Hydrolyzed egg yolk powder preparation as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of small molecule polypeptides, and particularly relates to a hydrolyzed yolk powder preparation as well as a preparation method and application thereof. The hydrolyzed egg yolk powder preparation comprises at least one egg yolk polypeptide, and the amino acid sequence of the egg yolk polypeptide is one of GFRPR, TTSFPHAS, MGWVR and ASWPKFL. The preparation method of the hydrolyzed egg yolk powder preparation comprises the following steps: S1, carrying out enzymolysis on lecithin-removed egg yolk powder to obtain an enzymolysis product; and S2, separating and extracting the zymolyte to obtain the hydrolyzed yolk powder preparation. The hydrolyzed egg yolk powder preparation is applied to preparation of antioxidant products, products for improving survival resilience, products for improving athletic ability or nerve protection products. The hydrolyzed egg yolk powder in the hydrolyzed egg yolk powder preparation is small in molecular weight and easy to absorb, has better oxidation resistance, can play an important role in improving survival resilience, exercise ability and neuroprotection, and widens the application range of the hydrolyzed egg yolk powder.

Description

Technical Field

[0001] The present invention belongs to the technical field of small molecule polypeptides, and particularly relates to a hydrolyzed egg yolk powder preparation, a preparation method thereof, and an application thereof. Technical Background

[0002] Hydrolyzed egg yolk powder, also known as enzymatically hydrolyzed egg yolk powder and egg yolk polypeptide powder, is a polypeptide powder derived from egg yolk. It has the advantages of being easily absorbed and promoting calcium absorption. It is widely used in the food field and in the health product field for preventing and treating osteoporosis, promoting bone growth and toughness, etc., and is beneficial to children, adults, and the elderly.

[0003] Chinese Patent CN117137090B discloses a hydrolyzed egg yolk powder complex with enriched sialic acid glycopeptide and a preparation method thereof. The main steps include: pretreatment of egg yolk powder, non-specific purification and enrichment, and specific purification and enrichment using sialic acid structural fragments as molecular imprinting template molecules. It also discloses that the hydrolyzed egg yolk powder prepared by this invention can not only promote bone growth and prevent osteoporosis, but also has functions such as good antibacterial and antiviral effects and enhanced immunity.

[0004] Chinese Invention CN118388602A discloses a calcium-absorbing promoting egg yolk peptide, a preparation method thereof, and an application thereof. The present invention relates to a calcium-absorbing promoting egg yolk peptide, a preparation method thereof, and an application thereof. The calcium-absorbing promoting egg yolk peptides are G1, G2, G3, G4, and G5. G1 has the amino acid sequence shown in SEQ ID NO.1, G2 has the amino acid sequence shown in SEQ ID NO.2, G3 has the amino acid sequence shown in SEQ ID NO.3, G4 has the amino acid sequence shown in SEQ ID NO.4, and G5 has the amino acid sequence shown in SEQ ID NO.5. It also discloses that this invention can prepare hatched egg yolk peptides with good calcium chelating activity, improve the utilization rate of egg yolk protein, and has a high calcium binding ability.

[0005] In summary, the existing research on the application of hydrolyzed egg yolk powder mainly focuses on aspects such as improving calcium binding ability, promoting calcium absorption, preventing and treating osteoporosis, promoting bone growth and toughness, etc., while there is relatively little research on antioxidant, improving survival recovery ability, exercise ability, and neuroprotection. Summary of the Invention

[0006] Aiming at the above problems, the purpose of the present invention is to provide a hydrolyzed egg yolk powder preparation, which contains a brand-new egg yolk polypeptide, has excellent antioxidant ability, and can play an important role in improving survival recovery ability, exercise ability, and neuroprotection, providing a raw material basis for preparing antioxidant products, products for improving survival recovery ability, products for improving exercise ability, or neuroprotection products.

[0007] To achieve the above purpose, the present invention adopts the following technical solutions:

[0008] A hydrolyzed egg yolk powder preparation, wherein the hydrolyzed egg yolk powder preparation comprises at least one egg yolk polypeptide, and the amino acid sequence of the egg yolk polypeptide is one of GFRPR, TTSFPHAS, MGWVR, and ASWPKFL.

[0009] Preferably, the hydrolyzed egg yolk powder preparation comprises egg yolk polypeptides with amino acid sequences of GFRPR and MGWVR.

[0010] Another object of the present invention is to provide a method for preparing the hydrolyzed egg yolk powder preparation, comprising the following steps:

[0011] S1. Degreasing lecithin egg yolk powder is enzymatically hydrolyzed to obtain an enzymatic hydrolysate;

[0012] S2. The enzymatic hydrolysate is separated and extracted to obtain the hydrolyzed egg yolk powder preparation.

[0013] Preferably, in step S1, the hydrolytic enzyme used for the enzymatic hydrolysis is at least one of papain and chymotrypsin.

[0014] Preferably, the hydrolytic enzyme used for the enzymatic hydrolysis is papain and chymotrypsin.

[0015] Preferably, in step S1, papain is first added for hydrolysis; after the hydrolysis of papain is completed, chymotrypsin is added to continue the hydrolysis.

[0016] Preferably, for the hydrolysis by chymotrypsin, the pH value is 7.0 - 9.0, the temperature is 25 - 40 °C, and the time is 3 - 5 h; for the hydrolysis by papain, the pH value is 5.0 - 7.0, the temperature is 50 - 65 °C, and the time is 3 - 5 h.

[0017] Another object of the present invention is to provide the application of the hydrolyzed egg yolk powder preparation in the preparation of antioxidant products.

[0018] Another object of the present invention is to provide the application of the hydrolyzed egg yolk powder preparation in the preparation of products for improving survival and recovery ability.

[0019] Another object of the present invention is to provide the application of the hydrolyzed egg yolk powder preparation in the preparation of products for improving exercise ability.

[0020] Another object of the present invention is to provide the application of the hydrolyzed egg yolk powder preparation in the preparation of neuroprotective products.

[0021] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0022] (1) The hydrolyzed egg yolk powder preparation of the present invention contains a brand-new egg yolk polypeptide, which can be isolated and purified from lecithin-free egg yolk powder or obtained by chemical synthesis.

[0023] (2) The egg yolk polypeptide in the hydrolyzed egg yolk powder preparation of the present invention has a small molecular weight, is easy to absorb, has good antioxidant ability, can improve cell damage induced by oxidative stress and enhance survival recovery ability, and reduce the risk of neurodegenerative diseases; on the other hand, it can protect nerves by reducing the toxic aggregation of pathogenic proteins, can effectively improve the movement disorders mediated by the toxicity aggregation of aging-related proteins, and reduce the body paralysis caused by the toxic aggregation of pathogenic proteins in the body. The hydrolyzed egg yolk powder preparation of the present invention has broad application prospects in the preparation of antioxidant, survival recovery ability, motor ability and neuroprotective products.

[0024] (3) The preparation method of the hydrolyzed egg yolk powder preparation provided by the present invention uses chymotrypsin and papain in combination for enzymatic hydrolysis, and the degree of hydrolysis and protein recovery rate of the egg yolk polypeptide are relatively high, which is beneficial to the preparation of the hydrolyzed egg yolk powder preparation. The stepwise enzymatic hydrolysis enables chymotrypsin and papain to react under the most suitable conditions respectively, better exerts the specificity of the two enzymes, and thus improves the enzymatic hydrolysis efficiency and the yield of specific polypeptides as a whole. Detailed implementation mode

[0025] In order to better present the present invention, it is illustrated by specific implementation cases. These implementation cases belong to the protection scope of the present invention but do not limit the protection scope of the present invention.

[0026] Example 1

[0027] The lecithin-free egg yolk powder was purchased from Guangzhou Yingzhu Biotechnology Co., Ltd., pulverized by a pulverizer, sieved through an 80-mesh sieve, and stored frozen at -20°C in a refrigerator for later use. Papain (10U / mg) and chymotrypsin (10U / mg) were both purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd.

[0028] A preparation method of a hydrolyzed egg yolk powder preparation includes the following steps:

[0029] S1. Weigh 10 g of delecithinated egg yolk powder and disperse it in 90 mL of distilled water. Select at least one of papain and chymotrypsin for enzymatic hydrolysis. The enzyme addition amount is 10 mg / g (based on the mass of delecithinated egg yolk powder). Hydrolyze the egg yolk protein under the optimal pH conditions of each enzyme. After 3 - 5 hours of hydrolysis, take it out, inactivate the enzyme (95 °C, 10 min), centrifuge (10000×g, 10 min), and then take the supernatant. Measure the degree of hydrolysis and protein recovery rate of the enzymatic hydrolysis supernatant. The supernatant is subjected to rotary evaporation under reduced pressure and freeze-dried to obtain hydrolyzed egg yolk powder dry powder. Among them, the pH value for chymotrypsin hydrolysis is 7.0 - 9.0, and the temperature is 25 - 40 °C; the pH value for papain hydrolysis is 5.0 - 7.0, and the temperature is 50 - 65 °C;

[0030] S2. Prepare the hydrolyzed egg yolk powder dry powder into a 20 mg / mL solution with deionized water. Use the centrifugal filter of Amicon Ultra 2 equipped with Ultracel 3 and Ultracel 10 membranes of Millipore for ultrafiltration, and collect the components with a molecular weight less than 3 kDa; then pass through a 0.45 μm filter membrane to remove impurities, and use a desalting column to remove the salts in the polypeptide sample to obtain the ultrafiltration intermediate component for freeze-drying;

[0031] S3. Use the packed Sephadex G-25 dextran gel chromatography column to separate the intermediate component. Elute with primary deionized water at a flow rate of 1 mL / min, and monitor with a 785UV / VIS detector at 280 nm while collecting the fractions of each absorption peak, and then freeze-dry to obtain polypeptide dry powder; Identify the polypeptide composition and amino acid sequence by LC MS / MS, screen out the peptide segments with amino acid sequence numbers GFRPR, TTSFPHAS, MGWVR, and ASWPKFL, and synthesize the polypeptide by solid-phase synthesis method by Shanghai Taopu Biotechnology Co., Ltd., with a purity > 95%.

[0032] In this example, taking the type of hydrolytic enzyme, whether the enzymatic hydrolysis process is stepwise, and the addition order of the hydrolytic enzyme as variables, different treatment groups are set (as shown in Table 1) to investigate the effects of different enzymatic hydrolysis conditions on the degree of hydrolysis and protein recovery rate of hydrolyzed egg yolk powder.

[0033] Table 1 Settings of each treatment group in Example 1

[0034] Group Type of hydrolase Whether in steps Adding order of hydrolase Treatment 1 Chymotrypsin No / Treatment 2 Papain No / Treatment 3 Chymotrypsin, Papain No / Treatment 4 Chymotrypsin, Papain Yes Chymotrypsin → Papain Treatment 5 Chymotrypsin, Papain Yes Papain → Chymotrypsin

[0035] Example 2

[0036] A hydrolyzed egg yolk powder preparation, the hydrolyzed egg yolk powder preparation includes at least one egg yolk polypeptide, and the amino acid sequence of the egg yolk polypeptide is one of GFRPR, TTSFPHAS, MGWVR, and ASWPKFL.

[0037] In this embodiment, the types or compositions of egg yolk polypeptides are used as variables to set different treatment groups (as shown in Table 2) to investigate the effects of different egg yolk polypeptides or combinations of egg yolk polypeptides on their application effects.

[0038] Table 2 Settings of each treatment group in Example 2

[0039]

[0040]

[0041] Control group

[0042] A commercially available neuroprotective product.

[0043] I. Product performance determination experiment

[0044] 1. Determination of degree of hydrolysis

[0045] The degree of hydrolysis of protein refers to the percentage of the number of peptide bonds broken during the protein hydrolysis process in the total number of peptide bonds of the original protein. The degree of hydrolysis of the egg yolk protein enzymatic hydrolysis supernatant was determined by the formaldehyde titration method. Accurately weigh 5 g of the enzymatic hydrolysis supernatant (5 g of ultrapure water as the blank), add 75 g of ultrapure water, adjust the pH of the solution to 8.2 with 0.1 mol / L NaOH standard solution, then add 10.0 mL of formaldehyde solution, mix well, and titrate the pH of the sample solution to 9.2 with 0.1 mol / L NaOH standard solution. Record the consumption of the standard solution, and calculate the ammonia nitrogen content in the enzymatic hydrolysis supernatant according to the following formula:

[0046]

[0047] In the formula, C is the concentration of the NaOH standard solution, mol / L; V is the volume of the NaOH standard solution consumed by the sample group, mL; V0 is the volume of the NaOH standard solution consumed by the blank group, mL; m1 is the mass of the enzymatic hydrolysis supernatant taken for determination, g; m is the total mass of the enzymatic hydrolysis supernatant, g.

[0048] The total nitrogen content of the enzymatic hydrolysis system was determined by the Kjeldahl method, and the degree of hydrolysis of the enzymatic hydrolysis supernatant was calculated according to the following formula:

[0049]

[0050] 2. Determination of protein recovery rate

[0051] The protein recovery rate refers to the ratio of the total nitrogen content in the enzymatic hydrolysis supernatant to the total nitrogen content in the enzymatic hydrolysis system. The total nitrogen content was determined by the Kjeldahl method, and the protein recovery rate of the enzymatic hydrolysis supernatant was calculated according to the following formula:

[0052]

[0053] In the formula, m1 represents the mass of the enzymolysis supernatant, in g; N1 refers to the nitrogen content of the enzymolysis supernatant, in %; m2 refers to the mass of the dephosphatidylated egg yolk powder added, in g; N2 refers to the nitrogen content of the dephosphatidylated egg yolk powder, in %.

[0054] 3. The results are shown in Table 3.

[0055] Table 3 Degree of hydrolysis and protein recovery rate of hydrolyzed egg yolk powder in each treatment group of Example 1

[0056] Group Degree of hydrolysis (%) Protein recovery rate (%) Treatment 1 10.21 49.59 Treatment 2 5.30 61.20 Treatment 3 17.06 65.58 Treatment 4 17.89 65.91 Treatment 5 18.34 67.69

[0057] As can be seen from Table 3:

[0058] (1) Chymotrypsin and papain both have a certain decomposition effect on dephosphatidylated egg yolk powder. Among them, the degree of hydrolysis of papain is higher than that of chymotrypsin, while the protein recovery rate is lower than that of chymotrypsin.

[0059] (2) Since single proteases have fixed cleavage sites, when hydrolyzing proteins with single enzymes, hydrolysis can only occur at several specific amino acid residues, which greatly limits the hydrolysis efficiency, and the types of polypeptide fragments and small peptides produced by hydrolysis are relatively single; the double-enzyme hydrolysis method uses the specificity of different enzymes and combines the two enzymes. The complementary cleavage sites of different enzymes can improve the hydrolysis efficiency and simultaneously diversify the types of bioactive small peptides.

[0060] (3) Since the active centers, action mechanisms, and hydrolysis conditions of chymotrypsin and papain are different, when used in combination, their respective optimal hydrolysis conditions restrict each other, which will also affect the enzymolysis effect; therefore, using them step by step can give full play to their respective advantages. Papain first extensively hydrolyzes the proteins in the dephosphatidylated egg yolk powder meat paste, and chymotrypsin can further decompose the larger peptide segments produced after the action of papain. The sequential action of the two can decompose the proteins in food more thoroughly, forming smaller peptides and amino acids, thereby improving the digestion and absorption rate of proteins.

[0061] II. Application effect test experiment

[0062] The Caenorhabditis elegans models used in this experiment were all purchased from the CGC in the United States. The Caenorhabditis elegans models include: wild-type N2, Aβ model, and polyQ model.

[0063] 1. In vivo oxidation activity experiment of hydrolyzed egg yolk powder

[0064] 1.1 Treatment method:

[0065] For wild-type N2, distribute 20 worms per well into a 96-well plate, and sequentially add the test sample (concentration 2 mM), NA22 bacterial solution (OD 570nmAt 0.5) and 75 μg / mL FUDR, with 6 replicate wells set in each group (total number of worms > 100), the final volume of each well was made up to 100 μL with S medium solution. After adding the lid, the plate was sealed with Parafilm and incubated in a shaker at 20 °C and 120 rpm for 24 h. A 50 mM paraquat solution was added to each well to induce oxidative damage in Caenorhabditis elegans. The number of surviving worms in each well was counted every half day under an inverted biological microscope until they all died. The Kaplan–Meier survival curve was plotted based on the experimental results, and the log-rank test was used to compare the differences between the drug-administered group and the blank control group. When p < 0.05, the results were considered to be significantly different.

[0066] 1.1.1 The in vivo antioxidant capacity of marine polypeptides was represented by the relative total survival increment ΔAUC%. The definition of ΔAUC% was as follows: Under oxidative stress conditions, the area under the survival curve (AUC sample ) of Caenorhabditis elegans in the polypeptide-administered group control ) and the area under the survival curve (AUC control ) of the control group, and the difference was divided by the AUC of the blank control group

[0067] ΔAUC% = 100% × (AUC sample - AUC control ) / / AUC control (4)

[0068] 1.1.2 Survival resilience (S / L) was defined as the ratio of the cumulative survival duration of Caenorhabditis elegans under stress conditions to the cumulative lifespan duration under normal growth conditions (S / L). It could be quantitatively expressed by calculating the ratio of the area under the survival curve (AUC stressspan ) of the stress-exposed population to the area under the lifespan curve (AUC life span ) of the normal growth population, as shown in formula (5).

[0069] Survival resilience (S / L) = AUC stressspan / AUC life span (5)

[0070] 1.2 Experimental group: The hydrolyzed egg yolk powder preparation from treatment 6 - 20 and the commercial product in Comparative Example 1 were used as the test samples to administer drugs to wild-type N2 by feeding.

[0071] Blank control group: Wild-type N2 of Caenorhabditis elegans without any treatment;

[0072] Model control group: Wild-type N2 of Caenorhabditis elegans that was only exposed to paraquat and not fed with drugs.

[0073] 1.3 The results are shown in Table 4.

[0074] Table 4 Antioxidant activities and survival recovery abilities of each treatment

[0075]

[0076]

[0077] As can be seen from Table 4, compared with the model control group and the comparative example, the antioxidant activity index ΔAUC% of the hydrolyzed egg yolk powder preparation in the drug administration treatment groups 6 - 20 was relatively high. Among them, the combined application effect of treatments 10 - 20 was better than that of the single application of treatments 6 - 9. And in the combined application, the combined application effect of APFVYI and AADGVGFR in treatment 11 was the best. This is because the combined application of polypeptides can provide a more powerful and comprehensive antioxidant effect compared with the application of single polypeptides through aspects such as synergistic effects, multiple targets, enhanced stability, improved bioavailability, and reduced tolerance. And low peptides and high peptides have different antioxidant mechanisms. Low peptides generally play an antioxidant stress role through relatively simple and direct ways such as directly scavenging free radicals, such as inhibiting the generation of free radicals and directly combining with free radicals to form stable compounds; while high peptides play a role through relatively complex mechanisms such as regulating cell signaling pathways and increasing the activity of antioxidant enzymes. And low peptides have better application effects than high peptides because of their small molecular weight and easier absorption. The mixed use of low peptides and high peptides sometimes cannot achieve the expected antioxidant effect due to the problem of interfering with each other's action mechanisms.

[0078] 2. Effects of hydrolyzed egg yolk powder preparation on protein aggregation toxicity-mediated dyskinesia

[0079] 2.1 Treatment method

[0080] 2.1.1 Drug administration treatment

[0081] Prepare the wild N2 model, polyQ model, and Aβ model of Caenorhabditis elegans respectively. Distribute them into 48-well plates at a quantity of 200 per well. Each well contains 0.5 mL of S medium with the sample solution to be tested, Escherichia coli NA22 (OD570 nm is 0.5), 75 μg / mL FUDR, and 100 μg / mL ampicillin antibiotic. Set 3 parallel replicates for each group. After sealing the edges with Parafilm membrane, place the 48-well plates on a shaker at 20 °C and 120 rpm for oscillatory culture, which is recorded as Day 0 of the adult stage. For the N2 model, samples are taken and tested on the 5th and 10th days of the adult stage respectively; for the polyQ model, samples are taken and tested on the 5th and 10th days of the adult stage respectively; for the Aβ model, after culturing at 20 °C for 12 h, the temperature is raised to 25 °C (to induce Aβ expression) and continue culturing, and samples are taken and tested on the 3rd and 5th days after the temperature increase for culturing respectively.

[0082] 2.1.2 Detection of locomotor ability

[0083] Wash the Caenorhabditis elegans collected from the above sampling with M9 solution repeatedly for 3 times to reduce the influence of residual food on the locomotor behavior of Caenorhabditis elegans. To detect the locomotor ability of Caenorhabditis elegans on solid plates, transfer 30 Caenorhabditis elegans to 3.5 cm NGM plates without food. Set 3 parallel plates for each group. After the Caenorhabditis elegans adapt to the new environment for 10 min, with the help of a behavioral tracking microscope, adjust the magnification to 4×, collect a 30-s crawling video, and then use the Movement Tracker population tracking software to analyze the locomotor ability of Caenorhabditis elegans in each group of videos. The obtained locomotor parameters include the paralysis rate (fractionparalyzed) and the number of directional shifts. During the tracking process, Caenorhabditis elegans with an instantaneous speed less than 0.015 mm / s for 80% is recorded as paralyzed nematodes. The direction rescue index is defined as the proportion of the number of Caenorhabditis elegans with fewer directional shifts than the average number of directional shifts in the control group in the drug-administered group, as shown in formula (6).

[0084] Direction rescue index = Nematode blow - mean / Nematode total (6)

[0085] 2.2 Experimental group: Use the hydrolyzed egg yolk powder preparations from Treatments 6 - 20 and the commercially available products in the control group as the samples to be tested for feeding and administering drugs to the polyQ model and Aβ model.

[0086] Control group: Wild-type N2 without any treatment and the polyQ model and Aβ model without feeding and administering drugs.

[0087] 2.3 The results are shown in Table 5.

[0088] Table 5 Motility ability and paralysis phenotype of Caenorhabditis elegans in each treatment group

[0089]

[0090]

[0091] From the observation of the experimental process, compared with the wild-type N2 of Caenorhabditis elegans, on the 5th day of the adult stage, the number of turns of the Caenorhabditis elegans polyQ model increased significantly. The number of turns of the Caenorhabditis elegans Aβ model also increased significantly on the 3rd day after the induction of the expression of toxic proteins by temperature increase. These experimental results indicate that the number of turns of Caenorhabditis elegans will increase with age and neurodegenerative changes.

[0092] As can be seen from Table 5, for the Caenorhabditis elegans polyQ model without drug administration, the turn rescue index on the 5th day of the adult stage was 50%, and the turn rescue index of the Aβ model on the 3rd day of Aβ expression was 48%. The hydrolyzed egg yolk powder preparations in Treatments 6 - 20 could increase the turn rescue index of the polyQ model on the 5th day of the adult stage to more than 61%, and also increase the turn rescue index of the Aβ model on the 3rd day of Aβ expression to more than 60%.

[0093] The toxic aggregation of pathogenic proteins in Caenorhabditis elegans will ultimately lead to body paralysis. For the Caenorhabditis elegans polyQ model without drug administration, more than 90% did not show paralysis symptoms on the 5th day of the adult stage, while the non-paralysis rate decreased to 29% on the 10th day of the adult stage; after being fed and administered with the marine polypeptide preparation of the present invention, the non-paralysis rate of the polyQ model increased to more than 40%; for the Caenorhabditis elegans Aβ model, on the 3rd day of the induction of Aβ expression by temperature increase, the Aβ non-paralysis rate decreased slightly, and on the 5th day of Aβ expression, its non-paralysis rate decreased to 74%. After being fed and administered with the hydrolyzed egg yolk powder preparation of the present invention, the non-paralysis rate of the Aβ model could be increased to more than 80%. These experimental results indicate that the hydrolyzed egg yolk powder preparation of the present invention can effectively improve the movement disorder of Caenorhabditis elegans mediated by the toxicity aggregation of aging-related proteins, thereby reducing the body paralysis caused by the toxicity aggregation of pathogenic proteins in Caenorhabditis elegans.

[0094] The above content is a further detailed description of the present invention in combination with specific / preferred embodiments, and it cannot be determined that the specific implementation of the present invention is only limited to these descriptions. For those of ordinary skill in the technical field to which the present invention belongs, without departing from the concept of the present invention, several alternatives or modifications can be made to these described embodiments, and these alternative or modified forms should be regarded as belonging to the protection scope of the present invention.

Claims

1. A hydrolyzed egg yolk powder preparation, characterized in that: It comprises at least one egg yolk polypeptide, and the amino acid sequence of the egg yolk polypeptide is one of GFRPR, TTSFPHAS, MGWVR and ASWPKFL.

2. The hydrolyzed egg yolk powder preparation according to claim 1, characterized in that: It includes egg yolk polypeptides with amino acid sequences of GFRPR and MGWVR.

3. The method for preparing the hydrolyzed egg yolk powder preparation according to claim 1 or 2, characterized in that: The following steps are involved: S1. performing enzymatic hydrolysis on the de-lecithinized egg yolk powder to obtain an enzymatic hydrolyzate; S2. The enzymatic hydrolysate is separated and extracted to obtain a hydrolyzed egg yolk powder preparation.

4. The method for preparing the hydrolyzed egg yolk powder preparation as claimed in claim 3, characterized in that: In step S1, the hydrolase used in the enzymatic hydrolysis is at least one of papain and chymotrypsin.

5. The method for preparing the hydrolyzed egg yolk powder preparation as claimed in claim 4, characterized in that: The hydrolytic enzymes used in the enzymolysis are papain and chymotrypsin.

6. The method for preparing the hydrolyzed egg yolk powder preparation as claimed in claim 5, characterized in that: In step S1, papain is first added for hydrolysis; after the hydrolysis by papain is completed, chymotrypsin is added to continue the hydrolysis.

7. Use of the hydrolyzed egg yolk powder preparation as claimed in claim 1 or 2 in the preparation of antioxidant products.

8. Use of the hydrolyzed egg yolk powder preparation as claimed in claim 1 or 2 in the preparation of a product for improving survival resilience.

9. Use of the hydrolyzed egg yolk powder preparation as claimed in claim 1 or 2 in the preparation of a product for improving athletic performance.

10. Use of the hydrolyzed egg yolk powder preparation according to claim 1 or 2 in the preparation of neuroprotective products.

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