Preparation method and application of high-activity SOD enzyme compound raw material

Abstract

The invention relates to a preparation method and application of a high-activity SOD enzyme compound raw material. The preparation method of the high-activity SOD enzyme compound raw material comprises the following steps: (1) adding SOD enzyme into a zinc glycinate solution, and carrying out ultrasonic treatment; (2) collecting the reaction solution, and performing dialysis and / or ultrafiltration to obtain filtrate; (3) adjusting the pH value of the filtrate, adding a hydrolyzed protein zinc solution into the filtrate, and fully stirring to obtain a mixed solution; (4) adding a sodium carbonate and / or sodium bicarbonate buffer solution into the mixed solution, and adjusting the pH value; and (5) performing ultrafiltration concentration and freeze-drying to obtain the SOD enzyme compound raw material. According to the preparation method, high activity of SOD can be kept in the preparation and storage processes of the cosmetics, so that the anti-aging effect and the whitening effect of the SOD applied to the cosmetics are enhanced.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and particularly to a preparation method and application of a high-activity SOD enzyme compound raw material. Background Art

[0002] Superoxide dismutase (SOD) is an antioxidant metalloenzyme widely present in organisms. Its main function is to catalyze the dismutation of superoxide anion radicals into hydrogen peroxide and oxygen, thereby eliminating the oxidative stress damage of superoxide anion radicals to cells, maintaining the body's metabolic balance, being able to scavenge and reduce excessive free radicals in the human body and delay aging, and being the most important free radical scavenger in organisms. In addition, SOD has a certain absorption effect on ultraviolet light with wavelengths of 258 - 320 nm. It can inhibit melanin production by inhibiting the proliferation of melanocytes and the generation of ROS, prevent skin darkening caused by oxygen free radical damage induced by light, and at the same time repair cells damaged by ultraviolet rays. Therefore, SOD enzyme is widely used in various cosmetics to provide antioxidant protection and skin repair effects, help delay skin aging, reduce wrinkles, freckles, and provide sun protection.

[0003] The activity of SOD enzyme has an important impact on the efficacy, stability, and safety of cosmetics. The exertion of its multiple effects such as antioxidant, anti-inflammatory, anti-wrinkle, freckle removal, and sun protection depends on the activity of SOD. The duration of maintaining the activity of SOD in cosmetics depends on the type of SOD, production process, storage conditions, etc.; therefore, how to make the SOD enzyme in cosmetics continuously maintain a high activity for a long time is a major problem in the cosmetics production process. Summary of the Invention

[0004] Based on this, the object of the present invention is to provide a preparation method and application of a high-activity SOD enzyme compound raw material. This preparation method can enable the SOD enzyme to maintain a high activity for a long time during the preparation and storage of cosmetics, thereby enhancing its anti-aging and whitening effects when applied to cosmetics.

[0005] A preparation method of a high-activity SOD enzyme compound raw material includes the following steps:

[0006] (1) Add SOD enzyme to a zinc glycinate solution and perform ultrasonic treatment;

[0007] (2) Collect the reaction solution and perform dialysis and / or ultrafiltration to obtain a filtrate;

[0008] (3) Adjust the pH value of the filtrate, add a hydrolyzed protein zinc solution to the filtrate, and stir well to obtain a mixed solution;

[0009] (4) Add a sodium carbonate and / or sodium bicarbonate buffer solution to the mixed solution and adjust the pH value;

[0010] (5) Ultrafiltration and concentration, followed by freeze-drying to obtain the SOD enzyme compound raw material.

[0011] Through the research of the inventor, it is found that the main reasons for the easy inactivation of superoxide dismutase during its use as a cosmetic raw material are as follows: 1. Chemical instability: SOD is a biological enzyme, and its activity is affected by various factors such as pH value, temperature, and ionic strength. When it is compounded with raw materials, the environment it is in is changed, resulting in a decrease or loss of activity. 2. Oxidation: Some raw materials (such as the preservative bromochlorophene, etc.) have oxidizing properties and can undergo redox reactions with the SOD enzyme, thereby destroying its structure or active center and causing the SOD enzyme to become ineffective. 3. Adsorption: Some raw materials (such as surfactants) may have adsorptivity and can adsorb on the surface of the SOD enzyme, thus hindering its binding to the substrate or affecting its catalytic action, resulting in a decrease in activity. 4. Enzyme activity inhibition: Some raw materials (such as the plant extract resveratrol, alcohols, ketone preservatives, etc.) have inhibitory effects on the SOD enzyme and can directly act on the active center of SOD, making it unable to exert its catalytic function.

[0012] Therefore, in the present invention, the SOD enzyme is added to the zinc glycinate solution. On the one hand, zinc ions can capture free radicals and reduce the oxidation rate of other components of the compound raw material to the SOD enzyme; after being absorbed by the skin, zinc ions replace the redox-active molecules and bind to cell membranes and proteins, thereby helping cells to resist oxidation; zinc ions can also induce the synthesis of metallothionein that can scavenge reactive oxygen molecules in cells and cooperate with the SOD enzyme to play an antioxidant role. On the other hand, glycine is used to react with the SOD enzyme to form amide bonds and ionic bonds, increasing the steric hindrance and blocking the active functional groups of the SOD enzyme, thereby achieving a bio-reversible closure of the SOD enzyme in vitro, avoiding other components of the compound raw material from adsorbing on the active groups of the SOD enzyme or undergoing redox reactions with the SOD enzyme, and avoiding other components of the compound raw material from directly acting on the active center of the SOD enzyme to inhibit the biological activity of the SOD enzyme; after entering the human body or the skin, the biological enzymes in the human body or the microorganisms on the skin surface have biological enzymes that can react with the amide bonds and can open the blocked active functional groups to exert the role of the SOD enzyme. In addition, hydrolyzed protein is used to further wrap and close the SOD enzyme, and the zinc ions complexed by the hydrolyzed protein are electrostatically adsorbed on the outermost layer of the SOD enzyme body wrapped by the hydrolyzed protein due to the zeta potential, which can reduce the influence of other oxidation factors on the SOD enzyme during the production and transportation of cosmetics. In addition, the present invention also adds a carbonate buffer system similar to the blood buffer system. On the one hand, it is beneficial for the SOD enzyme to maintain its activity in the peripheral blood circulation after entering the human body or the skin; on the other hand, since the human skin is weakly acidic, it can react with the weakly alkaline sodium carbonate and sodium bicarbonate to produce carbon dioxide, thereby activating the "Bohr effect" on the skin surface and further improving the skin microenvironment.

[0013] Further, the buffer solution in step (4) comprises sodium carbonate at 30 - 500 mM and / or sodium bicarbonate at 30 - 500 mM.

[0014] Further, the concentration of zinc glycinate in the zinc glycinate solution in step (1) is 0.01 - 145 mg / L.

[0015] Further, the ultrasonic power in step (1) is 100 - 600 W.

[0016] Further, the cut-off molecular weight of the dialysis bag used for dialysis in step (2) is 100 - 1000 kDa; the cut-off molecular weight of the filter membrane used for ultrafiltration is 100 - 1000 kDa.

[0017] Further, the pH value of the filtrate in step (3) is 5 - 6.5. Keeping the pH value of the filtrate within the above range is beneficial to promoting the ionization of hydrolyzed protein zinc and enhancing the encapsulation effect of hydrolyzed protein on SOD enzyme.

[0018] Further, the concentration of hydrolyzed protein zinc in the hydrolyzed protein zinc solution in step (3) is 0.01 - 1000 mg / L.

[0019] Further, the pH value in step (4) is 6.5 - 7.5. This pH value range is beneficial to keeping SOD enzyme at a high activity.

[0020] Use of any of the above SOD enzyme compounding raw materials in cosmetics; the cosmetics include but are not limited to the following forms: cream, hand cream, hand mask, foot mask, essence.

[0021] For better understanding and implementation, the present invention will be described in detail below. Description of the Drawings

[0022] Figure 1 Schematic diagram of the melatonin whitening effect of the SOD enzyme compounding raw materials of Examples 1 - 3 and untreated SOD enzyme in the zebrafish whitening efficacy test. Detailed Embodiments

[0023] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the embodiments of the present application. It should be clear that the described embodiments are only a part of the embodiments of the present application, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without creative efforts belong to the scope of protection of the embodiments of the present application.

[0024] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the embodiments of the present application. The singular forms "a", "the" and "said" used in the embodiments of the present application and the appended claims are also intended to include the plural forms, unless the context clearly indicates otherwise. It should also be understood that the term " / and" used herein refers to and includes any and all possible combinations of one or more of the associated listed items.

[0025] In addition, in the description of the present application, unless otherwise specified, "a plurality" means two or more. " / and" describes the association relationship of associated objects and indicates that three relationships may exist. For example, A / and B may represent three situations: A exists alone, A and B exist simultaneously, and B exists alone. The character " / " generally represents an "or" relationship between the associated objects before and after.

[0026] It should be understood that the embodiments of the present application are not limited to the content described above and various modifications and changes can be made without departing from its scope. The scope of the embodiments of the present application is only limited by the appended claims.

[0027] A preparation method of a high-activity SOD enzyme compound raw material comprises the following steps:

[0028] (1) Add SOD enzyme into a zinc glycinate solution and perform ultrasonic treatment. The concentration of zinc glycinate in the zinc glycinate solution is 0.01 - 145 mg / L, preferably 140 mg / L. The ultrasonic power is 100 - 600 W, preferably 400 W.

[0029] (2) Collect the reaction solution and perform dialysis and / or ultrafiltration to obtain a filtrate. The cut-off molecular weight of the dialysis bag used for dialysis is 100 - 1000 kDa, preferably 500 kDa; the cut-off molecular weight of the filter membrane used for ultrafiltration is 100 - 1000 kDa, preferably 500 kDa.

[0030] (3) Adjust the pH value of the filtrate, add a zinc hydrolyzed protein solution to the filtrate, and stir well to obtain a mixed solution. The pH value of the filtrate is 5 - 6.5, preferably 5.5. The concentration of zinc hydrolyzed protein in the zinc hydrolyzed protein solution is 0.01 - 1000 mg / L, preferably 0.1 mg / L.

[0031] (4) Add a sodium carbonate and / or sodium bicarbonate buffer solution to the mixed solution and adjust the pH value. The buffer solution comprises 30 - 500 mM of sodium carbonate and / or 30 - 500 mM of sodium bicarbonate, both preferably 200 mM; the pH value is 6.5 - 7.5, preferably 7.25.

[0032] (5) Perform ultrafiltration concentration and lyophilize to obtain the SOD enzyme compound raw material.

[0033] Application of the above SOD enzyme compound raw material in cosmetics; the cosmetics include but are not limited to the following forms: cream, hand cream, hand mask, foot mask, essence, capsule, etc.

[0034] Example 1

[0035] A preparation method of a high-activity SOD enzyme compound raw material, comprising the following steps:

[0036] (1) Add SOD enzyme to a glycine zinc solution of 0.01 mg / L and perform ultrasonic treatment. The ultrasonic power is 100 W.

[0037] (2) Collect the reaction solution and perform dialysis and ultrafiltration to obtain a filtrate. The cut-off molecular weights of the dialysis bag used for dialysis and the filter membrane used for ultrafiltration are both 100 kDa.

[0038] (3) Adjust the pH value of the filtrate to 5, add a hydrolyzed protein zinc solution to the filtrate, and stir well to obtain a mixed solution. The concentration of hydrolyzed protein zinc in the hydrolyzed protein zinc solution is 0.01 mg / L.

[0039] (4) Add a sodium carbonate and sodium bicarbonate buffer solution to the mixed solution and adjust the pH value to 6.5. The buffer solution includes 30 mM of sodium carbonate and 30 mM of sodium bicarbonate.

[0040] (5) Perform ultrafiltration concentration and lyophilize to obtain the SOD enzyme compound raw material.

[0041] Example 2

[0042] A preparation method of a high-activity SOD enzyme compound raw material, comprising the following steps:

[0043] (1) Add SOD enzyme to a glycine zinc solution of 140 mg / L and perform ultrasonic treatment. The ultrasonic power is 400 W.

[0044] (2) Collect the reaction solution and perform dialysis and ultrafiltration to obtain a filtrate. The cut-off molecular weights of the dialysis bag used for dialysis and the filter membrane used for ultrafiltration are both 500 kDa.

[0045] (3) Adjust the pH value of the filtrate to 5.5, add a hydrolyzed protein zinc solution to the filtrate, and stir well to obtain a mixed solution. The concentration of hydrolyzed protein zinc in the hydrolyzed protein zinc solution is 0.1 mg / L.

[0046] (4) Add a sodium carbonate and sodium bicarbonate buffer solution to the mixed solution and adjust the pH value to 7.25. The buffer solution includes 200 mM of sodium carbonate and 200 mM of sodium bicarbonate.

[0047] (5) Ultrafiltration and concentration, followed by freeze-drying to obtain the SOD enzyme compound raw material.

[0048] Example 3

[0049] A preparation method of a high-activity SOD enzyme compound raw material, comprising the following steps:

[0050] (1) Add the SOD enzyme to a glycine zinc solution of 0.01 mg / L and perform ultrasonic treatment. The ultrasonic power is 600 W.

[0051] (2) Collect the reaction solution and perform dialysis and ultrafiltration to obtain a filtrate. The cut-off molecular weight of the dialysis bag used for dialysis and the filter membrane used for ultrafiltration is both 1000 kDa.

[0052] (3) Adjust the pH value of the filtrate to 6.5, add a hydrolyzed protein zinc solution to the filtrate, and stir well to obtain a mixed solution. The concentration of hydrolyzed protein zinc in the hydrolyzed protein zinc solution is 1000 mg / L.

[0053] (4) Add a sodium carbonate and sodium bicarbonate buffer solution to the mixed solution and adjust the pH value to 6.5. The buffer solution includes 500 mM of sodium carbonate and 500 mM of sodium bicarbonate.

[0054] (5) Ultrafiltration and concentration, followed by freeze-drying to obtain the SOD enzyme compound raw material.

[0055] Example 4

[0056] A cream containing the high-activity SOD enzyme compound raw material, comprising at least the following components:

[0057] Deionized water: As a solvent, providing the basis for the cream.

[0058] Glycerol: A moisturizer that can absorb and retain moisture, keeping the skin moist.

[0059] The high-activity SOD enzyme compound raw material according to any one of Examples 1 to 3: The core active ingredient, having antioxidant and anti-aging effects.

[0060] Shea butter: A natural vegetable oil, having the effects of moisturizing, nourishing and soothing the skin.

[0061] Glyceryl stearate: An emulsifier that helps to mix the oil phase and the water phase together.

[0062] Coconut oil: Providing additional moisturizing and nourishing effects.

[0063] Phenoxyethanol: A preservative to ensure the hygiene and safety of the cream.

[0064] Retinol A (Vitamin A): It has the function of anti-wrinkle and skin repair.

[0065] Vitamin E: An antioxidant that protects the skin from damage by free radicals.

[0066] Triethanolamine: Regulates the pH value and consistency of the cream.

[0067] Other auxiliary ingredients: Such as non-irritating fragrance (optional), pigment (optional), etc.

[0068] The formulation steps of the above-mentioned cream containing the high-activity SOD enzyme compound raw material are as follows:

[0069] (1) Prepare the aqueous phase solution: Mix the deionized water, the glycerol and the high-activity SOD enzyme compound raw material together, and heat to an appropriate temperature (usually about 70 - 80 °C) to ensure that the high-activity SOD enzyme compound raw material can be uniformly dissolved.

[0070] (2) Prepare the oil phase solution: Mix the shea butter, the glyceryl stearate and the coconut oil together, and also heat to an appropriate temperature.

[0071] (3) Emulsification: Under stirring, slowly pour the aqueous phase solution into the oil phase solution, and continue stirring until a uniform emulsion is formed.

[0072] (4) Adjust the pH value and consistency: Add the triethanolamine to adjust the pH value and consistency of the cream to the required level.

[0073] (5) Add other ingredients: After the cream is cooled to room temperature, add the retinol A, the vitamin E, the phenoxyethanol and other auxiliary ingredients (such as fragrance, pigment, etc.).

[0074] (6) Homogenization: Use a homogenizer to homogenize the cream to ensure its texture is delicate and uniform.

[0075] (7) Filling and packaging: Fill the cream into appropriate containers, and seal and package it.

[0076] Please note that this is just a basic cream formulation example, and actual production may need to be adjusted according to specific requirements and conditions.

[0077] The physical property indexes and their test methods of the examples or comparative examples of the present invention are specifically as follows:

[0078] The SOD enzyme compound raw materials obtained from Examples 1 - 3 are respectively labeled as Samples A, B, and C in sequence, and the untreated SOD enzyme is taken as Sample D. Take Samples A, B, C, and D for the following test experiments:

[0079] (1) Zebrafish Toxicity Evaluation

[0080] a. Sexually mature zebrafish are separately reared in male and female tanks in the zebrafish culture unit. Water temperature: 26 ± 2 °C;

[0081] b. Healthy zebrafish embryos at 9 hpf (hours post fertilization) after fertilized eggs are selected and randomly placed in a 6-well cell culture plate, 50 embryos per well, and 5 mL of the sample solution is added to each well. A solvent control group and a sample group are set up for the experiment, and each group has 3 replicate wells. Incubate in a curve-controlled biochemical incubator (pH: 6.7; conductivity: 520 us / cm; light / dark cycle: 14 h / 10 h). One day before the start of the exposure experiment, males and females are paired at a ratio of 1:2 and allowed to mate and lay eggs naturally.

[0082] c. After exposure to 24 hpf, 48 hpf, and 72 hpf, the growth and development of zebrafish are observed with a stereomicroscope and photographed respectively, and the mortality rate is calculated.

[0083] d. Statistical analysis: The Origin9.0 software is used for data processing and analysis, and the mortality rates of each test material at different concentrations are summarized in Table 1 below.

[0084] Table 1

[0085]

[0086]

[0087] (2) Zebrafish Whitening Efficacy Evaluation

[0088] a. Healthy zebrafish embryos at 9 hpf after fertilization are randomly placed in a 6-well cell culture plate, 50 embryos per well, and 5 mL of the solution is added to each well. A solvent control group and a sample group are set up for the experiment, and each group has 3 replicate wells. Incubate in a curve-controlled biochemical incubator.

[0089] b. After exposure to 72 hpf, the pigmentation of zebrafish is observed with a stereomicroscope and photographed.

[0090] c. 40 larvae are selected from each well and placed in a culture dish, rinsed twice with culture water, and the larvae are transferred to a 1.5 mL EP tube. 150 μL of sodium deoxycholate solution is added, and a homogenate is prepared by ultrasonic treatment at low temperature.

[0091] d. Determination of melanin content: Add 150 μL of NaOH solution to the EP tube, heat in a water bath at 100 °C for 10 min, and shake to fully dissolve the melanin in the EP tube. Take 100 μL of the solution and place it in a 96-well plate, and measure the absorbance at a wavelength of 405 nm with an enzyme-linked immunosorbent assay (ELISA) reader. Calculate the melanin content of the sample according to the following formula:

[0092]

[0093] e. Determination of tyrosinase activity: The homogenate was centrifuged at 10,000 rpm for 5 min, and 100 μL of the supernatant was taken and placed in a 96-well plate. At the same time, 100 μL of L-dopa solution was added, and the mixture was incubated at 37 °C for 1 h. The absorbance value (OD475 value) of each experimental group was detected using an enzyme-linked immunosorbent assay (ELISA) reader. The tyrosinase activity of the sample was calculated according to the following formula:

[0094]

[0095] f. Statistical analysis: The data were statistically processed using SPSS 19.0 software to obtain Table 2 below; the melatonin whitening effect in zebrafish is shown in the appendix Figure 1 .

[0096] Table 2

[0097]

[0098]

[0099] (Compared with the blank control group: *p < 0.05, **p < 0.01, ***p < 0.001)

[0100] (3) SOD activity detection test (riboflavin-NBT method):

[0101] The SOD activity detection test (riboflavin-NBT method) was performed on the samples stored for one month and the samples stored for one year respectively. A certain amount of the initial sample was weighed and dispersed in 8 mL of PBS buffer solution containing 0.05 mol / L methionine, 1.16×10 -3 mol / L NBT, 8.50×10 -5 mol / L riboflavin and 2.55×10 -6 mol / L EDTA. The operation is as follows: First, air was bubbled into the mixture and stirred for 5 minutes under dark conditions, and then the reaction was carried out for 10 minutes under light conditions. The absorbance before and after light irradiation was measured using a UV-visible spectrophotometer. Taking the activity of the just-prepared sample as 100%, the inactivation inhibition rates for one month and one year were calculated according to the following formula:

[0102]

[0103] (4) DPPH free radical scavenging experiment:

[0104] First, prepare the sample into a solution with methanol, and prepare the DPPH into a 0.2 mM solution with absolute ethanol, and store it in the dark at 4 °C. Take 100 μL of the sample solution (SOD concentration is 0.5%), add 100 μL of the DPPH solution, mix well, react in the dark for 30 min, measure the absorbance at 517 nm, and record it as Ai; perform the same operation, measure the absorbance of 100 μL of the sample solution and 100 μL of the absolute ethanol solution at 517 nm, and record it as Aj; perform the same operation, measure the absorbance of 100 μL of methanol and 100 μL of the DPPH solution at 517 nm, and record it as A0. Use BHA as the positive control, set three parallel experiments for each group, and calculate the DPPH· scavenging activity according to the following formula:

[0105] DPPH· scavenging rate (%) = [A0 - (Ai - Aj)] / A0 × 100.

[0106] (5) Elastase inhibition rate test:

[0107] Mix 85 μL of 50 mmol / L Tris-HCl buffer (pH 8.0) with 15 μL of the reaction solution, then add 25 μL of the elastase solution (60 mU / mL), incubate at 25 °C for 15 min, then add 25 μL of 1.015 mmol / L AAAPAN solution, measure the absorbance at 410 nm after 15 min. The elastase solution and the AAAPAN solution are both prepared with 50 mmol / L Tris-HCl buffer (pH 8.0). Use EGCG as the positive control and measure in parallel 3 times. Calculate the inhibition rate of the sample on elastase according to the following formula:

[0108]

[0109] In the formula: C is the absorbance of the reaction solution without the sample; D is the absorbance of the reaction solution without the sample and the enzyme; A is the absorbance of the reaction solution containing the sample and the enzyme, and the SOD enzyme concentration is 5%; B is the absorbance of the reaction solution without the enzyme.

[0110] The test results of the SOD enzyme activity, DPPH free radical scavenging and elastase inhibition ability of the high-activity SOD enzyme compound raw materials described in Examples 1 to 3 and the untreated SOD enzyme are shown in Table 3 below:

[0111] Table 3

[0112]

[0113] As can be seen from Table 3, the inactivation inhibition rates of the SOD enzymes contained in the high-activity SOD enzyme compounding raw materials described in Examples 1 to 3 after one month and one year of storage are far lower than those of the untreated SOD enzymes, indicating that the preparation method of the high-activity SOD enzyme compounding raw materials of the present invention can keep the SOD active at a high level for a long time and has excellent stability. In addition, the DPPH free radical scavenging ability and elastase inhibitory ability of the high-activity SOD enzyme compounding raw materials described in Examples 1 to 3 also far exceed those of the untreated SOD enzymes, showing stronger antioxidant properties; combined with Table 2 and Figure 1 it can be seen that the whitening effects of the high-activity SOD enzyme compounding raw materials described in Examples 1 to 3 are better than those of the untreated SOD enzymes.

[0114] The above-described embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can be made, and the present invention is also intended to include these modifications and improvements.

Claims

1. A method for preparing a high-activity SOD enzyme composite raw material, characterized in that: The following steps are involved: (1) adding SOD enzyme to zinc glycine solution and performing ultrasound; (2) collecting the reaction solution and performing dialysis and / or ultrafiltration to obtain a filtrate; (3) adjusting the pH value of the filtrate, adding hydrolyzed protein zinc to the solution, and stirring thoroughly to obtain a mixed solution; (4) adding sodium carbonate and / or sodium bicarbonate buffer solution to the mixed solution and adjusting the pH value; (5) ultrafiltration concentration and freeze-drying to obtain the SOD enzyme compound raw material.

2. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The buffer solution in step (4) comprises 30-500 mM sodium carbonate and / or 30-500 mM sodium bicarbonate.

3. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The concentration of zinc glycinate in the zinc glycinate solution in step (1) is 0.01 to 145 mg / L.

4. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The ultrasonic power of step (1) is 100-600W.

5. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The molecular weight cutoff of the dialysis bag used in step (2) dialysis is 100-1000 kDa; the molecular weight cutoff of the filter membrane used in ultrafiltration is 100-1000 kDa.

6. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The pH value of the filtrate in step (3) is 5 to 6.

5.

7. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The concentration of the hydrolyzed protein zinc in the hydrolyzed protein zinc solution in step (3) is 0.01 to 1000 mg / L.

8. The method for preparing the composite raw material of high-activity SOD enzyme according to claim 1, characterized in that: The pH value in step (4) is 6.5 to 7.

5.

9. Use of the SOD enzyme compound raw material according to any one of claims 1 to 8 in cosmetics; the cosmetics include but are not limited to the following forms: face cream, hand cream, hand mask, foot mask, essence.

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