CN102115505A - Method for extracting crude polysaccharide of agaricus blazei - Google Patents
Method for extracting crude polysaccharide of agaricus blazei Download PDFInfo
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- CN102115505A CN102115505A CN2009102452271A CN200910245227A CN102115505A CN 102115505 A CN102115505 A CN 102115505A CN 2009102452271 A CN2009102452271 A CN 2009102452271A CN 200910245227 A CN200910245227 A CN 200910245227A CN 102115505 A CN102115505 A CN 102115505A
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- agaricus blazei
- blazei murrill
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Abstract
The invention relates to a method for extracting the crude polysaccharide of the agaricus blazei, which comprises the steps of: frying by water, extracting by ultrasound, frying by water, centrifuging, precipitating by alcohol, and drying. The extraction ratio and the bioactivity of the polysaccharide of the agaricus blazei are obviously improved, the raw material is free of deformation and pollution during extracting, and the raw material can be abundantly utilized. The method is simple in technology and low in cost, thereby being capable of being industrially produced, popularized and applied.
Description
Technical field
The invention belongs to protective foods and medical technical field, be specially a kind of extracting method of Agaricus Blazei Murrill coarse polysaccharide, this method has been optimized extraction process, significantly improves the extraction yield and the biological activity of Agaricus blazei Murrill active polysaccharide, and raw material does not have sex change, pollution-free in the leaching process, the utilization ratio height.
Background technology
Agaricus blazei Murrill (Agaricus blazei Muril1) has another name called Xiao Song mushroom, Brazilian mushroom, belongs to Basidiomycotina, Hymenomycetes, and Agaricales, Agaricus edibilis, Agaricus originates in Brazil, is that a kind of food (medicine) of delicious food is used fungi.Protein and polysaccharide content are more than 2 times of mushroom in the Agaricus blazei Murrill sporophore, are a kind of contain saccharic and profuse edible mushroomss of protein, and wherein polysaccharide content is 6.55%.
Recent study finds, Agaricus blazei Murrill have the growth that suppresses tumour cell, antitumor, hypoglycemic, improve diabetes, decreasing cholesterol, the arteriosclerotic effect of improvement.Wherein α-(1,6) ramose β-(1,3) dextran can directly work to kill tumour by inducing apoptosis of tumour cell.Along with the further investigation of Agaricus Blazei Murrill polysaccharide function, more and more people study the extracting method of polysaccharide.Extracting method to polysaccharide mainly contains hot water extraction, solvent-extraction process, enzyme extraction method etc. at present.
1, hot water extraction
The polysaccharide great majority of biologically active are water miscible, can directly extract with the method for hot water lixiviate.Hot water extraction adopts hot water, the Different concentrations of alcohol precipitator method of different ratios.But, have much room for improvement because the hot water extraction time is long and efficient is low.
2, solvent-extraction process
Solvent-extraction process can be divided into diluted acid method, diluted alkaline method and organic solvent extraction method.When diluted acid extracted, the time should lack, and temperature should not surpass 50 ℃, but at present most researchers think in the extraction of polysaccharide, should avoid as far as possible using sour as vat liquor except that albumen.Alkaline process extracts and is better than acid system and conventional hot water extraction. and experimental results show that with oxalic acid, ammonium oxalate solution lixiviate Agaricus blazei Murrill, can improve the extraction yield of polysaccharide, but glycosidic link and the three-dimensional arrangement and the activity of soda acid extraction method destructible polysaccharide.
3, prozyme extraction method
Combined-enzyme method whole process is divided into enzymatic reaction cell wall breaking process and two stages of lixiviate. and it is enzymolysis structure of cell surface and intercellular connector that the fs mainly acts on, and with the stripping of part glucide; Subordinate phase had both had the enzyme of going out effect by improving temperature, made thing leaching yield increase in the born of the same parents that dissolve in hot water simultaneously.Prozyme extraction method a certain proportion of polygalacturonase, cellulase and the neutral proteases of adopting, this method has mild condition, the easy removal of impurity and extraction yield advantages of higher more.The major cause that the combined-enzyme method extraction yield is high is owing to help the stripping of water-soluble polysaccharide behind the sporophore cell wall breaking, and prozyme can make the cellulose degradation of cell walls, make and formerly serve water-fast polyose and be degraded into water-soluble polysaccharide, thereby can improve extraction yield. adopting combined-enzyme method to extract in addition can shorten extraction time. and combined-enzyme method lixiviate whole process only needs about 1.5h, be half of hot water lixiviate required time, and the water logging of extraction rate specific heat improves, also apparently higher than acid, alkali extraction.Adopt combined-enzyme method, adding entry, to extract the back effect fine.Shen Aiying, Zhang Yun insults etc. and prozyme-water extract method to be studied. and the former adopts the better simply enzyme process that directly adds, earlier with the fragmentation of sporophore dry powder, pH transfers to 6.5, add 2% prozyme, place 45 ℃ of insulation 40min, be warming up to 85 ℃ of enzymes that go out immediately and live, adding 85 ℃ of water to the ratio of dry powder and water is 1: 20, and insulation lixiviate 1h, centrifugal, concentrate, Sevage method isolating protein finally obtains 15.67% polysaccharide. and this method had both obtained higher extraction yield, simultaneously also shortened a lot of times. the latter then earlier with water-soluble polysaccharide by lixiviate 3h, centrifugation extracts, separate the residue polygalacturonase, cellulase and proteolytic enzyme carry out enzymolysis 1h, again by centrifugal, alcohol precipitation, drying is though obtain 16.67% polysaccharide at last. obtained very high extraction yield, but this method is time-consuming, complexity, the extraction cost height.
Patent ZL 200410089523.4 Agaricus blazei Murrill aqueous solution polysaccharide and preparation technology thereof and purposes have been introduced a kind of water-soluble polysaccharide extracting method, one is removed soluble small molecular impurity with extraction using alcohol earlier, use the water extraction water-soluble polysaccharide again, the water extract directly concentrates promptly; It two is the water body ultrafiltration technology.Polysaccharide content is about 85% in the last medicinal extract, and total extraction yield is 6.8%, though polysaccharide content is higher relatively in this method medicinal extract, total extraction yield is low, and this method time spent is longer.
Summary of the invention
For these reasons, the invention provides a kind of extracting method of Agaricus Blazei Murrill coarse polysaccharide, adopt after the pulverizing of Agaricus blazei Murrill sporophore, boiling, the method for supersound extraction is extracted Agaricus Blazei Murrill polysaccharide, has increased extraction efficiency.
The present invention realizes by technical scheme under passing through:
1, getting the Agaricus blazei Murrill sporophore cleans, dries, pulverizes;
2, take by weighing above-mentioned Agaricus blazei Murrill sporophore powder, add distilled water, decoct;
3, above-mentioned solution supersound extraction is carried out broken wall treatment to Agaricus blazei Murrill sporophore cell;
4, solution continues to decoct after the said extracted, and is centrifugal;
5, get above-mentioned centrifugal back supernatant concentration to 1/4 of original volume, add 2.5 times of amount ethanol sedimentations, centrifugal, the precipitation oven dry promptly gets Crude polysaccharides.
Sporophore powder after the above-mentioned pulverizing, powder diameter are controlled between the 1000-1500 μ m, preferred mistake 20 mesh sieves.
The ratio that adds distilled water in the above-mentioned technological step 2 is 1: 10-1: between 40, and preferred 1: 25.
Above-mentioned technological step 3 adopts ultrasonicly carries out broken wall treatment to the Agaricus blazei Murrill sporophore, ultrasonic power between 100-200W, preferred 140W.
6, in the above-mentioned technological step 3, the supersound extraction time is 10-20min, preferred 15min.
7, in the above-mentioned technological step 3, it is 60-100 ℃ that Crude polysaccharides extracts temperature, preferred 80 ℃.
8, the related decocting time of above-mentioned technological step is 1-2h, preferred 1.5h.
9, related centrifugal rotational speed is 2000-6000r/min in the above-mentioned technological step, preferred 4000r/min, and centrifugation time is 5-20min, preferred 10min.
Beneficial effect of the present invention:
1, raw material does not have sex change, pollution-free in the technology leaching process, and raw material can be fully utilized.
2, production technique is simple, and is with low cost, but industrialization promotion is used.
Embodiment
The preparation of embodiment one, Agaricus Blazei Murrill coarse polysaccharide:
1, gets Agaricus blazei Murrill sporophore 10kg, clean, dry, pulverize, cross 20 mesh sieves;
2, get powder after the pulverizing, add 250L, decoct 1.5h;
3, the solution supersound extraction after the above-mentioned decoction (extracting power 140W, extraction time 15min);
4, solution continues to decoct 1.5h after the said extracted;
5, the solution 4000r/min after will decocting, centrifugal 10min;
6, get above-mentioned centrifugal back supernatant concentration to 1/4 of original volume, add 2.5 times of amount ethanol sedimentations, centrifugal, the precipitation oven dry promptly gets Crude polysaccharides 1.68kg
The preparation of embodiment two, Agaricus Blazei Murrill coarse polysaccharide:
1, gets Agaricus blazei Murrill sporophore 10kg, clean, dry, pulverize, cross 20 mesh sieves;
2, get powder after the pulverizing, add water 200L, decoct 1.5h;
3, the solution supersound extraction after the above-mentioned decoction (extracting power 120W, extraction time 20min);
4, solution continues to decoct 1.5h after the said extracted;
5, the solution 4000r/min after will decocting, centrifugal 15min;
6, get above-mentioned centrifugal back supernatant concentration to 1/4 of original volume, add 2.5 times of amount ethanol sedimentations, centrifugal, the precipitation oven dry promptly gets Crude polysaccharides 1.62kg.
Embodiment three, be explanation effect of the present invention, adopt anthrone method to carry out assay to the prepared polysaccharide of the embodiment of the invention one, the polysaccharide that extracts of the prozyme extraction method of being introduced in the application background technology carries out assay relatively simultaneously.
3.1 test sample: sample 1: the Agaricus Blazei Murrill coarse polysaccharide of the embodiment of the invention one preparation
Sample 2: the Agaricus Blazei Murrill coarse polysaccharide that adopts 200410089523.4 introduction method preparations of patent ZL
3.2 reagent preparation
3.2.1 anthrone reagent: take by weighing anthrone 0.5g and be dissolved in the 25ml ethyl acetate, jolting dissolving, it is standby to put in the brown bottle close plug.
3.2.2 the preparation of reference substance solution is taken at 105 ℃ of dextrose anhydrous 0.1g that are dried to constant weight, accurate claims surely, puts in the 100ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, precision is measured 10ml, puts in the 100ml measuring bottle, and thin up shakes up to scale, promptly.
The preparation of need testing solution sampling I, sample 2 are an amount of, and precision takes by weighing 1.5g, puts in the 100ml measuring bottle, and it is an amount of to add water, supersound process adds water to scale, shakes up, and filters, precision is measured subsequent filtrate 50ml, puts in the 100ml measuring bottle, adds 30% solution of zinc sulfate 5ml, and heating is after 5 minutes in water-bath, under jolting, add yellow prussiate of potash test solution 5ml immediately, add water to scale after the cooling, shake up, filter, discard filtrate just, precision is measured subsequent filtrate 10ml, puts in the 100ml measuring bottle, add water to scale, shake up, promptly.
The assay method precision is measured reference substance solution and each 2ml of need testing solution, puts respectively in the 10ml measuring bottle, respectively adds 2% phenol solution 0.5ml, sulfuric acid 5ml, shake up, put in the water-bath and heated 15 minutes, put cold, be diluted with water to scale, shake up, the place measures optical density respectively at the 490nm wavelength, calculates, that is Agaricus Blazei Murrill coarse polysaccharide content.Measurement result sees Table 1.
Table 1, Agaricus Blazei Murrill coarse polysaccharide assay be table as a result
As can be seen from the above table, the Agaricus Blazei Murrill polysaccharide that adopts the inventive method to extract, polysaccharide content is suitable with the polysaccharide content that adopts combined-enzyme method to extract in the Crude polysaccharides, and extraction yield is suitable.Adopt extracting method of the present invention to extract Agaricus Blazei Murrill coarse polysaccharide, method is easy, saves time, and saves cost.
Claims (8)
1. the extracting method of an Agaricus Blazei Murrill coarse polysaccharide is characterized in that this method is to adopt following technological step to realize:
A, get that the Agaricus blazei Murrill sporophore is cleaned, oven dry, pulverize;
B, take by weighing above-mentioned Agaricus blazei Murrill sporophore powder, add distilled water, decoct;
C, above-mentioned solution supersound extraction are carried out broken wall treatment to Agaricus blazei Murrill sporophore cell;
Solution continues to decoct after d, the said extracted, and is centrifugal;
E, get above-mentioned centrifugal back supernatant concentration to 1/4 of original volume, add 2.5 times of amount ethanol sedimentations, centrifugal, the precipitation oven dry promptly gets Crude polysaccharides.
2. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1 is characterized in that pulverizing the sporophore powder, and powder diameter is controlled between the 1000-1500 μ m, and preferred mistake 20 mesh sieves, particle diameter are 1250 μ m.
3. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1, the ratio that it is characterized in that adding distilled water were at 1: 1 0-1: between 40, and preferred 1: 25.
4. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1, the ultrasonic power that it is characterized in that adopting between 100-200W, preferred 140W.
5. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1 is characterized in that ultrasonic time at 10-20min, preferred 15min.
6. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1 is characterized in that extracting temperature at 60-100 ℃, preferred 80 ℃.
7. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1 is characterized in that decocting time at 1-2h, preferred 1.5h.
8. the extracting method of a kind of Agaricus Blazei Murrill coarse polysaccharide according to claim 1 is characterized in that centrifugal rotational speed is 2000-6000r/min, and preferred 4000 r/min, centrifugation time are 5-20min, preferred 10min.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102838686A (en) * | 2012-09-28 | 2012-12-26 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN102924619A (en) * | 2012-11-09 | 2013-02-13 | 中海科创(北京)生物医药科技有限公司 | Agaricus blazei murill extract and preparation method thereof |
| CN112062873A (en) * | 2020-11-02 | 2020-12-11 | 石家庄飞龙饲料有限公司 | Method for extracting agaricus blazei murill polysaccharide under ultrahigh pressure and application thereof |
| CN112079939A (en) * | 2020-08-25 | 2020-12-15 | 福州康来生物科技有限公司 | Preparation method of agaricus blazei polysaccharide extract |
-
2009
- 2009-12-31 CN CN2009102452271A patent/CN102115505A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102838686A (en) * | 2012-09-28 | 2012-12-26 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN102838686B (en) * | 2012-09-28 | 2014-11-12 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN102924619A (en) * | 2012-11-09 | 2013-02-13 | 中海科创(北京)生物医药科技有限公司 | Agaricus blazei murill extract and preparation method thereof |
| CN102924619B (en) * | 2012-11-09 | 2015-02-18 | 中海科创(北京)生物医药科技有限公司 | Agaricus blazei murill extract and preparation method thereof |
| CN112079939A (en) * | 2020-08-25 | 2020-12-15 | 福州康来生物科技有限公司 | Preparation method of agaricus blazei polysaccharide extract |
| CN112062873A (en) * | 2020-11-02 | 2020-12-11 | 石家庄飞龙饲料有限公司 | Method for extracting agaricus blazei murill polysaccharide under ultrahigh pressure and application thereof |
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Application publication date: 20110706 |