Micromolecular protein peptide for enhancing transdermal absorption and application thereof

Abstract

The invention relates to the technical field of cosmetics, and aims to provide a micromolecular protein peptide for enhancing transdermal absorption, which is prepared by the following steps: cleaning a salmon skin raw material, and performing water extraction to obtain a salmon collagen stock solution; papain is added into the salmon collagen stock solution for primary enzymolysis, carboxypeptidase B is added for secondary enzymolysis, enzyme deactivation is performed, and a salmon collagen peptide solution is obtained; and sequentially carrying out centrifugation and ultrafiltration treatment on the salmon collagen peptide solution, and freeze-drying to obtain the micromolecular protein peptide for enhancing transdermal absorption. According to the preparation method disclosed by the invention, the salmon collagen is decomposed into larger peptide fragments by adopting papain, and then the peptide fragments are further hydrolyzed by using carboxypeptidase B, so that the extracted salmon collagen peptide is low in bitter taste degree and has high transdermal absorbability through cooperative use of the papain and the carboxypeptidase B.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and more specifically to a small molecule protein peptide for enhancing transdermal absorption and application thereof. Background Art

[0002] Fish protein peptides are rich in a variety of small molecule nutrients such as polypeptides, small peptides, amino acids, trace elements, etc. It has the advantages of high bioavailability and also has antioxidant, blood pressure lowering, anti-aging, immune enhancement and other activities. It can be widely used as a base for seasonings, puffed foods, convenience and health foods, and can also be used as a recovery food for patients. Its products can exist in a variety of forms such as powder, liquid and paste, which is convenient for people with different physiological states, nutritional characteristics of people under different working conditions and market demand.

[0003] For the above reasons, salmon collagen belongs to type I collagen in structure, has good biocompatibility and biodegradability. In the field of cosmetics, salmon collagen is used in skin care products to help moisturize and improve skin elasticity. Enzymatic hydrolysis is currently the most commonly used method for industrial preparation of salmon collagen. This method has mild conditions, strong controllability and stable products. However, during the enzymatic hydrolysis process, hydrophobic amino acids are exposed when proteins are broken into polypeptides, which often makes polypeptides with hydrophobic amino acids at the C-terminus bitter, resulting in reduced sensory quality of peptide products.

[0004] Therefore, a low-bitter salmon collagen peptide is urgently needed to solve the above problems. Summary of the invention

[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a small molecule protein peptide with enhanced transdermal absorption, by first using papain to decompose salmon collagen into larger peptide segments, and then using carboxypeptidase B to further hydrolyze these peptide segments. The combined use of the two makes the extracted salmon collagen peptide have a low bitterness.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0008] (1) cleaning the salmon skin raw material and then extracting it with water to obtain salmon collagen stock solution;

[0009] (2) adding papain to the salmon collagen stock solution for primary enzymolysis, then adding carboxypeptidase B for secondary enzymolysis, inactivating the enzyme, and obtaining a salmon collagen peptide solution;

[0010] (3) The salmon collagen peptide solution is centrifuged and ultrafiltered in sequence, and then freeze-dried to obtain a small molecule protein peptide with enhanced transdermal absorption.

[0011] Preferably, the method for obtaining the salmon collagen stock solution in step (1) is as follows: the washed salmon skin is soaked in a NaOH solution at a solid-liquid ratio of 1:10-15 for 2-3 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in an ethanol solution at a solid-liquid ratio of 1:10-15 for 4-6 hours, and the salmon skin is rinsed until the washing solution is neutral; the mixture of salmon skin and deionized water is subjected to collagen extraction at 55-65° C. for 7-10 hours, the impurities in the solution are filtered to remove, centrifuged, and the supernatant is concentrated to obtain the salmon collagen stock solution.

[0012] Preferably, the salmon skin is mixed with deionized water at a material-to-liquid ratio of 1:5-10.

[0013] Preferably, the conditions for the primary enzymatic hydrolysis in step (2) are to adjust the pH of the salmon collagen stock solution to 6-9, the enzymatic hydrolysis temperature to 45-55° C., and the enzymatic hydrolysis time to 1-3 h.

[0014] Preferably, the mass percentage of papain in the salmon collagen stock solution is 0.5-2%.

[0015] Preferably, the conditions for the secondary enzymolysis in step (2) are to adjust the pH of the primary enzymolysis solution to 7-7.5, the enzymolysis temperature to 45-55° C., and the enzymolysis time to 1-3 h.

[0016] Preferably, the mass percentage of carboxypeptidase B in the primary enzymatic hydrolysis solution is 0.5-1%.

[0017] Preferably, the enzyme inactivation condition in step (2) is to adjust the temperature of the secondary enzymatic hydrolysis solution to 85-95° C., heat for 15-20 minutes, and terminate the enzymatic hydrolysis reaction.

[0018] Preferably, the ultrafiltration cutoff in step (3) is 3000-6000Da and the pressure is 0.4-0.6MPa.

[0019] Another object of the present invention is to provide an application of the aforementioned small molecule protein peptide for enhancing transdermal absorption, by adding the aforementioned small molecule protein peptide for enhancing transdermal absorption into cosmetics to prepare cosmetics with anti-aging effects.

[0020] Compared with the prior art, the present invention has the following beneficial effects: the present invention first uses papain to decompose salmon collagen into larger peptide segments, and then uses carboxypeptidase B to further hydrolyze these peptide segments. Through the coordinated use of the two, the extracted salmon collagen peptides have low bitterness and high transdermal absorbability. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a fluorescence image of the small molecule protein peptide prepared in Example 4 of the present invention entering human keratinocytes; red represents cytoskeleton protein, blue represents cell nucleus, and green represents small molecule protein peptide;

[0022] Figure 2 This is a relative fluorescence intensity diagram of the small molecule protein peptide prepared in Example 4 of the present invention entering human keratinocytes;

[0023] Figure 3 It is the fluorescence image of each sample group of Test Example 3 of the present invention entering the abdominal skin of rats;

[0024] Figure 4 It is a fluorescence intensity diagram of each sample group of Test Example 3 of the present invention entering the abdominal skin of rats. DETAILED DESCRIPTION

[0025] Example 1

[0026] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0027] (1) The cleaned salmon skin is soaked in a 10 wt % NaOH solution at a solid-liquid ratio of 1:15 for 2.5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in a 45 wt % ethanol solution at a solid-liquid ratio of 1:15 for 5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin and deionized water are mixed at a solid-liquid ratio of 1:10, and collagen is extracted at 60° C. for 8.5 hours, impurities in the solution are removed by filtration, centrifugation is performed, and the supernatant is concentrated until the volume is reduced to 40% of the original volume to obtain a salmon collagen stock solution.

[0028] (2) adjusting the pH of the salmon collagen stock solution to 6-9, adding papain to the salmon collagen stock solution, performing a primary enzymolysis at 50° C. for 2 hours, and obtaining a primary enzymolysis solution; adjusting the pH of the primary enzymolysis solution to 7-7.5, adding carboxypeptidase B to the primary enzymolysis solution, performing a secondary enzymolysis at 50° C. for 2 hours, and obtaining a secondary enzymolysis solution; adjusting the temperature of the secondary enzymolysis solution to 90° C., keeping the temperature for 15 minutes, terminating the enzymolysis reaction, and obtaining a salmon collagen peptide solution; wherein the mass percentage of papain in the salmon collagen stock solution is 2%, and the mass percentage of carboxypeptidase B in the primary enzymolysis solution is 1%.

[0029] (3) The salmon collagen peptide solution is subjected to centrifugation and ultrafiltration treatments in sequence, and then freeze-dried to obtain a powdered small molecule protein peptide with enhanced transdermal absorption; wherein the centrifugal speed is 8000 r / min and the time is 10 min; the polypeptides and insoluble proteins can be separated as much as possible, thereby increasing the polypeptide yield; the ultrafiltration retention capacity is 3000-6000 Da, and the pressure is 0.4-0.6 Mpa, which can remove a small amount of residual lipids and large molecular proteins and polypeptides as much as possible, effectively remove impurities, and obtain a clear and pure peptide solution.

[0030] Example 2

[0031] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0032] (1) The cleaned salmon skin is soaked in a 10 wt % NaOH solution at a solid-liquid ratio of 1:15 for 2.5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in a 45 wt % ethanol solution at a solid-liquid ratio of 1:15 for 5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin and deionized water are mixed at a solid-liquid ratio of 1:10, and collagen is extracted at 60° C. for 8.5 hours, impurities in the solution are removed by filtration, centrifugation is performed, and the supernatant is concentrated until the volume is reduced to 40% of the original volume to obtain a salmon collagen stock solution.

[0033] (2) adjusting the pH of the salmon collagen stock solution to 6-9, adding papain to the salmon collagen stock solution, performing a primary enzymolysis at 50° C. for 2 hours, and obtaining a primary enzymolysis solution; adjusting the pH of the primary enzymolysis solution to 7-7.5, adding carboxypeptidase B to the primary enzymolysis solution, performing a secondary enzymolysis at 50° C. for 2 hours, and obtaining a secondary enzymolysis solution; adjusting the temperature of the secondary enzymolysis solution to 90° C., keeping the temperature for 15 minutes, terminating the enzymolysis reaction, and obtaining a salmon collagen peptide solution; wherein the mass percentage of papain in the salmon collagen stock solution is 2%, and the mass percentage of carboxypeptidase B in the primary enzymolysis solution is 0.5%.

[0034] (3) The salmon collagen peptide solution is subjected to centrifugation and ultrafiltration treatments in sequence, and then freeze-dried to obtain a powdered small molecule protein peptide with enhanced transdermal absorption; wherein the centrifugal speed is 8000 r / min and the time is 10 min; the polypeptides and insoluble proteins can be separated as much as possible, thereby increasing the polypeptide yield; the ultrafiltration retention capacity is 3000-6000 Da, and the pressure is 0.4-0.6 Mpa, which can remove a small amount of residual lipids and large molecular proteins and polypeptides as much as possible, effectively remove impurities, and obtain a clear and pure peptide solution.

[0035] Example 3

[0036] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0037] (1) The cleaned salmon skin is soaked in a 10 wt % NaOH solution at a solid-liquid ratio of 1:15 for 2.5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in a 45 wt % ethanol solution at a solid-liquid ratio of 1:15 for 5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin and deionized water are mixed at a solid-liquid ratio of 1:10, and collagen is extracted at 60° C. for 8.5 hours, impurities in the solution are removed by filtration, centrifugation is performed, and the supernatant is concentrated until the volume is reduced to 40% of the original volume to obtain a salmon collagen stock solution.

[0038] (2) adjusting the pH of the salmon collagen stock solution to 6-9, adding papain to the salmon collagen stock solution, performing a primary enzymolysis at 50° C. for 2 hours, and obtaining a primary enzymolysis solution; adjusting the pH of the primary enzymolysis solution to 7-7.5, adding carboxypeptidase B to the primary enzymolysis solution, performing a secondary enzymolysis at 50° C. for 2 hours, and obtaining a secondary enzymolysis solution; adjusting the temperature of the secondary enzymolysis solution to 90° C., keeping the temperature for 15 minutes, terminating the enzymolysis reaction, and obtaining a salmon collagen peptide solution; wherein the mass percentage of papain in the salmon collagen stock solution is 1%, and the mass percentage of carboxypeptidase B in the primary enzymolysis solution is 0.5%.

[0039] (3) The salmon collagen peptide solution is subjected to centrifugation and ultrafiltration treatments in sequence, and then freeze-dried to obtain a powdered small molecule protein peptide with enhanced transdermal absorption; wherein the centrifugal speed is 8000 r / min and the time is 10 min; the polypeptides and insoluble proteins can be separated as much as possible, thereby increasing the polypeptide yield; the ultrafiltration retention capacity is 3000-6000 Da, and the pressure is 0.4-0.6 Mpa, which can remove a small amount of residual lipids and large molecular proteins and polypeptides as much as possible, effectively remove impurities, and obtain a clear and pure peptide solution.

[0040] Example 4

[0041] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0042] (1) The cleaned salmon skin is soaked in a 10 wt % NaOH solution at a solid-liquid ratio of 1:15 for 2.5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in a 45 wt % ethanol solution at a solid-liquid ratio of 1:15 for 5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin and deionized water are mixed at a solid-liquid ratio of 1:10, and collagen is extracted at 60° C. for 8.5 hours, impurities in the solution are removed by filtration, centrifugation is performed, and the supernatant is concentrated until the volume is reduced to 40% of the original volume to obtain a salmon collagen stock solution.

[0043] (2) adjusting the pH of the salmon collagen stock solution to 6-9, adding papain to the salmon collagen stock solution, performing a primary enzymolysis at 50° C. for 2 hours, and obtaining a primary enzymolysis solution; adjusting the pH of the primary enzymolysis solution to 7-7.5, adding carboxypeptidase B to the primary enzymolysis solution, performing a secondary enzymolysis at 50° C. for 2 hours, and obtaining a secondary enzymolysis solution; adjusting the temperature of the secondary enzymolysis solution to 90° C., keeping the temperature for 15 minutes, terminating the enzymolysis reaction, and obtaining a salmon collagen peptide solution; wherein the mass percentage of papain in the salmon collagen stock solution is 1%, and the mass percentage of carboxypeptidase B in the primary enzymolysis solution is 1%.

[0044] (3) The salmon collagen peptide solution is subjected to centrifugation and ultrafiltration treatments in sequence, and then freeze-dried to obtain a powdered small molecule protein peptide with enhanced transdermal absorption; wherein the centrifugal speed is 8000 r / min and the time is 10 min; the polypeptides and insoluble proteins can be separated as much as possible, thereby increasing the polypeptide yield; the ultrafiltration retention capacity is 3000-6000 Da, and the pressure is 0.4-0.6 Mpa, which can remove a small amount of residual lipids and large molecular proteins and polypeptides as much as possible, effectively remove impurities, and obtain a clear and pure peptide solution.

[0045] Example 5

[0046] A small molecule protein peptide for enhancing transdermal absorption is prepared by the following steps:

[0047] (1) The cleaned salmon skin is soaked in a 10 wt % NaOH solution at a solid-liquid ratio of 1:15 for 2.5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in a 45 wt % ethanol solution at a solid-liquid ratio of 1:15 for 5 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin and deionized water are mixed at a solid-liquid ratio of 1:10, and collagen is extracted at 60° C. for 8.5 hours, impurities in the solution are removed by filtration, centrifugation is performed, and the supernatant is concentrated until the volume is reduced to 40% of the original volume to obtain a salmon collagen stock solution.

[0048] (2) adjusting the pH of the salmon collagen stock solution to 6-9, adding papain to the salmon collagen stock solution, performing a primary enzymolysis at 50° C. for 2 hours, and obtaining a primary enzymolysis solution; adjusting the pH of the primary enzymolysis solution to 7-7.5, adding carboxypeptidase B to the primary enzymolysis solution, performing a secondary enzymolysis at 50° C. for 2 hours, and obtaining a secondary enzymolysis solution; adjusting the temperature of the secondary enzymolysis solution to 90° C., keeping the temperature for 15 minutes, terminating the enzymolysis reaction, and obtaining a salmon collagen peptide solution; wherein the mass percentage of papain in the salmon collagen stock solution is 0.5%, and the mass percentage of carboxypeptidase B in the primary enzymolysis solution is 1%.

[0049] (3) The salmon collagen peptide solution is subjected to centrifugation and ultrafiltration treatments in sequence, and then freeze-dried to obtain a powdered small molecule protein peptide with enhanced transdermal absorption; wherein the centrifugal speed is 8000 r / min and the time is 10 min; the polypeptides and insoluble proteins can be separated as much as possible, thereby increasing the polypeptide yield; the ultrafiltration retention capacity is 3000-6000 Da, and the pressure is 0.4-0.6 Mpa, which can remove a small amount of residual lipids and large molecular proteins and polypeptides as much as possible, effectively remove impurities, and obtain a clear and pure peptide solution.

[0050] Comparative Example 1

[0051] A small molecule protein peptide for enhancing transdermal absorption, the preparation steps of which are different from those of Example 4, except that an equal amount of carboxypeptidase A is used to replace carboxypeptidase B.

[0052] Comparative Example 2

[0053] A small molecule protein peptide for enhancing transdermal absorption, the preparation steps of which are different from those of Example 4, except that an equal amount of aminopeptidase is used to replace carboxypeptidase B.

[0054] Comparative Example 3

[0055] A small molecule protein peptide for enhancing transdermal absorption, the preparation steps of which are different from those of Example 4, except that an equal amount of pepsin is used to replace papain.

[0056] Comparative Example 4

[0057] A small molecule protein peptide for enhancing transdermal absorption, the preparation steps of which are different from those of Example 4, except that an equal amount of chymosin is used to replace papain.

[0058] Test Example 1

[0059] Bitterness test: The small molecule protein peptides prepared in Examples 1-5 and Comparative Examples 1-4 were prepared into two concentration solutions of 3wt% and 6wt% for sensory evaluation. Eight assessors (4 males and 4 females, aged 25-35 years) with sensory evaluation skills were selected to judge the bitterness of the solutions, wherein the bitterness value was evaluated based on tannin solutions of different concentrations, with 10 mmol / L tannin solution being non-bitter, 50 mmol / L tannin solution being slightly bitter, and 100 mmol / L tannin solution being significantly bitter. The test results are shown in Table 1.

[0060] Table 1: Bitterness test results of small molecule protein peptides of Examples 1-5 and Comparative Examples 1-4

[0061]

[0062]

[0063] It can be seen from Table 1 that the bitterness of the small molecule protein peptides provided by Examples 1-5 of the present invention is significantly lower than that of the small molecule protein peptides provided by Comparative Examples 1-4. Analysis shows that the bitterness of the extracted salmon collagen peptides can be effectively reduced only when the salmon collagen is hydrolyzed by papain and carboxypeptidase B at specific contents. When other proteases (such as pepsin and chymosin) are used instead of papain, or carboxypeptidase A or aminopeptidase is used instead of carboxypeptidase B, the bitterness of the salmon collagen peptides cannot be significantly reduced.

[0064] Test Example 2

[0065] Permeability test: The small molecule protein peptide prepared in Example 4 was prepared into a solution with a concentration of 3 wt%, and the solution was used to treat human keratinocytes after green fluorescence labeling to observe the time when the small molecule protein peptide appeared in the cells. The test results are as follows: Figure 1-2 shown.

[0066] from Figure 1-2 It can be seen that small molecule protein peptides can be observed in the cells 10 minutes after treatment; as time goes by, the number of small molecule protein peptides entering the cells continues to increase, and the fluorescence intensity is observed to be maintained at a relatively balanced level from 60 to 120 minutes, indicating that the salmon collagen peptides in the cytoplasm continue to play a role.

[0067] Test Example 3

[0068] Transdermal absorption test: A fluorescently labeled experimental group (small molecule protein peptide prepared in Example 4), a fluorescently labeled rhEGF group, and a control group (normal saline) were set up. Each group of samples was prepared into a solution with a concentration of 3wt% and applied to the abdominal skin of rats. After 4 hours, the absorption of each sample was observed (the higher the relative fluorescence intensity value, the better the transdermal effect). The test results are as follows: Figure 3-4 shown.

[0069] from Figure 3-4 It can be seen that the green fluorescent labeled salmon collagen peptide can quickly penetrate the stratum corneum of the skin and be absorbed by the skin, and the penetration absorption amount is about twice that of the rhEGF group.

[0070] Test Example 4

[0071] Skin irritation safety test: refer to the human skin patch test in the "2015 Cosmetic Safety Technical Specifications". The method of skin occlusion patch test is: select 30 people aged 18-60 years old (divided into 6 groups, 5 people in each group), select an area not exceeding 50mm 2 , a qualified spot test device with a depth of about 1mm, take 0.020mL of the solution (concentration of 6wt%) prepared by the small molecule protein peptide prepared in Examples 1-5 above and put it into the small chamber of the spot tester, the control well is a blank control (no substance is placed), and the spot tester with the composition is applied to the curved side of the forearm of the subject with a low-allergenic tape, and gently press it with the palm to evenly apply it to the skin, and continue for 24 hours. Observe the skin reaction 30 minutes after removing the spot tester containing the composition (after the indentation disappears), and record the observation results. The results show that the five test groups and the control group are all negative reactions, indicating that the small molecule protein peptides prepared in Examples 1-5 passed the human patch safety test.

[0072] The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.

Claims

1. A small molecule protein peptide for enhancing transdermal absorption, characterized in that: Prepared by the following steps: (1) cleaning the salmon skin raw material and then extracting it with water to obtain salmon collagen stock solution; (2) adding papain to the salmon collagen stock solution for primary enzymolysis, then adding carboxypeptidase B for secondary enzymolysis, inactivating the enzyme, and obtaining a salmon collagen peptide solution; (3) The salmon collagen peptide solution is centrifuged and ultrafiltered in sequence, and then freeze-dried to obtain a small molecule protein peptide with enhanced transdermal absorption.

2. The small molecule protein peptide for enhancing transdermal absorption according to claim 1, characterized in that: The method for obtaining the salmon collagen stock solution in step (1) is as follows: the washed salmon skin is soaked in a NaOH solution at a solid-liquid ratio of 1:10-15 for 2-3 hours, and the salmon skin is rinsed until the washing solution is neutral; the salmon skin treated with the NaOH solution is soaked in an ethanol solution at a solid-liquid ratio of 1:10-15 for 4-6 hours, and the salmon skin is rinsed until the washing solution is neutral; the mixture of salmon skin and deionized water is subjected to collagen extraction at 55-65° C. for 7-10 hours, the impurities in the solution are filtered to remove, the solution is centrifuged, and the supernatant is concentrated to obtain the salmon collagen stock solution.

3. The small molecule protein peptide for enhancing transdermal absorption according to claim 2, characterized in that: The salmon skin is mixed with deionized water at a material-to-liquid ratio of 1:5-10.

4. The small molecule protein peptide for enhancing transdermal absorption according to claim 1, characterized in that: The conditions for the primary enzymatic hydrolysis in step (2) are to adjust the pH of the salmon collagen stock solution to 6-9, the enzymatic hydrolysis temperature to 45-55° C., and the enzymatic hydrolysis time to 1-3 h.

5. The small molecule protein peptide for enhancing transdermal absorption according to claim 4, characterized in that: The mass percentage of papain in the salmon collagen stock solution is 0.5-2%.

6. The small molecule protein peptide for enhancing transdermal absorption according to claim 1, characterized in that: The conditions for the secondary enzymolysis in step (2) are to adjust the pH of the primary enzymolysis solution to 7-7.5, the enzymolysis temperature to 45-55° C., and the enzymolysis time to 1-3 h.

7. The small molecule protein peptide for enhancing transdermal absorption according to claim 6, characterized in that: The mass percentage of carboxypeptidase B in the primary enzymolysis solution is 0.5-1%.

8. The small molecule protein peptide for enhancing transdermal absorption according to claim 1, characterized in that: The conditions for inactivating the enzyme in step (2) are to adjust the temperature of the secondary enzymatic hydrolysis solution to 85-95° C., heat for 15-20 minutes, and terminate the enzymatic hydrolysis reaction.

9. The small molecule protein peptide for enhancing transdermal absorption according to claim 1, characterized in that: The ultrafiltration cutoff in step (3) is 3000-6000Da and the pressure is 0.4-0.6MPa.

10. An application of a small molecule protein peptide for enhancing transdermal absorption, characterized in that: The small molecule protein peptide for enhancing transdermal absorption according to any one of claims 1 to 9 is added to cosmetics to prepare cosmetics with anti-aging effects.

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