CN115028749B - Process for extracting polysaccharide by utilizing stropharia rugoso-annulata - Google Patents

Process for extracting polysaccharide by utilizing stropharia rugoso-annulata Download PDF

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CN115028749B
CN115028749B CN202210639817.8A CN202210639817A CN115028749B CN 115028749 B CN115028749 B CN 115028749B CN 202210639817 A CN202210639817 A CN 202210639817A CN 115028749 B CN115028749 B CN 115028749B
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annulata
stropharia rugoso
polysaccharide
solution
extracting
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CN115028749A (en
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刘利
李秀杰
冯桂萍
韩真
朱自果
李勃
陈广霞
马玉姣
吴玉森
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Shandong Grape Research Institute
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    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
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Abstract

The invention discloses a process for extracting polysaccharide by utilizing stropharia rugoso-annulata, which comprises the following steps: (1) Adding the stropharia rugoso-annulata after wall breaking treatment into hot water for heat preservation and extraction, and carrying out solid-liquid separation after the wall breaking treatment is finished to obtain first leaching solution and mushroom residues. (2) And adding the mushroom residues into alkali liquor for heating and extracting, and carrying out solid-liquid separation after completion to obtain secondary leaching liquor. (3) And combining the first leaching solution and the second leaching solution, heating and concentrating, then carrying out alcohol precipitation, carrying out solid-liquid separation after the completion of the alcohol precipitation, and washing to remove residual alkali liquor in the precipitate, thus obtaining the stropharia rugoso-annulata polysaccharide crude product. (4) And (3) dissolving the crude stropharia rugoso-annulata polysaccharide, adding a decoloring agent for decoloring, performing solid-liquid separation, deproteinizing the obtained decoloring liquid, performing solid-liquid separation again after the process is completed, performing alcohol precipitation on the obtained liquid phase, and drying after the solid-liquid separation to obtain the stropharia rugoso-annulata polysaccharide product. The process can decolorize the leaching solution, and effectively improves the purity of the obtained polysaccharide target product.

Description

Process for extracting polysaccharide by utilizing stropharia rugoso-annulata
Technical Field
The invention relates to the technical field of stropharia rugoso-annulata, in particular to a process for extracting polysaccharide by utilizing stropharia rugoso-annulata.
Background
Stropharia rugoso-annulata is commonly referred to as Stropharia rugoso-annulata, and is named after the upper part of the fungus ring. The stropharia rugoso-annulata is rich in nutrition, crisp and tender in taste and rich in delicate flavor, and contains a large amount of protein, mineral substances, vitamins, polysaccharide and other nutritional ingredients, so that the stropharia rugoso-annulata also has higher nutritional and medicinal values, and can prevent coronary heart disease, promote digestion, relieve mental fatigue and the like. The stropharia rugoso-annulata contains abundant polysaccharide active substances, mainly comprising D-glucose, D-xylose, D-fructose and the like. Research shows that the polysaccharide active substances can play roles in clearing cholesterol in blood, purifying blood, softening blood vessels and preventing coronary heart disease after entering human bodies. At present, polysaccharide in stropharia rugoso-annulata is mainly extracted by a leaching method, and the obtained leaching solution is concentrated and then extracted by an alcohol precipitation method and the like. However, since stropharia rugoso-annulata has bright color and contains a large amount of pigments, part of pigments enter the leaching solution in the extraction process, and the purity and quality of the finally obtained polysaccharide target product are affected.
Disclosure of Invention
The invention provides a process for extracting polysaccharide by utilizing stropharia rugoso-annulata, which can decolorize leaching solution and effectively improve the purity of the obtained polysaccharide target product. In order to achieve the aim, the invention also provides a preparation process of the decoloring agent. Specifically, the invention discloses the following technical scheme.
A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata after wall breaking treatment into hot water for heat preservation and extraction, and carrying out solid-liquid separation after the wall breaking treatment to obtain first leaching solution and mushroom residues for later use.
(2) And adding the mushroom residues into alkali liquor for heating and extracting, and carrying out solid-liquid separation after the completion of the extraction to obtain secondary leaching liquor for later use.
(3) And combining the first leaching solution and the second leaching solution, heating and concentrating, then carrying out alcohol precipitation, carrying out solid-liquid separation after completion, and washing to remove residual alkali liquor in the precipitate, thus obtaining a stropharia rugoso-annulata polysaccharide crude product for later use.
(4) And (3) dissolving the crude stropharia rugoso-annulata polysaccharide, adding a decoloring agent for decoloring, performing solid-liquid separation, deproteinizing the obtained decoloring liquid, performing solid-liquid separation again after the process is completed, performing alcohol precipitation on the obtained liquid phase to precipitate stropharia rugoso-annulata polysaccharide, and drying after the solid-liquid separation to obtain the stropharia rugoso-annulata polysaccharide product.
Further, in the step (2), the decoloring agent is composed of a porous core and a flexible shell wrapped on the surface of the core. Wherein: the porous inner core comprises active carbon and alumina particles, the flexible shell comprises silica gel and ferroferric oxide coated in the silica gel, and the flexible shell is of a porous structure, and a transitional connecting layer which is mutually penetrated is formed between the flexible shell and the surface of the porous inner core.
Further, in the step (2), the preparation method of the decoloring agent comprises the following steps:
s1, uniformly mixing carbon particles, alumina particles and polyethylene glycol, granulating by a wet method, and drying to obtain a core precursor.
S2, adding ferroferric oxide particles, sodium chloride particles and polyethylene glycol into the silicone glue solution, and uniformly mixing to obtain the coated glue solution.
S3, immersing the core precursor into the coating glue solution, and then taking out the core precursor coated with the coating glue solution, and obtaining the decoloring agent precursor after curing is completed.
And S4, placing the decolorant precursor into hot water for washing to remove polyethylene glycol in the decolorant precursor, forming a porous inner core and a porous flexible shell wrapped on the surface of the porous inner core, and drying after finishing to obtain the decolorant.
Further, in step S1, the weight ratio of the carbon particles, the alumina particles, and the polyethylene glycol is 5 to 7.5 parts: 1.2-2 parts: 2-3 parts.
In step S1, the drying temperature is 70-90 ℃ and the time is 40-70 min, so that the core precursor obtained by granulation is solidified and molded.
Optionally, in step S1, the particle size of the carbon particles and the alumina particles is between 100 and 170 mesh, and other suitable particle sizes may be selected.
Further, in step S2, the weight ratio of the ferroferric oxide particles, the sodium chloride particles, the polyethylene glycol and the silicone solution is 1.5 to 2 parts in sequence: 1 to 1.8 portions: 2.5-3 parts: 9-11 parts. The silica gel is preferably food grade, medical grade silica gel. The sodium chloride particles have a particle size between 80 and 100 mesh to form a larger pore size in the flexible housing.
In step S2, the silica gel is AB-type silica gel, and the solution a and the solution B are mixed and cured at normal temperature or heated to accelerate curing. The AB type silica gel can directly adopt commercial products
Further, in step S2, the ferroferric oxide particles are 200-500 mesh. The ferroferric oxide particles can realize magnetic recovery of the decoloring agent, and are coated in the flexible shell to avoid oxidative deterioration, so that the decoloring agent can be recycled for multiple times. The recovered decolorizer can be regenerated by ethanol, etc.
Further, in step S3, the curing temperature is 70-80 ℃ and the time is 20-35 min, and the heating helps to accelerate the curing rate.
Further, in step S4, the temperature of the hot water is kept at 50-65 ℃, and the washing time is not less than 40min, such as 40min, 60min, 90min, 120min, etc., so as to remove polyethylene glycol components in the decolorizing agent precursor, thereby forming the porous inner core and the porous flexible shell.
Further, in step S4, the drying temperature is 50 to 60 ℃ and the time is 2 to 3 hours.
Further, in the step (1), the feed liquid ratio of the stropharia rugoso-annulata to the hot water is 1:30-40, the temperature of the hot water is 50-80 ℃, and the extraction time is 20-60 min. The extraction condition cost performance is higher by adopting the extraction conditions.
Further, in the step (1), the wall breaking treatment mode includes any one of mechanical grinding wall breaking, mechanical shearing wall breaking, ultrasonic wall breaking and the like.
Further, in the step (2), the alkali solution comprises sodium hydroxide or potassium hydroxide solution containing sodium borohydride to prevent the problem of degradation of the reducing end of the polysaccharide molecules under the condition of high pH value. Optionally, the pH value of the alkali liquor is 8-10, and the mass fraction of sodium borohydride in the alkali liquor is 2-4%.
Further, in the step (2), the feed liquid ratio of the mushroom residues to the alkali liquor is 1:50-70, the heating temperature is 60-80 ℃, and the extraction time is 40-70 min. The alkali-soluble polysaccharide in the stropharia rugoso-annulata can be extracted by alkali extraction, which is beneficial to improving the extraction rate of the polysaccharide.
In the step (3), the first leaching solution and the second leaching solution are combined and then heated and concentrated to 40-55% of the initial volume, then ethanol with the volume of 3-5 times of that of the concentrated solution is added, and the mixture is stood for 20-50 min for alcohol precipitation, so that stropharia rugoso-annulata polysaccharide is precipitated and separated out, and the stropharia rugoso-annulata polysaccharide precipitated and separated out is washed by ethanol after the completion of the alcohol precipitation, so that alkali liquor remained in the stropharia rugoso-annulata polysaccharide is removed, and a crude stropharia rugoso-annulata polysaccharide product is obtained.
Further, in the step (4), the volume ratio of the addition amount of the decoloring agent to the crude stropharia rugoso-annulata polysaccharide solution is 1-2.5 g:1L, and the decoloring time is 20-45 min. Preferably, the decoloring treatment is performed under stirring.
Further, in the step (4), the deproteinizing method includes at least one of a Sevag method, a trichloroacetic acid method, an ammonium sulfate precipitation method, and the like.
Further, in the step (4), ethanol with the volume 3 to 5 times of that of the liquid phase is added into the liquid phase, and the mixture is kept stand for 20 to 50 minutes for alcohol precipitation.
Further, in the step (4), the drying is vacuum drying, the temperature is 40-50 ℃, and the time is 60-80 min.
Compared with the prior art, the invention has the following beneficial effects:
firstly, the process adopts the extraction process of water extraction and then alkali extraction, so that the water-soluble polysaccharide in the stropharia rugoso-annulata can be extracted, and the non-aqueous polysaccharide in the stropharia rugoso-annulata can be extracted, thereby improving the extraction rate of the polysaccharide in the stropharia rugoso-annulata, and avoiding the problem that the extraction effect of a single extraction means is effective. Experiments prove that the extraction rate of polysaccharide by adopting the water and alkali combined extraction process is improved by more than 28% compared with that of polysaccharide by adopting a single extraction process.
Secondly, the stropharia rugoso-annulata contains a large amount of pigment, and the pigment easily enters the leaching solution in the extraction process, so that the purity and quality of the polysaccharide product are affected. Therefore, the invention provides the decoloring agent with a special structure, which removes the pigment in the stropharia rugoso-annulata polysaccharide, and effectively improves the purity and quality of the stropharia rugoso-annulata polysaccharide. The decoloring agent prepared by the process adopts a structure that a porous silica gel layer coats a porous inner core, and has the characteristics of:
(1) The porous core consists of the active carbon particles and the alumina particles, and the active carbon particles and the alumina particles have the decoloring function, and meanwhile, the alumina particles also serve as propping agents of the porous core, so that the problem of collapse caused by losing connection and support among the carbon particles in the subsequent process of washing and removing polyethylene glycol in the core is solved, and the decoloring capacity of the core is influenced.
(2) The surface of the inner core is coated with the silica gel layer, so that the decoloring agent has excellent anti-collision capability in the use process, and the problems of shell damage and falling and inner core abrasion are avoided. On the other hand, the silica gel shell has excellent hydrophilicity, so that the decoloring agent disclosed by the invention can be fully contacted with pigment in a solution, and the decoloring effect and the decoloring efficiency are improved. In addition, after the silica gel coating layer is gradually aged and hardened, the silica gel coating layer becomes a hard shell to be coated on the surface of the inner core, and the silica gel coating layer still plays a role in protecting the inner core well.
(3) In the preparation process of the decoloring agent, after polyethylene glycol in the inner core is removed, silica gel liquid more easily enters the surface layer of the inner core to form a transitional connecting layer which is mutually penetrated, carbon particles and alumina particles entering the transitional connecting layer play a role in reinforcing silica gel, so that the transitional connecting layer is reinforced, delamination between the inner core and the flexible shell is prevented, and the overall stability of the volume of the decoloring agent is better ensured.
(4) The polyethylene glycol added into the silica gel solution not only plays a role in forming a porous structure on the whole silica gel coating layer, but also has a role in enabling sodium chloride particles to be in faster contact with water through a channel formed after washing and removal, and removing sodium chloride to form a larger through hole. In addition, the polyethylene glycol in the silicone gum solution also has the function of improving the crosslinking density, thereby improving the volume stability of the decoloring agent.
(5) The invention also adds ferroferric oxide (Fe) into the silica gel flexible shell 3 O 4 ) The decolorizer is wrapped and sealed in a silica gel flexible shell, so that most of Fe can be avoided in the use process of the decolorizer 3 O 4 Is oxidized to Fe under the action of water and oxygen 2 O 3 The decoloring agent is unfavorable for recovery and affects the reutilization of the decoloring agent due to the loss of magnetism. In another aspect, the Fe 3 O 4 The strength of the flexible shell is increased, so that the volume stability of the decoloring agent is improved.
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.
In the following examples, the silica gel liquid is a commercially available AB silica gel (brand GINCHO), and when in use, the A liquid and the B liquid are mixed according to a weight ratio of 1:1, and uniformly stirred to obtain the silica gel liquid. It should be noted that, the skilled person may also use other suitable silica gel products as required.
Example 1
1. A preparation method of a decoloring agent comprises the following steps:
s1, uniformly mixing 120-mesh carbon particles, 150-mesh alumina particles and polyethylene glycol 2000 powder according to a weight ratio of 6:1.5:2, performing wet granulation in a granulator to obtain particles with a particle size of 3-5 mm, and drying in an oven at 85 ℃ for 50min to obtain a core precursor for later use.
S2, mixing 400 mesh Fe 3 O 4 Particulate, 100 mesh sodium chloride particlesAnd adding polyethylene glycol 2000 powder into the mixed silica gel liquid according to the weight ratio of 2:1.5:2.5:10, and rapidly stirring for 10min to uniformly mix the raw materials to obtain the coated glue liquid.
S3, immersing the kernel precursor obtained in the S1 into the coating glue solution of the S2, so that the glue solution is coated on the surface of the kernel precursor. And then taking out the core precursor, and preserving heat for 30min at 80 ℃ to solidify the glue solution coated on the surface of the core precursor to form a flexible shell, thus obtaining the decoloring agent precursor after completion.
And S4, placing the decolorant precursor into hot water at 60 ℃ and stirring for 40min, washing to remove polyethylene glycol in the decolorant precursor, forming a porous inner core and a porous flexible shell coated on the surface of the porous inner core, and drying in an oven at 60 ℃ for 2 hours after the completion of the porous inner core and the porous flexible shell, thus obtaining the decolorant.
2. A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into hot water with the temperature of 70 ℃ and the feed-liquid ratio of 1:35, then carrying out heat preservation and extraction for 40min, filtering after completion, carrying out solid-liquid separation, and respectively collecting filtrate and filter residues to obtain first leaching liquor and mushroom residues for later use.
(2) Adding the mushroom residues into a sodium hydroxide solution (pH=10), extracting at 70 ℃ for 50min with a feed-liquid ratio of 1:65, filtering after completion, performing solid-liquid separation, and collecting filtrate to obtain a second leaching solution for later use.
(3) And (3) mixing the first leaching solution and the second leaching solution, heating to 70 ℃ to concentrate to 50% of the initial volume, adding ethanol (the mass concentration is 95%) of 4 times of the volume of the concentrate, standing for 40min, precipitating and separating out stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion of the precipitation, and washing for 3 times by using ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(4) The stropharia rugoso-annulata polysaccharide crude product is dissolved in deionized water according to 2g:1L of the decoloring agent prepared in the embodiment is added into the solution of the stropharia rugoso-annulata polysaccharide crude product according to the weight-volume ratio, and the decoloring treatment is carried out for 30min under the stirring condition. Filtering and separating the decoloring agent after completion, collecting filtrate to obtain decoloring liquid, deproteinizing the decoloring liquid by adopting a Sevag method, filtering again after completion to perform solid-liquid separation to remove precipitated protein, adding ethanol with the volume of 4 times of the collected filtrate, standing for 40min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and vacuum drying at 50 ℃ for 60min to obtain stropharia rugoso-annulata polysaccharide products.
Example 2
1. A preparation method of a decoloring agent comprises the following steps:
s1, uniformly mixing 100-mesh carbon particles, 100-mesh alumina particles and polyethylene glycol 2000 powder according to the weight ratio of 7.5:2:3, performing wet granulation in a granulator to obtain granules with the particle size of 3-5 mm, and drying in an oven at 70 ℃ for 70min to obtain a core precursor for later use.
S2, 200 meshes of Fe 3 O 4 And adding the particles, 80-mesh sodium chloride particles and polyethylene glycol 2000 powder into the mixed silica gel liquid according to the weight ratio of 1.5:1.0:2.5:9, and rapidly stirring for 10min to uniformly mix the raw materials to obtain the coated glue liquid.
S3, immersing the kernel precursor obtained in the S1 into the coating glue solution of the S2, so that the glue solution is coated on the surface of the kernel precursor. And then taking out the core precursor, and preserving heat at 70 ℃ for 35min to solidify the glue solution coated on the surface of the core precursor to form a flexible shell, thus obtaining the decoloring agent precursor after completion.
And S4, placing the decolorant precursor into hot water at 65 ℃ and stirring for 60min, washing to remove polyethylene glycol in the decolorant precursor, forming a porous inner core and a porous flexible shell coated on the surface of the porous inner core, and drying in an oven at 55 ℃ for 3 hours after the completion of the porous inner core and the porous flexible shell, thus obtaining the decolorant.
2. A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into hot water with the temperature of 80 ℃ and the feed-liquid ratio of 1:40, then carrying out heat preservation and extraction for 60min, filtering after completion, carrying out solid-liquid separation, and respectively collecting filtrate and filter residues to obtain first leaching liquor and mushroom residues for later use.
(2) The mushroom residue is added into a sodium hydroxide solution (ph=8) with a feed-to-liquid ratio of 1:70, wherein the sodium hydroxide solution contains 4% sodium borohydride. Then extracting at 80 ℃ for 40min, filtering after completion, carrying out solid-liquid separation, and collecting filtrate to obtain secondary leaching liquor for later use.
(3) And (3) mixing the first leaching solution and the second leaching solution, heating to 70 ℃ to concentrate to 55% of the initial volume, adding ethanol (the mass concentration is 95%) which is 5 times of the volume of the concentrate, standing for 20min, precipitating and separating out stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion of the precipitation, and washing for 3 times by using ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(4) The stropharia rugoso-annulata polysaccharide crude product is dissolved in deionized water according to 2.5g:1L of the decoloring agent prepared in the embodiment is added into the solution of the stropharia rugoso-annulata polysaccharide crude product according to the weight-volume ratio, and the decoloring treatment is carried out for 20min under the stirring condition. Filtering and separating the decoloring agent after completion, collecting filtrate to obtain decoloring liquid, deproteinizing the decoloring liquid by adopting a Sevag method, filtering again after completion to perform solid-liquid separation to remove precipitated proteins, adding ethanol with the volume of 5 times of the filtrate into the collected filtrate, standing for 20min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and vacuum drying at 50 ℃ for 70min to obtain stropharia rugoso-annulata polysaccharide products.
Example 3
1. A preparation method of a decoloring agent comprises the following steps:
s1, uniformly mixing 170-mesh carbon particles, 150-mesh alumina particles and polyethylene glycol 2000 powder according to a weight ratio of 5:1.2:2, performing wet granulation in a granulator to obtain particles with a particle size of 3-5 mm, and drying the particles in an oven at 90 ℃ for 40min to obtain a core precursor for later use.
S2, mixing 500 mesh Fe 3 O 4 The particles, 90-mesh sodium chloride particles and polyethylene glycol 2000 powder are added into the silica gel liquid according to the weight ratio of 2:1.8:3:11, and the silica gel liquid is medium and fast after being mixedStirring for 10min to mix the raw materials uniformly to obtain the coating glue solution.
S3, immersing the kernel precursor obtained in the S1 into the coating glue solution of the S2, so that the glue solution is coated on the surface of the kernel precursor. And then taking out the core precursor, and preserving heat at 80 ℃ for 20min to solidify the glue solution coated on the surface of the core precursor to form a flexible shell, thus obtaining the decoloring agent precursor after completion.
And S4, placing the decolorant precursor into hot water at 50 ℃ and stirring for 90min, washing to remove polyethylene glycol in the decolorant precursor, forming a porous inner core and a porous flexible shell coated on the surface of the porous inner core, and drying in an oven at 50 ℃ for 3 hours after the completion of the porous inner core and the porous flexible shell, thus obtaining the decolorant.
2. A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into hot water with the temperature of 50 ℃, the feed-liquid ratio of 1:30, then carrying out heat preservation and extraction for 20min, filtering after completion, carrying out solid-liquid separation, and respectively collecting filtrate and filter residues to obtain first leaching liquor and mushroom residues for later use.
(2) The mushroom residue is added into sodium hydroxide solution (pH=9) with a feed liquid ratio of 1:50, wherein the sodium hydroxide solution contains 2% sodium borohydride. Then extracting at 60 ℃ for 70min, filtering after completion, carrying out solid-liquid separation, and collecting filtrate to obtain secondary leaching liquor for later use.
(3) And (3) mixing the first leaching solution and the second leaching solution, heating to 75 ℃ to concentrate to 40% of the initial volume, adding 3 times of ethanol (the mass concentration is 95%) of the concentrate, standing for 50min, precipitating and separating out stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion, and washing for 3 times by using ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(4) The stropharia rugoso-annulata polysaccharide crude product is dissolved in deionized water according to 1g:1L of the decoloring agent prepared in the embodiment is added into the solution of the stropharia rugoso-annulata polysaccharide crude product according to the weight-volume ratio, and the decoloring treatment is carried out for 45min under the stirring condition. Filtering and separating the decoloring agent after completion, collecting filtrate to obtain decoloring liquid, deproteinizing the decoloring liquid by adopting a Sevag method, filtering again after completion to perform solid-liquid separation to remove precipitated protein, adding 3 times of ethanol into the collected filtrate, standing for 20min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and vacuum drying at 40 ℃ for 80min to obtain stropharia rugoso-annulata polysaccharide products.
Example 4
A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into hot water with the temperature of 70 ℃ and the feed-liquid ratio of 1:35, then carrying out heat preservation and extraction for 40min, filtering after completion, carrying out solid-liquid separation, and collecting filtrate to obtain leaching liquor for later use.
(2) Heating the leaching solution to 70 ℃ for concentration to 50% of the initial volume, adding ethanol (the mass concentration is 95%) which is 4 times of the volume of the concentrated solution, standing for 40min for alcohol precipitation, precipitating stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion of the precipitation, and washing 3 times with ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(3) The stropharia rugoso-annulata polysaccharide crude product is dissolved in deionized water according to 2g:1L of decoloring agent prepared in the method of example 1 is added into the solution of the stropharia rugoso-annulata polysaccharide crude product according to the weight-volume ratio, and decoloring is carried out for 30min under the condition of stirring. Filtering and separating the decoloring agent after completion, collecting filtrate to obtain decoloring liquid, deproteinizing the decoloring liquid by adopting a Sevag method, filtering again after completion to perform solid-liquid separation to remove precipitated protein, adding ethanol with the volume of 4 times of the collected filtrate, standing for 40min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and vacuum drying at 50 ℃ for 60min to obtain stropharia rugoso-annulata polysaccharide products.
Example 5
A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into sodium hydroxide solution (pH=10) with a feed-liquid ratio of 1:65, extracting at 70 ℃ for 50min, filtering after completion, performing solid-liquid separation, and collecting filtrate to obtain leaching liquor for later use.
(2) Heating the leaching solution to 70 ℃ for concentration to 50% of the initial volume, adding ethanol (the mass concentration is 95%) which is 4 times of the volume of the concentrated solution, standing for 40min for alcohol precipitation, precipitating stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion of the precipitation, and washing 3 times with ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(3) The stropharia rugoso-annulata polysaccharide crude product is dissolved in deionized water according to 2g:1L of the decoloring agent prepared in the embodiment is added into the solution of the stropharia rugoso-annulata polysaccharide crude product according to the weight-volume ratio, and the decoloring treatment is carried out for 30min under the stirring condition. Filtering and separating the decoloring agent after completion, collecting filtrate to obtain decoloring liquid, deproteinizing the decoloring liquid by adopting a Sevag method, filtering again after completion to perform solid-liquid separation to remove precipitated protein, adding ethanol with the volume of 4 times of the collected filtrate, standing for 40min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and vacuum drying at 50 ℃ for 60min to obtain stropharia rugoso-annulata polysaccharide products.
Example 6
A process for extracting polysaccharide by utilizing stropharia rugoso-annulata comprises the following steps:
(1) Adding the stropharia rugoso-annulata subjected to wall breaking treatment by a wall breaking machine into hot water with the temperature of 70 ℃ and the feed-liquid ratio of 1:35, then carrying out heat preservation and extraction for 40min, filtering after completion, carrying out solid-liquid separation, and respectively collecting filtrate and filter residues to obtain first leaching liquor and mushroom residues for later use.
(2) Adding the mushroom residues into a sodium hydroxide solution (pH=10), extracting at 70 ℃ for 50min with a feed-liquid ratio of 1:65, filtering after completion, performing solid-liquid separation, and collecting filtrate to obtain a second leaching solution for later use.
(3) And (3) mixing the first leaching solution and the second leaching solution, heating to 70 ℃ to concentrate to 50% of the initial volume, adding ethanol (the mass concentration is 95%) of 4 times of the volume of the concentrate, standing for 40min, precipitating and separating out stropharia rugoso-annulata polysaccharide in the leaching solution, separating stropharia rugoso-annulata polysaccharide precipitate after the completion of the precipitation, and washing for 3 times by using ethanol to obtain a stropharia rugoso-annulata polysaccharide crude product.
(4) Dissolving the stropharia rugoso-annulata polysaccharide crude product in deionized water, deproteinizing the decolored liquid by adopting a Sevag method, filtering again after the deproteinization treatment is completed, performing solid-liquid separation to remove precipitated proteins, adding ethanol with the volume of 4 times of that of the filtrate into the collected filtrate, standing for 40min for alcohol precipitation to precipitate stropharia rugoso-annulata polysaccharide, filtering and separating stropharia rugoso-annulata polysaccharide precipitate, and performing vacuum drying at 50 ℃ for 60min to obtain stropharia rugoso-annulata polysaccharide products.
Example 7
1. A preparation method of a decoloring agent comprises the following steps:
s1, uniformly mixing 100-mesh carbon particles and polyethylene glycol 2000 powder according to the weight ratio of 7.5:2:3, performing wet granulation in a granulator to obtain particles with the particle size of 3-5 mm, and drying the particles in an oven at 70 ℃ for 70min to obtain a core precursor for later use.
S2, 200 meshes of Fe 3 O 4 And adding the particles, 80-mesh sodium chloride particles and polyethylene glycol 2000 powder into the mixed silica gel liquid according to the weight ratio of 1.5:1.0:2.5:9, and rapidly stirring for 10min to uniformly mix the raw materials to obtain the coated glue liquid.
S3, immersing the kernel precursor obtained in the S1 into the coating glue solution of the S2, so that the glue solution is coated on the surface of the kernel precursor. And then taking out the core precursor, and preserving heat at 70 ℃ for 35min to solidify the glue solution coated on the surface of the core precursor to form a flexible shell, thus obtaining the decoloring agent precursor after completion.
2. A process for extracting polysaccharide by utilizing stropharia rugoso-annulata, which is similar to example 2, and is characterized in that: the decolorizing agent used in step (4) was prepared in this example.
Example 8
1. A preparation method of a decoloring agent comprises the following steps:
s1, uniformly mixing 170-mesh carbon particles, 150-mesh alumina particles and polyethylene glycol 2000 powder according to a weight ratio of 5:1.2:2, performing wet granulation in a granulator to obtain particles with a particle size of 3-5 mm, and drying the particles in an oven at 90 ℃ for 40min to obtain a core precursor for later use.
S2, placing the core precursor in hot water at 50 ℃ and stirring for 90min to wash to remove polyethylene glycol in the decolorant precursor, wherein the core precursor is scattered due to dissolution of the polyethylene glycol in the process and cannot obtain the decolorant, so that the problem that the core precursor is scattered due to dissolution of the polyethylene glycol can also occur when the core precursor is directly used as the decolorant for decoloring the stropharia rugoso-annulata polysaccharide crude product solution.
And (3) effect test:
the yield of stropharia rugoso-annulata polysaccharide of examples 1 to 7 above (phenol-sulfuric acid colorimetry, based on glucose, calculated with the measured content) was tested, and the purity of the stropharia rugoso-annulata polysaccharide products finally obtained in examples 1 to 7 was also tested, and the results are shown in table 1.
TABLE 1
Example sequence number 1 2 3 4 5 6 7
Polysaccharide yield/% 10.23 11.44 10.86 7.37 6.92 10.33 12.31
Purity/% 91.77 93.38 93.02 91.56 91.42 60.82 87.58
It can be seen from table 1 that example 4 and example 5 only use a single extraction mode, whereas the water-base combined extraction process used in example 1 increases polysaccharide yield by more than 28% over a single extraction process. Meanwhile, the polysaccharide yield of other examples is obviously improved compared with that of examples 4 and 5. In addition, as can be seen from the purity results of the stropharia rugoso-annulata polysaccharide product, the purity of the stropharia rugoso-annulata polysaccharide product can be effectively improved by adopting the decoloring agent.
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 (23)

1. A process for extracting polysaccharide by utilizing stropharia rugoso-annulata is characterized by comprising the following steps:
(1) Adding the stropharia rugoso-annulata after wall breaking treatment into hot water for heat preservation and extraction, and carrying out solid-liquid separation after the wall breaking treatment is finished to obtain first leaching solution and mushroom residues;
(2) Adding the mushroom residues into alkali liquor for heating and extracting, and carrying out solid-liquid separation after completion to obtain secondary leaching liquor;
(3) Combining the first leaching solution and the second leaching solution, heating and concentrating, then carrying out alcohol precipitation, carrying out solid-liquid separation after completion, and washing to remove residual alkali liquor in the precipitate, thus obtaining a stropharia rugoso-annulata polysaccharide crude product;
(4) Dissolving the crude stropharia rugoso-annulata polysaccharide, adding a decoloring agent for decoloring, then carrying out solid-liquid separation, deproteinizing the obtained decoloring liquid, carrying out solid-liquid separation again after the process is completed, carrying out alcohol precipitation on the obtained liquid phase, and drying after the solid-liquid separation to obtain a stropharia rugoso-annulata polysaccharide product;
the decoloring agent consists of a porous inner core and a flexible shell wrapped on the surface of the inner core; wherein: the porous inner core comprises active carbon and alumina particles, the flexible shell comprises silica gel and ferroferric oxide coated in the silica gel, and the flexible shell is of a porous structure, and a transitional connecting layer which is mutually penetrated is formed between the flexible shell and the surface of the porous inner core.
2. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 1, wherein in step (4), the preparation method of the decoloring agent comprises the following steps:
s1, uniformly mixing carbon particles, alumina particles and polyethylene glycol, and then granulating and drying by a wet method to obtain a kernel precursor;
s2, adding ferroferric oxide particles, sodium chloride particles and polyethylene glycol into the silicone glue solution, and uniformly mixing to obtain a coated glue solution;
s3, immersing the core precursor into the coating glue solution, and then taking out the core precursor coated with the coating glue solution, and obtaining a decoloring agent precursor after curing is completed;
and S4, placing the decolorant precursor into hot water for washing to remove polyethylene glycol in the decolorant precursor, forming a porous inner core and a porous flexible shell wrapped on the surface of the porous inner core, and drying after finishing to obtain the decolorant.
3. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S1, the weight ratio of the carbon particles, the alumina particles and the polyethylene glycol is 5-7.5 parts: 1.2-2 parts: 2-3 parts.
4. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S1, the drying temperature is 70-90 ℃ and the time is 40-70 min, so that the kernel precursor obtained by granulation is solidified and molded.
5. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S1, the particle size of the carbon particles and the alumina particles is 100-170 mesh.
6. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S2, the weight ratio of the ferroferric oxide particles, sodium chloride particles, polyethylene glycol and silica gel solution is 1.5-2: 1 to 1.8 portions: 2.5-3 parts: 9-11 parts.
7. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein the particle size of the sodium chloride particles is between 80 and 100 mesh.
8. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S2, the silica gel is AB-type silica gel, and the solution a and the solution B are mixed and cured at normal temperature or heated to accelerate curing.
9. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S2, the ferroferric oxide particles are 200-500 mesh.
10. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S3, the curing temperature is 70-80 ℃ and the time is 20-35 min.
11. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S4, the temperature of the hot water is kept at 50-65 ℃ and the washing time is not less than 40min.
12. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step S4, the drying temperature is 50-60 ℃ for 2-3 hours.
13. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (1), the feed liquid ratio of stropharia rugoso-annulata to hot water is 1:30-40, the temperature of the hot water is 50-80 ℃, and the extraction time is 20-60 min.
14. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (1), the wall breaking treatment comprises any one of mechanical grinding, mechanical shearing and ultrasonic wall breaking.
15. The process for extracting polysaccharides from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (2) the alkaline solution comprises sodium hydroxide or potassium hydroxide solution containing sodium borohydride.
16. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 15, wherein the pH of the lye is 8-10.
17. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 15, wherein the mass fraction of sodium borohydride in the alkaline solution is 2-4%.
18. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (2), the feed liquid ratio of the mushroom residue to the alkali liquid is 1:50-70, the heating temperature is 60-80 ℃, and the extraction time is 40-70 min.
19. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (3), the first leaching solution and the second leaching solution are combined, heated and concentrated to 40 to 55% of the initial volume, then ethanol with 3 to 5 times of the volume of the concentrated solution is added, and the mixture is left to stand for 20 to 50 minutes for alcohol precipitation, and after the completion, stropharia rugoso-annulata polysaccharide precipitated and separated out is separated and washed by ethanol, and a crude stropharia rugoso-annulata polysaccharide product is obtained.
20. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (4), the volume ratio of the addition amount of the decoloring agent to the crude stropharia rugoso-annulata polysaccharide solution is 1 to 2.5g:1L, and the decoloring time is 20-45 min; and (3) decoloring under stirring.
21. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step (4), the deproteinizing treatment comprises at least one of Sevag method, trichloroacetic acid method, and ammonium sulfate precipitation method.
22. The process for extracting polysaccharide from stropharia rugoso-annulata according to any one of claims 1 to 12, wherein in step (4), ethanol 3 to 5 times the volume of the solution is added to the solution, and the solution is allowed to stand for 20 to 50 minutes for alcohol precipitation.
23. The process for extracting polysaccharide from stropharia rugoso-annulata according to claim 2, wherein in step (4), the drying is vacuum drying at 40-50 ℃ for 60-80 min.
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