CN114774493A - A kind of extraction and purification method of Phellinus linteus active substance polysaccharide - Google Patents

Abstract

The invention discloses a method for extracting and purifying phellinus igniarius active substance polysaccharide, which comprises the following steps: freeze-drying and crushing at low temperature, performing enzymolysis after ultraviolet irradiation, performing hot water-ultrasonic extraction, extracting crude phellinus igniarius polysaccharides, degreasing, decoloring, deproteinizing, and dialyzing to finally obtain high-purity phellinus igniarius active substance polysaccharides. The invention extracts phellinus igniarius polysaccharides to the maximum extent by ultraviolet irradiation treatment and combined extraction of enzymolysis and ultrasound, and finally obtains the high-purity phellinus igniarius polysaccharides by degreasing, depigmentation and deproteinization treatment. The method provided by the invention has the advantages of high extraction rate, short extraction time, convenient operation, high purity of the obtained phellinus igniarius polysaccharides and great medicinal value.

Description

A kind of extraction and purification method of Phellinus linteus active substance polysaccharide

technical field

The invention belongs to the technical field of active substance extraction, and in particular relates to a method for extracting and purifying polysaccharide of Phellinus linteus active substance.

Background technique

Phellinus igniarius is a large and rare medicinal fungus belonging to the class Basidiomycetes, with sweet, pungent and slightly bitter taste. Modern research shows that Phellinus linteus has anti-tumor, antioxidant and other effects. This effect of Phellinus linteus is mainly due to the active substances in Phellinus linteus, and the polysaccharide is the most abundant in its active substances. At present, Phellinus linteus polysaccharide products are used for cancer treatment, but they are expensive in the market. The use of Phellinus linteus polysaccharide as an immune drug in clinic is a new direction for the treatment of tumors in recent years. It participates in the body's metabolic regulation activities with various functions. In recent years, more and more attention has been paid to it, and the research on Phellinus linteus polysaccharides has gradually increased.

At present, the research on Phellinus linteus polysaccharides mainly focuses on Phellinus linteus fruiting bodies. Phellinus linteus has a long growth cycle. It takes time and raw materials to obtain fruiting bodies. It is expensive for production and medical treatment, and it is not widely used. If the content of the polysaccharide in the mycelium of Phellinus linteus differs little from the composition and the fruiting body, the polysaccharide in the mycelium can be extracted and processed, which can reduce a lot of economic costs and can be applied to the market. The traditional sample pretreatment mainly adopts the boiling method, the extraction time is more than 6 hours, the energy consumption is high, and the yield of crude polysaccharide is low, only 2% of the crude drug. Then the crude Phellinus linteus polysaccharide is purified by gel filtration technology, which is cumbersome and time-consuming, and the product purity is low, generally about 25%. As a pharmaceutical preparation, a higher purity Phellinus linteus polysaccharide product is required.

SUMMARY OF THE INVENTION

In view of the deficiencies of the prior art, the present invention provides a method for extracting and purifying Phellinus linteus active substance polysaccharide to overcome the technical defects of low yield and low purity of Phellinus linteus polysaccharide in the prior art.

To achieve the above object, the present invention provides the following technical solutions:

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: the washed Phellinus linteus mycelium is pulverized at low temperature, and after sieving, Phellinus linteus powder is obtained;

(2) Enzymatic hydrolysis after ultraviolet irradiation: put the Phellinus linteus powder obtained in step (1) into an ultraviolet irradiation field for irradiation, then add deionized water and compound enzymes, and filter after enzymatic hydrolysis to obtain enzymatic hydrolysis filtrate and filter residue ;

(3) leaching: then adding the filter residue obtained in step (2) into deionized water, carrying out ultrasonic leaching, centrifuging after the extraction is completed, and taking the supernatant to obtain the leaching solution and the residue;

(4) Extraction of Phellinus linteus crude polysaccharide: combine the enzymatic hydrolysis filtrate obtained in step (2) and the extract obtained in step (3), heat and concentrate, centrifuge, remove the precipitate, add 95% ethanol to the supernatant, and let stand 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting and deproteinizing treatment: adding the crude polysaccharide of Phellinus linteus obtained in step (4) into petroleum ether, centrifuging, sucking and discarding the petroleum ether, repeating twice to remove lipid substances; Add hydrogen peroxide to the concentration of 20%, react in a water bath, remove the pigment, adjust the pH of the polysaccharide solution to 7.0 with ammonia water; add trichloroacetic acid to the crude polysaccharide sample after depigmentation to a concentration of 5%, stand, centrifuge, Discard the precipitation and repeat 3 times to obtain a defatted, depigmented and deproteinized polysaccharide solution;

(6) dialysis: select the dialysis bag of 7000D, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with the dialysis bag, obtain the purified Phellinus linteus polysaccharide solution, then concentrate under reduced pressure to obtain purification after the Phellinus linteus polysaccharide.

Preferably, the pulverization temperature of the step (1) is -65～-45°C, and the sieving mesh number is 40～80 meshes.

Preferably, the ultraviolet irradiation dose in step (2) is 1000-1500 mJ/cm ² ; the composite enzyme is prepared by mixing pectinase, cellulase and papain, and the mass ratio of the three enzymes is 1:1: 1.

Preferably, in step (2), the solid-liquid ratio of Phellinus linteus powder to deionized water is 1 g: 45-50 mL; the mass ratio of Phellinus linteus powder and compound enzyme is 10:1, and the enzymatic hydrolysis temperature is 25-30° C. , the enzymatic hydrolysis time is 5 ~ 10min.

Preferably, in step (3), the solid-liquid ratio of the filter residue to deionized water is 1 g: 45 mL, the ultrasonic extraction temperature is 50-70° C., the ultrasonic extraction time is 15-30 min, and the centrifugal speed is 4000-5000 r/ min, and the centrifugation time is 15-30 min.

Preferably, in step (5), the centrifugal speed is 1800-2000 r/min, and the centrifugal time is 15-20 min.

Preferably, in step (5), the concentration of hydrogen peroxide is 5%, the temperature of the water bath is 45-50° C., and the time of the water bath is 1-2 hours.

Preferably, in step (5), the concentration of trichloroacetic acid is 1.5 mol/L, the standing temperature is 60° C., and the standing time is 30 min.

Preferably, the dialysis time in step (6) is 24 hours, and the water is changed every 2-3 hours.

Compared with the prior art, the present invention has the following beneficial effects:

(1) the present invention firstly carries out low-temperature freeze-drying and pulverization to Phellinus linteus mycelium, under the low temperature state, the cell liquid in the cell wall can be made to be in a frozen state, and will not remain on the inner wall of the machine during pulverization; The yellow mycelium is brittle, easier to crush, and effectively improves the crushing efficiency; the crushed Phellinus linteus powder is also treated at a lower temperature, and it is immediately put into the low eutectic solvent aqueous solution at room temperature, under the action of thermal expansion and cold contraction and penetration , destroy the cell wall of Phellinus linteus mycelium, and extract the active substance polysaccharide of Phellinus linteus faster.

(2) In this application, the active substance polysaccharide in Phellinus linteus mycelium is extracted by applying ultraviolet irradiation technology, which improves the extraction rate of polysaccharide. At the same time, under the combined extraction effect of enzymatic hydrolysis and ultrasonic wave, the extraction time is greatly shortened, and the Extraction efficiency; finally, by defatting, depigmenting, and deproteinizing the polysaccharide, the purity of the polysaccharide is improved, and the prepared Phellinus linteus active substance polysaccharide has high purity and has great medicinal value.

Detailed ways

The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Example 1

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: pulverize the washed Phellinus linteus mycelium at -65 ° C, and obtain Phellinus linteus powder after passing through a 40-mesh sieve;

(2) Enzymatic hydrolysis after ultraviolet irradiation: put the Phellinus linteus powder (10 g) obtained in step (1) into an ultraviolet irradiation field for irradiation, and the irradiation dose is 1000 mJ/cm ² , and then add 450 mL of deionized water, 1 g of complex enzymes (the mass ratio of pectinase, cellulase, and papain is 1:1:1), enzymatic hydrolysis at 25°C for 5 min, and filtered to obtain enzymatic hydrolysis filtrate and filter residue;

(3) Extraction: then add the filter residue (10 g) obtained in step (2) into 450 mL of deionized water, perform ultrasonic extraction at 50° C. for 15 minutes, and centrifuge at a rate of 4000 r/min for 15 minutes after extraction, and take the supernatant. liquid to obtain extract and residue;

(4) extraction of Phellinus linteus crude polysaccharide: the enzymolysis filtrate obtained in step (2) and the extract obtained in step (3) are combined, heated and concentrated, centrifuged to remove the precipitate, and an equal volume of 95% ethanol is added to the supernatant. , stand for 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting, and deproteinizing treatment: adding 50 mL of petroleum ether to the crude polysaccharide of Phellinus linteus obtained in step (4), centrifuging at 1800 r/min for 15 min, sucking and discarding petroleum ether, and repeating 2 times to remove lipids; Add 5% hydrogen peroxide to the defatted polysaccharide until the polysaccharide concentration is 20%, react in a water bath at 45°C for 1 h, remove the pigment, and adjust the pH of the polysaccharide solution to 7.0 with ammonia; add the depigmented crude polysaccharide sample Add 1.5mol/L trichloroacetic acid until the polysaccharide concentration is 5%, stand at 60°C for 30min, centrifuge, discard the precipitate, repeat 3 times to obtain a defatted, depigmented, and deproteinized polysaccharide solution;

(6) dialysis: select a 7000D dialysis bag, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with a dialysis bag, the dialysis time is 24h, and the water is changed every 2h to obtain the purified mulberry The yellow polysaccharide solution was then concentrated under reduced pressure to obtain the purified Phellinus linteus polysaccharide.

Example 2

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: pulverize the washed Phellinus linteus mycelium at -55°C, and obtain Phellinus linteus powder after passing through a 60-mesh sieve;

(2) Enzymatic hydrolysis after ultraviolet irradiation: put the Phellinus linteus powder (10 g) obtained in step (1) into the ultraviolet irradiation field for irradiation, and the irradiation dose is 1200 mJ/cm ² , and then add 480 mL of deionized water, 1 g of complex enzymes (the mass ratio of pectinase, cellulase, and papain is 1:1:1), enzymatic hydrolysis at 27°C for 8 min, and filtered to obtain enzymatic hydrolysis filtrate and filter residue;

(3) leaching: then add the filter residue (10 g) obtained in step (2) into 450 mL of deionized water, perform ultrasonic leaching at 60° C. for 20 min, and centrifuge at a rate of 4500 r/min for 20 min after the extraction, and take the supernatant. liquid to obtain extract and residue;

(4) extraction of Phellinus linteus crude polysaccharide: the enzymolysis filtrate obtained in step (2) and the extract obtained in step (3) are combined, heated and concentrated, centrifuged to remove the precipitate, and an equal volume of 95% ethanol is added to the supernatant. , stand for 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting, and deproteinizing treatment: adding 50 mL of petroleum ether to the crude polysaccharide of Phellinus linteus obtained in step (4), centrifuging at 1900 r/min for 18 min, sucking and discarding petroleum ether, repeating 2 times to remove lipids; Add 5% hydrogen peroxide to the defatted polysaccharide until the polysaccharide concentration is 20%, react in a water bath at 48°C for 1.5h, remove the pigment, and adjust the pH of the polysaccharide solution to 7.0 with ammonia; add the depigmented crude polysaccharide sample Add 1.5mol/L trichloroacetic acid to the polysaccharide concentration to 5%, stand at 60°C for 30min, centrifuge, discard the precipitation, repeat 3 times to obtain a defatted, depigmented and deproteinized polysaccharide solution;

(6) dialysis: select a 7000D dialysis bag, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with a dialysis bag, the dialysis time is 24h, and the water is changed every 3h to obtain the purified mulberry The yellow polysaccharide solution was then concentrated under reduced pressure to obtain the purified Phellinus linteus polysaccharide.

Example 3

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: pulverize the washed Phellinus linteus mycelium at -45 ° C, and obtain Phellinus linteus powder after passing through an 80-mesh sieve;

(2) Enzymatic hydrolysis after ultraviolet irradiation: put the Phellinus linteus powder (10 g) obtained in step (1) into an ultraviolet irradiation field for irradiation, and the irradiation dose is 1500 mJ/cm ² , and then add 500 mL of deionized water, 1 g of complex enzymes (the mass ratio of pectinase, cellulase, and papain is 1:1:1), enzymatic hydrolysis at 30°C for 10 min, and filtered to obtain enzymatic hydrolysis filtrate and filter residue;

(3) leaching: the filter residue (10 g) obtained in step (2) was then added to 450 mL of deionized water, and ultrasonically leached at 70° C. for 30 min. liquid to obtain extract and residue;

(4) extraction of Phellinus linteus crude polysaccharide: the enzymolysis filtrate obtained in step (2) and the extract obtained in step (3) are combined, heated and concentrated, centrifuged to remove the precipitate, and an equal volume of 95% ethanol is added to the supernatant. , stand for 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting, and deproteinizing treatment: adding 50 mL of the crude polysaccharide of Phellinus linteus obtained in step (4), centrifuging at 2000 r/min for 20 min, sucking and discarding the petroleum ether, repeating 2 times to remove lipids; Add 5% hydrogen peroxide to the defatted polysaccharide until the polysaccharide concentration is 20%, react in a water bath at 50°C for 2 hours, remove the pigment, and adjust the pH of the polysaccharide solution to 7.0 with ammonia; add the depigmented crude polysaccharide sample Add 1.5mol/L trichloroacetic acid until the polysaccharide concentration is 5%, stand at 60°C for 30min, centrifuge, discard the precipitate, repeat 3 times to obtain a defatted, depigmented, and deproteinized polysaccharide solution;

(6) dialysis: select a 7000D dialysis bag, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with a dialysis bag, the dialysis time is 24h, and the water is changed every 3h to obtain the purified mulberry The yellow polysaccharide solution was then concentrated under reduced pressure to obtain the purified Phellinus linteus polysaccharide.

Comparative Example 1

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: pulverize the washed Phellinus linteus mycelium at -65°C, and obtain Phellinus linteus powder after passing through a 40-mesh sieve;

(2) Enzymatic hydrolysis: add the Phellinus linteus powder (10 g) obtained in step (1) into 450 mL of deionized water and 1 g of compound enzymes (the mass ratio of pectinase, cellulase, and papain is 1:1:1), Enzymatic hydrolysis was carried out at 25°C for 5 min, and the enzymatic hydrolysis filtrate and filter residue were obtained after filtration;

(3) Extraction: then add the filter residue (10 g) obtained in step (2) into 450 mL of deionized water, perform ultrasonic extraction at 50° C. for 15 minutes, and centrifuge at a rate of 4000 r/min for 15 minutes after extraction, and take the supernatant. liquid to obtain extract and residue;

(4) extraction of Phellinus linteus crude polysaccharide: the enzymolysis filtrate obtained in step (2) and the extract obtained in step (3) are combined, heated and concentrated, centrifuged to remove the precipitate, and an equal volume of 95% ethanol is added to the supernatant. , stand for 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting, and deproteinizing treatment: adding 50 mL of petroleum ether to the crude polysaccharide of Phellinus linteus obtained in step (4), centrifuging at 1800 r/min for 15 min, sucking and discarding petroleum ether, and repeating 2 times to remove lipids; Add 5% hydrogen peroxide to the defatted polysaccharide until the polysaccharide concentration is 20%, react in a water bath at 45°C for 1 h, remove the pigment, and adjust the pH of the polysaccharide solution to 7.0 with ammonia; add the depigmented crude polysaccharide sample Add 1.5mol/L trichloroacetic acid until the polysaccharide concentration is 5%, stand at 60°C for 30min, centrifuge, discard the precipitate, repeat 3 times to obtain a defatted, depigmented, and deproteinized polysaccharide solution;

(6) dialysis: select a 7000D dialysis bag, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with a dialysis bag, the dialysis time is 24h, and the water is changed every 2h to obtain the purified mulberry The yellow polysaccharide solution was then concentrated under reduced pressure to obtain the purified Phellinus linteus polysaccharide.

Comparative Example 2

An extraction and purification method of Phellinus linteus active substance polysaccharide, comprising the following steps:

(1) low-temperature freeze-drying pulverization: pulverize the washed Phellinus linteus mycelium at -65°C, and obtain Phellinus linteus powder after passing through a 40-mesh sieve;

(2) Ultraviolet irradiation: put the Phellinus linteus powder (10 g) obtained in step (1) into an ultraviolet irradiation field for irradiation, and the irradiation dose is 1000 mJ/cm ² , then add 450 mL of deionized water, and stir at 25° C. Filter after 5min to obtain filtrate and filter residue;

(3) Extraction: then add the filter residue (10 g) obtained in step (2) into 450 mL of deionized water, perform ultrasonic extraction at 50° C. for 15 minutes, and centrifuge at a rate of 4000 r/min for 15 minutes after extraction, and take the supernatant. liquid to obtain extract and residue;

(4) extraction of Phellinus linteus crude polysaccharide: the filtrate obtained in step (2) and the leaching solution obtained in step (3) were combined, heated and concentrated, centrifuged to remove the precipitation, an equal volume of 95% ethanol was added to the supernatant, and the Set for 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide;

(5) Degreasing, depigmenting, and deproteinizing treatment: adding 50 mL of petroleum ether to the crude polysaccharide of Phellinus linteus obtained in step (4), centrifuging at 1800 r/min for 15 min, sucking and discarding petroleum ether, and repeating 2 times to remove lipids; Add 5% hydrogen peroxide to the defatted polysaccharide until the polysaccharide concentration is 20%, react in a water bath at 45°C for 1 h, remove the pigment, and adjust the pH of the polysaccharide solution to 7.0 with ammonia; add the depigmented crude polysaccharide sample Add 1.5mol/L trichloroacetic acid until the polysaccharide concentration is 5%, stand at 60°C for 30min, centrifuge, discard the precipitate, repeat 3 times to obtain a defatted, depigmented, and deproteinized polysaccharide solution;

(6) dialysis: select a 7000D dialysis bag, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with a dialysis bag, the dialysis time is 24h, and the water is changed every 2h to obtain the purified mulberry The yellow polysaccharide solution was then concentrated under reduced pressure to obtain the purified Phellinus linteus polysaccharide.

The numerical values of the extraction rate and the purified polysaccharide purity in Examples 1 to 3 and Comparative Examples 1 to 2 were calculated respectively. The calculation formula is as follows, and the summary of the calculation results is shown in Table 1.

Extraction rate % (g/g) = Phellinus linteus crude polysaccharide / Phellinus linteus powder amount × 100%.

Take 10 mL of the purified polysaccharide solution, and obtain the polysaccharide quality by means of alcohol precipitation, centrifugation and drying. The concentration of Phellinus linteus polysaccharide was obtained by the phenol-sulfuric acid color development method, and then the purity of the Phellinus linteus crude polysaccharide after purification was calculated.

Purified polysaccharide purity (%)=C1/C2*100%

In the formula: C1 is the concentration of pure polysaccharide in the polysaccharide after purification; C2 is the concentration of polysaccharide before purification.

Table 1 Extraction rate and purity test results of Phellinus linteus polysaccharides

Extraction rate of Phellinus linteus crude polysaccharide (%) Phellinus linteus polysaccharide purity (%) Example 1 6.53 73.6 Example 2 6.21 68.5 Example 3 6.48 72.1 Comparative Example 1 3.19 36.1 Comparative Example 2 2.94 34.9

As shown in the results of the above table, the extraction rate of Phellinus linteus crude polysaccharide in Comparative Example 1-2 is lower than that of Example 1-3, indicating that the extraction method of Phellinus linteus polysaccharide can improve the extraction rate of Phellinus linteus polysaccharide in Example 1-3. The purity is higher than that of Comparative Examples 1-2, indicating that the purification method of the present invention can improve the purity of Phellinus linteus polysaccharides.

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, and substitutions can be made in these embodiments without departing from the principle and spirit of the invention and modifications, the scope of the present invention is defined by the appended claims and their equivalents.

Claims (9)

1. an extraction and purification method of Phellinus linteus active substance polysaccharide, is characterized in that, comprises the following steps: (1) low-temperature freeze-drying pulverization: the washed Phellinus linteus mycelium is pulverized at low temperature, and after sieving, Phellinus linteus powder is obtained; (2) Enzymatic hydrolysis after ultraviolet irradiation: put the Phellinus linteus powder obtained in step (1) into an ultraviolet irradiation field for irradiation, then add deionized water and compound enzymes, and filter after enzymatic hydrolysis to obtain enzymatic hydrolysis filtrate and filter residue ; (3) hot water-ultrasonic extraction: then add the filter residue obtained in step (2) into deionized water, carry out ultrasonic extraction, centrifuge after the extraction is completed, and get supernatant to obtain extract and residue; (4) Extraction of Phellinus linteus crude polysaccharide: combine the enzymatic hydrolysis filtrate obtained in step (2) and the extract obtained in step (3), heat and concentrate, centrifuge, remove the precipitate, add 95% ethanol to the supernatant, and let stand 24h, centrifuge, collect the precipitate, and dry to obtain Phellinus linteus crude polysaccharide; (5) Degreasing, depigmenting and deproteinizing treatment: adding the crude polysaccharide of Phellinus linteus obtained in step (4) into petroleum ether, centrifuging, sucking and discarding the petroleum ether, repeating twice to remove lipid substances; Add hydrogen peroxide to the concentration of 20%, react in a water bath, remove the pigment, adjust the pH of the polysaccharide solution to 7.0 with ammonia water; add trichloroacetic acid to the crude polysaccharide sample after depigmentation to a concentration of 5%, stand, centrifuge, Discard the precipitation and repeat 3 times to obtain a defatted, depigmented and deproteinized polysaccharide solution; (6) dialysis: select the dialysis bag of 7000D, dialyze the degreasing, depigmented, and deproteinized polysaccharide solution obtained in step (5) with the dialysis bag, obtain the purified Phellinus linteus polysaccharide solution, then concentrate under reduced pressure to obtain purification after the Phellinus linteus polysaccharide. 2 . The extraction method according to claim 1 , wherein the pulverization temperature of the step (1) is -65～-45° C., and the sieving mesh number is 40～80 meshes. 3 . 3. The extraction method according to claim 1, wherein in step (2), the ultraviolet irradiation dose is 1000-1500 mJ/cm ² ; the composite enzyme is a mixture of pectinase, cellulase and papain. Therefore, the mass ratio of the three enzymes is 1:1:1. 4. extraction method according to claim 1, is characterized in that, described in step (2), the solid-liquid ratio of Phellinus linteus powder and deionized water is 1g: 45～50mL; The mass ratio of described Phellinus linteus powder and compound enzyme It is 10:1, the enzymatic hydrolysis temperature is 25～30℃, and the enzymatic hydrolysis time is 5～10min. 5. extraction method according to claim 1, is characterized in that, the solid-liquid ratio of described filter residue and deionized water in step (3) is 1g: 45mL, described ultrasonic extraction temperature is 50～70 ℃, ultrasonic extraction The time is 15 to 30 minutes, the centrifugation rate is 4000 to 5000 r/min, and the centrifugation time is 15 to 30 minutes. 6. The extraction method according to claim 1, characterized in that, in step (5), the centrifugal speed is 1800-2000 r/min, and the centrifugal time is 15-20 min. 7 . The extraction method according to claim 1 , wherein the concentration of hydrogen peroxide in step (5) is 5%, the temperature of the water bath is 45～50° C., and the time of the water bath is 1～2h. 8 . 8. extraction method according to claim 1, is characterized in that, in step (5), the concentration of trichloroacetic acid is 1.5mol/L, and the standing temperature is 60 ℃, and the standing time is 30min. 9 . The extraction method according to claim 1 , wherein the dialysis time in step (6) is 24 hours, and the water is changed every 2 to 3 hours. 10 .