Active peptide with oxidative stress regulation effect as well as preparation and application thereof

Abstract

The invention discloses an active peptide with an oxidative stress regulation effect as well as preparation and application of the active peptide. 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 the following steps: collecting pearl shells, obtaining an extract, collecting supernate, selecting, screening and synthesizing polypeptide. The active peptide with the oxidative stress regulation function has high activity, free radicals can be well removed, and damage of H2O2 to skin fibroblasts is relieved; the skin wound healing can be promoted. The active peptide with the oxidative stress regulation effect has a relatively good application prospect in drugs or instruments for relieving cell oxidative stress.

Description

Technical Field

[0001] The invention belongs to the technical field of biological polypeptides, and specifically relates to an active peptide with oxidative stress regulating effect and a preparation and application thereof. Background Art

[0002] Oxidative stress refers to a pathological process in which the body's oxidation and antioxidant states are unbalanced due to excessive production of reactive oxygen species (ROS) or insufficient function of the antioxidant system. Oxidative stress not only damages cell membranes, proteins and DNA molecules, but also inhibits tissue repair and regeneration capabilities. This process is closely related to the occurrence and development of a variety of chronic diseases, especially playing a vital role in diabetes and its complications.

[0003] In recent years, a large number of studies have focused on alleviating cell damage by regulating oxidative stress to promote diabetic wound healing. As a class of functionally diverse biological molecules, active peptides have become an important direction of antioxidant research due to their structural specificity and multiple biological activities. Specific active peptides with oxidative stress regulating effects can remove excess reactive oxygen species, inhibit inflammatory responses, and promote cell proliferation and angiogenesis by targeting and regulating oxidative stress signaling pathways. Broadening the sources of active peptides with oxidative stress regulating effects and developing active peptides with specific and efficient oxidative stress regulating effects are of great significance for the preparation of drugs or devices that reduce cellular oxidative stress.

[0004] Chinese patent CN114031669A (publication date: February 11, 2022) discloses a thick-shelled mussel antioxidant active peptide and its preparation and application, which relates to the field of polypeptide technology. A thick-shelled mussel antioxidant peptide Gln Glu ThrTyr and Tyr GluLeuHis Asp can reduce the production of intracellular reactive oxygen species ROS and form a protective effect. It activates the Keap1 / Nrf2 signaling pathway, exerts an antioxidant effect 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, heart failure, etc.

[0005] Chinese patent CN114317654A (publication date: April 12, 2022) discloses a method for preparing marine antioxidant active peptides, which relates to the technical field of antioxidant active peptides. The method comprises: S1) extracting collagen from crushed squid tissue; S2) mixing the extracted collagen, trypsin, papain and flavor protease in water for enzymolysis to obtain an enzymolysis solution; S3) subjecting the enzymolysis solution to salting out precipitation and dialysis treatment to obtain marine antioxidant active peptides. Compared with the prior art, the extraction of antioxidant active peptides using squid as raw material and the adoption of different enzymolysis methods can effectively and thoroughly catalyze the protein, thereby obtaining active polypeptides 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, which relates to the technical field of comprehensive utilization of oyster shells. This patent is used to solve the problem that the lack of research on antioxidant active peptides based on oyster shells has resulted in the comprehensive utilization of oyster shells being still not ideal, limiting the development and utilization of oyster shell resources; the preparation method first fully refines the oyster shells, making it easy to extract the protein on the oyster shells, and then dissolves the protein to achieve excellent protein extraction effects, and then uses a variety of proteases for testing to screen out proteases suitable for proteins on oyster shells, so that the proteins can be fully enzymolyzed to form active peptides, and then the oyster shell enzymatic hydrolysate is subjected to step-by-step ultrafiltration, so that the components with antioxidant capacity are concentrated, the antioxidant properties of the antioxidant active peptides are improved, and the oyster shell resources are fully utilized, which is of great significance to the development and utilization of oyster shell resources.

[0007] After searching, it was found that although some patents on the preparation of peptides are based on shellfish as raw materials, their research direction is mainly on how to use shellfish to prepare antioxidant peptides, and the preparation process is complicated. The Hepu pearl oyster is a common shellfish used in pearl production, and its body mucus is an underutilized biological resource. At present, there are few studies on active peptides with oxidative stress regulating effects derived from the surface mucus of the Hepu pearl oyster. The amino acid sequence of its peptide is unclear, and the preparation method and application need to be further developed. Summary of the invention

[0008] In order to solve the above technical problems, the present invention provides an active peptide having oxidative stress regulating effect and its preparation and application.

[0009] An active peptide with oxidative stress regulating effect, the amino acid sequence of the active peptide is

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

[0011] The active peptide is capable of binding to a protease that regulates oxidative stress;

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

[0013] A method for preparing an active peptide having an oxidative stress regulating effect comprises the following steps:

[0014] S1: obtaining an extract, collecting pearl oysters, opening the oysters, placing the oyster meat and mucus in a gauze and filtering to obtain a mucus extract;

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

[0016] S3: Select peptides, and select peptides with oxidative stress regulation effects based on proteomics data;

[0017] S4: Screening peptides, using molecular docking technology to predict and screen peptides from the supernatant;

[0018] S5: Synthesize peptides using solid phase synthesis and verify the activity of the peptides in regulating oxidative stress.

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

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

[0021] Furthermore, the step S3 includes the following steps:

[0022] Step S301: Based on the proteomics data, the peptides were scored using the deep learning-based online prediction tool PeptideRanker. The peptide VGARVVGCCAHVASVLWYLGYFR scored 0.9889 points, which is close to 1 point, indicating potential biological activity.

[0023] Step S302: further predicting the activity of the peptides using the Katedra Biochemii Zywnosci online tool, and discovering the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro with the potential to regulate oxidative stress.

[0024] Furthermore, in step S4, the three-dimensional structure model of the polypeptide is established using Discovery Studio, and the binding ability of the active peptide with the antioxidant enzymes SOD, CAT and Keap1 is simulated using AutoDock Vina.

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

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

[0027] The invention discloses an application of an active peptide with oxidative stress regulating effect in preparing a drug or device for alleviating cellular oxidative stress.

[0028] Compared with the prior art, the advantages and effects of the present invention are as follows:

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

[0030] 2. The present invention provides a method for preparing active peptides with oxidative stress regulating effects. The preliminary separation of pearl oyster surface mucus is helpful for rapid protein and peptide identification; peptides with oxidative stress regulating potential are discovered through proteomics data; antioxidant peptides are screened through molecular docking and then solid-phase synthesis, which helps to improve the efficiency of developing marine-derived active peptides.

[0031] 3. The present invention provides an application of an active peptide with oxidative stress regulating effect. The antioxidant active peptide has good free radical scavenging ability and reduces the damage of H2O2 to cells. The antioxidant active peptide has good application prospects in drugs or devices for reducing cell oxidative stress.

[0032] The above description is only an overview of the technical solution of the present application. In order to more clearly understand the technical means of the present application so that it can be implemented in accordance with the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the following is a detailed description of the preferred embodiments of the present application in conjunction with the accompanying drawings as follows.

[0033] Based on the detailed description of the specific embodiments of the present application in combination with the accompanying drawings below, those skilled in the art will become more aware of the above and other objects, advantages and features of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can also be obtained based on these drawings without creative work. In all drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale.

[0035] in:

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

[0037] Figure 2 Schematic diagram of the 2D simulated structure of the polypeptide prepared by the present invention

[0038] Figure 3 Schematic diagram of the simulated combination of the polypeptide prepared by the present invention and the antioxidant enzyme

[0039] Figure 3 (a) is a schematic diagram of the simulated binding of the polypeptide prepared by the present invention with the antioxidant enzyme SOD

[0040] Figure 3 (b) is a schematic diagram of the simulated binding of the polypeptide prepared by the present invention with the antioxidant enzyme CAT

[0041] Figure 3 (c) is a schematic diagram of the simulated binding of the polypeptide prepared by the present invention with the antioxidant enzyme Keap1

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

[0043] Figure 4 (a) is a statistical diagram of the results of the polypeptide prepared by the present invention scavenging DPPH free radicals

[0044] Figure 4 (b) is a statistical graph showing the results of the polypeptide prepared by the present invention scavenging ABTS free radicals

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

[0046] Figure 5 (a) The results of the control group fibroblasts before migration

[0047] Figure 5 (b) is the result of fibroblast migration before the peptide composition

[0048] Figure 5(c) Results of fibroblast migration in the control group

[0049] Figure 5 (d) The result of fibroblast migration after peptide composition

[0050] Figure 6 Comparison of detection of the polypeptide prepared by the present invention protecting cells from H2O2 damage

[0051] Figure 6 (a) The control group protected cells from H2O2 damage.

[0052] Figure 6 (b) The detection results of the control group's protected cells from H2O2 damage

[0053] Figure 6 (c) Detection results of peptide group protecting cells from H2O2 damage

[0054] Figure 7 Comparison of the results of the polypeptide prepared by the present invention promoting wound healing in diabetic rats

[0055] Figure 7 (a) The wound healing results of diabetic rats in the control group

[0056] Figure 7 (b) The wound healing results of diabetic rats in the peptide group DETAILED DESCRIPTION

[0057] To make the purpose, technical scheme and advantages of the embodiment of the present application clearer, the technical scheme in the embodiment of the present application will be clearly and completely described below in conjunction with the accompanying drawings in the embodiment of the present application. Obviously, the described embodiment is a part of the embodiment of the present application, rather than all of the embodiments. In the following description, specific details such as specific configuration and components are provided only to help fully understand the embodiments of the present application. Therefore, it should be clear to those skilled in the art 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, for clarity and brevity, the description of known functions and structures is omitted in the embodiment.

[0058] It should be understood that the references to "one embodiment" or "this embodiment" throughout the specification mean that the specific features, structures, or characteristics associated with the embodiment are included in at least one embodiment of the present application. Therefore, the references to "one embodiment" or "this embodiment" appearing throughout the specification do not necessarily refer to the same embodiment. In addition, these specific features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0059] In addition, the present application may repeat reference numerals and / or letters in different examples. This repetition is for the purpose of simplicity and clarity, and does not in itself indicate the relationship between the various embodiments and / or settings discussed.

[0060] The term "and / or" in this article is only a description of the association relationship of associated objects, indicating that there can be three relationships. For example, A and / or B can mean: A exists alone, B exists alone, and A and B exist at the same time. The term " / and" in this article describes another type of association object relationship, indicating that there can be two relationships. For example, A / and B can mean: A exists alone, and A and B exist alone. In addition, the character " / " in this article generally indicates that the previous and next associated objects are in an "or" relationship.

[0061] The term "at least one" in this article is merely a description of the association relationship of associated objects, indicating that there may be three relationships. For example, at least one of A and B can mean: A exists alone, A and B exist at the same time, and B exists alone.

[0062] It should also be noted that, in this article, relational terms such as first and second, etc. 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 these entities or operations. Moreover, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusions.

[0063] Example 1

[0064] This example is a method for preparing an active peptide having an oxidative stress regulating effect.

[0065] S1: obtaining an extract, collecting pearl oysters, opening the oysters, placing the oyster meat and mucus in a gauze and filtering to obtain a mucus extract;

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

[0067] S3: Select peptides, and select peptides with oxidative stress regulation effects based on proteomics data;

[0068] S4: Screening peptides, using molecular docking technology to predict and screen peptides from the supernatant;

[0069] S5: Synthesize peptides using solid phase synthesis and verify the activity of the peptides in regulating oxidative stress.

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

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

[0072] Furthermore, the step S3 includes the following steps:

[0073] Step S301: Based on the proteomics data, the peptides were scored using the deep learning-based online prediction tool PeptideRanker. The peptide VGARVVGCCAHVASVLWYLGYFR scored 0.9889 points, which is close to 1 point, indicating potential biological activity.

[0074] Step S302: further predicting the activity of the peptides using the Katedra Biochemii Zywnosci online tool, and discovering the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro with the potential to regulate oxidative stress.

[0075] Furthermore, in step S4, the three-dimensional structure model of the polypeptide is established using Discovery Studio, and the binding ability of the active peptide with the antioxidant enzymes SOD, CAT and Keap1 is simulated using AutoDock Vina.

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

[0077] The technical effect of this embodiment is that active peptides with oxidative stress regulating effects are prepared by a simple method, which helps to improve the efficiency of developing marine-derived active peptides.

[0078] Example 2

[0079] Based on Example 1, this example is a method for identifying synthetic polypeptides.

[0080] like Figure 1 As shown, Figure 1 The LC-MS spectrum of the polypeptide prepared by the present invention shows that the synthesized polypeptide has obvious characteristic peaks, indicating that the synthesized polypeptide has a high purity.

[0081] The technical effect of this embodiment: the active peptide with oxidative stress regulating effect prepared by the present invention has a higher purity.

[0082] Example 3

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

[0084] The peptide molecules predicted to be highly active were modeled using Discovery Studio software. Figure 2 , Figure 2 The 2D simulated structure of the polypeptide prepared by the present invention is shown in Figure 1. The predicted structure of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro shows a ring structure with two turns, which may help it to conduct signal transduction in cells. The repetitive pattern of A and V residues is related to its specific biological activity.

[0085] Technical effect of this embodiment: The present invention models a polypeptide molecule predicted to be highly active by using Discovery Studio software, and through structural prediction, it can be determined that this structure is conducive to signal transduction in cells and has specific biological activity.

[0086] Example 4

[0087] Based on Examples 1-3, this example is a verification of the ability of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro to regulate oxidative stress.

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

[0089] Before docking simulation, the peptide and target protein structures are preprocessed, including adding hydrogen atoms, calculating charges and assigning flexibility. Then, a docking algorithm is selected for docking simulation to reveal the potential biological activity and mechanism of action of the docked peptide by evaluating binding energy and analyzing interaction patterns.

[0090] like Figure 3 As shown, Figure 3 The schematic diagram of the simulated binding of the polypeptide prepared by the present invention and the antioxidant enzyme is shown in FIG. Figure 3 (a) is a schematic diagram of the simulated binding of the polypeptide prepared by the present invention with the antioxidant enzyme SOD. 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 repetitive pattern of A and V residues may be related to its specific biological activity and may regulate its antioxidant activity through interaction 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 simulated binding between the polypeptide prepared by the present invention and the antioxidant enzyme CAT. The results show that the binding energy between the polypeptide and the CAT enzyme is 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 by the present invention with the antioxidant enzyme Keap1. The results show that the binding energy between the polypeptide and the Keap1 enzyme is less than -5 kcal / mol, indicating that it has the potential to regulate oxidative stress.

[0093] Technical effect of this embodiment: Through molecular docking simulation and analysis of the simulation results, it can be found that the polypeptide has the ability to regulate oxidative stress.

[0094] Example 5

[0095] Based on Example 4, this example is a test of the ability of the polypeptide 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 polypeptides prepared by the present invention (P<0.01). Figure 4 (a) is a statistical graph of the results of scavenging DPPH free radicals by the polypeptide prepared in the present invention. The results show that the DPPH free radical scavenging rate increases with the increase of the concentration of the polypeptide solution. When the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 0.5 mg / mL, the DPPH free radical scavenging rate reaches 45%; and the scavenging rate increases with the increase of concentration, showing a dose-dependent manner; when the concentration of Ala-Val-Pro-Ile-Cys-Ala-Val-Pro is 1.5 mg / mL, the DPPH free radical scavenging rate is 55%.

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

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

[0099] Example 6

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

[0101] like Figure 5 As shown, Figure 5 This is a comparison chart of the results of the polypeptide prepared by the present invention promoting fibroblast migration. Figure 5 (a) is the result of the control group (no polypeptide added) before fibroblast migration. Figure 5 (b) is the result of the fibroblasts in the polypeptide group before migration. It can be seen from the figure that the distance between the fibroblasts in the control group and the polypeptide group before migration is 250 μm. Figure 5 (c) is the result of the migration of fibroblasts in the control group. It can be seen from the figure that after 12 hours, the distance between fibroblasts in the control group is about 200 μm, indicating that the migration of fibroblasts is not obvious; Figure 5 (d) is the result of fibroblast migration after the polypeptide composition. It can be seen from the figure that after 12 hours, the distance between the polypeptide composition fibroblasts is about 30μm, indicating that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro significantly promotes the migration of fibroblasts.

[0102] The technical effect of this embodiment is as follows: it is verified that the polypeptide 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 polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro in protecting cells from damage by H2O2.

[0105] like Figure 6 As shown, Figure 6 This is a comparison chart of the detection of the polypeptide prepared by the present invention protecting cells from H2O2 damage. Figure 6 (a) is a test result of the control group (no polypeptide added) protecting cells from H2O2 damage. The results show that in the absence of H2O2, the cells exhibit lower fluorescence intensity and lower ROS production.

[0106] Figure 6 (b) is the detection result of the control group's protected cells damaged by H2O2. The results show that after H2O2 treatment, the fluorescence intensity is increased and the cellular ROS production increases;

[0107] Figure 6 (c) is a test result of the peptide group protecting cells from H2O2 damage. The results show that after incubation with the peptide, the fluorescence intensity of the cells decreased, indicating that the peptide can reduce the production of ROS in cells.

[0108] The technical effect of this embodiment is as follows: it is verified that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro can protect cells from damage by H2O2.

[0109] Example 8

[0110] Based on Example 7, this example verifies the effect of the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro in promoting wound healing in diabetic rats.

[0111] like Figure 7 As shown, Figure 7 This is a comparison chart of the results of the polypeptide prepared by the present invention promoting wound healing in diabetic rats; wherein, Figure 7 (a) is the wound healing results of diabetic rats in the control group (no peptide added). Figure 7 (b) is the wound healing result of diabetic rats in the peptide group;

[0112] The results showed that the wound healing rate of diabetic rats was slow, while the wound healing rate was significantly accelerated after peptide treatment; and 18 days later, the wounds of diabetic rats in the control group still had obvious wounds, while the wounds of diabetic rats in the peptide group were basically healed after peptide treatment, and obvious new skin could be observed.

[0113] The technical effect of this embodiment is to verify that the polypeptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro can effectively promote the healing of diabetic wounds.

[0114] In summary, the present invention provides an active peptide with oxidative stress regulating effect, a preparation method and application thereof. The amino acid sequence of the active peptide is Ala-Val-Pro-Ile-Cys-Ala-Val-Pro, which has high activity, can better scavenge free radicals, and protect H2O2 from damaging cell activity; it can promote skin wound healing; the application of the antioxidant active peptide in drugs or devices for reducing cell oxidative stress has good prospects, and the invention is innovative.

[0115] The specific implementation methods described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

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

2. The active peptide having oxidative stress regulating effect according to claim 1, characterized in that: The active peptide is capable of binding to antioxidant proteases.

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

4. The method for preparing an active peptide having oxidative stress regulating effect according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1: obtaining an extract, collecting pearl oysters, opening the oysters, placing the oyster meat and mucus in a gauze and filtering to obtain a mucus extract; S2: collecting the supernatant, centrifuging the mucus extract described in S1, collecting the supernatant and detecting protein and peptide proteomics; S3: Select peptides, and select 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 peptides using solid phase synthesis.

5. The method for preparing an active peptide having oxidative stress regulating effect according to claim 4, characterized in that: The centrifugal treatment in step S2 has a centrifugal speed of 8000-10000 rpm and a centrifugal time of 10-15 min.

6. The method for preparing an active peptide having oxidative stress regulating effect according to claim 4 or 5, characterized in that: The step S3 comprises the following steps: Step S301: Based on the proteomics data, the peptides are scored using the deep learning-based online prediction tool PeptideRanker, and a score close to 1 indicates potential biological activity; Step S302: further predicting the activity of the peptides using the Katedra Biochemii Zywnosci online tool, and discovering the peptide Ala-Val-Pro-Ile-Cys-Ala-Val-Pro with the potential to regulate oxidative stress.

7. Use of an active peptide having oxidative stress regulating effect according to any one of claims 1 to 3 in the preparation of drugs or devices for reducing cellular oxidative stress.

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