CN110592167B - Purslane polypeptide extract and preparation method thereof - Google Patents
Purslane polypeptide extract and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a purslane polypeptide extract and a preparation method thereof, wherein the preparation method comprises the following steps: (1) Pulverizing dried herba Portulacae into coarse powder, adding petroleum ether, reflux extracting, filtering, and volatilizing solvent; (2) Adding water, adding neutral protease, warm-soaking and extracting, adjusting the pH to 9-10 with an alkali solution, adding alkaline protease, warm-soaking and extracting, boiling to inactivate enzyme, filtering with filter cloth, concentrating under reduced pressure, centrifuging and retaining supernatant; (3) Passing the supernatant through D101 macroporous resin, collecting eluate, washing with water, collecting water eluate, mixing eluate with water eluate, and concentrating under reduced pressure; (4) Adding ethanol, standing, filtering, and concentrating under reduced pressure to remove ethanol; (5) Dialyzing at room temperature by using a dialysis bag with molecular weight cutoff of 2kDa, concentrating under reduced pressure, and vacuum drying and pulverizing to obtain the purslane polypeptide extract. The extraction rate of the protein of the purslane polypeptide extract prepared by the invention is more than or equal to 53.5%, the protein content is more than or equal to 73.8%, and the ratio of the molecular weight of the polypeptide to the molecular weight of 2kDa is more than or equal to 92.7%.
Description
Technical Field
The invention belongs to the field of plant extraction, and particularly relates to a purslane polypeptide extract and a preparation method thereof.
Background
Purslane, also called longevity herb, is a whole plant of Portulacaceae, and is mostly used as a medicine by using dry overground parts thereof, and grows in wet and fertile places in mountainous areas and is distributed all over the country. Sour in taste and cold in nature. It belongs to liver and large intestine channels, has effects of clearing heat and detoxicating, cooling blood and arresting hemorrhage, and relieving dysentery, and can be used for treating heat toxin bloody dysentery, carbuncle swelling and sore, eczema, erysipelas, snake and insect bite, hematochezia, hemorrhoidal bleeding and metrorrhagia and metrostaxis. Pharmacological research in recent years shows that the purslane has the effects of reducing blood fat, reducing blood sugar, resisting atherosclerosis and the like.
The purslane contains various nutrients and chemical components, and has rich nutritive value and special medical health-care effect. According to the measurement, every 100g of fresh purslane contains 2.3g of protein, 0.5g of fat, 3g of carbohydrate, 0.7g of crude fiber, 23mg of VC, 12.2mg of VE12, 0.7mg of nicotinic acid, 2.23mg of carotene, 300-400 mg of alpha-linolenic acid, 18 amino acids (8 of which are essential amino acids for human bodies), vitamin A, B, D and inorganic salt. The purslane can be used as a medicine, can be eaten, can also be used as a cosmetic raw material, and is one of 101 medicinal and edible wild plants defined by the Ministry of health of China.
Compared with protein, the polypeptide has lower molecular weight, is beneficial to the absorption of human bodies, and a plurality of researches show that the hydrolyzed polypeptide from plant sources has a plurality of special biological activities and has wide application prospect.
In the prior art, CN108276472A extracts selenoprotein in purslane by using a method of extracting pure water, a 95% ethanol solution, a phosphate buffer solution and an ultrasonic generator phosphate buffer solution, wherein the extraction rate of selenoprotein in the ultrasonic generator phosphate buffer solution is 60.24% at most; the process has high requirements on equipment, is difficult to apply to large-scale production, mostly contains high molecular weight protein in the extract, and has a limited product application range.
In the prior art, CN103554237A is extracted by NaCl solution, concentrated and desalted by an ultrafilter, precipitated by absolute ethyl alcohol and finally freeze-dried to obtain purslane active glycoprotein; the process uses an ultrafilter to purify the protein in the purslane extraction, the protein in the extract is not hydrolyzed, the protein molecular weight is larger, the purification effect is insufficient, and the protein purity of the final product is lower.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the technical defects of the background technology and provide a purslane polypeptide extract and a preparation method thereof. The extraction rate of the protein of the purslane polypeptide extract prepared by the invention is more than or equal to 53.5%, the protein content is more than or equal to 73.8%, and the ratio of the molecular weight of the polypeptide to the molecular weight of 2kDa is more than or equal to 92.7%. The invention has the advantages of simple process, easy operation, low polypeptide molecular weight and the like, and the process is easy for industrial production.
The technical means adopted by the invention for solving the technical problems is as follows:
a preparation method of a purslane polypeptide extract comprises the following steps:
(1) Degreasing: crushing the dried purslane medicinal material into coarse powder, adding petroleum ether with the boiling range of 60-90 ℃ in an amount which is 8-10 times that of the medicinal material, refluxing and extracting for 2-3 h, filtering, and volatilizing the solvent in filter residues to obtain the medicinal material degreased by the petroleum ether;
(2) And (3) extraction and concentration: adding water into the degreased medicinal materials of petroleum ether, adding neutral protease accounting for 0.3-0.5% of the medicinal materials, and extracting by warm immersion; after warm-soaking, adjusting the pH of the extraction solution to 9-10 by using an alkali solution, adding alkaline protease accounting for 0.3-0.5% of the amount of the medicinal materials into the extraction solution, and performing warm-soaking extraction; after warm immersion, boiling to inactivate enzyme, filtering with filter cloth, concentrating under reduced pressure, adjusting pH of the concentrated solution to neutrality with acid solution, centrifuging, and retaining supernatant;
(3) D101, purification: enabling the supernatant to pass through D101 macroporous resin, wherein the dosage of the D101 macroporous resin is 1/10-1/8 of the amount of the medicinal materials, collecting lower column liquid, washing the column with water, collecting water washing liquid, combining the lower column liquid and the water washing liquid, and concentrating under reduced pressure to obtain first concentrated solution;
(4) Alcohol precipitation: slowly adding 90-100% ethanol in an amount which is 4-6 times that of the first concentrated solution while stirring, standing for 2-5 h at the temperature of 2-6 ℃, then carrying out suction filtration, and carrying out reduced pressure water supplement concentration on the suction filtration until no ethanol exists to obtain a second concentrated solution;
(5) And (3) dialysis: dialyzing the second concentrated solution for 5-10 h at room temperature by using a dialysis bag with the molecular weight cutoff of 2kDa, and carrying out vacuum drying and crushing on the dialyzate after decompressing and concentrating the dialyzate to obtain the purslane polypeptide extract.
Preferably, in the step (1), the mesh number of the coarse powder is 20-30 meshes.
Preferably, in the step (2), the amount of the water added is 10-12 times (v/m) of the medicinal materials.
Preferably, in the step (2), the neutral protease is papain.
Preferably, in the step (2), the temperature of the warm leaching extraction is 45-60 ℃ and the time is 1.5-2 h.
Preferably, in the step (2), the alkali solution is a NaOH solution.
Preferably, in the step (2), the time for boiling to inactivate the enzyme is 10-30 min.
Preferably, in the step (2), the mesh number of the filter cloth is 300 meshes.
Preferably, in the step (2), the vacuum degree during the reduced pressure concentration is-0.07 to-0.09 Mpa, the temperature is 60 to 80 ℃, and the volume of the concentrated solution is 1/3 to 1/2 (v/m) of the amount of the medicinal materials.
Preferably, in the step (2), the acid solution is an aqueous HCl solution.
Preferably, in the step (3), the amount of the water is 1 to 1.5BV of the resin volume.
Preferably, in the step (3), the concentration under reduced pressure is performed until the concentration is 1/15 to 1/10 (v/m) of the amount of the medicinal materials.
Preferably, in the step (4), the concentration of the ethanol is 95%.
Preferably, in the step (4), the volume of the concentration is 1/15-1/10 (v/m) of the amount of the medicinal materials.
A herba Portulacae polypeptide extract is prepared by the above preparation method.
In the above technical scheme, in the step (1), the addition amount of the petroleum ether is a volume-to-mass ratio of the amount of the medicinal material, i.e., v/m.
In the above technical scheme, in the step (2), the addition amount of the neutral protease is a mass ratio of the neutral protease to the amount of the medicinal material, that is, m/m.
In the above technical scheme, in the step (2), the adding amount of the alkaline protease is a mass ratio of the adding amount to the amount of the medicinal material, namely m/m.
In the above technical scheme, in the step (3), the dosage of the D101 macroporous resin is a mass ratio to the amount of the medicinal material, i.e., m/m.
In the above technical solution, in the step (4), the adding amount of the ethanol is a volume ratio of the ethanol to the first concentrated solution, that is, v/v.
The basic principle of the invention is as follows:
the purslane contains more protein, which accounts for about 19.1% of the dry weight of the purslane plant, and fat-soluble and small molecular substances in the purslane can be removed by using petroleum ether for degreasing before extraction, so that the extraction of the protein is facilitated; by using a method of extracting with the assistance of proteolytic enzyme, the proteolysis can be promoted to be polypeptide with low molecular weight, so that the dissolution of the polypeptide is promoted, and the extraction rate is improved; adsorbing nonpolar or partially low-polar impurities by using nonpolar macroporous resin D101, and allowing the polypeptide to flow out along with the lower column liquid and the water washing liquid; by using an ethanol precipitation mode, a large amount of polysaccharides and alcohol-insoluble substances can be precipitated, so that the purity of the purslane polypeptide extract is improved; ultrafiltration can separate high molecular weight substances from low molecular weight polypeptides to obtain a high purity, low molecular weight purslane polypeptide extract.
The method takes dried purslane as a raw material, uses petroleum ether (with a boiling range of 60-90 ℃) for reflux to degrease, uses a certain amount of aqueous solution of neutral protease as an extraction solvent to carry out warm-immersion extraction after volatilizing the solvent, adjusts the pH of the solution to be alkaline after the warm-immersion is finished, adds a proper amount of alkaline protease to carry out warm-immersion extraction, boils the extracting solution to inactivate the enzyme, and then filters the extracting solution; passing the extract through D101 macroporous resin, washing with water of appropriate volume, collecting combined lower column liquid and water washing liquid, combining, concentrating to a certain volume, slowly adding a certain amount of 95% ethanol under stirring, standing at low temperature in a refrigerator, filtering, concentrating the filtrate until no alcohol exists, dialyzing by using a dialysis bag with a certain molecular weight cut-off, concentrating the dialysate under reduced pressure, and vacuum drying to obtain the final product, wherein the protein extraction rate is not less than 53.5%, the protein content is not less than 73.8%, and the ratio of polypeptide molecular weight not more than 2kDa is not less than 92.7%.
Compared with the prior art, the technical scheme of the invention has the following advantages:
the extraction rate of the protein of the purslane polypeptide extract prepared by the invention is more than or equal to 53.5%, the protein content is more than or equal to 73.8%, and the ratio of the molecular weight of the polypeptide to the molecular weight of 2kDa is more than or equal to 92.7%. The invention has the advantages of simple process, easy operation, low polypeptide molecular weight and the like, and the process is easy for industrial production.
Detailed Description
For a better understanding of the present invention, reference is made to the following examples. It is to be understood that these examples are for further illustration of the invention and are not intended to limit the scope of the invention. Moreover, it should be understood that the invention is not limited to the above-described embodiments, but is capable of various modifications and changes within the scope of the invention.
Example 1
Crushing a dried purslane medicinal material into 30-mesh coarse powder, weighing 500g, adding 5L petroleum ether (boiling range is 60-90 ℃) for reflux extraction for 2h, filtering, and volatilizing the petroleum ether in filter residues; adding 5L of water into the defatted medicinal materials, adding 1.5g of papain, and soaking at 50 deg.C for 2 hr; adjusting pH of the extractive solution to 9 with NaOH aqueous solution, adding 1.5g alkaline protease, soaking at 50 deg.C for 2 hr, boiling for 20min to inactivate enzyme, filtering with 300 mesh filter cloth, concentrating the extractive solution under reduced pressure at-0.07 Mpa under 65 deg.C to 200ml, adjusting pH of the concentrated solution to neutral with HCl aqueous solution, centrifuging, and collecting supernatant; passing through 50g D101 macroporous resin column, collecting eluate, washing with 80ml water, and mixing eluate with water. Concentrating the lower column solution and water washing solution to 50ml, slowly adding 200ml 95% ethanol under stirring, standing at 4 deg.C for 3 hr, filtering, and collecting supernatant. Concentrating the supernatant until no alcohol exists, adding water to a constant volume of 50ml, dialyzing at room temperature for 10h by using a dialysis bag with the molecular weight cutoff of 2kDa, concentrating the dialysate under reduced pressure, and drying in vacuum to obtain the purslane polypeptide extract product.
The detection calculation shows that the protein extraction rate of the purslane polypeptide extract is 53.5%, the protein content is 75.4%, and the ratio of the polypeptide molecular weight less than or equal to 2kDa is 92.7%.
Example 2
Crushing the dried purslane medicinal material into 30-mesh coarse powder, weighing 500g, adding 4L petroleum ether (boiling range is 60-90 ℃) for reflux extraction for 3h, filtering, and volatilizing the petroleum ether in filter residues. Adding 6L water into the defatted medicinal materials, adding 2g papain, and soaking at 55 deg.C for 2 hr. Adjusting pH of the extractive solution to 10 with NaOH aqueous solution, adding 2g alkaline protease, soaking at 55 deg.C for 2h, boiling for 30min to inactivate enzyme, filtering with 300 mesh filter cloth, concentrating the extractive solution under reduced pressure at vacuum degree of-0.07 Mpa and temperature of 65 deg.C to 250ml, adjusting pH of the concentrated solution to neutral with HCl aqueous solution, centrifuging, and retaining supernatant; passing through 60g D101 macroporous resin column, collecting lower column liquid, washing with 90ml water, and mixing the lower column liquid and the water washing liquid. Concentrating the lower column solution and water washing solution to 40ml, slowly adding 200ml 95% ethanol under stirring, standing at 4 deg.C for 2 hr, filtering, and collecting supernatant. Concentrating the supernatant until no alcohol exists, adding water to a constant volume of 40ml, dialyzing at room temperature for 5h by using a dialysis bag with the molecular weight cutoff of 2kDa, concentrating the dialysate under reduced pressure, and drying in vacuum to obtain the purslane polypeptide extract product.
The detection calculation shows that the extraction rate of protein in the purslane polypeptide extract is 56.2%, the protein content is 73.8%, and the ratio of polypeptide molecular weight less than or equal to 2kDa is 94.1%.
Example 3
Crushing the dried purslane medicinal material into 30-mesh coarse powder, weighing 500g, adding 5L petroleum ether (boiling range is 60-90 ℃) for reflux extraction for 2h, filtering, and volatilizing the petroleum ether in filter residues. Adding 6L water into the defatted medicinal materials, adding 2.5g papain, and soaking at 55 deg.C for 1.5h. Adjusting pH of the extractive solution to 9.5 with NaOH aqueous solution, adding 2.5g alkaline protease, soaking at 60 deg.C for 1.5h, boiling for 20min to inactivate enzyme, filtering with 300 mesh filter cloth, concentrating the extractive solution under reduced pressure at-0.07 Mpa and 65 deg.C to 200ml, adjusting pH of the concentrated solution to neutral with HCl aqueous solution, centrifuging, and collecting supernatant; passing through 50g D101 macroporous resin column, collecting eluate, washing with 80ml water, and mixing eluate with water. Concentrating the lower column solution and water washing solution to 50ml, slowly adding 300ml 95% ethanol under stirring, standing at 6 deg.C for 4 hr, filtering, and collecting supernatant. Concentrating the supernatant until no alcohol exists, adding water to a constant volume of 50ml, dialyzing at room temperature for 8h by using a dialysis bag with the molecular weight cutoff of 2kDa, concentrating the dialysate under reduced pressure, and drying in vacuum to obtain the purslane polypeptide extract product.
The detection calculation shows that the extraction rate of protein in the purslane polypeptide extract is 58.5%, the content of protein is 78.3%, and the ratio of the molecular weight of the polypeptide to the molecular weight of the polypeptide is not more than 2kDa is 95.9%.
Comparative example 1
Pulverizing dried herba Portulacae into 30 mesh coarse powder, weighing 500g, adding 5L 95% ethanol, reflux extracting for 2 hr, filtering, and volatilizing ethanol in the residue. Adding 6L water into the defatted medicinal materials, adding 2.5g papain, and soaking at 55 deg.C for 1.5h. Adjusting the pH of the extracting solution to 9.5 by using NaOH aqueous solution, adding 2.5g of alkaline protease, soaking at 60 ℃ for 1.5h, boiling for 20min to inactivate enzyme, filtering by using 300-mesh filter cloth, extracting the residue by using 4L of water under reflux for 1.5h, filtering by using 300-mesh filter cloth, concentrating the extracting solution under reduced pressure to 200ml at the temperature of 65 ℃ under the vacuum degree of-0.07 Mpa, adjusting the pH of the concentrated solution to be neutral by using HCl aqueous solution, and centrifuging to retain supernatant; passing through 50g AB-8 macroporous resin column, collecting lower column liquid, washing with 100ml water, and mixing the lower column liquid and the water washing liquid. Concentrating the lower column solution and water washing solution to 60ml, slowly adding 350ml 95% ethanol under stirring, standing at 6 deg.C for 4 hr, filtering, and collecting supernatant. Concentrating the supernatant under reduced pressure, and vacuum drying to obtain the final product.
The detection calculation shows that the protein extraction rate of the purslane polypeptide extract is 32.5%, the protein content is 43.9%, and the ratio of the molecular weight of the polypeptide to the molecular weight of the polypeptide is less than or equal to 2kDa is 72.8%.
Comparative example 2
Crushing a dried purslane medicinal material into coarse powder of 30 meshes, weighing 500g, adding 6L of water, adjusting the pH of an extracting solution to 9 by using NaOH aqueous solution, adding 2.5g of alkaline protease, soaking for 2h at 60 ℃, boiling for 20min to inactivate enzyme, filtering by using 300-mesh filter cloth, refluxing and extracting medicinal residues for 1.5h by using 4L of water, filtering by using 300-mesh filter cloth, concentrating the extracting solution under reduced pressure, adjusting the pH of a concentrated solution to be neutral by using HCl aqueous solution at 65 ℃ of-0.07 Mpa, centrifuging and reserving supernate; passing through 60g active carbon (200 mesh) column, collecting lower column liquid, washing with 100ml water, and mixing the lower column liquid and the water washing liquid. Concentrating the lower column solution and water washing solution to 60ml, slowly adding 200ml 95% ethanol under stirring, standing at 4 deg.C for 2 hr, filtering, and collecting supernatant. Concentrating the supernatant under reduced pressure to 60ml, dialyzing with 3.5kDa dialysis bag at room temperature for 8h, concentrating the dialysate under reduced pressure, and vacuum drying to obtain the final product.
The detection calculation shows that the protein extraction rate of the purslane polypeptide extract is 36.5%, the protein content is 38.1%, and the ratio of the polypeptide molecular weight not more than 2kDa is 64.8%.
Comparative example 3
Pulverizing dried herba Portulacae into 30 mesh coarse powder, weighing 500g, adding 5L ethyl acetate, reflux extracting for 2 hr, filtering, and volatilizing ethyl acetate in the residue. Adding 6L water into the defatted medicinal materials, adding 2.5g papain, and soaking at 55 deg.C for 1.5h. Adjusting pH of the extractive solution to 9.5 with NaOH aqueous solution, adding 2.5g alkaline protease, soaking at 60 deg.C for 1.5h, boiling for 20min to inactivate enzyme, filtering with 300 mesh filter cloth, concentrating the extractive solution under reduced pressure at-0.07 Mpa and 65 deg.C to 200ml, adjusting pH of the concentrated solution to neutral with HCl aqueous solution, centrifuging, and collecting supernatant; passing through 50g D101 macroporous resin column, collecting eluate, washing with 80ml water, and mixing eluate with water. Concentrating the lower column solution and water washing solution to 50ml, slowly adding 300ml 95% ethanol under stirring, standing at 6 deg.C for 4 hr, filtering, and collecting supernatant. Concentrating the supernatant until no alcohol exists, adding water to a constant volume of 50ml, dialyzing at room temperature for 8h by using a dialysis bag with the molecular weight cutoff of 2kDa, concentrating the dialysate under reduced pressure, and drying in vacuum to obtain the purslane polypeptide extract product.
The detection calculation shows that the protein extraction rate of the purslane polypeptide extract is 41.0 percent, the protein content is 53.7 percent, and the ratio of the molecular weight of the polypeptide to the molecular weight of the polypeptide is less than or equal to 2kDa is 60.1 percent.
Comparative example 4
Crushing the dried purslane medicinal material into 30-mesh coarse powder, weighing 500g, adding 5L petroleum ether (boiling range is 60-90 ℃) for reflux extraction for 2h, filtering, and volatilizing the petroleum ether in filter residues. Adding 6L water into the defatted medicinal materials, adding 2.5g papain, and soaking at 55 deg.C for 1.5h. Adjusting pH of the extractive solution to 9.5 with NaOH aqueous solution, adding 2.5g alkaline protease, soaking at 60 deg.C for 1.5h, boiling for 20min to inactivate enzyme, filtering with 300 mesh filter cloth, concentrating the extractive solution under reduced pressure at-0.07 Mpa and 65 deg.C to 200ml, adjusting pH of the concentrated solution to neutral with HCl aqueous solution, centrifuging, and collecting supernatant; passing through 50g D101 macroporous resin column, collecting eluate, washing with 80ml water, and mixing eluate with water. Concentrating the lower column solution and water washing solution to 50ml, slowly adding 400ml 95% ethanol under stirring, standing at 6 deg.C for 4 hr, filtering, and collecting supernatant. Concentrating the supernatant until no alcohol exists, adding water to a constant volume of 50ml, dialyzing at room temperature for 8h by using a dialysis bag with the molecular weight cutoff of 2kDa, concentrating the dialysate under reduced pressure, and drying in vacuum to obtain the purslane polypeptide extract product.
The detection calculation shows that the extraction rate of protein in the purslane polypeptide extract is 43.1%, the protein content is 43.9%, and the ratio of the molecular weight of the polypeptide to the molecular weight of the polypeptide is not more than 2kDa is 55.3%.
Effects of the embodiment
1. Related detection method for purslane polypeptide extract
The protein detection method comprises the following steps: the determination method is based on GB 5009.5-2016
The molecular weight of the polypeptide is less than or equal to 10 kDa: the determination method is based on GB 31645-2018
2. Formula for calculation
Protein extraction rate = (protein mass/medicinal material mass) × 100%
The indexes of the purslane polypeptide extracts prepared in examples 1-3 and comparative examples 1-4 are shown in table 1.
TABLE 1 indices of purslane polypeptide extracts prepared in examples 1-3 and comparative examples 1-4
Case(s) | Protein content% | The extraction rate% | The molecular weight of the polypeptide is less than or equal to 2kDa |
Example 1 | 53.5 | 75.4 | 92.7 |
Example 2 | 56.2 | 73.8 | 94.1 |
Example 3 | 58.5 | 78.3 | 95.9 |
Comparative example 1 | 32.5 | 43.9 | 72.8 |
Comparative example 2 | 36.5 | 38.1 | 64.8 |
Comparative example 3 | 41.0 | 53.7 | 60.1 |
Comparative example 4 | 43.1 | 43.9 | 55.3 |
The purslane polypeptide extract is prepared from purslane as a raw material through degreasing, enzymolysis, macroporous resin purification, alcohol precipitation and dialysis, wherein the extraction rate of protein in the purslane polypeptide extract is not less than 53.5%, the content of protein is not less than 73.8%, and the ratio of polypeptide molecular weight not more than 2kDa is not less than 92.7%. The process has the advantages of simple operation, high extraction rate, high protein purity, low polypeptide molecular weight and the like; meanwhile, the process is easy for industrial production.
The above description is not intended to limit the invention, nor is the invention limited to the examples described above. Those skilled in the art should also realize that changes, modifications, additions and substitutions can be made without departing from the spirit of the invention.
Claims (2)
1. A preparation method of a purslane polypeptide extract is characterized by comprising the following steps:
(1) Degreasing: crushing the dried purslane medicinal material into coarse powder, adding petroleum ether with the boiling range of 60-90 ℃ in an amount which is 8-10 times that of the medicinal material, refluxing and extracting for 2-3 h, filtering, and volatilizing the solvent in the filter residue to obtain the medicinal material degreased by the petroleum ether;
(2) And (3) extraction and concentration: adding water into the degreased medicinal materials of petroleum ether, adding neutral protease accounting for 0.3-0.5% of the medicinal materials, and extracting by warm immersion; after warm-soaking, adjusting the pH of the extraction solution to 9-10 by using an alkali solution, adding alkaline protease accounting for 0.3-0.5% of the amount of the medicinal materials into the extraction solution, and performing warm-soaking extraction; after warm immersion, boiling to inactivate enzyme, filtering with filter cloth, concentrating under reduced pressure, adjusting pH of the concentrated solution to neutrality with acid solution, centrifuging, and retaining supernatant;
(3) D101 purification: enabling the supernatant to pass through D101 macroporous resin, wherein the dosage of the D101 macroporous resin is 1/10-1/8 of the amount of the medicinal materials, collecting lower column liquid, washing the column with water, collecting water washing liquid, combining the lower column liquid and the water washing liquid, and concentrating under reduced pressure to obtain first concentrated solution;
(4) Alcohol precipitation: slowly adding 90-100% ethanol in an amount which is 4-6 times that of the first concentrated solution while stirring, standing for 2-5 h at the temperature of 2-6 ℃, then performing suction filtration, and performing reduced pressure water replenishing and concentration on the suction filtered solution until no ethanol exists to obtain a second concentrated solution;
(5) And (3) dialysis: dialyzing the second concentrated solution for 5 to 10 hours at room temperature by using a dialysis bag with the molecular weight cutoff of 2kDa, and carrying out vacuum drying and crushing on the concentrated solution after carrying out reduced pressure concentration on the dialyzate to obtain the purslane polypeptide extract;
in the step (2), the addition amount of the water is 10-12 times of the amount of the medicinal materials;
in the step (2), the neutral protease is papain;
in the step (2), the temperature of the warm leaching extraction is 45-60 ℃, and the time is 1.5-2 h;
in the step (2), the time for boiling and enzyme deactivation is 10-30 min;
in the step (2), the vacuum degree during the reduced pressure concentration is-0.07 to-0.09 Mpa, the temperature is 60 to 80 ℃, and the volume of the concentrated solution is 1/3 to 1/2 of the amount of the medicinal materials;
in the step (3), the using amount of the water is 1-1.5 BV of the volume of the resin;
in the step (3), the vacuum concentration is carried out until the amount of the medicinal materials is 1/15-1/10;
in the step (4), the volume of the concentration is 1/15-1/10 of the amount of the medicinal materials.
2. A purslane polypeptide extract, which is prepared by the preparation method of the purslane polypeptide extract as claimed in claim 1.
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