CN115043951B - Extraction method and application of polysaccharide in stropharia rugoso-annulata - Google Patents
Extraction method and application of polysaccharide in stropharia rugoso-annulata Download PDFInfo
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Abstract
The invention relates to the technical field of stropharia rugoso-annulata, in particular to a method for extracting polysaccharide from stropharia rugoso-annulata and application thereof. The method comprises the following steps: (1) Crushing fresh stropharia rugoso-annulata into powder, soaking the powder in an ammonium carbonate solution, and carrying out solid-liquid separation to obtain leaching liquor I and stropharia rugoso-annulata extraction residues, and respectively collecting the leaching liquor I and stropharia rugoso-annulata extraction residues for later use. (2) And freezing and preserving the stropharia rugoso-annulata extraction residues, then heating in vacuum, and quickly recovering to normal pressure to obtain the wall-broken stropharia rugoso-annulata. (3) And (3) placing the wall-broken stropharia rugoso-annulata in the leaching solution I, leaching again under the airtight condition, heating and leaching, carrying out solid-liquid separation after the completion of the heating and leaching, and collecting the leaching solution II. (4) Concentrating, precipitating with ethanol, and purifying the leaching solution II to obtain stropharia rugoso-annulata polysaccharide. The method disclosed by the invention combines the extraction and wall breaking of polysaccharide, lays a foundation for the rupture of stropharia rugoso-annulata cells by utilizing the leaching process, effectively improves the extraction effect of polysaccharide and improves the utilization rate of stropharia rugoso-annulata.
Description
Technical Field
The invention relates to the technical field of stropharia rugoso-annulata, in particular to a method for extracting polysaccharide from stropharia rugoso-annulata and application thereof.
Background
The stropharia rugoso-annulata is an introduced edible fungus variety in China, has the characteristics of big mushroom and reddish wine, and is deeply favored by consumers due to the unique delicate flavor of the stropharia rugoso-annulata. The stropharia rugoso-annulata contains a large amount of high-quality natural polysaccharide, various minerals and vitamins, and the substances are beneficial components of human organ tissues. The edible stropharia rugoso-annulata has the advantages of promoting body metabolism, nourishing body, relieving asthenia, etc. In addition, the research shows that the polysaccharide in the stropharia rugoso-annulata is beneficial to clearing cholesterol in blood, so that the intake of the polysaccharide in the stropharia rugoso-annulata is beneficial to preventing coronary heart disease, hyperlipidemia and arteriosclerosis, and also beneficial to promoting digestion and relieving neurasthenia.
However, the polysaccharide content in stropharia rugoso-annulata is limited, and it is difficult to ingest sufficient polysaccharide by a method of eating stropharia rugoso-annulata in large quantities, and extraction of polysaccharide in stropharia rugoso-annulata is an effective measure to overcome the above-mentioned problems. Most of polysaccharide in the stropharia rugoso-annulata is positioned in cells, but the polysaccharide in the stropharia rugoso-annulata cells is not easy to release due to the protection of cell walls, so that the extraction rate of the stropharia rugoso-annulata polysaccharide is limited, and the utilization rate is lower.
Disclosure of Invention
The invention provides a method for extracting polysaccharide from stropharia rugoso-annulata and application thereof, which fuses the extraction and wall breaking of polysaccharide, lays a foundation for the rupture of stropharia rugoso-annulata cells by utilizing the leaching process, effectively improves the extraction effect of polysaccharide and improves the utilization rate of stropharia rugoso-annulata. In order to achieve the above purpose, the present invention discloses the following technical solutions.
In a first aspect, a method for extracting polysaccharide from stropharia rugoso-annulata is provided, comprising the following steps:
(1) Crushing fresh stropharia rugoso-annulata into powder, soaking the powder in an ammonium carbonate solution, and carrying out solid-liquid separation to obtain leaching liquor I and stropharia rugoso-annulata extraction residues, and respectively collecting the leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And freezing and preserving the stropharia rugoso-annulata extraction residues, then heating in vacuum, and quickly recovering to normal pressure to obtain the wall-broken stropharia rugoso-annulata.
(3) And (3) placing the wall-broken stropharia rugoso-annulata in the leaching solution I, leaching again under a closed condition, heating and leaching under an open ventilation state, carrying out solid-liquid separation after the completion of the leaching, and collecting the leaching solution II for later use.
(4) Concentrating, precipitating with ethanol, and purifying the leaching solution II to obtain stropharia rugoso-annulata polysaccharide.
Further, in the step (1), the mass percentage of the ammonium carbonate solution is 0.15-0.3 mol/L. The ammonium carbonate solution is alkaline, and the water-insoluble polysaccharide in the cell wall of the stropharia rugoso-annulata is extracted, so that the cell wall is weakened, and the stropharia rugoso-annulata cells are convenient to break in the subsequent process.
Further, in the step (1), the feed liquid ratio of the stropharia rugoso-annulata powder to the ammonium carbonate solution is 1:30 to 60 percent. Preferably, the mixture is continuously stirred in the soaking process, so that the stropharia rugoso-annulata fragments are fully contacted with the ammonium carbonate solution, and the floating influence of the stropharia rugoso-annulata fragments on the extraction is prevented.
Further, in the step (1), the soaking time is controlled between 65 and 110 minutes. Preferably, the soaking is performed under aeration conditions of not more than 20 ℃. Experiments show that the extraction effect is not obviously improved due to the overlong soaking time, the soaking time is more efficient, and other suitable soaking time can be selected according to actual needs.
Further, in the step (2), the stropharia rugoso-annulata extraction residues are frozen to be below 20 ℃ below zero and then kept for 10 to 30 minutes. The solution entering the stropharia rugoso-annulata cells can form a solid structure by quick freezing, so that the cell wall breaking can be conveniently carried out by sublimating into a gaseous state in the subsequent process.
Further, in the step (2), the heating temperature is 60-80 ℃ for 20-30 min, so that solid water in stropharia rugoso-annulata cells is quickly sublimated into a gaseous state, and simultaneously, ammonium carbonate is decomposed to generate gas, so that the pressure in the stropharia rugoso-annulata cells is quickly increased, and the cells are broken.
Further, in the step (3), the ratio of the feed liquid for re-leaching is 1: 40-65, leaching time is 40-60 min, and leaching is carried out at room temperature and below. The polysaccharide substances in the cells can be extracted by re-leaching the wall-broken stropharia rugoso-annulata.
Further, in the step (3), the temperature of the heating leaching is 72-85 ℃ and the time is 1.5-2 hours, so that the ammonium carbonate component in the solution is decomposed and released, and the alkali extraction is switched to the water extraction, thereby realizing one-solution and two-extraction.
Further, in the step (4), the leaching solution II is heated and concentrated to 50% or less of the original volume under vacuum condition. Preferably, the heating temperature is not higher than 85 ℃ to reduce the decomposition of stropharia rugoso-annulata polysaccharide.
In the step (4), ethanol with the volume of 3-5 times of that of the concentrated solution obtained by concentration is added for alcohol precipitation, so that the stropharia rugoso-annulata polysaccharide dissolved in the concentrated solution is precipitated and separated out, and a crude stropharia rugoso-annulata polysaccharide product is obtained.
Further, in step (4), the purifying includes: and (3) dissolving the crude stropharia rugoso-annulata polysaccharide in water, decoloring, deproteinizing, concentrating again, precipitating with alcohol, and vacuum drying the precipitated product to obtain stropharia rugoso-annulata polysaccharide.
In a second aspect, the application of the stropharia rugoso-annulata sugar obtained by the extraction method of the polysaccharide in the stropharia rugoso-annulata in the fields of biology, medicine, rehabilitation and the like is provided.
Compared with the prior art, the invention has the following beneficial effects:
the extraction method disclosed by the invention combines the extraction and wall breaking of polysaccharide, lays a foundation for the rupture of stropharia rugoso-annulata cells by utilizing the leaching process, has high wall breaking efficiency, and solves the problem that impurities are easy to additionally introduce in the traditional wall breaking technology. The reason for this is that: firstly, the invention utilizes the ammonium carbonate solution to initially extract stropharia rugoso-annulata, and the ammonium carbonate solution with alkaline characteristics not only can extract the water-insoluble polysaccharide in the cell wall of stropharia rugoso-annulata, so that the cell wall is weakened, and the cell wall of stropharia rugoso-annulata is convenient to rupture in the subsequent process; and after the ammonium carbonate solution enters the stropharia rugoso-annulata cells, the stropharia rugoso-annulata cells can be effectively broken by serving as an expanding agent. On the basis, the method comprises the steps of rapidly freezing the initially extracted stropharia rugoso-annulata to low temperature to solidify and freeze the solution therein, then heating in vacuum, rapidly sublimating ice in stropharia rugoso-annulata cells into a gaseous state in the process, meanwhile, decomposing solute ammonium carbonate into a gas under the conditions of heating and negative pressure, and breaking the stropharia rugoso-annulata cells due to internal pressure mutation when the stropharia rugoso-annulata cells are rapidly recovered to normal pressure, so as to finish the wall breaking of the stropharia rugoso-annulata cells. Further, the invention uses the initial leaching solution to leach the wall-broken stropharia rugoso-annulata again, and starts to heat and extract after leaching for a certain time, thereby realizing the alkali-water combined extraction process of one solution and two extractions, thereby not only extracting the water-insoluble polysaccharide in the stropharia rugoso-annulata, but also extracting the water-soluble polysaccharide in the stropharia rugoso-annulata, and improving the extraction rate of the polysaccharide. This is because when the ammonium carbonate solution is heated to a certain temperature, the solute ammonium carbonate therein starts to decompose into gas to be released, so that the ammonium carbonate solution gradually changes to water, thereby conveniently switching from alkaline extraction to water extraction. According to the extraction method disclosed by the invention, the extraction and wall breaking of polysaccharide are combined, and the leaching solution is used as an expanding agent to realize wall breaking treatment, so that compared with the traditional wall breaking methods, the process disclosed by the invention does not introduce extra impurities, such as the introduction of abrasive materials caused by mechanical grinding, the introduction of chemical reagents caused by chemical wall breaking and the like, and is beneficial to improving the purity of polysaccharide products and reducing the difficulty of purifying the polysaccharide products.
Detailed Description
It is to be noted that all terms of art and science used herein have the same meanings as those familiar to those skilled in the art unless otherwise defined. The reagents or materials used in the present invention may be purchased in conventional manners, and unless otherwise indicated, they may be used in conventional manners in the art or according to the product specifications.
In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred methods and materials described herein are presented for illustrative purposes only. The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention.
Example 1
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Fresh stropharia rugoso-annulata is placed in a stirrer to be crushed into powder, then placed in an ammonium carbonate solution with the concentration of 0.2mol/L, the feed-liquid ratio is 1:50, and then soaked for 90min at the ambient temperature of 20 ℃. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And (3) quickly freezing the stropharia rugoso-annulata extraction residues to-35 ℃ and keeping the temperature for 20min, then heating to 75 ℃ in vacuum and keeping the temperature for 30min, and then quickly recovering to normal pressure to obtain the wall-broken stropharia rugoso-annulata for later use.
(3) And (3) the wall-broken stropharia rugoso-annulata and leaching liquor I are mixed according to a feed liquid ratio of 1:55 are mixed in a reaction kettle, and then the reaction kettle is closed and leached at room temperature for 50min. After completion, the reaction vessel was opened and heated to 85℃for 1.5 hours, and this process was carried out in a fume hood to discharge the gas generated by decomposition of ammonium carbonate. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(4) Heating the leaching solution II to 80 ℃ and concentrating to 50% of the initial volume, cooling the concentrated solution to room temperature, adding ethanol (the mass concentration is 95%) with the volume of 4 times of the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(5) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (4) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
Example 2
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Fresh stropharia rugoso-annulata is placed in a stirrer to be crushed into powder, then placed in an ammonium carbonate solution with the concentration of 0.15mol/L, the feed-liquid ratio is 1:60, and then soaked for 110min at the ambient temperature of 20 ℃. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And (3) quickly freezing the stropharia rugoso-annulata extraction residues to-20 ℃ and keeping the temperature for 30min, then heating the stropharia rugoso-annulata extraction residues to 80 ℃ in vacuum and keeping the temperature for 20min, and then quickly recovering the stropharia rugoso-annulata extraction residues to normal pressure to obtain the wall-broken stropharia rugoso-annulata for later use.
(3) And (3) the wall-broken stropharia rugoso-annulata and leaching liquor I are mixed according to a feed liquid ratio of 1:40 are mixed in a reaction kettle, and then the reaction kettle is closed for leaching for 60min at room temperature. After completion, the reaction vessel was opened and heated to 75℃for 2 hours, and this process was carried out in a fume hood to discharge the gas generated by decomposition of ammonium carbonate. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(4) Heating the leaching solution II to 85 ℃ and concentrating to 45% of the initial volume, cooling the concentrated solution to room temperature, adding 3 times of ethanol (the mass concentration is 95%) into the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(5) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (4) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
Example 3
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Fresh stropharia rugoso-annulata is placed in a stirrer to be crushed into powder, then placed in an ammonium carbonate solution with the concentration of 0.3mol/L, the feed-liquid ratio is 1:30, and then soaked for 65min at the ambient temperature of 15 ℃. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And (3) quickly freezing the stropharia rugoso-annulata extraction residues to-40 ℃ and keeping the temperature for 10min, then heating the stropharia rugoso-annulata extraction residues to 60 ℃ in vacuum and keeping the temperature for 30min, and then quickly recovering the stropharia rugoso-annulata extraction residues to normal pressure to obtain the wall-broken stropharia rugoso-annulata for later use.
(3) And (3) the wall-broken stropharia rugoso-annulata and leaching liquor I are mixed according to a feed liquid ratio of 1:65 are mixed in a reaction kettle, and then the reaction kettle is closed and leached at room temperature for 40min. After completion, the reaction vessel was opened and heated to 72℃for 2 hours, and this process was carried out in a fume hood to discharge the gas generated by decomposition of ammonium carbonate. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(4) Heating the leaching solution II to 85 ℃ and concentrating to 50% of the initial volume, cooling the concentrated solution to room temperature, adding ethanol (the mass concentration is 95%) which is 5 times of the volume of the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(5) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (4) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
Example 4
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Fresh stropharia rugoso-annulata is placed in a stirrer to be crushed into powder, then placed in an ammonium carbonate solution with the concentration of 0.2mol/L, the feed-liquid ratio is 1:50, and then soaked for 90min at the ambient temperature of 20 ℃. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And (3) quickly freezing the stropharia rugoso-annulata extraction residues to-35 ℃ and keeping the temperature for 20min, then heating to 75 ℃ in vacuum and keeping the temperature for 30min, and then quickly recovering to normal pressure to obtain the wall-broken stropharia rugoso-annulata for later use.
(3) Mixing the wall-broken stropharia rugoso-annulata with clear water according to a feed liquid ratio of 1:55 are mixed in a reaction kettle, and then the reaction kettle is closed and leached at room temperature for 50min. After completion the reactor was opened and heated to 85 ℃ for 1.5 hours, the process was carried out in a fume hood. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(4) Heating the leaching solution II to 80 ℃ and concentrating to 50% of the initial volume, cooling the concentrated solution to room temperature, adding ethanol (the mass concentration is 95%) with the volume of 4 times of the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(5) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (4) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
Example 5
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Fresh stropharia rugoso-annulata is placed in a stirrer to be crushed into powder, then placed in an ammonium carbonate solution with the concentration of 0.2mol/L, the feed-liquid ratio is 1:50, and then soaked for 90min at the ambient temperature of 20 ℃. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) Extracting residues from stropharia rugoso-annulata and leaching solution I according to a feed liquid ratio of 1:55 are mixed in a reaction kettle, and then the reaction kettle is closed and leached at room temperature for 50min. After completion, the reaction vessel was opened and heated to 85℃for 1.5 hours, and this process was carried out in a fume hood to discharge the gas generated by decomposition of ammonium carbonate. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(3) Heating the leaching solution II to 80 ℃ and concentrating to 50% of the initial volume, cooling the concentrated solution to room temperature, adding ethanol (the mass concentration is 95%) with the volume of 4 times of the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(4) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (3) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
Example 6
A method for extracting polysaccharide from stropharia rugoso-annulata comprises the following steps:
(1) Pulverizing fresh stropharia rugoso-annulata in a pulverizer, placing into clear water with a feed-liquid ratio of 1:60, and soaking at 20deg.C for 110min. And (3) filtering and separating the feed liquid, and respectively collecting liquid phase and filter residues to obtain leaching liquor I and stropharia rugoso-annulata extraction residues for later use.
(2) And (3) quickly freezing the stropharia rugoso-annulata extraction residues to the temperature of minus 20 ℃ and keeping the temperature for 30min, then heating the stropharia rugoso-annulata extraction residues to the temperature of 80 ℃ in vacuum and keeping the temperature for 20min, and repeating the above process for 5 times. And then quickly recovering to normal pressure to obtain the wall-broken stropharia rugoso-annulata for standby.
(3) And (3) the wall-broken stropharia rugoso-annulata and leaching liquor I are mixed according to a feed liquid ratio of 1:40 are mixed in a reaction kettle, and then the reaction kettle is closed for leaching for 60min at room temperature. After completion the reactor was opened and heated to 75 ℃ for 2 hours, the process was carried out in a fume hood. And (3) filtering and separating the feed liquid after the completion, and collecting the leaching liquid II for later use.
(4) Heating the leaching solution II to 85 ℃ and concentrating to 45% of the initial volume, cooling the concentrated solution to room temperature, adding 3 times of ethanol (the mass concentration is 95%) into the concentrated solution, uniformly stirring, standing for 40min to precipitate stropharia rugoso-annulata polysaccharide in the concentrated solution, and filtering to separate polysaccharide precipitate to obtain a stropharia rugoso-annulata polysaccharide crude product for later use.
(5) The crude stropharia rugoso-annulata polysaccharide product was dissolved in water and then mixed according to 1L:2.0g of active carbon is added into the solution for decoloring for 30min, then the active carbon is filtered and separated, and the obtained decoloring solution is deproteinized by a Sevag method. Filtering to remove precipitated protein after completion, then treating the collected filtrate by adopting the method of the step (4) of the embodiment, and drying the obtained stropharia rugoso-annulata polysaccharide in vacuum at 50 ℃ for half an hour after completion to obtain stropharia rugoso-annulata polysaccharide.
And (3) effect test:
the yield of the polysaccharide and the wall breaking rate of the wall-broken stropharia rugoso-annulata obtained in each example are tested, and the test method adopts a phenol-sulfuric acid colorimetric method, uses glucose as a standard and calculates the yield by measuring the content. The results are shown in the following table. From the test results, the yield of the polysaccharide and the wall breaking rate of the wall-broken stropharia rugoso-annulata obtained in examples 1 to 3 are obviously improved compared with other examples.
The foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (15)
1. The extraction method of polysaccharide in stropharia rugoso-annulata is characterized by comprising the following steps:
(1) Crushing fresh stropharia rugoso-annulata into powder, soaking the powder in an ammonium carbonate solution, and carrying out solid-liquid separation to obtain leaching liquor I and stropharia rugoso-annulata extraction residues, and respectively collecting the leaching liquor I and stropharia rugoso-annulata extraction residues for later use;
(2) Freezing and preserving the stropharia rugoso-annulata extraction residues, then heating in vacuum, and quickly recovering to normal pressure to obtain wall-broken stropharia rugoso-annulata;
(3) Placing the wall-broken stropharia rugoso-annulata in leaching liquor I, leaching again under a closed condition, heating and leaching under an open ventilation state, carrying out solid-liquid separation after completion, and collecting leaching liquor II for later use;
(4) Concentrating, precipitating with ethanol, and purifying the leaching solution II to obtain stropharia rugoso-annulata polysaccharide.
2. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in the step (1), the mass percentage of the ammonium carbonate solution is 0.15-0.3 mol/L.
3. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in the step (1), the feed liquid ratio of the stropharia rugoso-annulata powder to the ammonium carbonate solution is 1:30 to 60 percent.
4. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (1), the soaking time is controlled between 65 and 110min.
5. The method for extracting polysaccharide from stropharia rugoso-annulata as claimed in claim 4, wherein the soaking is performed under ventilation at 20 ℃ or less.
6. The method for extracting polysaccharide from stropharia rugoso-annulata as claimed in claim 4, wherein stirring is continued during the soaking process.
7. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (2), the stropharia rugoso-annulata extraction residue is frozen to below-20 ℃ and then kept for 10-30 min.
8. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (2), the heating temperature is 60-80 ℃ for 20-30 min.
9. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (3), the ratio of the re-extracted feed solution in step (3) is 1: 40-65, leaching time is 40-60 min, and leaching is carried out at room temperature and below.
10. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (3), the temperature of the heating leaching is 72-85 ℃ for 1.5-2 h.
11. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (4), the leaching solution II is concentrated by heating under vacuum to 50% or less of the original volume.
12. The method for extracting polysaccharides from stropharia rugoso-annulata according to claim 11, wherein the heating temperature is not higher than 85 ℃.
13. The method for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (4), ethanol 3-5 times the volume of the concentrated solution obtained by concentration is added for ethanol precipitation.
14. The method for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 13, wherein in step (4), the purification comprises: and (3) dissolving the crude stropharia rugoso-annulata polysaccharide in water, decoloring, deproteinizing, concentrating again, precipitating with alcohol, and vacuum drying the precipitated product to obtain stropharia rugoso-annulata polysaccharide.
15. Use of stropharia rugoso-annulata polysaccharide obtained by the method for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1-14 in the fields of biology, medicine or rehabilitation.
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