Active peptide with oxidative stress regulation function and preparation and application thereof

Abstract

The application discloses an active peptide with an oxidative stress regulation function and preparation and application thereof, and the amino acid sequence of the active peptide is Ala-Val-Pro-Ile-Cys-Ala-Val-Pro. The preparation method of the active peptide comprises collecting pearl mussels, obtaining an extract, collecting supernatant, selecting, screening and synthesizing polypeptides. The active peptide with the oxidative stress regulation function has high activity, can better remove free radicals, reduce the damage of H2O2 to skin fibroblasts, and can promote skin wound healing. The active peptide with the oxidative stress regulation function has a good application prospect in drugs or instruments for reducing cell oxidative stress.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological polypeptides, and specifically relates to an active peptide with an oxidative stress regulation function and preparation and application thereof. BACKGROUND

[0002] Oxidative stress refers to a pathological process in which the imbalance between oxidation and antioxidant states in the body is caused by excessive production of reactive oxygen species (ROS) or insufficient antioxidant system function. Oxidative stress not only damages cell membranes, proteins and DNA molecules, but also inhibits tissue repair and regeneration capacity. This process is closely related to the occurrence and development of various chronic diseases, especially in diabetes and its complications.

[0003] In recent years, a large number of studies have focused on relieving cell damage by regulating oxidative stress to promote diabetic wound healing. Active polypeptides, as a class of functionally diverse biological molecules, have become an important direction of antioxidant research due to their structural specificity and various biological activities. Specific active polypeptides with oxidative stress regulation function can target the regulation of oxidative stress signaling pathways, eliminate excessive active oxygen, inhibit inflammatory response, and promote cell proliferation and angiogenesis. Broadening the source of active peptides with oxidative stress regulation function and developing active peptides with specificity and high efficiency with oxidative stress regulation function are of great significance for the preparation of drugs or devices for reducing cell oxidative stress.

[0004] Chinese patent CN114031669A (published on February 11, 2022) discloses a Mytilus edulis antioxidant active peptide and its preparation and application, relating to the technical field of polypeptides. The Mytilus edulis antioxidant peptides Gln Glu Thr Tyr and Tyr Glu Leu His Asp can reduce the production of intracellular active oxygen ROS and form a protective effect. By activating the Keap1 / Nrf2 signaling pathway, it plays an antioxidant role and protects vascular endothelial cells, and can provide candidate drugs for the prevention and treatment of cardiovascular and cerebrovascular diseases such as hypertension, coronary heart disease, cerebral thrombosis, myocardial infarction, atherosclerosis, and heart failure.

[0005] Chinese patent CN114317654A (publication date: April 12, 2022) discloses a preparation method of marine organism antioxidant active peptide, relating to the technical field of antioxidant active peptide. The method comprises: S1) extracting collagen from broken squid tissue; S2) mixing the extracted collagen, trypsin, papain and flavor protease in water for enzymolysis to obtain an enzymolysis solution; S3) after the enzymolysis solution is treated by salting-out precipitation and dialysis, marine organism antioxidant active peptide is obtained. Compared with the prior art, the antioxidant active peptide is extracted from squid as raw material, and different enzymolysis methods are adopted, which can effectively and completely catalyze the protein, so as to obtain active polypeptide with good quality, reasonable amino acid composition, stable efficacy and reliable quality.

[0006] Chinese patent CN116904542A (publication date: October 20, 2023) discloses an antioxidant active peptide based on oyster shell and a preparation method thereof, relating to the technical field of comprehensive utilization of oyster shell. The present application solves the problem that there is a lack of research on antioxidant active peptide based on oyster shell, which leads to unsatisfactory comprehensive utilization of oyster shell and limits the development and utilization of oyster shell resources. The preparation method first finely grinds the oyster shell, which makes it easy to extract the protein on the oyster shell. Then the protein is dissolved to obtain excellent protein extraction effect. Then a variety of proteases are tested to screen out the protease suitable for the protein on the oyster shell. Thus, the protein can be fully enzymolyzed to form active peptide. Then the oyster shell enzymolysis solution is subjected to step-by-step ultrafiltration to concentrate the components with antioxidant capacity, improve the antioxidant performance of the antioxidant active peptide, and fully utilize the oyster shell resources, which is of great significance for the development and utilization of oyster shell resources.

[0007] After searching, it is found that although some patents on polypeptide preparation use shellfish as raw material, the research direction is mainly how to use shellfish to prepare antioxidant peptides, and the preparation process is complex. Pinctada fucata is a common shellfish used for pearl production, and its body surface mucus is a kind of biological resource that has not been fully utilized. At present, there are few studies on the active peptide with oxidation stress regulation function derived from Pinctada fucata body surface mucus, and its polypeptide amino acid sequence is not clear, and its preparation method and application need to be further developed. SUMMARY

[0008] To solve the above technical problems, the present application provides an active peptide with oxidation stress regulation function and its preparation and application.

[0009] An active peptide with oxidation stress regulation function, the amino acid sequence of the active peptide is

[0010] Ala-Val-Pro-Ile-Cys-Ala-Val-Pro.

[0011] The active peptide can be combined with a proteinase that regulates oxidative stress.

[0012] The proteinase is any one of SOD, CAT and keap1.

[0013] A preparation method of an active peptide with oxidative stress regulation effect, comprising the following steps:

[0014] S1: obtaining an extract, collecting pearl mussels, and filtering the mucus extract after opening the mussels and placing the meat together with the mucus in a gauze net;

[0015] S2: collecting the supernatant, centrifuging the mucus extract in S1, collecting the supernatant and detecting proteomics;

[0016] S3: selecting polypeptides, selecting polypeptides with oxidative stress regulation effect according to proteomics data;

[0017] S4: screening polypeptides, predicting and screening polypeptides from the supernatant by using molecular docking technology;

[0018] S5: synthesizing polypeptides, synthesizing polypeptides by using solid-phase synthesis method, and verifying the activity of the polypeptides in regulating oxidative stress.

[0019] Further, the pore size of the gauze net in step S1 is 0.5-1 cm

[0020] Further, the centrifugal speed of the centrifugal treatment in step S2 is 8000-10000 rpm, and the centrifugal time is 10-15 min.

[0021] Further, the step S3 comprises the following steps:

[0022] Step S301, based on proteomics data, using an online prediction tool PeptideRanker based on deep learning to score polypeptides, the polypeptide VGARVVGCCAHVASVLWYLGYFR scores 0.9889 points, close to 1 point, indicating that it has potential biological activity;

[0023] Step S302, further using Katedra Biochemii Zywnosci online tool to predict polypeptide activity, and finding that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro has the potential of oxidative stress regulation.

[0024] Further, the step S4 uses Discovery Studio to establish a three-dimensional structure model of the polypeptide, and uses AutoDock Vina to simulate the binding ability of the active peptide with antioxidant enzymes SOD, CAT and Keap1.

[0025] Further, the activity of regulating oxidative stress in step S5 is verified by taking free radical scavenging rate as an evaluation index; the free radicals include DPPH and ABTS.

[0026] Further, the activity of regulating oxidative stress of the polypeptide is verified by co-incubating the polypeptide with H2O2 treated cells and detecting cell viability in step S5.

[0027] The application of the active peptide with the function of regulating oxidative stress in the preparation of a medicine or device for reducing cell oxidative stress.

[0028] Compared with the prior art, the application has the following advantages and effects:

[0029] 1. The application provides an active peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro with the function of regulating oxidative stress, which can promote the proliferation of human fibroblasts and can be applied to promote skin wound healing and can be used to find and develop new wound healing drugs.

[0030] 2. The application provides a preparation method of the active peptide with the function of regulating oxidative stress, which is prepared from the preliminarily separated pearl mussel surface mucus, helps to quickly realize protein polypeptide identification, finds polypeptides with the potential of regulating oxidative stress through proteomics data, screens antioxidant polypeptides through molecular docking, and then is synthesized by solid phase synthesis, and helps to improve the development efficiency of marine source active peptides.

[0031] 3. The application provides an application of the active peptide with the function of regulating oxidative stress, the antioxidant active peptide has the advantages of good free radical scavenging and reducing the damage of H2O2 to cells, and the antioxidant active peptide has a good application prospect in a medicine or device for reducing cell oxidative stress.

[0032] The above description is only a summary of the technical scheme of the application, in order to more clearly understand the technical means of the application, so as to implement the content of the description, and in order to make the above and other purposes, characteristics and advantages of the application more obvious and easy to understand, the following will be described in detail with the preferred embodiments of the application and the accompanying drawings.

[0033] According to the detailed description of the specific embodiments of the application in combination with the drawings below, those skilled in the art will more clearly understand the above and other purposes, advantages and characteristics of the application. BRIEF DESCRIPTION OF DRAWINGS

[0034] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the accompanying drawings needed to be used in the embodiments or prior art description will be briefly introduced. Obviously, the accompanying drawings in the following description are some embodiments of the present application, and other drawings can be obtained by those skilled in the art without any creative effort. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn according to the actual proportions.

[0035] wherein:

[0036] Figure 1 LC-MS spectrum of the polypeptide prepared by the present application

[0037] Figure 2 2D simulation structure diagram of the polypeptide prepared by the present application

[0038] Figure 3 Schematic diagram of the polypeptide prepared by the present application and the simulation binding of antioxidant enzyme

[0039] Figure 3 (a) Schematic diagram of the polypeptide prepared by the present application and the simulation binding of antioxidant enzyme SOD

[0040] Figure 3 (b) Schematic diagram of the polypeptide prepared by the present application and the simulation binding of antioxidant enzyme CAT

[0041] Figure 3 (c) Schematic diagram of the polypeptide prepared by the present application and the simulation binding of antioxidant enzyme Keap1

[0042] Figure 4 Comparison chart of the free radical scavenging results of the polypeptide prepared by the present application

[0043] Figure 4 (a) Statistical chart of the DPPH free radical scavenging results of the polypeptide prepared by the present application

[0044] Figure 4 (b) Statistical chart of the ABTS free radical scavenging results of the polypeptide prepared by the present application

[0045] Figure 5 Comparison chart of the results of the polypeptide prepared by the present application promoting fibroblast migration

[0046] Figure 5 (a) Result chart of the control group of fibroblast migration before migration

[0047] Figure 5 (b) Result chart of the polypeptide group of fibroblast migration before migration

[0048] Figure 5(c) result chart of fibroblast migration after treatment of control group

[0049] Figure 5 (d) result chart of fibroblast migration after treatment of polypeptide group

[0050] Figure 6 result chart of cell protection from H2O2 damage by polypeptide prepared in the present application

[0051] Figure 6 (a) result chart of cell protection from H2O2 damage by control group

[0052] Figure 6 (b) result chart of cell protection from H2O2 damage by control group

[0053] Figure 6 (c) result chart of cell protection from H2O2 damage by polypeptide group

[0054] Figure 7 result chart of wound healing of diabetic rats promoted by polypeptide prepared in the present application

[0055] Figure 7 (a) result chart of wound healing of diabetic rats of control group

[0056] Figure 7 (b) result chart of wound healing of diabetic rats of polypeptide group DETAILED DESCRIPTION

[0057] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some but not all of the embodiments of the present application. In the following description, specific details such as specific configurations and components are provided only to help a comprehensive understanding of the embodiments of the present application. Therefore, those skilled in the art should understand that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present application. In addition, descriptions of known functions and configurations are omitted in the embodiments for clarity and conciseness.

[0058] It should be understood that the "one embodiment" or "the embodiment" mentioned throughout the specification means that the specific features, structures or characteristics related to the embodiment are included in at least one embodiment of the present application. Therefore, "one embodiment" or "the embodiment" appearing throughout the specification does not necessarily mean the same embodiment. In addition, these specific features, structures or characteristics can be combined in one or more embodiments in any suitable manner.

[0059] In addition, reference numerals and / or letters in the various examples maybe repeated usethroughout the specification and drawing figures in order to more easily illustrate similar or related elements, structures, modules, or functions. Such repetition is for illustrative simplicity and clarity and does not itself dictate a relationship between the described or illustrated corresponding items or methods.

[0060] The term "and / or", merely an associative relationship to describe the associated object, can exist three kinds of relationship, for example, A and / or B, can represent: the existence of A alone, B alone, A and B exist at the same time three cases, the term "and" in this paper is to describe another kind of relationship between the associated object, can exist two kinds of relationship, for example, A and B, can represent: the existence of A alone, A and B exist at the same time two cases, in addition, the character " / " in this paper, generally speaking, the associated object before and after is a kind of "or" relationship.

[0061] The term "at least one" in this paper, merely an associative relationship to describe the associated object, can exist three kinds of relationship, for example, A and B at least one, can represent: the existence of A alone, A and B exist at the same time, B alone three cases.

[0062] It should also be noted that the relationship terms such as first and second in this paper are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between the entities or operations. Moreover, the terms "include", "contain" or any other variants thereof are intended to cover non-exclusive inclusion.

[0063] Embodiment 1

[0064] The embodiment is a preparation method of an active peptide with oxidative stress regulation effect.

[0065] S1: obtaining the extract, collecting the pearl mussel, and filtering the mucus extract by placing the mussel meat together with the mucus in a gauze after opening the shell;

[0066] S2: collecting the supernatant, centrifuging the mucus extract in S1, collecting the supernatant and detecting the proteomics;

[0067] S3: selecting the polypeptide, selecting the polypeptide with oxidative stress regulation effect according to the proteomics data;

[0068] S4: screening the polypeptide, predicting and screening the polypeptide from the supernatant by using the molecular docking technology;

[0069] S5: synthesizing the polypeptide, synthesizing the polypeptide by using the solid-phase synthesis method, and verifying the activity of the polypeptide in regulating oxidative stress.

[0070] Further, the pore size of the gauze in step S1 is 0.5-1 cm

[0071] Further, the centrifugal speed of the centrifugal treatment in step S2 is 8000-10000 rpm, and the centrifugal time is 10-15 min.

[0072] Further, the step S3 comprises the following steps:

[0073] Step S301, based on proteomics data, using an online prediction tool PeptideRanker based on deep learning to score polypeptides, the polypeptide VGARVVGCCAHVASVLWYLGYFR scores 0.9889 points, close to 1 point, indicating that it has potential biological activity;

[0074] Step S302, further using Katedra Biochemii Zywnosci online tool to predict polypeptide activity, and finding that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro has the potential of oxidative stress regulation.

[0075] Further, the step S4 uses Discovery Studio to establish a three-dimensional structure model of the polypeptide, and uses AutoDock Vina to simulate the binding ability of the active peptide to antioxidant enzymes SOD, CAT and Keap1.

[0076] The amino acid sequence of the polypeptide is Ala-Val-Pro-Ile-Cys-Ala-Val-Pro.

[0077] Technical effect of the embodiment: The active peptide with oxidative stress regulation prepared by a simple method helps to improve the development efficiency of marine source active peptide.

[0078] Embodiment 2

[0079] Based on embodiment 1, this embodiment is a method for identifying a synthetic polypeptide.

[0080] As Figure 1 shown, Figure 1 LC-MS spectrum of the polypeptide prepared by the present application, the results show that the synthesized polypeptide has obvious characteristic peaks, indicating that the synthesized polypeptide has high purity.

[0081] Technical effect of the embodiment: The active peptide with oxidative stress regulation prepared by the present application has high purity.

[0082] Embodiment 3

[0083] Based on embodiment 2, this embodiment is to establish a molecular model of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro.

[0084] The polypeptide molecule with high predicted activity was modeled using Discovery Studio software. Please refer to Figure 2 , Figure 2 The 2D analog structure of the polypeptide prepared by the present application is shown in the figure. The predicted structure of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro presents a cyclic structure with two turns, which may help it to signal in cells. The repeating pattern of A and V residues is related to its specific biological activity.

[0085] Technical effect of the embodiment: The polypeptide molecule with high predicted activity is modeled by Discovery Studio software. The structure prediction structure can determine that this structure is conducive to signal transmission in cells and has specific biological activity.

[0086] Example 4

[0087] Based on examples 1-3, this embodiment verifies the oxidative stress regulation ability of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro.

[0088] The open source molecular docking tool AutoDockVina software is used to perform molecular docking simulation of the interaction between the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro and the antioxidant proteins SOD, CAT and Keap1.

[0089] Before docking simulation, the polypeptide and target protein structure are pretreated, including adding hydrogen atoms, calculating charges and assigning flexibility. Then, the docking algorithm is selected for docking simulation, and by evaluating the binding energy and analyzing the interaction mode, the potential biological activity and mechanism of action of the docked polypeptide are revealed.

[0090] As shown in Figure 3 , Figure 3 The polypeptide prepared by the present application and the antioxidant enzyme simulation binding schematic diagram. Among them, Figure 3 (a) is a schematic diagram of the polypeptide prepared by the present application and the antioxidant enzyme SOD simulation binding, the results show that the cyclic structure and two turns of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro may enable it to flexibly adapt to the surface of SOD. The repeating pattern of A and V residues may be related to its specific biological activity and may regulate its antioxidant activity by interacting with SOD. This structure may help to enhance the stability and function of SOD, thereby improving the resistance of cells to oxidative stress.

[0091] Figure 3(b) is a schematic diagram of the peptide prepared in this invention and the antioxidant enzyme CAT. The results show that the peptide and the CAT enzyme have a binding energy of less than -5 kcal / mol, indicating that it has the potential to regulate oxidative stress.

[0092] Figure 3 (c) is a schematic diagram of the simulated binding of the polypeptide prepared in this invention with the antioxidant enzyme Keap1. The results show that the binding energy of the polypeptide and the Keap1 enzyme is less than -5 kcal / mol, indicating that it has the potential to regulate oxidative stress.

[0093] The technical effect of this embodiment is that by analyzing the simulation results through molecular docking simulation, it can be found that peptides have the ability to regulate oxidative stress.

[0094] Example 5

[0095] Based on Example 4, this example is an evaluation of the ability of the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro to scavenge DPPH and ABTS free radicals.

[0096] like Figure 4 As shown, Figure 4 This is a comparison chart of the free radical scavenging results of the peptides prepared in this invention (P<0.01). Among them, Figure 4 (a) is a statistical graph showing the DPPH radical scavenging results of the peptides prepared in this invention. The results show that the DPPH radical scavenging rate increases with the increase of peptide solution concentration. When the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 0.5 mg / mL, the DPPH radical scavenging rate reaches 45%. Moreover, the scavenging rate increases with increasing concentration, showing a dose-dependent relationship. When the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 1.5 mg / mL, the DPPH radical scavenging rate is 55%.

[0097] Figure 4 (b) is a statistical graph showing the ABTS radical scavenging results of the peptides prepared in this invention. The results show that the ABTS radical scavenging rate increases with the increase of peptide solution concentration. When the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 0.5 mg / mL, the ABTS radical scavenging rate is 40%; when the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 1.5 mg / mL, the ABTS radical scavenging rate is 63%; showing a dose-dependent relationship.

[0098] The technical effect of this embodiment: By comparing the DPPH and ABTS free radical scavenging rates of the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro at different concentrations, it can be found that the peptide has a significant ability to scavenge DPPH and ABTS free radicals (P<0.01), and this ability is dose-dependent.

[0099] Example 6

[0100] Based on Example 5, this example verifies the promoting effect of the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro on fibroblast migration.

[0101] like Figure 5 As shown, Figure 5 This is a comparison chart showing the results of the polypeptides prepared in this invention promoting fibroblast migration. Among them, Figure 5 (a) shows the results of fibroblasts in the control group (without peptides) before migration. Figure 5 (b) is a diagram showing the results of fibroblasts before migration of polypeptide-based fibroblasts. As can be seen from the diagram, the distance between fibroblasts in both the control group and the polypeptide group before migration was 250 μm. Figure 5 (c) is a diagram showing the migration results of fibroblasts in the control group. As can be seen from the diagram, after 12 hours, the distance between fibroblasts in the control group was about 200 μm, indicating that the migration of fibroblasts was not obvious. Figure 5 (d) shows the results of fibroblast migration after peptide-based fibroblasts migrate. As can be seen from the figure, after 12 hours, the distance between peptide-based fibroblasts was about 30 μm, indicating that the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro significantly promotes fibroblast migration.

[0102] The technical effect of this embodiment is that it verifies that the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro promotes fibroblast migration.

[0103] Example 7

[0104] Based on Example 6, this example verifies the effect of the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro in protecting cells from H2O2 damage.

[0105] like Figure 6 As shown, Figure 6 This is a comparison chart showing the protection of cells from H2O2 damage by the polypeptide prepared in this invention. Figure 6 (a) shows the results of H2O2 damage detection for the control group (without added peptides). The results indicate that in the absence of H2O2, the cells exhibited lower fluorescence intensity and lower ROS production.

[0106] Figure 6 (b) is a result detection diagram of protecting cells from H2O2 damage of the control group, and the result shows that after H2O2 treatment, fluorescence intensity is provided, and the increase of ROS production of cells is indicated;

[0107] Figure 6 (c) is a result detection diagram of protecting cells from H2O2 damage of the polypeptide group, and the result shows that after polypeptide incubation, the fluorescence intensity of cells is reduced, and it is indicated that the polypeptide can reduce the ROS production of cells.

[0108] Technical effect of the embodiment: it is verified that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro can protect cells from H2O2 damage.

[0109] Example 8

[0110] Based on example 7, the embodiment is the verification of the effect of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro on promoting wound healing of diabetic rats.

[0111] As shown in Figure 7 , the result comparison diagram of the polypeptide prepared in the application on promoting wound healing of diabetic rats is shown; wherein, Figure 7 (a) is a result diagram of wound healing of the control group (without adding the polypeptide) of diabetic rats, Figure 7 (b) is a result diagram of wound healing of the polypeptide group of diabetic rats; Figure 7 The result shows that the wound healing rate of the diabetic rats is slow, and the wound healing rate is obviously accelerated after polypeptide treatment; and after 18 days, the wound of the control group of diabetic rats still exists obviously, and the wound of the polypeptide group of diabetic rats is basically healed after polypeptide treatment, and obvious new skin can be observed.

[0112] Technical effect of the embodiment: it is verified that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro can effectively promote the healing of diabetic wounds.

[0113] In summary, the application provides an active peptide with oxidative stress regulation function and a preparation method and application thereof, the amino acid sequence of the active peptide is Ala-Val-Pro-Ile-Cys-Ala-Val-Pro, has high activity, can better scavenge free radicals, and protect H2O2 damaged cell activity; can promote skin wound healing; the antioxidant active peptide has good prospects in reducing cell oxidative stress drugs or instruments, and the application has innovation.

[0114]

[0115] ​The above detailed description of the specific embodiments of the present application has been given to understand the purpose, technical solutions and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. An active peptide with oxidative stress regulating activity, characterized in that, The amino acid sequence of the active peptide is Ala-Val-Pro-Ile-Cys-Ala-Val-Pro.

2. The bioactive peptide with oxidative stress regulating effect according to claim 1, characterized in that, The active peptide can bind to antioxidant proteases.

3. The bioactive peptide with oxidative stress regulating effect according to claim 2, characterized in that, The antioxidant protease is any one of SOD, CAT, and keap1.

4. A method for preparing an active peptide with oxidative stress regulating effect according to any one of claims 1-3, characterized in that, Includes the following steps: S1: Obtain the extract by collecting pearl oysters, opening the oysters and filtering the oyster meat along with the mucus in a mesh to obtain the mucus extract. S2: Collect the supernatant, centrifuge the mucus extract described in S1, collect the supernatant and detect protein peptides and proteomics. S3: Select peptides, choosing peptides with oxidative stress regulation effects based on proteomics data; S4: Screening peptides, using molecular docking technology to predict and screen peptides from the supernatant; S5: Synthesize polypeptides using solid-phase synthesis.

5. The method for preparing an active peptide with oxidative stress regulating effect according to claim 4, characterized in that, The centrifugation speed in step S2 is 8000-10000 rpm, and the centrifugation time is 10-15 min.

6. A method for preparing an active peptide with oxidative stress regulating effect according to claim 4 or 5, characterized in that, Step S3 includes the following steps: Step S301: Based on proteomics data, use the deep learning-based online prediction tool PeptideRanker to score peptides. A score close to 1 indicates potential biological activity. Step S302: Further utilize the Kadara Biochemii Zywnosci online tool to predict peptide activity and discover the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro with oxidative stress regulation potential.

7. The use of an active peptide with oxidative stress regulation as described in any one of claims 1-3 in the preparation of a drug for promoting the healing of diabetic wounds.

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