An acetylcholinesterase inhibitory peptide from trachurus japonicus and a preparation method and application thereof

Abstract

This invention discloses an acetylcholinesterase inhibitory peptide derived from the oval pomfret, its preparation method, and its application, belonging to the field of bioactive peptide technology. The acetylcholinesterase inhibitory peptide is any polypeptide with an amino acid sequence as shown in SEQ ID NO.1-SEQ ID NO.4. This invention uses the oval pomfret as raw material and obtains an acetylcholinesterase inhibitory peptide with antioxidant activity and Ca2+ ion affinity chromatography through enzymatic hydrolysis and immobilized calcium ion affinity chromatography. 2+ The active heptapeptides DLTDYLM and QGPIGPR, hexapeptide APDPFR, and pentapeptide WGDAG were combined, and their chelation mechanism with acetylcholinesterase and Keap1 was clarified. The polypeptides derived from the oval pomfret provided by this invention have promising applications in the preparation of acetylcholinesterase inhibitors, antioxidants, and calcium supplements.

Description

Technical Field

[0001] This invention relates to the field of bioactive peptide technology, and in particular to an acetylcholinesterase inhibitory peptide derived from the oval pomfret, its preparation method, and its application. Background Technology

[0002] Acetylcholinesterase (AChE) is a key enzyme in the neurocholinergic system. It hydrolyzes the neurotransmitter acetylcholine (ACh), terminating nerve signal transmission and preventing excessive nerve excitation. However, in patients with neurodegenerative diseases such as Alzheimer's disease (AD), ACh deficiency leads to impaired signal transmission, resulting in memory and cognitive impairment. Therefore, using AChE inhibitors to reduce AChE activity can increase the ACh content in the brain and alleviate cognitive impairment. Acetylcholinesterase inhibitors have become the main drugs for treating Alzheimer's disease, with donepezil, gramamine, and neostigmine being used clinically. However, because small-molecule drugs often produce significant toxic side effects, finding natural, highly effective, and low-toxic active substances has become a new direction for the development of acetylcholinesterase inhibitors. Meanwhile, oxidative stress and excessive calcium ion influx are also important causes of AD pathogenesis; therefore, finding multi-target drugs is of great significance for neuroprotection.

[0003] Currently, substances with AChE inhibitory activity extracted from natural products include alkaloids, flavonoids, polyphenols, and polypeptides. Among these, food-derived bioactive peptides, due to their advantages of low toxicity, high efficiency, easy absorption, and ability to cross the blood-brain barrier, have become ideal raw materials for developing neuroprotective active foods and pharmaceuticals. The oval pomfret is rich in protein and amino acids, and studies have shown that its enzymatic hydrolysates possess bioactivities such as lowering uric acid and blood lipids, making it a high-quality raw material for preparing bioactive peptides. Summary of the Invention

[0004] The purpose of this invention is to provide an acetylcholinesterase inhibitory peptide derived from the oval pomfret, its preparation method, and its application, in order to solve the problems existing in the prior art. The polypeptide derived from the oval pomfret provided by this invention can be used in the preparation of acetylcholinesterase inhibitors, antioxidants, and calcium supplements, and has good application prospects.

[0005] To achieve the above objectives, the present invention provides the following solution:

[0006] The present invention provides an acetylcholinesterase inhibitory peptide derived from the oval pomfret, wherein the acetylcholinesterase inhibitory peptide is any one of the polypeptides shown in SEQ ID NO.1-SEQ ID NO.4.

[0007] This invention also provides a method for preparing the acetylcholinesterase inhibitory peptide, characterized by comprising the following steps:

[0008] Oval pomfret surimi was enzymatically hydrolyzed, and after enzymatic hydrolysis, the enzyme was inactivated, centrifuged, filtered, and freeze-dried under vacuum to obtain oval pomfret enzymatically hydrolyzed protein powder.

[0009] The enzymatically hydrolyzed protein powder of the oval pomfret was dissolved to prepare an enzymatic hydrolysate. The enzymatic hydrolysate was then passed through an immobilized calcium ion affinity chromatography column and freeze-dried to obtain the purified component of the enzymatically hydrolyzed protein of the oval pomfret.

[0010] The purified components of the enzymatically hydrolyzed pomfret protein were separated into peptides by liquid chromatography-tandem mass spectrometry.

[0011] The peptide was molecularly simulated and docked with acetylcholinesterase and Keap1 to screen and identify the acetylcholinesterase inhibitory peptide.

[0012] Optionally, the enzymatic hydrolysis conditions are: adding papain and reacting at pH 6.0 and 55°C for 4 hours.

[0013] Optionally, the centrifugation conditions are: centrifugation at 10000 r / min for 20 min.

[0014] Optionally, the liquid chromatography-tandem mass spectrometry is a liquid chromatography-tandem mass spectrometry equipped with an online nano-spray ion source;

[0015] The mobile phase A of the liquid chromatography is ultrapure water containing 0.1% formic acid, and the mobile phase B is acetonitrile containing 0.1% formic acid.

[0016] The tandem mass spectrometry is MS and HCD-MS / MS.

[0017] This invention also provides the use of the acetylcholinesterase inhibitory peptide in any of the following:

[0018] (1) Application in the preparation of acetylcholinesterase inhibitors;

[0019] (2) Application in the preparation of antioxidants;

[0020] (3) Application in the preparation of calcium supplements.

[0021] The present invention also provides an acetylcholinesterase inhibitor, characterized in that the active ingredient is the acetylcholinesterase inhibitory peptide.

[0022] The present invention also provides an antioxidant, characterized in that the active ingredient is the acetylcholinesterase inhibitory peptide described above.

[0023] The present invention also provides a calcium supplement, characterized in that the active ingredient is the acetylcholinesterase inhibitor peptide described above.

[0024] This invention also provides a neuroprotective oligopeptide oral solution, comprising the following components by weight:

[0025] The ingredients are: 40 parts acetylcholinesterase inhibitory peptide, 12 parts acerola cherry powder, 6 parts vitamin C, 6 parts citric acid, 10 parts erythritol, and purified water to make up to 100 parts.

[0026] The present invention discloses the following technical effects:

[0027] This invention uses oval pomfret as raw material and obtains a product through enzymatic hydrolysis and immobilized calcium ion affinity chromatography that can effectively exert acetylcholinesterase inhibitory activity, antioxidant activity, and Ca2+. 2+ By combining the active heptapeptides DLTDYLM and QGPIGPR, the hexapeptide APDPFR and the pentapeptide WGDAG, and clarifying their chelation mechanism with acetylcholinesterase and Keap1, a new approach is provided for the preparation of multi-target acetylcholinesterase inhibitory peptides, while promoting the high-value utilization of oval pomfret.

[0028] This invention utilizes immobilized calcium ion affinity chromatography to extract peptide components simultaneously possessing acetylcholinesterase inhibitory activity, antioxidant activity, and calcium ion binding activity, simplifying the purification process. The extracted peptides exhibit acetylcholinesterase inhibitory and antioxidant activities consistent with those obtained using traditional methods. Compared to chemically synthesized acetylcholinesterase inhibitors, bioactive peptides offer advantages such as greater safety, higher efficiency, and easier absorption.

[0029] The polypeptides APDPFR, DLTDYLM, QGPIGPR, and WGDAG obtained in this invention can inhibit acetylcholinesterase activity by chelating the active site of acetylcholinesterase, thereby reducing the degradation of acetylcholine. Furthermore, by binding to Keap1, they release the antioxidant factor Nrf2 (as is known, Nrf2 is a key transcription factor regulating oxidative stress in the Keap1-Nrf2 signaling pathway; binding to Keap1 releases Nrf2, activating the antioxidant pathway), thus exerting an antioxidant effect. They also possess a high calcium binding rate. Therefore, the polypeptides derived from the oval pomfret provided in this invention can be used in the preparation of acetylcholinesterase inhibitors, antioxidants, and calcium supplements, showing promising application prospects. Attached Figure Description

[0030] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0031] Figure 1This is a schematic diagram of the three-dimensional binding site of the polypeptide APDPFR to acetylcholinesterase.

[0032] Figure 2 This is a schematic diagram of the two-dimensional interaction between the polypeptide APDPFR and acetylcholinesterase.

[0033] Figure 3 This is a schematic diagram of the three-dimensional binding site of the polypeptide DLTDYLM to acetylcholinesterase.

[0034] Figure 4 This is a schematic diagram of the two-dimensional interaction between the polypeptide DLTDYLM and acetylcholinesterase.

[0035] Figure 5 This is a schematic diagram of the three-dimensional binding site of the polypeptide QGPIGPR to acetylcholinesterase.

[0036] Figure 6 This is a schematic diagram of the two-dimensional interaction between the polypeptide QGPIGPR and acetylcholinesterase.

[0037] Figure 7 This is a schematic diagram of the three-dimensional binding sites of the polypeptide WGDAG and acetylcholinesterase.

[0038] Figure 8 This is a schematic diagram of the two-dimensional interaction between the polypeptide WGDAG and acetylcholinesterase.

[0039] Figure 9 This is a schematic diagram of the three-dimensional binding sites of the peptide APDPFR and Keap1.

[0040] Figure 10 This is a schematic diagram of the two-dimensional interaction between the peptide APDPFR and Keap1.

[0041] Figure 11 This is a schematic diagram of the three-dimensional binding sites of the peptide DLTDYLM and Keap1.

[0042] Figure 12 This is a schematic diagram of the two-dimensional interaction between the peptide DLTDYLM and Keap1.

[0043] Figure 13 This is a schematic diagram of the three-dimensional binding sites of peptide QGPIGPR and Keap1.

[0044] Figure 14 This is a schematic diagram of the two-dimensional interaction between peptide QGPIGPR and Keap1;

[0045] Figure 15 This is a schematic diagram of the three-dimensional binding sites of the peptide WGDAG and Keap1;

[0046] Figure 16This is a schematic diagram of the two-dimensional interaction between the peptide WGDAG and Keap1.

[0047] Figure 17 This is a chromatogram of immobilized calcium ion affinity chromatography. Detailed Implementation

[0048] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0049] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0050] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0051] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This specification and embodiments are merely exemplary.

[0052] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0053] Example 1: Preparation of acetylcholinesterase inhibitory peptide from oval pomfret

[0054] (1) Take the meat of oval pomfret, grind it into fish paste using a meat grinder, dissolve it in deionized water at a ratio of 1:3 (m:V), add 0.3% papain, and react at a constant temperature of pH 6.0 and 55℃ for 4 hours with shaking. After the reaction, inactivate the enzyme in a boiling water bath for 15 minutes, centrifuge at 10000r / min for 20 minutes, take the supernatant, filter, and freeze dry under vacuum to obtain oval pomfret enzymatic protein powder.

[0055] (2) The enzymatic hydrolysate of oval pomfret was dissolved in buffer solution to prepare an enzymatic hydrolysate. An immobilized calcium ion affinity chromatography column was prepared using 0.2 mol / L calcium chloride solution. The enzymatic hydrolysate was passed through the immobilized calcium ion affinity chromatography column at a loading volume of 10 mg / time and a chelation time of 90 min. Unchelated components were washed away with buffer solution with a pH of 7.4. The chelated peptides were then eluted with a 0.5 mol / L NaCl solution with a pH of 6.0 at a elution rate of 4 mL / min. The eluted sample solution was dialyzed (100 Da) for 24 h and then freeze-dried to obtain the purified fraction of oval pomfret enzymatic hydrolysate. Figure 17 ).

[0056] (3) The purified protein fraction of oval pomfret was analyzed by liquid chromatography-tandem mass spectrometry equipped with an online nanospray ion source. A total of 5 μL of sample was loaded and the sample was separated by a gradient of 60 min. The column flow rate was controlled at 300 nL / min, the column temperature was 40℃, the electrospray voltage was 2 kV, and the gradient started from 4% of phase B (mobile phase A: ultrapure water containing 0.1% (volume fraction) formic acid, mobile phase B: acetonitrile containing 0.1% (volume fraction) formic acid) and increased to 50% in a nonlinear gradient within 53 min 40 seconds, increased to 95% within 40 seconds, and maintained for 5 min 40 seconds. The mass spectrometry parameters were set as follows: (1) MS: Scan range (m / z): 100-1500; Resolution: 120,000; Normalized AGC target: 200%; Maximum injection time: 100ms; (2) HCD-MS / MS: Resolution: 50,000; Normalized AGC target: 200%; Maximum injection time: 86ms; Collision energy: 25%, 30%, 35%; Dynamic exclusion time: 30s. The tandem mass spectra were analyzed using PEAKS Studio version 10.6, and the final components were selected based on the Uniprot-Trachinotus ovatus database.

[0057] (4) Molecular docking screening: The obtained peptides were simulated and docked using the molecular docking software Autodock Vina. Four peptides that chelated most tightly with acetylcholinesterase were screened out. The sequences of these four peptides are APDPFR (SEQ ID NO.1), DLTDYLM (SEQ ID NO.2), QGPIGPR (SEQ ID NO.3) and WGDAG (SEQ ID NO.4), respectively. The sequences were visualized using Pymol and Discovery Studio software.

[0058] The structures of acetylcholinesterase (PDB ID: 4M0E) and Keap1 (PDB ID: 2FLU) were obtained from the RCSB protein library. Before docking, all water molecules and co-crystallized ligands in the acetylcholinesterase were removed using Pymol, and hydrogen atoms were added. Secondary structures of the heptapeptides DLTDYLM and QGPIGPR, the hexapeptide APDPFR, and the pentapeptide WGDAG were plotted and energy minimization was performed. In AutoDock Vina software, a GridBox was set up, adjusting the X, Y, Z coordinates and the grid center, and the box size was set to cover the entire protein molecule. The "-Docker Energy" value was used as a screening criterion; a smaller value indicates a higher likelihood of binding.

[0059] Peptide sequences identified by liquid chromatography-tandem mass spectrometry (LC-MS / MS) were screened for molecular docking, yielding four peptides with the highest activity: APDPFR, DLTDYLM, QGPIGPR, and WGDAG. Their molecular weights and docking results are shown in Table 1. The docking results indicated that the minimum binding energies of all four peptides with acetylcholinesterase and Keap1 were less than -7 kcal / mol, demonstrating that both ligands and receptors can bind spontaneously. Furthermore, a binding free energy ≤ -5.0 kcal / mol is generally considered to indicate that the binding of the ligand molecule to the target protein is of practical significance.

[0060] Table 1. Molecular weight and molecular docking results of acetylcholinesterase inhibitory peptides from oval pomfret.

[0061]

[0062] Figure 1 , Figure 3 , Figure 5 and Figure 7 The binding sites of peptides APDPFR, DLTDYLM, QGPIGPR, and WGDAG after docking with acetylcholinesterase are shown, all of which bind to the active site of acetylcholinesterase. Figure 2 , Figure 4 and Figure 6 and Figure 8It can be seen that the four peptides form hydrogen bonds and hydrophobic interactions with the active site of acetylcholinesterase. Among them, APDPFR interacts with the amino acid residues Trp286, Gln291, Ser293 and Tyr341 of AChE through hydrogen bonds, forms a Π-alkyl force with Leu76 and a Π-Π stacking force with Trp286 and Tyr341. DLTDYLM forms hydrogen bonds with the amino acid residues Trp286, Ser293, His287 and Thr75 of AChE, forms a Π-Π stacking with Trp286 residue and forms an alkyl force with Tyr72 and Val365 residues. QGPIGPR forms hydrogen bonds with Pro368, Gln369, Asn233, His405, Gln413, Asn533, Pro235, Arg247, Thr238, and Trp236, exhibits electrostatic attraction with Glu313, and forms alkyl groups with His405, Trp532, Leu536, Leu540, and Val370. WGDAG forms hydrogen bonds with Pro235, Phe297, Trp236, Arg247, and Glu313 in AChE. The residues of each peptide that bind to acetylcholinesterase show some similarity.

[0063] Figure 9 , Figure 11 , Figure 13 and Figure 15 The binding sites of peptides APDPFR, DLTDYLM, QGPIGPR, and WGDAG after docking with Keap1 are shown. Figure 10 , Figure 12 and Figure 14 and Figure 16It can be seen that APDPFR forms conventional hydrogen bonds with amino acid residues Val418, Val463, Val465, Val467, Val512, Val561, and Ile416 in the Keap1 molecule, and forms alkyl and π-alkyl forces with amino acid residues Ala607, Cys368, Val369, and Val420; DLTDYLM forms conventional hydrogen bonds with amino acid residues Val369, Val561, Val608, Arg326, and Arg470 in the Keap1 molecule, and forms alkyl forces with amino acid residues Leu515, Val514, and Ala466; QGPIGPR forms conventional hydrogen bonds with amino acid residues Val369, Val561, Val608, Arg326, and Arg470 in the Keap1 molecule; and forms alkyl forces with amino acid residues Leu515, Val514, and Ala466. The amino acid residues Gly371, Gly372, Gly367, Val608, Val467, Val606, Val463, Ile559, and Ile416 of the Keap1 molecule form conventional hydrogen bonds, and form alkyl forces with amino acid residues Val514, Val561, Ala607, Cys368, and Cys513. WGDAG forms conventional hydrogen bonds with amino acid residues Val512, Val561, Val465, Val463, Val604, Ile559, and Leu365 of the Keap1 molecule, and forms Π-alkyl forces with amino acid residues Arg415 and Ala556.

[0064] Example 2: Detection of acetylcholinesterase inhibition rate of peptides

[0065] The inhibition rates of acetylcholinesterase in the above-mentioned enzymatically hydrolyzed protein powder of oval pomfret, purified enzymatically hydrolyzed protein fraction of oval pomfret, APDPFR, DLTDYLM, QGPIGPR, and WGDAG were determined by the following methods:

[0066] Add 30 μL of ATCI (7.5 mM), 125 μL of DTNB (3 mM), and 50 μL of sample solutions of different concentrations sequentially to a 96-well plate. After pre-incubation at 37°C for 15 min, add 30 μL of acetylcholinesterase (0.055 U / mL) and continuously measure at 412 nm for 15 min. The blank control was replaced with PBS solution (pH 8.0, 50 mM). The formula for calculating acetylcholinesterase inhibitory activity is as follows:

[0067] Acetylcholinesterase inhibitory activity (%) = [1 - (Sample A - Sample A blank) / (Control A - Control A blank)] × 100;

[0068] Wherein, Sample A is the absorbance value of the sample group, Sample A blank is the absorbance value of the sample group without enzyme, Control A is the absorbance value of the sample group with enzyme but without enzyme, and Control A blank is the absorbance value of the sample group without enzyme and without sample.

[0069] The acetylcholinesterase inhibitory activities of 10 mg / mL of enzymatically hydrolyzed pomfret protein powder and its purified fraction were 18.02 ± 0.78% and 25.17 ± 0.65%, respectively. Four peptides identified through screening—APDPFR, DLTDYLM, QGPIGPR, and WGDAG—exhibited good acetylcholinesterase inhibitory activity, with an IC50 inhibition rate of [missing value]. 50 The results of the value detection are shown in Table 2. As can be seen from Table 2, the acetylcholinesterase inhibitory activities of the four peptides are ranked as follows: DLTDYLM > APDPFR > QGPIGPR > WGDAG.

[0070] Table 2. Acetylcholinesterase half-inhibition rate (IC50) of the peptides 50

[0071]

[0072] Example 3: Detection of ABTS free radical scavenging rate of peptides

[0073] The ABTS clearance rates of the above-mentioned enzymatically hydrolyzed protein powder, purified enzymatically hydrolyzed protein fraction of oval pomfret, APDPFR, DLTDYLM, QGPIGPR, and WGDAG were determined using the following methods:

[0074] The ABTS radical scavenging capacity of each sample was determined according to the instructions of the Total Antioxidant Capacity (T-AOC) kit. Each sample was prepared into a 1 mg / mL solution using deionized water. Reagent 1 and Reagent 2 were mixed at a 1:1 ratio and reacted in the dark for 12 hours, then diluted 40 times to obtain the working solution. 10 μL of the sample solution and 190 μL of the working solution were added sequentially to a 96-well plate, mixed well, and incubated at room temperature in the dark for 6 minutes. The absorbance (A) was measured at 414 nm. Deionized water was used instead of the working solution in the control tubes, and deionized water was used instead of the sample solution in the blank tubes. The formula for calculating the ABTS radical scavenging capacity is as follows:

[0075] ABTS free radical scavenging rate (%) = [A blank - (A sample - A control) / A blank] × 100;

[0076] Where A represents the absorbance of the sample group, A blank represents the absorbance of the sample group without enzyme, and A control represents the absorbance of the sample group with and without enzyme.

[0077] The ABTS radical scavenging rate of 1 mg / mL of enzymatically hydrolyzed pomfret protein powder was 52.54 ± 0.89%, and the ABTS radical scavenging rate of the purified protein fraction was 62.78 ± 0.47%. The inhibition rates of peptides APDPFR, DLTDYLM, QGPIGPR, and WGDAG were further improved. Based on the molecular weight of each peptide, the IC50 was... 50 The values ​​of mass concentration units are converted to molar concentration units, as shown in Table 3, to obtain the IC values.50 As a result, the ABTS scavenging activity was ranked as follows: WGDAG > DLTDYLM > APDPFR > QGPIGPR.

[0078] Table 3. ABTS scavenging activity of the peptides (IC50) 50

[0079]

[0080] Example 4: Detection of Ca in peptides 2+ Binding activity

[0081] The Ca content of the above-mentioned enzymatically hydrolyzed protein powder of oval pomfret, purified enzymatically hydrolyzed protein fraction of oval pomfret, APDPFR, DLTDYLM, QGPIGPR and WGDAG was detected. 2+ The inhibition rate was measured using the following method:

[0082] The sample and Ca were determined using inductively coupled plasma mass spectrometry (ICP-MS). 2+ To determine the binding affinity, the sample was mixed with a metal salt at a mass ratio of 3:1 to prepare a solution of a specific concentration. The pH was adjusted to 5.0, and the sample was placed in a 37°C water bath with constant temperature and shaking for 90 minutes to ensure complete binding. The bound solution was then removed and dialyzed for 48 hours, with the water changed every 4 hours. The total expansion volume of the solution after dialysis was recorded. After microwave digestion, the solution was brought to a final volume of 50 mL, and the metal ion content was determined using ICP-MS. The binding rate formula is as follows:

[0083]

[0084] Among them, R T denoted as ...

[0085] Table 4 shows that the purified protein from the enzymatic hydrolysis of oval pomfret contains Ca... 2+ The binding rate was significantly improved, and the calcium binding rates of the peptides DLTDYLM, QGPIGPR, and WGDAG identified from them were further enhanced, demonstrating good calcium binding capacity. 2+ The ability.

[0086] Table 4. Ca content of each sample 2+ Binding rate

[0087]

[0088] Example 5: Neuroprotective Oligopeptide Oral Solution

[0089] Based on the above experimental results, the polypeptide identified in this invention can be used to prepare a neuroprotective oligopeptide oral solution, which, by weight, includes 40 parts of acetylcholinesterase inhibitory peptide (DLTDYLM), 12 parts of acerola cherry powder, 6 parts of vitamin C, 6 parts of citric acid, 10 parts of erythritol, and purified water to make up to 100 parts.

[0090] According to the above raw material formula, mix all raw materials evenly to obtain the neuroprotective oligopeptide oral solution.

[0091] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. An acetylcholinesterase inhibitory peptide derived from the oval pomfret, characterized in that, The acetylcholinesterase inhibitory peptide is a polypeptide with the amino acid sequence shown in SEQ ID NO.

4.

2. The use of the acetylcholinesterase inhibitory peptide as described in claim 1 in the preparation of antioxidants.

3. An antioxidant, characterized in that, The active ingredient is the acetylcholinesterase inhibitory peptide as described in claim 1.

Images