CN105367683A - Method for extracting water-soluble beta glucan from black fungus sporocarp - Google Patents
Method for extracting water-soluble beta glucan from black fungus sporocarp Download PDFInfo
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- CN105367683A CN105367683A CN201510948129.XA CN201510948129A CN105367683A CN 105367683 A CN105367683 A CN 105367683A CN 201510948129 A CN201510948129 A CN 201510948129A CN 105367683 A CN105367683 A CN 105367683A
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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 invention provides a method for extracting water-soluble beta glucan from black fungus sporocarp. The method includes the steps: 1, preparation of black fungus crude polysaccharide, wherein the black fungus sporocarp is picked and crushed, powder is obtained, by weight, 2-8% of activated carbon is added to the powder, extraction is conducted with alkali liquor, filtration is conducted to remove residues, ethyl alcohol is added to the filter liquor, the filter liquor stands still, filtration is conducted, sediment is obtained, and the black fungus crude polysaccharide is obtained; 2, preparation of black fungus beta sporocarp, wherein the black fungus crude polysaccharide is picked and dried, then water is added to the dried black fungus crude polysaccharide and stirred, alpha amylase is added for hydrolysis, hydrolysis liquor is filtered, filter liquor is removed, residues are obtained, and the black fungus beta sporocarp is obtained; 3, preparation of water-soluble beta sporocarp, wherein the black fungus beta sporocarp is picked, alkali liquor is added, stirring and extraction are conducted, the pH value of the extraction liquor is adjusted to be 2.0-6.8, heating and hydrolysis are conducted, hydrolysis liquor is filtered, ethyl alcohol is added to the filter liquor, the mixture stands still, separation, sediment and drying are conducted, and the water-soluble beta sporocarp is obtained. By means of the method, the yield of black fungus sporocarp is raised, water solubility is increased, and absorption by a human body is more facilitated.
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 black fungus sporophore.
Background technology
black fungus has another name called black dish, mulberry ear, this bacterium, tree chicken, wooden moth, wood young pilose antler etc., because of likeness in form ear, its color chocolate and gaining the name in addition, the former plant of black fungus is Mycophyta Basidiomycotina Auriculariaceae Auricularia plant auricularia auriculajudae Auriculariaauricula (L.exHook.) Underwood.The traditional Chinese medical science thinks black fungus, and the flat taste of its property is sweet, people's stomach, large intestine channel.There is nourishing, moisturize, the effect of nourish blood beneficial stomach, promoting blood circulation and hemostasis, moistening lung, ease constipation.
the polysaccharide kind contained in black fungus is more, adopts different extraction and isolation means, can obtain different types of polysaccharide.The experimental results shows, Auricularia polycose has reducing blood-fat, anticoagulation, anti-oxidant, the multiple bioactive functions such as radioprotective, Tumor suppression, has now become the study hotspot in the fields such as food, medical treatment, health care.
along with the application of black fungus is more and more extensive, the activeconstituents of black fungus is extracted in order to a more popular problem.Auricularia polycose is one of main active ingredient of black fungus, and the extracting solution for Auricularia polycose has had some to report, such as Xia Erning, the biological activity of the Auricularia polycose of Chen Qionghua, China Medicine University's journal, 1989, Han Chunran, Ma Yongqiang, the extraction of the Auricularia polycose of Tang Juan and hypoglycemic activity, food and biotechnology journal, 2006, all be studied the extraction of Auricularia polycose in document, extracting method is also different.
but these documents are all in laboratory conditions to the Hydrolysis kinetics of black fungus total polysaccharides, Auricularia polycose coexists with α and β two kinds of configurations in black fungus, the separation of these two kinds of configurations all do not mentioned by the current document for the technical study of black fungus total polysaccharides, only start with from extraction total polysaccharides itself, after with laboratory condition separation and purification Auricularia polycose, the usual inapplicable industrial production of these methods.
polysaccharide molecular weight from several thousand to millions of not etc., molecular weight is larger, and it is water-soluble poorer, and Auricularia polycose is no exception.Auricularia polycose 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 the macromole Auricularia polycose of this type of poorly water-soluble.
Summary of the invention
the object of the present invention is to provide a kind of method extracting water-soluble beta dextran from black fungus sporophore, solve the water solubility problems of beta glucan, more be conducive to absorbing and product development of black fungus beta glucan, improve the yield of Auricularia polycose, add water-soluble, be more conducive to absorption of human body.
object of the present invention is achieved through the following technical solutions:
from black fungus sporophore, extract a method for water-soluble beta dextran, comprise the following steps:
step 1, prepare black fungus Crude polysaccharides:
get black fungus sporophore, pulverize and obtain powder, powder adds 2 ~ 8wt% and enters gac, uses alkali lye lixiviate, crosses and filters residue, add ethanol in filtrate, and leave standstill, filtration is precipitated, and obtains black fungus Crude polysaccharides;
step 2, prepare black fungus beta glucan:
get black fungus Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain black fungus beta glucan;
step 3, prepare water-soluble beta dextran:
get black 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 60 object sieves after black fungus sporophore being pulverized, obtain powder.
in described step 1,1-6 hour is got in the alkali lye lixiviate adding 25-40 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 black 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 black 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 black 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 Auricularia polycose.Improve the yield of Auricularia polycose, add water-soluble, be more conducive to absorption of human body.
Embodiment
embodiment 1
take black fungus sporophore 1 kilogram pulverizing, cross 60 objects sieves, the gac of powder mixing 50g, add sodium hydroxide solution 25 liters of lixiviates 1 hour of 0.1 mol/L, the plate basket of 0.5 micron pore size filter cloth is crossed and is filtered residue, obtains filtrate 6.2 liters.Filtrate adds the dehydrated alcohol of 20 liters, leaves standstill after 2 hours and filters to obtain precipitation with the plate basket of 0.5 micron pore size cloth, weigh totally 152 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 0.1 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 1.2 liters, 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 5 liters of dehydrated alcohols, leave standstill precipitation separation after 2 hours and drying is weighed as 103 grams of infrared spectras shows that this product is beta glucan, it is 71% that Phenol-sulphate acid method surveys polysaccharide content, and it is 5606 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 black fungus sporophore 1 kilogram pulverizing, cross 60 objects sieves, the gac of powder mixing 30g, add sodium hydroxide solution 30 liters of lixiviates 2 hours of 0.7 mol/L, the plate basket of 0.5 micron pore size filter cloth is crossed and is filtered residue, obtains filtrate 11.2 liters.Filtrate adds the dehydrated alcohol of 35 liters, leaves standstill after 2 hours and filters to obtain precipitation with the plate basket of 0.5 micron pore size cloth, weigh totally 122 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 68% that Phenol-sulphate acid method surveys polysaccharide content, and it is 497506 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 black fungus sporophore 1 kilogram pulverizing, cross 50 objects sieves, the gac of powder mixing 40g, add sodium hydroxide solution 40 liters of lixiviates 3 hours of 0.5 mol/L, the plate basket of 0.2 micron pore size filter cloth is crossed and is filtered residue, obtains filtrate 22.1 liters.Filtrate adds the dehydrated alcohol of 60 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 65% that Phenol-sulphate acid method surveys polysaccharide content, and it is 100460 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, black fungus sporophore pulverize obtain powder, powder adds 2 ~ 8wt%(mass percent) the lixiviate of gac alkali lye, filter cleaner, adds ethanol in filtrate, leave standstill, filtration is precipitated; This precipitation and black fungus Crude polysaccharides.Black fungus beta glucan is a part for black fungus Crude polysaccharides, its quite a few be positioned on the cell walls of fungi, in order to extract black fungus beta glucan to greatest extent, the present invention selects alkali lye to extract, alkali lye dosage is 25-40 times of raw material weight, the concentration of alkali lye chooses arbitrary concentration in 0.1-1.0 mol/L, extraction time 1-6 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 black 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 black fungus beta glucan but water-soluble poor.This step adopts the precipitation (black 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 black fungus 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 black fungus 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 Auricularia polycose 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 Auricularia polycose, also water-soluble by improve it to the molecular weight control of Auricularia polycose.The present invention is by acid hydrolysis, and the molecular weight that the method that water redissolves controls Auricularia polycose enhances the water-soluble of Auricularia polycose within the specific limits, improves the yield of Auricularia polycose macromolecule section, is conducive to suitability for industrialized production black fungus β water-soluble polysaccharide.
extracting method of the present invention to obtain black 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 100000-1000000 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 black fungus sporophore, extract a method for water-soluble beta dextran, it is characterized in that: comprise the following steps:
Step 1, prepare black fungus Crude polysaccharides:
Get black fungus sporophore, pulverize and obtain powder, powder adds 2 ~ 8wt% and enters gac, uses alkali lye lixiviate, crosses and filters residue, add ethanol in filtrate, and leave standstill, filtration is precipitated, and obtains black fungus Crude polysaccharides;
Step 2, prepare black fungus beta glucan:
Get black fungus Crude polysaccharides, add water after drying stirring, and add α-amylase hydrolysis, hydrolyzed solution filters, and discards filtrate, obtains residue, obtain black fungus beta glucan;
Step 3, prepare water-soluble beta dextran:
Get black 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 black fungus sporophore according to claim 1, is characterized in that: in described step 1, crosses 60 object sieves, obtain powder after black fungus sporophore being pulverized.
3. a kind of method extracting water-soluble beta dextran from black fungus sporophore according to claim 1, it is characterized in that: in described step 1,1-6 hour is got in the alkali lye lixiviate adding 25-40 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 black 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 black 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 black 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 black fungus Crude polysaccharides stirs, and fully dissolves.
7. a kind of method extracting water-soluble beta dextran from black 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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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN101712727A (en) * | 2009-11-24 | 2010-05-26 | 中国计量学院 | Method for extracting black fungus polysaccharide |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN101712727A (en) * | 2009-11-24 | 2010-05-26 | 中国计量学院 | Method for extracting black fungus polysaccharide |
Non-Patent Citations (2)
| Title |
|---|
| 邵秀芝 等: "《食品化学实验》", 31 December 2013, 郑州：郑州大学出版社 * |
| 韩晓倩 等: "黑木耳多糖脱色方法的研究", 《CIFST-中国食品科学技术学会第六届年会暨第五届东西方食品业高层论坛论文摘要集》 * |
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Application publication date: 20160302 |