Trichosanthes peel enzyme and preparation method thereof

Abstract

The invention relates to a snakegourd peel enzyme and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine preparation. The method comprises the following steps: uniformly mixing snakegourd peel powder, cane sugar and water, then adjusting the pH value of a mixed solution obtained by mixing to a target pH value, then carrying out oscillation fermentation, after fermentation is completed, centrifuging, and taking a supernatant to obtain the snakegourd peel enzyme. The snakegourd peel enzyme is obtained by fermenting snakegourd peel, the subsequent antioxidant activity experiment of the snakegourd peel enzyme proves that the snakegourd peel enzyme has good antioxidant activity, and the content of effective components of the snakegourd peel enzyme obtained by the method disclosed by the invention is determined through an effective component determination experiment. And a support is provided for research and development of trichosanthes peel enzyme derivative products.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparation, and particularly relates to a trichosanthes peel enzyme and a preparation method thereof. Background Art

[0002] Trichosanthes kirilowii is a plant of the genus Trichosanthes in the Cucurbitaceae family. It is a plant that can be used as both medicine and food. Its mature seeds, Trichosanthes seeds, are both snack foods and traditional Chinese medicines that can moisten the lungs and resolve phlegm, and moisten the intestines and relieve constipation. Its mature peel, Trichosanthes kirilowii peel, has the effects of clearing away heat and resolving phlegm, promoting qi and relieving chest tightness, and is used to treat cough caused by phlegm and heat, and chest tightness and flank pain. Currently, Trichosanthes kirilowii peel injection, which uses Trichosanthes kirilowii peel as a raw material, is clinically used for the treatment of coronary heart disease and stable angina pectoris. Chemical composition studies have shown that Trichosanthes kirilowii peel contains polysaccharides, flavonoids, saponins, amino acids, and phenolic acids; Trichosanthes kirilowii peel has a variety of pharmacological activities, such as antioxidant, anti-inflammatory, and anti-atherosclerotic activities, especially in the protection of cardiovascular system, and has significant biological activity.

[0003] Enzymes are fermented products that use animals, plants, fungi, etc. as fermentation substrates and utilize the transformation of microorganisms to produce functional active substances required by the human body. Currently, there are many enzyme products, but there are not many medicinal and edible plant enzyme products. Since medicinal and edible plant enzymes contain rich active ingredients, such as flavonoids, polyphenols, polysaccharides, saponins and other ingredients, they are endowed with multiple functions. Therefore, medicinal and edible plant enzymes have important research value and broad market application prospects.

[0004] Trichosanthes peel is the mature peel of Trichosanthes. After the seeds are taken, the peel needs to be dried in time to prevent mildew. Due to weather or equipment restrictions, the peel is not dried in time and is often discarded as waste, which wastes resources and pollutes the environment. Dried Trichosanthes peel is currently mainly used for medicinal purposes. There is a lack of research and development of Trichosanthes peel enzyme products, which limits the application of Trichosanthes peel. How to increase its processing methods, improve its market added value, and expand its application range is a problem that needs to be solved at present. Summary of the invention

[0005] In view of the above problems, the present invention provides a trichosanthes peel enzyme and a preparation method thereof.

[0006] An object of the present invention is to provide a method for preparing Trichosanthes peel enzyme, comprising:

[0007] The powder of Trichosanthes peel, sucrose and water are mixed evenly, and then the pH of the mixed solution is adjusted to the target pH, and then the mixture is fermented by shaking. After the fermentation is completed, the mixture is centrifuged and the supernatant is taken to obtain the Trichosanthes peel enzyme.

[0008] Furthermore, the mass of the sucrose accounts for 4-16% of the mass of the Trichosanthes peel powder.

[0009] Preferably, the mass of the sucrose accounts for 4% of the mass of the Trichosanthes peel powder.

[0010] Furthermore, the ratio of the total mass of the Trichosanthes peel powder and sucrose to the volume of water is 1g:40-50mL.

[0011] Preferably, the ratio of the total mass of the Trichosanthes peel powder and sucrose to the volume of water is 1g:50mL.

[0012] Furthermore, the target pH is 5-6.

[0013] The target pH is 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 or 6.

[0014] Preferably, the target pH is 5.5.

[0015] Furthermore, the fermentation is natural fermentation or fermentation with inoculated bacteria.

[0016] Furthermore, the operation of accessing the bacterial strain for fermentation includes:

[0017] The mixed solution is sterilized by heat and ultraviolet successively, and then the bacteria are inoculated and then fermented by shaking.

[0018] Furthermore, the temperature of the heat sterilization is 110-130° C., and the time of the heat sterilization is 15-30 min.

[0019] The temperature of the heat sterilization is 110°C, 111°C, 112°C, 113°C, 114°C, 115°C, 116°C, 117°C, 118°C, 119°C, 120°C, 121°C, 122°C, 123°C, 124°C, 125°C, 126°C, 127°C, 128°C, 129°C or 130°C.

[0020] The time of the heat sterilization is 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, 21 min, 22 min, 23 min, 24 min, 25 min, 26 min, 27 min, 28 min, 29 min or 30 min.

[0021] Preferably, the temperature of the heat sterilization is 121° C., and the time of the heat sterilization is 20 min.

[0022] Furthermore, the ultraviolet sterilization time is 15-30 minutes.

[0023] The ultraviolet sterilization time is 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, 21 min, 22 min, 23 min, 24 min, 25 min, 26 min, 27 min, 28 min, 29 min or 30 min.

[0024] Preferably, the ultraviolet sterilization time is 20 minutes.

[0025] Furthermore, the bacterial species is one or a combination of yeast, lactic acid bacteria, acetic acid bacteria, and Rhizopus.

[0026] Furthermore, the bacteria are a mixture of yeast and lactic acid bacteria in a mass ratio of 1-4:1-4.

[0027] Preferably, the bacteria are a mixture of yeast and lactic acid bacteria in a mass ratio of 2:1.

[0028] Furthermore, when the bacteria are inoculated, the inoculation amount (the percentage of the mass of the bacteria to the mass of the Trichosanthes peel powder) is 5-10%.

[0029] The inoculation amount is 5%, 6%, 7%, 8%, 9% or 10%.

[0030] Preferably, the inoculation amount is 8%.

[0031] Furthermore, the shaking fermentation time is 4-10 days.

[0032] Preferably, the shaking fermentation time is 6 days.

[0033] Furthermore, the temperature of the shaking fermentation is 20-30°C.

[0034] The temperature of the shaking fermentation is 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C.

[0035] Preferably, the shaking fermentation temperature is 30°C.

[0036] Another object of the present invention is to provide a kind of Trichosanthes peel enzyme, prepared by the above method.

[0037] Beneficial effects of the present invention:

[0038] The present invention discloses a trichosanthes peel enzyme and a preparation method thereof. The trichosanthes peel enzyme is obtained by fermenting the trichosanthes peel. Subsequent antioxidant activity experiments of the trichosanthes peel enzyme prove that the trichosanthes peel enzyme obtained by the present invention has good antioxidant activity. The effective ingredient determination experiment clarifies the effective ingredient content of the trichosanthes peel enzyme obtained by the method of the present invention, which provides support for the research and development of trichosanthes peel enzyme derivative products.

[0039] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention can be realized and obtained by the structures pointed out in the description, claims and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0041] Figure 1 A bar graph showing the antioxidant activity of the supernatant obtained from each group in Example 1 of the present invention;

[0042] Figure 2 A bar graph showing the antioxidant activity during the fermentation time study in Example 2 of the present invention is shown;

[0043] Figure 3 A bar graph showing the antioxidant activity in the investigation of sucrose content in Example 2 of the present invention;

[0044] Figure 4 A bar graph showing the antioxidant activity in the mixed bacterial species ratio investigation in Example 2 according to an embodiment of the present invention;

[0045] Figure 5 The standard curve for determining the carbohydrate content of the Trichosanthes peel enzyme according to Example 4 of the present invention is shown;

[0046] Figure 6 The standard curve for determining the total phenol content of the Trichosanthes peel enzyme in Example 4 of the present invention is shown;

[0047] Figure 7 The standard curve for determining the total flavonoid content of the Trichosanthes peel enzyme in Example 4 of the present invention is shown;

[0048] Figure 8 The appearance of the Trichosanthes peel enzyme solution obtained according to Example 4 of the present invention is shown;

[0049] Fig. 9 A schematic diagram of the process according to Embodiment 1 to Embodiment 4 of the present invention is shown. DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0051] like Fig. 9 As shown, a schematic diagram of the flow chart of Examples 1 to 4 of the present invention is given. Specifically, the operations and results of Examples 1 to 4 are as follows:

[0052] Example 1

[0053] This example screened the best fermentation strains for Trichosanthes peel enzyme:

[0054] Weigh some medicinal material Trichosanthes peel and crush them with a pulverizer. Weigh 21g of Trichosanthes peel powder and divide it into 7 equal parts, namely a, b, c, d, e, f, and g groups. The strains in the fermentation process of each group are shown in Table 1. The mass percentage of sucrose added to each group accounts for 4% of the mass percentage of the Trichosanthes peel powder (hereinafter referred to as sucrose content), and distilled water is added according to the total mass of the Trichosanthes peel powder and sucrose and the volume of water (hereinafter referred to as the material-liquid ratio) of 1g:50mL, stir evenly at room temperature, and adjust the initial pH to 5.5 with sodium bicarbonate. Seal with biological breathable membrane, except for group f (the conical flask of this group was pasteurized at 65℃ before use), the rest of the groups were sterilized in a high temperature sterilizer at 121℃ for 20min, taken out after sterilization and cooled to room temperature, placed in a clean bench for ultraviolet sterilization for 20min, inoculated with bacteria as shown in Table 1 and sealed, the percentage of bacteria mass to the mass of Trichosanthes peel powder (hereinafter referred to as inoculation amount) was 8%, put into a constant temperature shaking incubator at 30℃ for 8d and then taken out. Centrifuge at 5000r / min for 10min, take the supernatant (i.e. Trichosanthes peel enzyme), measure the antioxidant activity with the scavenging of 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical as an indicator, and screen out the best active bacteria.

[0055] The bar graphs of the antioxidant activity of the supernatants obtained in examples a, b, c, d, e, f, and g are as follows: Figure 1 The data comparison table of the antioxidant activity of the supernatants obtained from a, b, c, d, e, f, and g is shown in Table 1.

[0056] Table 1

[0057]

[0058] from Figure 1From the data in Table 1, it can be seen that the mass ratio of yeast to lactic acid bacteria (1:1) has the best antioxidant activity, with a DPPH scavenging rate of 87.54%.

[0059] Example 2

[0060] This example optimizes the fermentation conditions of Trichosanthes peel enzyme:

[0061] Based on Example 1, this example optimizes the fermentation time of the access strain fermentation, the ratio of the mixed strains of yeast and lactic acid bacteria, and the sucrose content. The entire fermentation process is the same as the fermentation process of the access strain in Example 1. And the experimental results are based on the clearance rate of DPPH to screen out the best conditions. The same treatment is carried out according to the above-mentioned medicinal material processing method for screening the best strain, and 3g of Trichosanthes peel powder is added to each fermentation bottle, the solid-liquid ratio is 1:50g / mL, the inoculation amount is 8%, and the fermentation temperature is 30°C.

[0062] 2.1 Fermentation time investigation

[0063] Under fixed conditions, the sucrose content was 4%, the mass ratio of yeast and lactic acid bacteria was 1:1, and the scavenging ability of DPPH at different fermentation times (4d, 6d, 8d, 10d) was investigated. The results are as follows Figure 2 As shown;

[0064] from Figure 2 It can be seen that the enzyme solution obtained after 6 days of fermentation has the best DPPH scavenging ability, with a scavenging rate of 92.8%.

[0065] 2.2 Investigation of sucrose content

[0066] The fermentation was carried out under fixed conditions for 8 days. The mass ratio of yeast and lactic acid bacteria was 1:1. The scavenging ability of DPPH by different sucrose contents (4%, 8%, 12%, 16%) was investigated. The results are shown in the figure. Figure 3 As shown;

[0067] from Figure 3 It can be seen that the enzyme solution obtained by fermentation with a sucrose content (i.e., the mass percentage of sucrose in the Trichosanthes peel powder) of 4% has the best DPPH scavenging ability, with a scavenging rate of 74.46%.

[0068] 2.3 Investigation of the proportion of mixed strains

[0069] The fermentation was carried out under fixed conditions for 8 days, with a sucrose content of 4%. The scavenging ability of different ratios (1:4, 1:2, 1:1, 2:1, 4:1) of the mixed strains of yeast and lactic acid bacteria on DPPH was investigated. The results are shown in the figure. Figure 4 As shown;

[0070] from Figure 4It can be seen that the enzyme solution obtained by fermentation with a mass ratio of yeast to lactic acid bacteria of 2:1 has the best DPPH scavenging ability, with a scavenging rate of 85.63%.

[0071] Example 3

[0072] This example uses the best process conditions for fermentation to obtain Trichosanthes peel enzyme, and the antioxidant activity of the finally obtained Trichosanthes peel enzyme is determined:

[0073] Through Example 1 and Example 2, it was finally determined that the optimal fermentation conditions for the trichosanthes peel enzyme were a mixed strain ratio of yeast: lactic acid bacteria = 2:1, a solid-liquid ratio of 1g:50mL, an inoculation amount of 8%, a fermentation temperature of 30°C, a sucrose content of 4%, and a fermentation of 6d. According to this condition, the trichosanthes peel enzyme solution was prepared, and the antioxidant activity of the solution obtained before and after fermentation was determined, with vitamin C (Vc) as a positive control.

[0074] 3.1 DPPH free radical scavenging ability determination

[0075] Take an appropriate amount of DPPH powder, weigh it accurately, and prepare a DPPH solution with anhydrous ethanol solution to a concentration of 0.2mmol / L. Store it at 4℃ away from light. Use a pipette to accurately transfer 500μL of sample solution into a brown EP tube, then add 1000μL of DPPH solution, mix thoroughly and let stand at room temperature for 30min, and measure the absorbance value A2 at 517nm; use anhydrous ethanol instead of DPPH solution to measure the absorbance as A1; use distilled water instead of sample solution to measure the absorbance as A0 (measure three times and take the average value). DPPH free radical scavenging rate (%) = [1-(A2-A1) / A0]×100%.

[0076] 3.2 Hydroxyl radical scavenging ability determination

[0077] Accurately weigh 50.04 mg of FeSO4·7H2O and dissolve it in 30 mL of distilled water. Prepare it for use immediately (6 mmol / L). Use a pipette to draw 100 μL of 30% H2O2 solution into 29.9 mL of distilled water (0.1% H2O2). Protect from light. Weigh 24.84 mg of salicylic acid and dissolve it in 30 mL of anhydrous ethanol (6 mmol / L). Store it in a dark place. Use a pipette to accurately pipette 500 μL of sample solution into a brown EP tube, add equal volumes (500 μL) of FeSO4 solution and H2O2 solution in turn, shake well and let stand for 5 minutes, then add equal volumes of salicylic acid-ethanol solution, shake well and let stand at room temperature for 30 minutes, and measure the absorbance value A2 at 510 nm; use distilled water instead of H2O2 solution to measure the absorbance as A1; use distilled water instead of sample solution to measure the absorbance as A0 (measure three times and take the average value). Hydroxyl radical scavenging rate (%) = [1-(A2-A1) / A0] × 100%

[0078] 3.3 Determination of superoxide anion free radical scavenging ability

[0079] Use a pipette to take 500μL of sample solution, 500μL of reduced coenzyme I disodium (NaDH-2Na, 557μmol / L), 500μL of phenazine methyl sulfate (PMS, 45μmol / L), and 500μL of nitro blue tetrazolium chloride (NBT, 108μmol / L), mix them evenly, warm them at 25℃ for 5min, and adjust to zero with distilled water. The absorbance A2 is measured at a wavelength of 510nm; the absorbance A1 is measured by replacing the reaction solution with distilled water; the absorbance A0 is measured by replacing the sample solution with distilled water (the average value is taken by measuring three times). The formula for superoxide anion radical scavenging rate is: (%) = [1-(A2-A1) / A0] × 100%.

[0080] The above measurement results are shown in Table 2.

[0081] Table 2

[0082]

[0083] From the data in Table 2, it can be seen that the enzyme produced by fermentation of Trichosanthes peel (yeast: lactic acid bacteria = 2:1) has a scavenging rate of 82.78%, 67.68% and 91.40% for DPPH free radicals, hydroxyl free radicals and superoxide anion free radicals, respectively. The antioxidant capacity of Trichosanthes peel enzyme is significantly higher than that of the unfermented solution of Trichosanthes peel.

[0084] Example 4

[0085] This example uses the optimal process conditions for fermentation to obtain Trichosanthes peel enzyme, and the content of the effective ingredient of the finally obtained Trichosanthes peel enzyme is determined.

[0086] According to the optimal fermentation conditions of Trichosanthes peel enzyme: mixed strains (yeast: lactic acid = 2:1), solid-liquid ratio of 1:50 g / mL, inoculation amount of 8%, fermentation temperature of 30 ° C, sucrose content of 4%, fermentation for 6 days, the Trichosanthes peel enzyme solution was obtained. The Trichosanthes peel enzyme solution is shown in Figure 8 .

[0087] Then, the contents of total carbohydrates, total phenols and flavonoids in the obtained Trichosanthes peel enzyme solution were determined, wherein the specific test method is as follows:

[0088] 4.1 Determination of total carbohydrate content

[0089] Accurately weigh 5 mg of glucose reference substance and distilled water to a 50 ml volumetric flask (0.1 mg / mL reference substance solution). Pipette 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 ml into stoppered test tubes respectively, add 1 ml of 5% phenol and shake well, quickly add 5 ml of concentrated sulfuric acid, shake well and cool, place at room temperature for 30 minutes, and measure the absorbance at 490 nm with a spectrophotometer. Regress with glucose mass concentration as the horizontal axis (X) and absorbance as the vertical axis (A) to obtain the equation. Take 1 ml of enzyme solution, add 1 mL of 5% phenol and 5 mL of concentrated sulfuric acid, shake well and cool, place at room temperature for 30 minutes, measure the absorbance at 490 nm, and calculate the total carbohydrate content.

[0090] 4.2 Determination of total phenol content

[0091] Weigh 8 mg of gallic acid reference substance accurately and dilute to 100 mL (0.08 mg / mL reference substance solution) with distilled water. Accurately pipette 0, 0.04, 0.08, 0.12, 0.16, 0.20, 0.24, 0.28, and 0.32 mL into a 10 mL volumetric flask, add distilled water to make up the volume to 0.4 mL, add 0.8 mL of 12% sodium carbonate solution and 0.4 mL of folin phenol, dilute to 10 mL with distilled water, shake well and place in the dark for 2 hours, and measure the absorbance at a wavelength of 765 nm with a spectrophotometer. Perform regression with gallic acid concentration as the horizontal axis (X) and absorbance as the vertical axis (A) to obtain the regression equation. Take 0.4 mL of enzyme supernatant, add 0.8 mL of 12% sodium carbonate solution and 0.4 mL of folin phenol to make up to 10 mL, leave it in the dark for 2 h, measure the absorbance at 765 nm, and calculate the total phenol content.

[0092] 4.3 Determination of total flavonoid content

[0093] Accurately weigh 2.5 mg of rutin reference substance, add a few drops of ethanol to dissolve, and then dilute to a 250 ml volumetric flask with 70% ethanol to obtain a 0.01 mg / mL reference substance solution. Accurately pipette 0, 1.0, 2.0, 3.0, 4.0, and 5.0 ml into a 10 ml volumetric flask, add 70% ethanol to a total volume of 5 ml, add 0.3 ml of 5% sodium nitrite solution, shake and mix, and let stand for 6 min, add 0.3 ml of 10% aluminum nitrate solution, shake and mix, and let stand for 6 min, add 4 ml of 4% sodium hydroxide solution, dilute to the scale with 70% ethanol, shake and mix for 15 min, and measure the absorbance at 510 nm with a spectrophotometer.

[0094] The equation was obtained by regression with rutin mass concentration as the horizontal axis (X) and absorbance as the vertical axis (A). Take 5 mL of enzyme solution, add 0.3 mL of 5% sodium nitrite solution, shake well and let it stand for 6 minutes, then add 0.3 mL of 10% aluminum nitrate solution, shake well and let it stand for 6 minutes, add 4 mL of 4% sodium hydroxide solution and dilute to 10 mL with 70% ethanol, shake well, let it stand for 10 minutes, measure the absorbance at 510 nm, and calculate the total flavonoid content.

[0095] Wherein, the carbohydrate content determination standard curve of the Trichosanthes peel enzyme of this embodiment is as follows Figure 5 The standard curve for the determination of total phenol content of Trichosanthes peel enzyme is shown in Figure 6 As shown in the figure, the standard curve for the determination of the total flavonoid content of Trichosanthes peel enzyme is as follows Figure 7 shown.

[0096] according to Figure 5-Figure 7 The carbohydrate content, total phenol content and total flavonoid content of the Trichosanthes peel enzyme of this example were calculated, and the specific data are shown in Table 3.

[0097] Table 3

[0098]

[0099] From the data in Table 3, it can be seen that the contents of total carbohydrates, total phenols and flavonoids of the Trichosanthes peel enzyme obtained under the optimal process conditions are 804.49 mg / mL, 18.58 mg / mL and 122.21 mg / mL, respectively.

[0100] Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent substitutions for some of the technical features therein; and these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing trichosanthes peel enzyme, characterized in that, include: The powder of Trichosanthes peel, sucrose and water are mixed evenly, and then the pH of the mixed solution is adjusted to the target pH, and then the mixture is fermented by shaking. After the fermentation is completed, the mixture is centrifuged and the supernatant is taken to obtain the Trichosanthes peel enzyme.

2. The method for preparing the Trichosanthes peel enzyme according to claim 1, characterized in that: The mass of the sucrose accounts for 4-16% of the mass of the Trichosanthes peel powder.

3. The method for preparing the Trichosanthes peel enzyme according to claim 1, characterized in that: The ratio of the total mass of the Trichosanthes peel powder and sucrose to the volume of water is 1g:40-50mL.

4. The method for preparing the Trichosanthes peel enzyme according to claim 1, characterized in that: The target pH is 5-6.

5. The method for preparing the Trichosanthes peel enzyme according to claim 1, characterized in that: The fermentation is natural fermentation or fermentation with inoculated bacteria.

6. The method for preparing the Trichosanthes peel enzyme according to claim 5, characterized in that: The operation of accessing the bacterial strain for fermentation includes: The mixed solution is sterilized by heat and ultraviolet successively, and then the bacteria are inoculated and then fermented by shaking.

7. The method for preparing the Trichosanthes peel enzyme according to claim 6, characterized in that: The bacterial species is one or a combination of yeast, lactic acid bacteria, acetic acid bacteria, and Rhizopus.

8. The method for preparing the Trichosanthes peel enzyme according to claim 7, characterized in that: The bacterial species are a mixture of yeast and lactic acid bacteria in a mass ratio of 1-4:1-4.

9. A method for preparing a Trichosanthes peel enzyme according to any one of claims 1 to 8, characterized in that: The shaking fermentation time is 4-10 days, and the shaking fermentation temperature is 20-30°C.

10. A trichosanthes peel enzyme, characterized in that: The method is prepared by any one of claims 1 to 9.

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