Application of phyllanthus emblica essential oil microcapsule and / or phyllanthus emblica extract in fruit and vegetable preservation
By developing Yuganzi essential oil microcapsules and extracts, the problems of degradation in quality and short shelf life of freshly cut fruits and vegetables during processing are solved, and the antibacterial and antioxidant preservation effect without chemical additives is achieved, and the shelf life of fruits and vegetables is extended.
Patent Information
- Application Number
- CN202411601705.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-11
- Publication Date
- 2025-02-18
AI Technical Summary
During the processing process, freshly cut fruits and vegetables have reduced quality and short shelf life due to mechanical damage. In addition, chemical additives are often used for existing anti-corrosion and color protection treatments, which violate the natural characteristics of fruits and vegetables.
The Yuganzi essential oil microcapsules and Yuganzi extract were developed. The raw materials suitable for preservation of fruits and vegetables were selected through screening tests, and essential oils were extracted by supercritical method, and the extract was prepared by enzymatic treatment to form a complex suitable for preservation of fruits and vegetables.
It achieves antibacterial and antioxidant effects on freshly cut fruits and vegetables, delays the color deepening of the surface of fruits and vegetables, and appropriately extends the shelf life. In addition, this method does not require the use of chemical additives, maintaining the natural characteristics of fruits and vegetables.
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Figure CN119453294A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of fruit and vegetable preservation, and particularly relates to application of emblica essential oil microcapsules and / or emblica extracts in fruit and vegetable preservation. Background Art
[0002] With the improvement of human living standards and the accelerated pace of life, more and more fresh-cut fruits and vegetables are appearing in major supermarkets and e-commerce platforms. Fresh-cut fruits and vegetables are made from fresh fruits and vegetables. After a series of processing such as grading, cleaning, trimming, peeling, cutting, preservation, packaging, etc., they are transported at low temperature and sold in refrigerators. For example, yam, because peeling may cause allergies in consumers, many merchants now peel it and vacuum pack it for sale. Because it is troublesome to peel water chestnuts, many merchants currently also peel them and vacuum pack them for sale.
[0003] However, compared with raw materials, the mechanical damage caused by the cutting process of fresh-cut fruits and vegetables will cause a series of physiological and biochemical changes, such as discoloration, taste change, aging, softening, and deterioration due to microbial infection, and the shelf life is shorter. For example, yam, water chestnut, potato, apple, etc. Therefore, maintaining quality and extending the shelf life are the key to the processing technology of fresh-cut fruits and vegetables.
[0004] In addition, during the processing of fresh-cut fruits and vegetables, in order to maintain better quality and a longer shelf life, pretreatments such as preservation and color protection are carried out during the processing of the products. Preservation and color protection treatments sometimes inevitably involve chemical products, which runs counter to the naturalness, nutrition, freshness, convenience and high availability of fresh-cut fruits and vegetables. Therefore, further research is needed on preservation technologies without preservatives and chemical additives. Summary of the invention
[0005] The invention aims to provide application of emblica essential oil microcapsules and / or emblica extracts in preserving fruits and vegetables.
[0006] The invention selects the emblica with good preservation effect as a raw material through a series of screening tests, and develops the emblica essential oil microcapsules and the emblica extract suitable for preserving fruits and vegetables.
[0007] The above-mentioned object of the present invention can be achieved through the following technical scheme: application of emblica essential oil microcapsules and / or emblica extracts in the preservation of fruits and vegetables.
[0008] Preferably, the emblica emblica is a mixture of emblica emblica essential oil microcapsules and emblica emblica extract, and the mass ratio of the emblica emblica essential oil microcapsules to the emblica emblica extract is 1:1.
[0009] Preferably, the fruits and vegetables are fresh-cut fruits and vegetables.
[0010] Furthermore, the fruits and vegetables are yam, water chestnut and apple.
[0011] Preferably, the emblica essential oil microcapsules or emblica extract are first placed in a non-woven bag, and then the non-woven bag is placed in fruits and vegetables for contactless preservation; the amount of the emblica essential oil microcapsules and / or emblica extract is 40-60 g / kg of fruits and vegetables.
[0012] Preferably, the emblica essential oil microcapsules are prepared by the following method: fresh emblica fruits are taken, emblica essential oil is extracted by supercritical method, the emblica essential oil is diluted by anhydrous ethanol to obtain emblica essential oil diluent, a mixed aqueous solution of β-cyclodextrin and Tween-80 is prepared, the emblica essential oil diluent is added to the mixed aqueous solution of β-cyclodextrin and Tween-80, and then stirred, cooled, precipitated, filtered and dried to obtain emblica essential oil microcapsules.
[0013] Preferably, during supercritical extraction, the extraction temperature is 40-50° C., the pressure is 30-40 MPa, and the time is 1.5-2.5 h.
[0014] Preferably, the mass ratio of β-cyclodextrin, Tween-80 and water in the mixed aqueous solution of β-cyclodextrin and Tween-80 is 3-5:0.1-0.2:30-50.
[0015] Preferably, the emblica extract is prepared by the following method:
[0016] (1) get fresh emblica fruit, after pulverizing, add water to extract, in leaching process, carry out homogenizing pressure treatment at intervals, obtain emblica crude extract;
[0017] (2) Adding a complex enzyme to the crude extract of Phyllanthus emblica for enzymolysis to obtain a crude enzyme solution of Phyllanthus emblica, and then performing enzyme inactivation, concentration and pulverization to obtain a Phyllanthus emblica extract.
[0018] Preferably, the amount of the emblica raw material and water in step (1) is 1g:5-10mL; the extraction time is 2-4h; during the extraction process, a homogenization and pressure treatment is performed every 20-40min, wherein the homogenization pressure is 5-10MPa, and the homogenization and pressure treatment includes taking the material out of the extraction container every 20-40min, passing it through a homogenizer for pressure treatment, and then entering the extraction container.
[0019] Preferably, the complex enzyme in step (2) comprises pectinase, tannase and lipase, the added amount of the complex enzyme is 0.05-0.2% of the total mass of the crude extract of Phyllanthus emblica, the mass ratio of the pectinase, tannase and lipase is 1:1:1; the enzymolysis temperature is 40-55°C, and the enzymolysis time is 2-10h.
[0020] The invention has the following advantages: the prepared emblica microcapsules and emblica extracts are derived from emblica fruits, are safe and have no peculiar smell; the prepared emblica microcapsules and emblica extracts, especially the composite of emblica microcapsules and emblica extracts, have good antibacterial, antioxidant and fresh-keeping effects on fruits and vegetables, especially fresh-cut fruits and vegetables, and have low cost. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 The emblica extract prepared in Example 1;
[0022] Figure 2 The emblica essential oil microcapsule powder prepared in Example 4;
[0023] Figure 3 The compound obtained by mixing the emblica extract prepared in Example 1 and the emblica essential oil microcapsule powder prepared in Example 4 in a mass ratio of 1:1 in Example 7;
[0024] Figure 4 Schematic diagram of packaging the Phyllanthus emblica extract prepared in Example 1 and the Phyllanthus emblica essential oil microcapsule powder prepared in Example 4 in a non-woven bag;
[0025] Figure 5 Schematic diagram of the preservation of yam samples for 2 days in Example 8, wherein 1 is the group of emblica extract, 2 is the group of emblica microcapsules, 3 is the group of the compound of emblica extract and emblica microcapsules 1;1, and 4 is the blank control group;
[0026] Figure 6 Schematic diagram of the preservation of yam samples for 3 days in Example 8, wherein 1 is the group of emblica extract, 2 is the group of emblica microcapsules, 3 is the group of the compound of emblica extract and emblica microcapsules 1;1, and 4 is the blank control group;
[0027] Figure 7 Schematic diagram of the preservation of yam samples for 4 days in Example 8, wherein 1 is the group of emblica extract, 2 is the group of emblica microcapsules, 3 is the group of the compound of emblica extract and emblica microcapsules 1;1, and 4 is the blank control group;
[0028] Figure 8 Schematic diagram of the preservation of fresh-cut apple samples for 2 hours in Example 8, wherein 1 is the group of emblica extract, 2 is the group of emblica microcapsules, 3 is the group of the compound of emblica extract and emblica microcapsules 1:1, and 4 is the blank control group;
[0029] Fig. 9It is a schematic diagram of the preservation of fresh-cut apple samples for 48 hours in Example 8, wherein 1 is the group of emblica extract, 2 is the group of emblica microcapsules, 3 is the group of 1:1 compound of emblica extract and emblica microcapsules, and 4 is the blank control group. DETAILED DESCRIPTION
[0030] The present invention is further specifically described below by way of examples, but the embodiments of the present invention are not limited thereto.
[0031] Example 1
[0032] The emblica extract of this embodiment is prepared by the following method:
[0033] (1) 1 kg of fresh Phyllanthus emblica fruit was selected and pre-treated by removing the core and crushing;
[0034] (2) adding 8L of water to the pretreated emblica raw material for extraction, performing homogenization and pressure treatment every 30min during the extraction process, the homogenization pressure is 10MPa, and the total extraction time is 3h to obtain a crude extract of emblica;
[0035] (3) adding a composite enzyme of pectinase, tannase and lipase in a mass ratio of 1:1:1 to the crude extract of Phyllanthus emblica for enzymolysis, wherein the amount of the composite enzyme added is 0.1% of the total mass of the crude extract of Phyllanthus emblica to obtain a crude enzyme solution of Phyllanthus emblica, wherein the pectinase, tannase and lipase are all commercially available products, the enzymolysis temperature is 50° C., and the enzymolysis time is 6 h; then the extract of Phyllanthus emblica is obtained by inactivating the enzyme, concentrating and pulverizing. Figure 1 shown.
[0036] The extract of Phyllanthus emblica was packaged in non-woven bags, such as Figure 4 shown.
[0037] Example 2
[0038] The emblica extract of this embodiment is prepared by the following method:
[0039] (1) 1 kg of fresh Phyllanthus emblica fruit was selected and pre-treated by removing the core and crushing;
[0040] (2) adding 10L of water to the pretreated emblica raw material for extraction, performing homogenization and pressure treatment every 20min during the extraction process, the homogenization pressure is 8MPa, and the total extraction time is 2h to obtain a crude extract of emblica;
[0041] (3) Adding a composite enzyme of pectinase, tannase and lipase in a mass ratio of 1:1:1 to the crude extract of emblica emblica for enzymatic hydrolysis, wherein the amount of the composite enzyme added is 0.15% of the total mass of the crude extract of emblica emblica to obtain a crude enzyme solution of emblica emblica, wherein pectinase, tannase and lipase are all commercially available products, the enzymatic hydrolysis temperature is 45°C, and the enzymatic hydrolysis time is 4h; then the emblica emblica extract is obtained after enzyme inactivation, concentration and pulverization.
[0042] Example 3
[0043] The emblica extract of this embodiment is prepared by the following method:
[0044] (1) 1 kg of fresh Phyllanthus emblica fruit was selected and pre-treated by removing the core and crushing;
[0045] (2) adding 5 L of water to the pretreated emblica raw material for extraction, performing homogenization and pressure treatment every 40 min during the extraction process, the homogenization pressure was 5 MPa, and the total extraction time was 4 h to obtain a crude extract of emblica;
[0046] (3) Adding a composite enzyme of pectinase, tannase and lipase in a mass ratio of 1:1:1 to the crude extract of emblica emblica for enzymatic hydrolysis, wherein the amount of the composite enzyme added is 0.2% of the total mass of the crude extract of emblica emblica to obtain a crude enzyme solution of emblica emblica, wherein pectinase, tannase and lipase are all commercially available products, the enzymatic hydrolysis temperature is 50°C, and the enzymatic hydrolysis time is 6 h; then the emblica emblica extract is obtained by inactivating the enzyme, concentrating and pulverizing.
[0047] Example 4
[0048] The emblica essential oil microcapsules of the present embodiment are prepared by the following method:
[0049] Fresh emblica fruit was taken and emblica essential oil was obtained by supercritical extraction. During the supercritical extraction, the extraction temperature was 45°C, the pressure was 35 MPa, and the time was 2.0 h. The emblica essential oil was diluted with anhydrous ethanol to obtain the emblica essential oil diluent, and a mixed aqueous solution of β-cyclodextrin and Tween-80 (the mass ratio of β-cyclodextrin, Tween-80 and water was 4:0.15:40) was prepared. The emblica essential oil diluent was added to the mixed aqueous solution of β-cyclodextrin and Tween β-80, and then stirred, cooled, precipitated, filtered and dried to obtain emblica essential oil microcapsules. Figure 2 shown.
[0050] The emblica essential oil microcapsules were packaged in non-woven bags, such as Figure 4 shown.
[0051] Example 5
[0052] The emblica essential oil microcapsules of the present embodiment are prepared by the following method:
[0053] Fresh emblica fruit was taken and emblica essential oil was obtained by supercritical extraction. During the supercritical extraction, the extraction temperature was 40°C, the pressure was 45 MPa, and the time was 1.5 h. The emblica essential oil was diluted with anhydrous ethanol to obtain the emblica essential oil diluent. A mixed aqueous solution of β-cyclodextrin and Tween-80 (the mass ratio of β-cyclodextrin, Tween-80 and water was 3:0.1:30) was prepared. The emblica essential oil diluent was added to the mixed aqueous solution of β-cyclodextrin and Tween β-80, and then stirred, cooled, precipitated, filtered and dried to obtain the emblica essential oil microcapsules.
[0054] Example 6
[0055] The emblica essential oil microcapsules of the present embodiment are prepared by the following method:
[0056] Fresh emblica fruit was taken and emblica essential oil was obtained by supercritical extraction. During the supercritical extraction, the extraction temperature was 50°C, the pressure was 30 MPa, and the time was 2.5 h. The emblica essential oil was diluted with anhydrous ethanol to obtain the emblica essential oil diluent. A mixed aqueous solution of β-cyclodextrin and Tween-80 (the mass ratio of β-cyclodextrin, Tween-80 and water was 5:0.2:50) was prepared. The emblica essential oil diluent was added to the mixed aqueous solution of β-cyclodextrin and Tween β-80, and then stirred, cooled, precipitated, filtered and dried to obtain the emblica essential oil microcapsules.
[0057] Example 7
[0058] In this example, the emblica extract prepared in Example 1 and the emblica essential oil microcapsule powder prepared in Example 4 were mixed in a mass ratio of 1:1 to obtain a composite of the two. Figure 3 shown.
[0059] Example 8
[0060] The performance test was carried out on the emblica extract in Example 1, the emblica microcapsules in Example 4, and the composite of the emblica microcapsules in Example 1 and Example 4 in a mass ratio of 1:1:
[0061] 1.1 Antimicrobial testing
[0062] The antibacterial effects of the Phyllanthus emblica extract in Example 1, the Phyllanthus emblica microcapsules in Example 4, and the composite of the Phyllanthus emblica microcapsules in Example 1 and Example 4 in a mass ratio of 1:1 were measured by the Oxford cup method:
[0063] Weigh 2.0 g of beef extract, 4.0 g of peptone, 2.0 g of sodium chloride, and 8.0 g of agar powder, add them to 400 mL of distilled water, heat to dissolve and prepare a solid culture medium for later use.
[0064] After shaking the solid agar medium sterilized by high-pressure steam, pour it into the culture dish, about 20 mL per dish, and place it on the clean bench to wait for solidification.
[0065] Take the slant freeze-dried bacterial tubes of Escherichia coli and Staphylococcus aureus, pick typical colonies with a sterile inoculation loop and streak inoculate them on the solid agar slant culture medium, and place it in a 37°C constant temperature incubator for 24 hours.
[0066] Take the activated Escherichia coli and Staphylococcus aureus strains, pick a loop with a sterile inoculation loop and inoculate it into 500 mL of liquid culture medium, shake to obtain Escherichia coli and Staphylococcus aureus suspensions, place them in a shaker at 37°C and 180 rpm for shaking culture for 24 hours to obtain Escherichia coli and Staphylococcus aureus suspensions, and select the most appropriate bacterial solution concentration through a series of concentration dilutions as the bacterial solution concentration of the solid culture medium.
[0067] After the solid culture medium solidifies, use a sterile pipette to draw 0.1mL of Escherichia coli and Staphylococcus aureus suspension on its surface, and use a sterile triangular ring to evenly apply the bacterial solution on the surface of the culture medium. Place an Oxford cup on the solid culture medium. According to Table 1, the dilution of the compound of the emblica extract, the emblica microcapsules in Example 4, and the emblica microcapsules in Example 1 and Example 4 in a mass ratio of 1:1 (diluted to about 200μg / mL with sterile water) is added to the Oxford cup, and then the culture dish is placed in a 37°C constant temperature incubator for 24h, and finally the diameter of the inhibition zone is measured and the size of the inhibition zone is observed.
[0068] Table 1 Antibacterial diameters of Example 1, Example 4 and the combination of Example 1 and Example 4 against Escherichia coli and Staphylococcus aureus
[0069] Group Bacteria Diameter of inhibition ring (mm) Example 1 Phyllanthus emblica extract Escherichia coli 35 Embodiment 4 Emblica essential oil microcapsules Escherichia coli 26 The compound of embodiment 1 and embodiment 4 Escherichia coli 50 Example 1 Phyllanthus emblica extract Staphylococcus aureus 32 Embodiment 4 Emblica essential oil microcapsules Staphylococcus aureus 27 The compound of embodiment 1 and embodiment 4 Staphylococcus aureus 53
[0070] As can be seen from Table 1, all three have strong antibacterial ability, among which the antibacterial effect of the compound of emblica extract and emblica essential oil microcapsules is better.
[0071] 1.2 Antioxidant test
[0072] Take the emblica extract in Example 1, the emblica microcapsules in Example 4, and the composite of the emblica extract in Example 1 and the emblica microcapsules in Example 4 in a mass ratio of 1:1 (diluted with sterile water to about 200 μg / mL), take 0.2 mL of the dilution, add 0.2 mL of DPPH (2×10 -4mol / L) solution, mix well, and react in the dark for 30 minutes. Measure the absorbance at 517nm A sample. At the same time, measure the absorbance at 517nm of 0.2mL of the sample solution (standard solution Vc) and 0.2mL of ethanol A blank, and then measure the absorbance at 517nm of 0.2mL DPPH solution and 0.2mL ethanol A control. Repeat the same determination 3 times. Substitute the detected absorbance value into the calculation formula to obtain the clearance rate of each concentration.
[0073] The calculation formula of DPPH free radical scavenging rate is: inhibition rate = [1-(A 样品 —A 空白 ) / A 对照 ]×100%.
[0074] A 样品 :0.2mL DPPH solution + 0.2mL different sample solutions;
[0075] A 空白 :0.2mL standard sample solution of different concentrations + 0.2mL anhydrous ethanol solution;
[0076] A 对照 : 0.2mL DPPH solution + 0.2mL anhydrous ethanol solution.
[0077] Table 2 Antioxidant performance of Example 1, Example 4 and the composite of Example 1 and Example 4
[0078] Example 1 Example 4 1:1 compound of Example 1 and Example 4 DPPH clearance 90％ 85％ 95％
[0079] As can be seen from Table 2, the emblica extract in Example 1, the emblica microcapsules in Example 4, and the composite of the emblica extract in Example 1 and the emblica microcapsules in Example 4 in a mass ratio of 1:1 all have strong antioxidant capacity, and the composite of the emblica extract in Example 1 and the emblica microcapsules in Example 4 in a mass ratio of 1:1 has the best antioxidant effect.
[0080] 1.3 Freshness preservation test
[0081] Select a number of yam and apples with uniform color, uniform size and no damage, wash them and divide them equally for later use. In an environment of constant temperature of 20° C. and constant humidity of 70%, put Example 1, Example 4 and the composite of Example 1 and Example 4 (2 g) and fruits and vegetables (about 300 g) in the same space, and observe their morphological changes after packaging.
[0082] like Figure 5 As shown in the figure, the color of fresh-cut yam did not change significantly within 2 days after washing and refrigeration. Figure 6 This is the effect after refrigeration for 3 days. Figure 6It can be seen that the surface of some yam samples in group 1 with emblica extract and group 2 with emblica microcapsules began to turn yellow, and in control 4 (without adding any preservatives), the surface of almost every sample showed obvious color changes, and the color of some samples became obviously darker, appearing yellowish brown, while all yam samples in group 3 with a 1:1 mixture of emblica extract and emblica microcapsules showed no obvious color change. Figure 7 This is the effect after refrigeration for 4 days. Figure 7 It can be seen that the surface color of all samples in control group 4 further deepened, with black spots appearing, and some samples even became sticky and rotten on the surface.
[0083] like Figure 8 As shown, after 2 hours of washing and refrigerating fresh-cut apples, the color of the control group 4 changed rapidly compared with the group 1 of the emblica extract, the group 2 of the emblica microcapsules, and the group 3 of the 1:1 mixture of the emblica extract and the emblica microcapsules. Fig. 9 As shown, after refrigeration in the refrigerator for 48 hours, the emblica extract group 1, the emblica microcapsule group 2 and the 1:1 combination group 3 of the emblica extract and the emblica microcapsule did not show obvious color changes, while the color of the control group 4 deepened further, and the surface began to become sticky and rotten.
[0084] Therefore, the emblica extract, emblica microcapsules and especially the combination of the two can be used in the preservation of fruits and vegetables, especially fresh-cut fruits and vegetables. They not only have antibacterial and antioxidant effects, but also can slow down the deepening of the surface color of fresh-cut fruits, and further can appropriately extend the shelf life of fresh-cut fruits and vegetables.
[0085] The above examples are used to illustrate the present invention. It is necessary to point out that the above examples are only used to further illustrate the present invention and do not limit the protection scope of the present invention. Some non-essential modifications and adjustments made by others based on the present invention still fall within the protection scope of the present invention.
Claims
1. Application of emblica essential oil microcapsules and / or emblica extracts in the preservation of fruits and vegetables.
2. The use according to claim 1, characterized in that: The emblica emblica is a mixture of emblica emblica essential oil microcapsules and emblica emblica extract, and the mass ratio of the emblica emblica essential oil microcapsules to the emblica emblica extract is 1:
1.
3. The use according to claim 1, characterized in that: The fruits and vegetables are fresh-cut fruits and vegetables.
4. The use according to claim 1, characterized in that: The emblica essential oil microcapsules and / or emblica extract are first placed in a non-woven bag, and then the non-woven bag is placed in fruits and vegetables for contactless preservation; the amount of the emblica essential oil microcapsules and / or emblica extract is 40-60 g / kg of fruits and vegetables.
5. The use according to any one of claims 1 to 4, characterized in that: The emblica essential oil microcapsules are prepared by the following method: fresh emblica fruits are taken, emblica essential oil is extracted by supercritical method, the emblica essential oil is diluted by anhydrous ethanol to obtain emblica essential oil diluent, a mixed aqueous solution of beta-cyclodextrin and Tween-80 is prepared, the emblica essential oil diluent is added to the mixed aqueous solution of beta-cyclodextrin and Tween-80, and then stirring, cooling, precipitating, suction filtering and drying are performed to obtain emblica essential oil microcapsules.
6. The use according to claim 5, characterized in that: When using supercritical extraction, the extraction temperature is 40-50°C, the pressure is 30-40Mpa, and the time is 1.5-2.5h.
7. The use according to claim 5, characterized in that: The mass ratio of β-cyclodextrin, Tween-80 and water in the β-cyclodextrin and Tween-80 mixed aqueous solution is 3-5:0.1-0.2:30-50.
8. The use according to any one of claims 1 to 4, characterized in that: The emblica extract is prepared by the following method: (1) get fresh emblica fruit, after pulverizing, add water to extract, in leaching process, carry out homogenizing pressure treatment at intervals, obtain emblica crude extract; (2) Adding a complex enzyme to the crude extract of Phyllanthus emblica for enzymolysis to obtain a crude enzyme solution of Phyllanthus emblica, and then performing enzyme inactivation, concentration and pulverization to obtain a Phyllanthus emblica extract.
9. The use according to claim 8, characterized in that: The dosage relationship of the emblica raw material and water in step (1) is 1g:5-10mL; the extraction time is 2-4h; during the extraction process, a homogenization and pressure treatment is performed every 20-40min, wherein the homogenization pressure is 5-10MPa, and the homogenization and pressure treatment comprises taking the material out of the extraction container every 20-40min, passing it through a homogenizer for pressure treatment, and then entering the extraction container.
10. The use according to claim 8, characterized in that: The complex enzyme in step (2) comprises pectinase, tannase and lipase, the added amount of the complex enzyme is 0.05-0.2% of the total mass of the crude extract of Phyllanthus emblica, the mass ratio of the pectinase, tannase and lipase is 1:1:1; the enzymolysis temperature is 40-55°C, and the enzymolysis time is 2-10h.
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