Green synergistic extraction system as well as extraction method and application thereof

Abstract

The invention relates to the technical field of biological extraction, in particular to a green synergistic extraction system and an extraction method and application thereof.The green synergistic extraction system comprises a deep eutectic solvent system, an enzymolysis system and an ultrasonic-assisted extraction system, and the deep eutectic solvent system comprises a DES solvent; the DES solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor, and the hydrogen bond acceptor is choline, choline chloride or L-proline; the hydrogen bond donor is lactic acid, urea or ethylene glycol; enzyme preparations in the enzymolysis system are at least two of cellulase, pectinase and beta-glucosidase; the problems that a traditional organic solvent is high in toxicity and poor in biodegradability and synthetic deep eutectic solvent raw materials are non-renewable are solved, a green synergistic extraction system which is environmentally friendly, low in cost and excellent in performance is developed, the mass transfer efficiency and the extraction yield are improved, the used DES and enzyme preparations can be recycled, and the cost is reduced while the environment is protected.

Description

Technical Field

[0001] The invention relates to the field of biological extraction technology, and in particular to a green synergistic extraction system and an extraction method and application thereof. Background Art

[0002] As a medicinal and edible plant rich in bioactive ingredients such as inulin, polyphenols, and flavonoids, burdock root has high nutritional and health value and has attracted much attention in the fields of functional foods, medicines, and health products. However, traditional extraction processes are limited by technical defects when extracting active ingredients from burdock root. On the one hand, the lignin and cellulose in the plant cell walls form a dense structure, which seriously hinders the release of active ingredients, resulting in low extraction efficiency. Taking inulin as an example, the yield using traditional water extraction is only 8%-10%; on the other hand, traditional processes are highly dependent on organic solvents such as ethanol and methanol. These solvents are not only toxic and threaten the health of operators, but also cause serious environmental pollution. In addition, high-temperature extraction processes (such as the reflux method) promote the dissolution of active substances by increasing the temperature. During the extraction process, heat-sensitive components such as polyphenols and flavonoids are easily degraded. Prolonged high temperature can lead to molecular oxidation, ester bond cleavage or isomerization, destroying their effective activity. It can be seen from this that a single solvent is difficult to achieve the synergistic extraction of multiple components, and often requires step-by-step purification, which makes the entire extraction process cumbersome and complicated, the cost remains high, and it is difficult to achieve efficient, green, and high-value development and utilization.

[0003] In recent years, although deep eutectic solvents (DES) and enzymatic hydrolysis technology have been attempted for plant extraction, the application of a single technology has limitations. For example, traditional solvents (such as water or organic solvents) have poor compatibility with enzyme systems, resulting in reduced enzyme activity and difficulty in effectively destroying cell walls. DES is composed of hydrogen bond donors (HBDs) and hydrogen bond acceptors (HBAs), and has limited penetration into plant cell walls. It requires high temperature or long-term extraction, and there is still a risk of destroying heat-sensitive components. In addition, synthetic DES has poor biodegradability and is difficult to recycle. Summary of the Invention

[0004] In order to solve the problems existing in the prior art of extracting active ingredients from burdock roots, such as the active ingredients are easily destroyed, it is difficult to achieve synergistic extraction of multiple ingredients, the extraction efficiency is low, and the extraction process of the reagents used is complex and difficult to reuse, it is urgent to develop a green, efficient, multi-technology synergistic extraction system to break through the bottleneck of traditional processes and meet the needs of high-value utilization of burdock root resources. Based on the above problems, the present invention provides a green synergistic extraction system and its extraction method and application. Based on hydrogen bond donors and hydrogen bond acceptors from natural sources, a synergistic mechanism of DES, biocatalysis and physical enhancement technology is established to develop an environmentally friendly, low-cost and high-performance green synergistic extraction system, which has a high extraction rate of active ingredients in burdock roots and can achieve efficient targeted extraction of burdock roots.

[0005] A green synergistic extraction system includes a deep eutectic solvent system and an enzymatic hydrolysis system, wherein the deep eutectic solvent system includes a DES solvent, and the DES solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor, wherein the hydrogen bond acceptor is choline, choline chloride or L-proline; the hydrogen bond donor is lactic acid, glycerol or ethylene glycol; and the enzyme preparation in the enzymatic hydrolysis system is at least two of cellulase, pectinase and β-glucosidase.

[0006] Preferably, the enzyme activity of the cellulase is ≥1000 U / g, the enzyme activity of the pectinase is ≥500 U / g, and the enzyme activity of the β-glucosidase is ≥200 U / g.

[0007] Furthermore, it also includes an ultrasonic assisted extraction system, which uses an ultrasonic reactor.

[0008] Furthermore, the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:1 to 1:5, preferably 1:2.

[0009] Furthermore, the deep eutectic solvent system further includes deionized water, and the volume ratio of the DES solvent to deionized water is 1:1 to 1:5, preferably 1:4.

[0010] Preferably, the water content of the deep eutectic solvent system is 10% to 30%.

[0011] Furthermore, the molar ratio of the cellulase to the pectinase is 2:1 or the molar ratio of the cellulase, the pectinase and the β-glucosidase is 2:1:1.

[0012] The present invention also provides an extraction method of a green extraction system, comprising the following steps:

[0013] (1) pre-treating the burdock root raw material;

[0014] (2) further treating the burdock root pretreated in step (1) using a deep eutectic solvent system;

[0015] (3) adding an enzymatic hydrolysis system to the burdock root treated with the deep eutectic solvent system in step (2) for enzymatic hydrolysis to obtain an enzymatic hydrolyzate;

[0016] (4) transferring the enzymatic hydrolyzate obtained in step (3) to an ultrasonic reactor, and extracting the active ingredients in the burdock root using ultrasonic technology;

[0017] (5) The extract obtained after the ultrasonic-assisted extraction in step (4) is separated and purified to finally obtain high-purity inulin, saponin and flavonoids.

[0018] Furthermore, the pretreatment in step (1) is crushing and screening, preferably passing through a 40-mesh sieve.

[0019] Furthermore, in step (2), the solid-liquid ratio of burdock root to deep eutectic solvent system is 1:15-1:25 g / mL, the treatment temperature is 70-90° C., and the treatment time is 1-4 h.

[0020] Preferably, the treatment process of step (2) further includes stirring and centrifugation, the stirring speed is 150-250 rpm, the centrifugal speed is 5000 rpm, and the centrifugal time is 15 minutes.

[0021] Furthermore, the mass volume percentage concentration of the enzyme preparation in step (3) is 1% to 3%.

[0022] Furthermore, the temperature of the enzymatic hydrolysis treatment in step (3) is 40-60° C., the pH is 4.8-5.2, and the time is 4-12 hours.

[0023] Furthermore, the deep eutectic solvent system in step (2) can be reused for ≥5 times; and the enzyme preparation in step (3) can be reused after being immobilized for ≥7 times.

[0024] Furthermore, the ultrasonic power of the ultrasonic extraction process in step (3) is 200-600W, and the time is 20-30 minutes.

[0025] The present invention provides application of the green synergistic extraction system or the extraction method in extracting active ingredients from burdock roots.

[0026] Preferably, the green synergistic extraction system or extraction method of the present invention can extract inulin, saponin and flavonoids from burdock roots at extraction rates of 15% to 17%, 1.3% to 1.6% and 1.5% to 2.0%, respectively.

[0027] The advantages of the present invention are:

[0028] 1. The DES solvent used in the present invention has the advantages of low melting point, high solubility and low volatility. It can dissolve plant cell wall components, destroy cell structure, and promote the release of active ingredients. The enzymatic hydrolysis system uses the specific catalytic action of enzymes to target and degrade cell walls. When the two are used in combination, DES can provide a microenvironment close to physiological activity for the enzyme, maintain enzyme stability, and enable it to maintain ≥90% activity in the deep eutectic solvent system, realizing the integration of "solvent penetration-enzymatic degradation" and improving the extraction yield.

[0029] 2. The ultrasonic microwave technology used in the present invention can assist the extraction of DES and enzymatic hydrolysis through physical effects. Ultrasonic waves generate microbubbles in the liquid, forming local high temperature and high pressure, destroying the cell wall. The sonic vibration promotes the full mixing of DES and the substrate, which can promote the rapid dissolution of the active ingredient and improve the mass transfer efficiency.

[0030] 3. The present application provides a green synergistic extraction system and an extraction method thereof, which are used to extract active ingredients from burdock roots. During the ternary combined synergistic extraction of DES+enzymatic hydrolysis+ultrasound, the inulin macromolecules are dissolved by DES and released by enzymatic hydrolysis; small molecules such as flavonoids and saponin aglycones can be quickly dissolved with the help of ultrasonic cavitation effect. Compared with the existing technology, the DES-enzymatic hydrolysis-ultrasound ternary synergistic system of the present invention breaks through the bottleneck of traditional technology and targets the destruction of cell structure, showing significant advantages. At the same time, the extraction process is simple, and the DES solvent and enzyme preparation used can be reused more than 5 times, which protects the environment while improving economic benefits. It is a green extraction system. DETAILED DESCRIPTION

[0031] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0032] It should be noted that the various installation methods and technical terms mentioned in this invention are technical terms that have long been clearly known in the relevant technical field and therefore will not be further explained. In addition, the same reference numerals are used for the same components, but this does not affect nor constitute an accurate understanding of the technical solution by those skilled in the art.

[0033] Example 1

[0034] This embodiment provides a green synergistic extraction system, characterized in that it includes a deep eutectic solvent system and an enzymatic hydrolysis system, wherein the deep eutectic solvent system includes a DES solvent and water, and the DES solvent is composed of choline and lactic acid (screened by Experimental Example 1 of the present invention); the enzyme preparations in the enzymatic hydrolysis system are cellulase and pectinase.

[0035] The method for extracting the active ingredients from the burdock root raw material using the green collaborative extraction system of this embodiment is as follows:

[0036] (1) Pretreatment of burdock root raw materials: crush the burdock root to a particle size of ≤0.5 mm to increase its specific surface area, then soak and defatted in petroleum ether for 3 h to remove fat-soluble impurities, and then dry before use;

[0037] (2) 1.5 g of burdock root powder pretreated in step (1) was added with 15 mL of a DES reagent composed of choline-lactic acid and water in a molar ratio of 1:2 (the volume ratio of DES reagent to water was 1:4), and further ultrasonicated at 500 W for 15 min to promote DES penetration;

[0038] (3) adding cellulase and pectinase (total enzyme activity of 150 U / g) in a molar ratio of 2:1 to the burdock root treated in step (2), adjusting the pH to 6.0, and performing enzymatic hydrolysis in a constant temperature water bath at 55° C. and a rotation speed of 150 rpm for 2 h. Finally, terminating the reaction in a boiling water bath for 15 min to obtain an enzymatic hydrolyzate;

[0039] (4) transferring the enzymatic hydrolyzate obtained in step (3) to an ultrasonic reactor, using ultrasonic technology to assist extraction, and ultrasonically treating for 10 minutes under the conditions of 500W and 45kHz to further release the active ingredients in the burdock root;

[0040] (5) The extract after ultrasonic-assisted extraction in step (4) was centrifuged at 7000 rpm for 15 min, the supernatant was taken, the residue was extracted again once, and the supernatants were combined; then, the DES solvent was removed by rotary evaporation under reduced pressure below 50° C., and the solvent was recovered for recycling;

[0041] (6) Purifying and enriching the active ingredients after centrifugation in step (5):

[0042] Ethanol was added to the supernatant to a final concentration of 85%, and the mixture was allowed to stand for 12 hours. The inulin precipitate was collected by centrifugation and freeze-dried to obtain enriched inulin. The mixture was then adsorbed using a macroporous resin (D101) and eluted with a gradient of 35% to 75% ethanol to collect the flavonoid-enriched fraction. Finally, the saponin fraction was collected by silica gel column chromatography (petroleum ether-ethyl acetate gradient elution) and dried under reduced pressure. Finally, high-purity inulin, saponins, and flavonoids were obtained. The total extraction yield of the active ingredients from burdock root is shown in Table 1.

[0043] Example 2

[0044] The preparation process of this embodiment is the same as that of Example 1, except that the extraction system and process parameters used in the extraction process are different. The deep eutectic solvent system used in step (2) of this embodiment is a DES reagent of proline-ethylene glycol and water in a molar ratio of 1:2, and the volume ratio of DES reagent to water is 1:4;

[0045] The enzyme preparation used in step (3) of this example is cellulase, with a total enzyme activity of 150 U / g;

[0046] The conditions for ultrasonic technology-assisted extraction used in step (4) of this embodiment are 500W and 15 minutes.

[0047] The total extraction yield of the active ingredients from the burdock root in this example is shown in Table 1.

[0048] Example 3

[0049] The preparation process of this embodiment is the same as that of Example 1, except that the extraction system and process parameters used in the extraction process are different. The deep eutectic solvent system used in step (2) of this embodiment is a DES reagent and water with a molar ratio of 1:1 of choline chloride and lactic acid, and the volume ratio of DES reagent to water is 1:6;

[0050] The enzyme preparation used in step (3) of this example is β-glucosidase, with a total enzyme activity of 100 U / g;

[0051] The conditions for ultrasonic technology-assisted extraction used in step (4) of this embodiment are 300W and 20min.

[0052] The total extraction yield of the active ingredients from the burdock root in this example is shown in Table 1.

[0053] Example 4

[0054] The preparation process of this embodiment is the same as that of Example 1, except that the extraction system and process parameters used in the extraction process are different. The deep eutectic solvent system used in step (2) of this embodiment is a DES reagent of proline-ethylene glycol and water with a molar ratio of 1:5, and the volume ratio of DES reagent to water is 1:3;

[0055] The enzyme preparation used in step (3) of this example is cellulase, pectinase and β-glucosidase in a molar ratio of 4:1:1, and the total enzyme activity is 150 U / g.

[0056] The total extraction yield of the active ingredients from the burdock root in this example is shown in Table 1.

[0057] Comparative Example 1

[0058] In this comparative example, the active ingredients in burdock root were extracted using the traditional water extraction method with a water-to-material ratio of 15:1. The final total extraction yield is shown in Table 1.

[0059] Comparative Example 2

[0060] In this comparative example, only a deep eutectic solvent system (choline chloride: ethylene glycol = 1:3, solid-liquid ratio of 1:10) was used to extract the active ingredients from burdock root. The reaction temperature was 55°C, the processing time was 2.5 h, and the final total extraction yield was shown in Table 1.

[0061] Comparative Example 3

[0062] In this comparative example, only an enzymatic hydrolysis system (enzyme concentration 3%, enzyme preparation composition of cellulase: pectinase = 2:1) was used to extract the active ingredients from burdock root. The reaction temperature was 45°C, the treatment time was 2 h, the system pH was 4.8, and the final total extraction yield is shown in Table 1.

[0063] Comparative Example 4

[0064] In this comparative example, only ultrasonic assisted extraction (power 250 W, frequency 35 kHz) was used to extract the active ingredients from the burdock root. The reaction temperature was 45° C., the processing time was 20 min, the system pH was 6.5, and the final total extraction yield is shown in Table 1.

[0065] Comparative Example 5

[0066] In this comparative example, an enzymatic hydrolysis system combined with ultrasonic assistance (enzymatic hydrolysis temperature was 45°C, time was 2 h; ultrasonic conditions were 180W, 15 min) was used to extract the active ingredients from burdock roots. The reaction temperature was 45°C, the treatment time was 175 min, and the system pH was 4.8. The final total extraction yield is shown in Table 1.

[0067] Table 1 Extraction yield results of Example 1 and Comparative Example 1

[0068] temperature pH Extraction rate (%) Example 1 50 6.0 42.5±2.5 Example 2 55 5.8 43.7±2.0 Example 3 60 6.3 44.3±1.8 Example 4 65 5.9 44.8±2.2 Comparative Example 1 85 6.8 10.2±0.4 Comparative Example 2 55 — 32.5±0.9 Comparative Example 3 45 4.8 26.8±1.1 Comparative Example 4 45 6.5 19.7±0.7 Comparative Example 5 45 4.8 36.2±1.3

[0069] Experimental Example 1

[0070] In this experiment, choline chloride (ChCl), betaine (Bet), and L-proline (L-Pro) were used as hydrogen bond acceptors (HBAs); ethylene glycol (EG), glycerol (Gly), 1,4-butanediol (1,4-bu), urea (U), lactic acid (LA), and glucose (D-glu) were used as hydrogen bond donors (HBDs). Eighteen natural DES solvents were synthesized from the components of HBA and HBD by ultrasonic heating, and their physicochemical properties were measured. It was found that when the molar ratio was 1:1, the DES solvents composed of choline-lactic acid, choline-ethylene glycol, choline chloride-lactic acid, choline chloride-ethylene glycol, L-proline-lactic acid, L-proline-ethylene glycol, and choline chloride-urea had relatively good extraction yields. Among them, the DES solvent composed of choline-lactic acid had the best extraction effect, with a yield of 78% of the active ingredients in burdock root.

[0071] Experimental Example 2

[0072] Based on Experimental Example 1, choline-lactic acid DES solvents with component molar ratios of 1:1, 1:2, 1:3, 1:4, and 1:5 were synthesized by ultrasonic heating. The experimental process is as follows:

[0073] Accurately weigh 0.4 g of burdock root powder sample and place it in a 10 mL centrifuge tube. Add 8 mL of the prepared choline-lactic acid DES solvent and shake well. Then place the centrifuge tube in an ultrasonic cleaner and extract at 40 kHz, 44 ° C and 230 W for 45 minutes. The extract is centrifuged at 12000 rpm for 4 minutes, and the supernatant is diluted to an appropriate multiple and then filtered through a 0.22 μm microporous filter membrane for detection and analysis. Finally, it was concluded that the optimal molar ratio of choline-lactic acid of the DES solvent with the best extraction rate is 1:2.

[0074] Experimental Example 3

[0075] Based on Experimental Example 2, the prepared DES solvent with the best composition was selected, and 20%, 30%, 40%, 50%, and 60% deionized water were added thereto, respectively, to prepare deep eutectic solvent systems with different water contents for the optimal extraction system from burdock root. The yield of active ingredients in burdock root was used as an indicator, and the optimal aqueous solution concentration was screened to be 30%.

[0076] Experimental Example 4

[0077] This experimental example uses DES solvents composed of choline chloride-urea and choline chloride-lactic acid as examples to explore the extraction of active ingredients from burdock roots. The extraction method is as shown in Example 1, and the traditional ethanol method is used as the control group. The results are shown in Table 2. It can be seen that when using DES treatment, the extraction rate and yield of active ingredients from burdock roots are significantly improved compared with the traditional ethanol method. At the same time, it can provide a microenvironment close to physiological activity for the enzyme, maintaining enzyme stability:

[0078] Table 2 Extraction results of active ingredients from burdock root with different DES compositions

[0079]

[0080]

[0081] Experimental Example 5

[0082] Finally, in order to minimize the cost-effectiveness of the extraction method and increase its sustainability, this experimental example studied the reusability of the deep eutectic solvent of the present invention without any pre-extraction step. The process is as follows:

[0083] After the first extraction with the prepared DES solvent, the extraction mixture was filtered and the solvent was reused to re-extract the target compound without removing the target compound. After extraction under the same extraction conditions, the increase in the target compound in the solution was measured by ultra-performance liquid chromatography to determine the reusability of the deep eutectic solvent. It was found that the DES solvent of this example could be reused more than 5 times.

[0084] Test Example 1

[0085] In this test example, the content of active ingredients in the burdock roots extracted by the methods of Example 1 and Comparative Example 1 was detected. The reference substances of each active ingredient were accurately weighed and placed in a volumetric flask. 80% methanol was added to dilute them into standard solutions of known concentrations. An appropriate amount of each reference substance was pipetted into the bottle, and methanol was added to the volume. The solution was diluted into different concentration gradients and recorded. Different color developing reagents were then added to measure the absorbance. The phenol-sulfuric acid method was used to determine the content of inulin (with distilled water as a blank control, and its absorbance was measured at 490 nm), the Folin phenol method was used to determine the content of polyphenols (with distilled water as a blank control, and its absorbance was measured at 765 nm), the NaNO2-Al(NO3)3-NaOH colorimetric method was used to determine the content of flavonoids (with ethanol as a blank control, and its absorbance was measured at 500 nm), and the HPLC method was used to determine the content of burdock root saponins (C18 column, mobile phase acetonitrile-water, detection wavelength 203 nm). The results are shown in Table 3. It can be concluded that the extraction method of this embodiment can achieve efficient and green extraction of active ingredients from burdock roots. Compared with the traditional water extraction method in Comparative Example 1, the yield is significantly improved, and the extraction time is significantly shortened by 50% to 70%.

[0086] Table 3 Extraction rate of active ingredients in burdock root

[0087] Example 1 Comparative Example 1 Inulin yield 15％～17％ 7％～10％ polyphenols 1.5~2.0mgGAE / g 0.8~1.2mgGAE / g Saponin yield 1.3％～1.6％ 0.5％～0.8％ Total flavonoids yield 1.5％～2.0％ 0.8％～1.2％

[0088] Test Example 2

[0089] This test example uses ultra-high performance liquid chromatography combined with gel permeation chromatography (GPC) and ion exchange chromatography (IEC) to purify inulin and test its purity. The specific process is as follows:

[0090] (1) Sample pretreatment: First, take a crude inulin sample, add an appropriate amount of ethanol to the crude inulin solution to promote the precipitation of impurities, and then collect the precipitate by centrifugation;

[0091] (2) Gel Permeation Chromatography (GPC) Separation: The pretreated inulin precipitate was dissolved to a solution of appropriate concentration, and then injected into an ultra-high performance liquid chromatograph (UPLC) with an injection volume of 4 ml. A gel permeation chromatography column was packed with Sepharose CL-6B (Toyopearl HW-50 could also be used), and eluted with ultrapure water as the mobile phase (GPC columns can separate inulin oligosaccharides based on their molecular weight). The main peak in the eluate was collected, and the inulin oligosaccharide molecular weight corresponding to the main peak was in the range of 10 to 50 kDa.

[0092] (3) Purity detection: The collected main peak eluate is further concentrated and then freeze-dried to obtain the final high-purity inulin product. After HPLC (high performance liquid chromatography) detection, its purity can reach ≥95%, indicating that inulin that meets the purity requirements is obtained.

[0093] In summary, it can be concluded that the present invention breaks through the bottleneck of low efficiency and high pollution of traditional burdock root extraction through the DES-enzymatic hydrolysis-ultrasound ternary green synergistic extraction system, realizes efficient and green extraction of active ingredients from burdock root, has both economic efficiency and industrial feasibility, and provides an innovative solution for the development of natural products. In the extraction of natural products, zero pollution emissions, efficient extraction and activity retention are achieved, the functional synergistic effect between the three, and cross-domain adaptability are both efficient and sustainable, providing an innovative solution for the application of green chemistry.

[0094] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the foregoing description. Therefore, all changes that come within the meaning and range of equivalents of the claims are intended to be included in the present invention. No sign in a claim should be construed as limiting the claim.

[0095] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any minor modifications, equivalent replacements, and improvements made to the above embodiments based on the technical essence of the present invention shall be included in the scope of protection of the technical solution of the present invention.

Claims

1. A green collaborative extraction system, characterized in that: The invention comprises a deep eutectic solvent system and an enzymatic hydrolysis system, wherein the deep eutectic solvent system comprises a DES solvent, and the DES solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor, wherein the hydrogen bond acceptor is choline, choline chloride or L-proline; the hydrogen bond donor is lactic acid, urea or ethylene glycol; and the enzyme preparation in the enzymatic hydrolysis system is at least two of cellulase, pectinase and β-glucosidase.

2. The green collaborative extraction system according to claim 1, characterized in that: Also included is an ultrasonic-assisted extraction system, which uses an ultrasonic reactor.

3. The green collaborative extraction system according to claim 1 or 2, characterized in that: The molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:1 to 1:5; the molar ratio of the cellulase to the pectinase is 2:1, or the molar ratio of the cellulase, the pectinase and the beta-glucosidase is 2:1:

1.

4. The green collaborative extraction system according to claim 1 or 2, characterized in that: The deep eutectic solvent system further includes deionized water, and the volume ratio of the DES solvent to the deionized water is 1:3 to 1:

6.

5. An extraction method based on the green extraction system according to claim 1, characterized in that: The following steps are involved: (1) pre-treating the burdock root raw material; (2) further treating the burdock root pretreated in step (1) using a deep eutectic solvent system; (3) adding an enzymatic hydrolysis system to the burdock root treated with the deep eutectic solvent system in step (2) for enzymatic hydrolysis to obtain an enzymatic hydrolyzate; (4) transferring the enzymatic hydrolyzate obtained in step (3) to an ultrasonic reactor, and extracting the active ingredients in the burdock root using ultrasonic technology; (5) The extract obtained after the ultrasonic-assisted extraction in step (4) is separated and purified to finally obtain high-purity inulin, saponin and flavonoids.

6. The extraction method according to claim 5, characterized in that In the step (2), the solid-liquid ratio of the burdock root to the deep eutectic solvent system is 1:15 to 1:25; the treatment temperature is 70 to 90° C., and the treatment time is 1 to 4 hours.

7. The extraction method according to claim 5, characterized in that The mass volume percentage concentration of the enzyme preparation in step (3) is 1% to 3%; the temperature of the enzymatic hydrolysis treatment is 40 to 60° C., the pH is 4.8 to 5.2, and the time is 4 to 12 hours.

8. The extraction method according to claim 5, characterized in that The ultrasonic extraction process in step (3) has an ultrasonic power of 200 to 600 W and takes 20 to 30 minutes.

9. The extraction method according to claim 5, characterized in that The deep eutectic solvent system in step (2) can be reused, with a repetition number of ≥5 times; the enzyme preparation in step (3) can be reused after immobilization, with a repetition number of ≥7 times.

10. Use of the green synergistic extraction system according to claim 1 or the extraction method according to any one of claims 5 to 9 in extracting active ingredients from burdock roots.

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