CN114276408A - Extraction method and application of acanthopanax senticosus glycoprotein - Google Patents
Extraction method and application of acanthopanax senticosus glycoprotein Download PDFInfo
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Abstract
The invention provides an extraction method and application of acanthopanax senticosus glycoprotein, wherein the extraction method comprises the step of heating and extracting acanthopanax senticosus coarse powder, wherein the extraction solution is PBS (phosphate buffer solution) with the pH value of 5-9, the extraction temperature is 30-40 ℃, and a table concentrator is also adopted for assisting in the heating and extraction process. The invention adopts PBS buffer solution with pH value of 5-9 as leaching solution, and the extraction is carried out at specific leaching temperature with the aid of a table concentrator, so that the extraction rate of glycoprotein can be improved, the obtained acanthopanax glycoprotein has stable structure, and the activity of glycoprotein is ensured.
Description
Technical Field
The invention relates to the technical field of biochemistry, in particular to an extraction method and application of acanthopanax glycoprotein.
Background
Glycoproteins are substances with biological activity in which oligosaccharide chains and polypeptides are covalently linked in various forms, and are one of important biological macromolecules in organisms, are widely found in animals, plants and microorganisms, even in unicellular organisms and viruses, and are distributed in various forms and types in intracellular and extracellular fluids and tissues of organisms to constitute various active substances in organisms. The natural glycoprotein has physiological activities of resisting oxidation, resisting tumor, improving immunity, etc. The glycoprotein plays an important role in growth and development, information transmission, immune system, nervous system and other various life activities, and is related to the occurrence of various diseases (such as cardiovascular diseases, liver and kidney diseases, diabetes, tumor and cancer).
Acanthopanax senticosus, also known as hovenia dulcis thunb, acanthopanax senticosus, acanthopanax bark and the like, is a plant of the genus acanthopanax of the family araliaceae, and is distributed in Heilongjiang, Jilin, Liaoning, Hebei and Shanxi in China, and is a rhizome medicinal material which is rich in nutrition and has various physiological functions and development potentials. Acanthopanax has wide application, has the effects of tonifying qi and spleen, tonifying kidney and soothing nerves and the like, and is mainly used for treating symptoms such as spleen-lung qi deficiency, body weakness and hypodynamia, inappetence, lung-kidney deficiency, chronic cough and asthma, kidney deficiency and soreness of waist and knees, heart and spleen deficiency, insomnia and dreaminess and the like.
However, no method for extracting acanthopanax senticosus glycoprotein is available, and the acanthopanax senticosus extract in the prior art is a crude extract, has complex components and lacks deep research significance, wherein the content of the glycoprotein is low and the purity is not enough, so that the acanthopanax senticosus glycoprotein can not be further applied at high level.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an extraction method and application of acanthopanax senticosus glycoprotein.
In a first aspect, the invention provides a method for extracting acanthopanax senticosus glycoprotein, which comprises the step of heating and extracting acanthopanax senticosus coarse powder, wherein the extracting solution is a PBS (phosphate buffer solution) with the pH of 5-9, the extracting temperature is 30-40 ℃, and a shaking table is adopted for assisting the heating and extracting process.
In the prior art, active ingredients are generally extracted from acanthopanax senticosus medicinal materials by a water extraction and alcohol precipitation method, and then the acanthopanax senticosus extract with acceptable purity is obtained through a series of purification steps, namely, the existing method is complicated, and the obtained product has common quality. The research of the invention finds that the extraction method adopts PBS buffer solution with pH of 5-9 as extraction solution, and the extraction is carried out at a specific extraction temperature with the aid of a table concentrator, so that the extraction rate of glycoprotein can be improved, the obtained acanthopanax glycoprotein has stable structure, and the activity of glycoprotein is ensured.
Further, in the heating leaching process, the shaking speed of the shaking table is 40-300 r/min.
Furthermore, in the heating leaching process, the material-liquid ratio is 1:10-1:20g/mL, the leaching time is 6-48h, and the leaching times are 2-4.
In a preferred embodiment of the invention, in the heating leaching process, the leaching solution is a PBS buffer solution with the pH value of 8, the leaching temperature is 30 ℃, the shaking rate of a shaking table is 160r/min, the material-liquid ratio is 1:15g/mL, the leaching time is 12h, and the leaching times are 3 times.
Further, after the heating extraction is carried out to obtain the acanthopanax glycoprotein crude extract, the steps of removing free protein, dialyzing and freeze-drying are also included.
In a specific embodiment provided by the present invention, the extraction method comprises the steps of:
(1) pretreatment: pulverizing radix Acanthopanacis Senticosi into coarse powder;
(2) table-assisted and heating leaching: uniformly mixing the acanthopanax powder obtained in the step (1) with a PBS (phosphate buffer solution) with the pH value of 5-9 at a feed-liquid ratio of 1:10-1:20g/mL, heating and leaching at 30-40 ℃ for 6-48h at a shaking table shaking speed of 40-300r/min for 1-4 times, filtering, and combining filtrates to obtain an acanthopanax glycoprotein crude extract;
(3) removing free protein: removing free protein from the coarse extract of the acanthopanax glycoprotein obtained in the step (2) by adopting a Sevage method to obtain an acanthopanax glycoprotein solution;
(4) and (3) dialysis and freeze-drying: and (4) filling the acanthopanax senticosus glycoprotein solution obtained in the step (3) into a dialysis bag, putting the dialysis bag into distilled water, dialyzing for 48 hours, and freeze-drying the dialyzed retentate to obtain acanthopanax senticosus glycoprotein powder.
The invention is assisted by a shaking table, uses PBS solution for extraction to obtain glycoprotein crude extract with protein content of about 15 percent, then removes free protein to obtain residual protein connected with sugar, namely glycoprotein, and then further purifies to obtain acanthopanax glycoprotein component with higher purity. The extraction method of the acanthopanax senticosus glycoprotein has the advantages of reasonable design, simple process, convenient operation, high product extraction rate, high purity, stable property, no need of using a large amount of organic reagents, low preparation cost, easy popularization and application and the like.
The coarse powder in the invention means that the powder passes through the No. one sieve totally, but the powder mixed with the coarse powder can pass through the No. three sieve by no more than 20%.
Further, the removal of free protein by the Sevage method specifically includes: adding Sevage reagent into the coarse extract of the acanthopanax senticosus glycoprotein, shaking for 1h at 4 ℃, then centrifuging for 15-20min at the rotating speed of 9000r/min, collecting supernatant, and repeating the step until no protein layer is separated out.
Further preferably, the dosage of the Sevage reagent is 1/4 volume of the acanthopanax senticosus glycoprotein crude extract, and the Sevage reagent is prepared by mixing n-butanol and chloroform according to the ratio of 1: 4.
Further, the dialysis was carried out at 4 ℃ using a dialysis bag with a molecular weight cut-off of 0.1kDa, and the dialysate was changed every 6 to 7 hours during the dialysis.
In a second aspect, the invention provides an acanthopanax senticosus glycoprotein extracted by the extraction method of the acanthopanax senticosus glycoprotein.
Further, the sum of the sugar content and the protein content of the acanthopanax senticosus glycoprotein is 50-80%, namely the purity of the acanthopanax senticosus glycoprotein is 50-80%.
In a third aspect, the invention provides an application of the acanthopanax senticosus glycoprotein in preparing a functional product with skin oxidation resistance and/or aging resistance.
The invention proves that the acanthopanax senticosus glycoprotein has obvious effects on oxidation resistance and skin photodamage through in-vivo and in-vitro experiments, and provides reference basis for the application of the acanthopanax senticosus glycoprotein in the research and development of skin care products.
The invention provides an extraction method and application of acanthopanax senticosus glycoprotein, wherein PBS buffer solution with pH of 5-9 is used as extraction solution, and the extraction is carried out at a specific extraction temperature in a way of being matched with a shaking table, so that the extraction rate of glycoprotein can be improved, the obtained acanthopanax senticosus glycoprotein has a stable structure, and the activity of glycoprotein is ensured.
Drawings
FIG. 1 shows the effect of Acanthopanax senticosus glycoprotein of example 1 on SOD activity in mouse skin tissue;
FIG. 2 is a graph showing the effect of Acanthopanax senticosus glycoprotein of example 1 on MDA activity in mouse skin tissue;
FIG. 3 is a graph showing the effect of Acanthopanax senticosus glycoprotein of example 1 of the present invention on GSH-Px activity in mouse skin tissue;
FIG. 4 shows the effect of the Acanthopanax senticosus glycoprotein of example 1 on HYP activity in mouse skin tissue.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless otherwise specified, the test reagents and materials used in the examples of the present invention are commercially available.
Unless otherwise specified, the technical means used in the examples of the present invention are conventional means well known to those skilled in the art.
Wherein the purity is determined as follows:
the glycoprotein is a macromolecular compound, the purity standard of which cannot be measured by the common purity standard of a small molecular compound, because even pure glycoprotein is not uniform in a microscopic scale, and therefore, the purity of the acanthopanax senticosus glycoprotein prepared by the invention is characterized by the following modes: the acanthopanax senticosus glycoprotein has a purity of sugar content + protein content.
Determination of sugar content
The determination method comprises the following steps: determining the absorbance of glucose with different concentrations by a phenol-sulfuric acid method to prepare a concentration-absorbance standard curve, preparing a solution with a certain concentration from the acanthopanax senticosus glycoprotein sample, measuring the absorbance of the solution according to the standard curve term, substituting the solution into a concentration-absorbance standard curve equation, and calculating to obtain the sugar content in the obtained glycoprotein.
Preparation of a standard curve: accurately weighing 2mg of anhydrous glucose standard substance, adding a proper amount of distilled water for dissolving, fixing the volume to a scale mark by using a 50mL volumetric flask, shaking up, and preparing into a standard solution of 40 mu g/mL.
0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1.0mL of glucose standard solution were aspirated, each supplemented with 1.0mL of deionized water. Then, 0.5mL of 6% phenol and 2.5mL of concentrated sulfuric acid were added, shaken, and after standing at room temperature for 30min, absorbance was measured at 490nm, and a standard curve was drawn with 1.0mL of distilled water, 0.5mL of phenol and 2.5mL of concentrated sulfuric acid as blanks, the polysaccharide concentration as abscissa, and the absorbance as ordinate. From the measurement results, a linear regression equation was prepared: y is 0.0077X +0.223, R2Is 0.9994, wherein y is the absorbance value and x is the polysaccharide concentration mg/mL.
Accurately weighing 2mg of acanthopanax senticosus glycoprotein, adding distilled water to dissolve the acanthopanax senticosus glycoprotein, and fixing the volume to 10mL, wherein the final concentration of a sample is 0.2 mg/mL; accurately sucking 1mL of each sample solution, and then measuring the light absorption value according to a standard curve preparation method.
Measurement of protein content
The protein content in the acanthopanax senticosus glycoprotein is determined by adopting a BCA method. Preparing the protein standard preparation solution into 0.5mg/mL protein standard solution, adding 0, 1, 2, 4, 8, 12, 16 and 20 mu L of each protein standard solution into a standard well of a 96-well plate, adding the standard dilution solution to make up to 20 mu L, adding 200 mu LBCA working solution into each well, standing at 37 ℃ for 20-30min, and measuring absorbance at the wavelength of 562 nm. And (3) drawing a standard curve by taking the absorbance as a vertical coordinate and the concentration of the protein standard solution as a horizontal coordinate, and fitting a regression equation.
The results show that the regression equation for the protein standard curve is: y-0.8369X +0.1588 (r-0.9964) where X is the concentration of protein (in mg-1) And y is the absorbance value.
The acanthopanax senticosus glycoprotein is determined by a BCA kit on an enzyme labeling instrument or an ultraviolet-visible spectrophotometer, and the operations are strictly carried out according to the kit instructions.
In summary, the acanthopanax senticosus glycoprotein prepared by the invention has the purity of sugar content plus protein content.
Example 1
The embodiment provides an extraction method of acanthopanax senticosus glycoprotein, which comprises the following specific steps:
(1) pretreatment: pulverizing radix Acanthopanacis Senticosi into coarse powder;
(2) table-assisted and heating leaching: uniformly mixing 3g of the acanthopanax powder obtained in the step (1) with a PBS (phosphate buffer solution) with the pH value of 8 according to a material-liquid ratio of 1:15g/mL, heating and leaching for 12 hours at 30 ℃ at a shaking table shaking speed of 160r/min for 3 times, filtering, and combining filtrates to obtain an acanthopanax glycoprotein crude extract;
(3) removing free protein: removing free protein from the coarse extract of the acanthopanax glycoprotein obtained in the step (2) by adopting a Sevage method to obtain an acanthopanax glycoprotein solution;
(4) and (3) dialysis and freeze-drying: and (4) filling the acanthopanax senticosus glycoprotein solution obtained in the step (3) into a dialysis bag with the molecular weight cutoff of 0.1kDa, putting the dialysis bag into distilled water, dialyzing for 48 hours, and freeze-drying the dialyzed retentate to obtain the acanthopanax senticosus glycoprotein powder.
Through detection, in the embodiment, the yield of the acanthopanax senticosus glycoprotein (the yield refers to the amount of the obtained glycoprotein freeze-dried powder/3 g of acanthopanax senticosus coarse powder) is 8.61%, and the purity is 73.16%.
Example 2
The embodiment provides an extraction method of acanthopanax senticosus glycoprotein, which comprises the following specific steps:
(1) pretreatment: pulverizing radix Acanthopanacis Senticosi into coarse powder;
(2) table-assisted and heating leaching: uniformly mixing 3g of the acanthopanax powder obtained in the step (1) with a PBS (phosphate buffer solution) with the pH value of 8 according to a material-liquid ratio of 1:15g/mL, heating and leaching for 12 hours at 60 ℃ at a shaking table shaking speed of 160r/min for 3 times, then filtering, and combining filtrates to obtain an acanthopanax glycoprotein crude extract;
(3) removing free protein: removing free protein from the coarse extract of the acanthopanax glycoprotein obtained in the step (2) by adopting a Sevage method to obtain an acanthopanax glycoprotein solution;
(4) and (3) dialysis and freeze-drying: and (4) filling the acanthopanax senticosus glycoprotein solution obtained in the step (3) into a dialysis bag with the molecular weight cutoff of 0.1kDa, putting the dialysis bag into distilled water, dialyzing for 48 hours, and freeze-drying the dialyzed retentate to obtain the acanthopanax senticosus glycoprotein powder.
Through detection, the yield of the acanthopanax senticosus glycoprotein in the embodiment is 7.86%, and the purity is 57.35%.
Example 3
The embodiment provides an extraction method of acanthopanax senticosus glycoprotein, which comprises the following specific steps:
(1) pretreatment: pulverizing radix Acanthopanacis Senticosi into coarse powder;
(2) table-assisted and heating leaching: uniformly mixing 3g of the acanthopanax powder obtained in the step (1) with a PBS (phosphate buffer solution) with the pH value of 8 according to a material-liquid ratio of 1:15g/mL, heating and leaching for 12 hours at 30 ℃ at a shaking table shaking speed of 400r/min for 3 times, filtering, and combining filtrates to obtain an acanthopanax glycoprotein crude extract;
(3) removing free protein: removing free protein from the coarse extract of the acanthopanax glycoprotein obtained in the step (2) by adopting a Sevage method to obtain an acanthopanax glycoprotein solution;
(4) and (3) dialysis and freeze-drying: and (4) filling the acanthopanax senticosus glycoprotein solution obtained in the step (3) into a dialysis bag with the molecular weight cutoff of 0.1kDa, putting the dialysis bag into distilled water, dialyzing for 48 hours, and freeze-drying the dialyzed retentate to obtain the acanthopanax senticosus glycoprotein powder.
Through detection, the yield of the acanthopanax senticosus glycoprotein in the embodiment is 9.12%, and the purity is 68.56%.
Example 4
The embodiment provides an extraction method of acanthopanax senticosus glycoprotein, which comprises the following specific steps:
(1) pretreatment: pulverizing radix Acanthopanacis Senticosi into coarse powder;
(2) table-assisted and heating leaching: uniformly mixing 3g of the acanthopanax powder obtained in the step (1) with a PBS (phosphate buffer solution) with the pH value of 7 according to a material-to-liquid ratio of 1:20g/mL, heating and leaching for 24 hours at 40 ℃ at a shaking table shaking speed of 200r/min, filtering, and combining filtrates to obtain an acanthopanax glycoprotein crude extract;
(3) removing free protein: removing free protein from the coarse extract of the acanthopanax glycoprotein obtained in the step (2) by adopting a Sevage method to obtain an acanthopanax glycoprotein solution;
(4) and (3) dialysis and freeze-drying: and (4) filling the acanthopanax senticosus glycoprotein solution obtained in the step (3) into a dialysis bag with the molecular weight cutoff of 0.1kDa, putting the dialysis bag into distilled water, dialyzing for 48 hours, and freeze-drying the dialyzed retentate to obtain the acanthopanax senticosus glycoprotein powder.
Through detection, the yield of the acanthopanax senticosus glycoprotein in the embodiment is 8.21%, and the purity is 62.36%.
Skin antioxidation test using acanthopanax senticosus glycoprotein
Experiments prove that the acanthopanax senticosus glycoprotein prepared in the embodiment 1 of the invention has antioxidation, and the experimental method and the results are as follows:
1. subject: SD rats.
2. Index measuring and analyzing method
The determination of SOD, MDA, GSH-Px and HYP in skin tissue is carried out on an enzyme labeling instrument or an ultraviolet-visible spectrophotometer, and the operation is strictly carried out according to the kit instruction.
Superoxide dismutase (SOD) kit (cat # A001-1, production lot # 20210423), Malondialdehyde (MDA) determination box (cat # A003-1-2, production lot # 20201203), glutathione peroxidase (GSH-Px) test box (cat # A005-1-1, production lot # 20201222) and hydroxyproline (alkaline hydrolysis) kit (cat # A030-2-1, production lot # 20201223) are all provided by Nanjing institute of bioengineering.
3. Grouping, modeling and administration of animals
30 SD-grade female rats are selected and randomly divided into blanksGroup, model group, positive control group, acanthopanax glucoprotein high, medium and low dose group, each group contains 5. Removing the back hair of the area of about 3cm multiplied by 4cm on the back of all the mice by using electric pushing, not being suitable for ultraviolet irradiation on the day of hair removal, and removing the hair of all the mice by using the same method every 3 days after the experiment is started. Then ultraviolet irradiation is carried out, the radiation intensity of UVA and UVB lamp tubes is measured before irradiation, and the minimum red spot quantity (minimum erythema quality) is measured to be 37.34J/cm by an ultraviolet irradiation measuring instrument2. The irradiation time is set according to the minimum erythema dose. The shaved rats were placed in a home-made ultraviolet lamp box, and ultraviolet (UVA + UVB) irradiation was performed with a fixed irradiation distance of 10 cm. 0.5 h/day in the first week, 1 h/day in the second week, and 2 h/day in the third week, and maintaining for 2 h/day thereafter until the skin of the back of the rat is dry, peeled, and even has local ulceration, namely the molding is finished. By adopting a method of administration after molding, except a blank group, the back of a positive control group is coated with 10mg/mL of vitamin E solution 0.5mL, and the back of groups with high, medium and low dosages of acanthopanax senticosus glycoprotein is coated with 25mg/mL, 10mg/mL and 1mg/mL of total acanthopanax senticosus glycoprotein 0.5mL, and each group freely takes food and drinks for 4 weeks.
4. Skin treatment
After the last dose, the cervical vertebrae were dislocated and sacrificed, skin tissue was cut off, surrounding connective tissue was stripped off, fresh weighed and data recorded.
5. Preparation of tissue homogenate
Adding 9 times volume of physiological saline into the accurately weighed skin tissue, treating in a homogenizer to prepare homogenate, centrifuging the homogenate for 20min at 4000r/min, taking the supernatant to obtain 10% skin tissue homogenate, and freezing for later use.
6. Data processing
The experimental data were statistically analyzed using the SPSS16.0 software, expressed as mean and standard deviation (x S), and compared between groups using the t-test, with differences of P <0.05 or P <0.01, indicating that the differences between groups were statistically significant.
7. Results of the experiment
7.1 Effect of Acanthopanax Senticosus glycoprotein on SOD Activity in mouse skin tissue
As shown in FIG. 1, it was found that the SOD activity was significantly reduced in the skin of the model rats as compared with that of the blank rats. Compared with the model group, the SOD activity of the acanthopanax senticosus glycoprotein administration group is obviously increased and is positively correlated with the dosage. The SOD activity of the acanthopanax senticosus glycoprotein group is higher than that of the positive group, which shows that the acanthopanax senticosus glycoprotein and vitamin E have similar antioxidant effect and the effect of the acanthopanax senticosus glycoprotein is probably better than that of the vitamin E.
7.2 Effect of Acanthopanax Senticosus glycoprotein on MDA Activity in mouse skin tissue
As shown in FIG. 2, it is clear that the MDA content in the skin of the model group rats is significantly increased compared to that of the blank group. Compared with the model group, the MDA content of the acanthopanax glycoprotein administration group is obviously reduced, and the MDA content is gradually reduced along with the increase of the acanthopanax glycoprotein dose. The MDA content of the acanthopanax senticosus glycoprotein group is lower than that of the acanthopanax senticosus glycoprotein group, which shows that the effect of inhibiting oxidative stress from generating malondialdehyde of the acanthopanax senticosus glycoprotein is better than that of VE.
7.3 Effect of Acanthopanax Senticosus glycoprotein on GSH-Px Activity in mouse skin tissue
As shown in FIG. 3, it is clear that the GSH-Px activity in the skin of the model group rats is significantly reduced compared to that of the blank group. Compared with the model group, the GSH-Px activity of the acanthopanax glycoprotein administration group is obviously increased and is positively correlated with the dosage. The GSH-Px activity of the acanthopanax senticosus glycoprotein group is higher than that of the positive group, which shows that the acanthopanax senticosus glycoprotein can promote the production of glutathione peroxidase in vivo, has the functions of scavenging free radicals and derivatives, reducing the formation of lipid peroxides and enhancing the capability of resisting oxidative damage of organisms.
7.4 Effect of Acanthopanax senticosus glycoprotein on HYP Activity in mouse skin tissue
The results are shown in FIG. 4, which shows that HYP content in the skin of the model group rats is significantly reduced compared to the blank group. Compared with the model group, the HYP content of the acanthopanax senticosus glycoprotein administration group is obviously increased and is positively correlated with the dosage. Hydroxyproline is a main amino acid contained in collagen, and the collagen is a main component forming interstitial collagen fibers of skin connective tissue cells, so that the acanthopanax senticosus glycoprotein can promote the damaged skin to generate the collagen and repair the damage.
In conclusion, the acanthopanax senticosus glycoprotein has obvious antioxidation and better repair effect on skin photoaging, and provides reference for the maximum development and utilization of acanthopanax senticosus in the research and development of antioxidant drugs, health-care foods and acanthopanax senticosus skin care products.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. The extraction method of the acanthopanax senticosus glycoprotein is characterized by comprising the step of heating and extracting acanthopanax senticosus coarse powder, wherein an extracting solution is a PBS (phosphate buffer solution) with the pH value of 5-9, the extracting temperature is 30-40 ℃, and a shaking table is adopted for assisting in the heating and extracting process.
2. The method of claim 1, wherein the shaking speed of the shaking table is 40-300r/min during the heat extraction.
3. The method of claim 1 or 2, wherein the material-to-liquid ratio in the heat extraction is 1:10-1:20g/mL, the extraction time is 6-48h, and the extraction times are 2-4.
4. The method of claim 3, wherein the temperature of the extraction is 30 ℃, the shaking rate of the shaking table is 160r/min, the material-to-liquid ratio is 1:15g/mL, the extraction time is 12h, and the extraction times are 3 times.
5. The method of claim 1, further comprising the steps of removing free protein and lyophilizing after the heat leaching to obtain a crude extract of Acanthopanax senticosus glycoprotein.
6. The method of claim 5, wherein the Sevage method is used to remove free protein.
7. The method of claim 5, wherein the dialysis is performed at 4 deg.C, the cut-off molecular weight of the dialysis bag is 0.1kDa, and the dialysate is changed every 6-7 hours during dialysis.
8. An Acanthopanax senticosus glycoprotein characterized by being extracted by the method for extracting Acanthopanax senticosus glycoprotein according to any one of claims 1 to 7.
9. The Acanthopanax senticosus glycoprotein according to claim 8, wherein the sum of the sugar content and the protein content is 50-80%.
10. Use of the acanthopanax senticosus glycoprotein according to claim 9 for the preparation of functional products with skin oxidation and/or aging resistance.
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