycan sidechain, cut off side chain and become β (1 → 3) glucose glycan, claim pachymaran (pachymaran), often be called Pachymose (PPS), tool anti-tumor activity. carboxymethyl pachymose tool immunological enhancement and antitumor action.
along with the application of Poria cocos is more and more extensive, the activeconstituents of Poria cocos is extracted in order to a more popular problem.Pachymose is one of main active ingredient of Poria cocos, and the extracting solution for Pachymose has had some to report.But these documents are all the extraction to Poria cocos total polysaccharides, Pachymose coexists with α and β two kinds of configurations in Poria cocos, the separation of these two kinds of configurations all do not mentioned by the current document for the technical study of Poria cocos 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 Pachymose is no exception.Pachymose 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 Pachymose.
Summary of the invention
the object of the present invention is to provide a kind of method extracting water-soluble beta dextran from Poria fungus sporophore, solve the water solubility problems of beta glucan, more be conducive to absorbing and product development of Poria fungus beta glucan, improve the yield of Pachymose, add water-soluble, be more conducive to absorption of human body.
object of the present invention is achieved through the following technical solutions:
from Poria fungus sporophore, extract a method for water-soluble beta dextran, comprise the following steps:
step 1, prepare Poria fungus Crude polysaccharides:
get Poria fungus sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains Poria fungus Crude polysaccharides;
step 2, prepare Poria fungus beta glucan:
get Poria fungus Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain Poria fungus beta glucan;
step 3, prepare water-soluble beta dextran:
get Poria fungus 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 Poria fungus 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 Poria fungus 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 Poria fungus 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 Poria fungus 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 Pachymose; Improve the yield of Pachymose, add water-soluble, be more conducive to absorption of human body.
Embodiment
embodiment 1
take Poria fungus 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 4.2 liters.Filtrate adds the dehydrated alcohol of 16.8 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 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 2
take Poria fungus 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 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 3
take Poria fungus 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 17.2 liters.Filtrate adds the dehydrated alcohol of 68.8 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.
embodiment 4
the specific descriptions of extracting method of the present invention:
a, Poria fungus 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 Poria fungus Crude polysaccharides.Poria fungus beta glucan is a part for Poria fungus Crude polysaccharides, its quite a few be positioned on the cell walls of fungi, in order to extract Poria fungus 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 Poria fungus 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 Poria fungus beta glucan but water-soluble poor.This step adopts the precipitation (Poria fungus 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 Poria cocos 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 Poria cocos 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 Poria 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 Pachymose, also water-soluble by improve it to the molecular weight control of Pachymose.The present invention is by acid hydrolysis, and the molecular weight that the method that water redissolves controls Pachymose enhances the water-soluble of Pachymose within the specific limits, improves the yield of Pachymose macromolecule section, is conducive to suitability for industrialized production Poria cocos β water-soluble polysaccharide.
extracting method of the present invention to obtain Poria fungus 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.
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 Poria fungus sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
Step 1, prepare Poria fungus Crude polysaccharides:
Get Poria fungus sporophore, pulverize and obtain powder, powder alkali lye lixiviate, cross and filter residue, add ethanol in filtrate, leave standstill, filtration is precipitated, and obtains Poria fungus Crude polysaccharides;
Step 2, prepare Poria fungus beta glucan:
Get Poria fungus Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain Poria fungus beta glucan;
Step 3, prepare water-soluble beta dextran:
Get Poria fungus 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 Poria fungus sporophore according to claim 1, is characterized in that: in described step 1, crosses 50 object sieves, obtain powder after Poria fungus sporophore being pulverized.
3. a kind of method extracting water-soluble beta dextran from Poria fungus 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 Poria fungus 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 Poria fungus 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 Poria fungus 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 Poria fungus Crude polysaccharides stirs, and fully dissolves.
7. a kind of method extracting water-soluble beta dextran from Poria fungus 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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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106562203A (en) * | 2016-11-13 | 2017-04-19 | 合肥学院 | Preparation method of modified poria cocos noodles |
| CN112390899A (en) * | 2020-11-11 | 2021-02-23 | 天津科技大学 | Preparation method of blend of pregelatinized starch and pachyman alkaline hydrolysis fragment and application of blend in weight-reducing meal replacement food |
| CN115176890A (en) * | 2022-06-29 | 2022-10-14 | 瑞普高科(天津)生物技术有限公司 | Application of poria beta-1,3-D-glucan in preparation of feed additive for improving growth performance |
| CN115850530A (en) * | 2022-12-24 | 2023-03-28 | 杭州时光肌生物科技有限公司 | Preparation method of poria polysaccharide |
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| CN102603916A (en) * | 2012-03-21 | 2012-07-25 | 丁友玲 | Refining method of (1-3)-beta-D-glucan |
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| CN101757045A (en) * | 2008-11-25 | 2010-06-30 | 天津太平洋制药有限公司 | Preparation method of pachyman |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106562203A (en) * | 2016-11-13 | 2017-04-19 | 合肥学院 | Preparation method of modified poria cocos noodles |
| CN112390899A (en) * | 2020-11-11 | 2021-02-23 | 天津科技大学 | Preparation method of blend of pregelatinized starch and pachyman alkaline hydrolysis fragment and application of blend in weight-reducing meal replacement food |
| CN115176890A (en) * | 2022-06-29 | 2022-10-14 | 瑞普高科(天津)生物技术有限公司 | Application of poria beta-1,3-D-glucan in preparation of feed additive for improving growth performance |
| CN115850530A (en) * | 2022-12-24 | 2023-03-28 | 杭州时光肌生物科技有限公司 | Preparation method of poria polysaccharide |
| CN115850530B (en) * | 2022-12-24 | 2024-04-16 | 杭州时光肌生物科技有限公司 | Preparation method of poria cocos polysaccharide |
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Application publication date: 20160413 |