Albumin peptide capable of enhancing body immunity and resisting allergy and preparation method of albumin peptide

Abstract

The invention discloses an anti-allergic albumin peptide capable of improving body immunity and a preparation method thereof, and relates to the field of small molecule active peptides, and the preparation method comprises the following steps: taking egg white powder, adding pure water, stirring for dissolving, adjusting pH, and standing for precipitating ovalbumin; adding pure water into the ovalbumin precipitate, stirring and dissolving, adjusting pH and temperature, and adding compound protease for enzymolysis; after the enzymolysis is finished, raising the temperature of the feed liquid for enzyme deactivation; centrifuging the material liquid after enzyme deactivation, and taking clear liquid for desalting; separating the desalted feed liquid through membrane equipment, and collecting permeate; and concentrating and drying the separated permeate to obtain the micromolecular albumin peptide. According to the albumin peptide capable of improving the immunity of the organism and resisting allergy, the preparation method of the albumin peptide and the anti-allergy active peptide fragment Ala-Leuu-Ala-Met, the allergenicity of ovalbumin is eliminated, and the albumin peptide has anti-allergy activity, has a good application prospect and has a good application prospect. The method has important significance on further application of ovalbumin related products in the fields of food and medicine.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of small molecule active peptides, and particularly relates to an albumin peptide for increasing immunity and resisting allergy and a preparation method thereof. BACKGROUND

[0002] Ovalbumin is the main protein in egg white, accounting for about 65% of egg white. It contains all the essential amino acids required by the human body, and its amino acid composition is close to the composition mode of the human body, with a biological value of more than 95. It is the most ideal high-quality protein in food. Due to its good gelation properties, foaming properties and emulsifying properties, ovalbumin is widely used in traditional food industry. In addition, it also has various biological activities, such as protecting liver, improving immunity, resisting oxidation, promoting brain development, etc.

[0003] However, ovalbumin also has the problem of allergy. Egg allergy is a common food allergy reaction disease, and often occurs in children, which can cause great harm to the body. It not only can cause a series of skin diseases, but also can cause functional disorders of the respiratory system and the gastrointestinal system. Studies have shown that ovalbumin is one of the main allergens in eggs, which can bind to IgE in human serum and trigger an immune response.

[0004] Studies have found that enzymatic hydrolysis can significantly reduce the allergenicity of macromolecular proteins, and even some peptides obtained by enzymatic hydrolysis have anti-allergy activity. Anti-allergy peptides are a class of small molecule peptide substances with significant anti-allergy activity. Compared with traditional anti-allergy drugs, they have the advantages of good curative effect, few side effects, high safety, etc. At present, many studies have reported anti-allergy peptides of different sources, such as soybean peptides, wheat peptides, oyster peptides, salmon skin collagen peptides, etc. In addition, there are also related patents reported, such as the Chinese patent with the publication number CN110452287A, which discloses an anti-allergy peptide and a preparation method thereof. The patent mainly uses Atlantic salmon viscera to prepare an anti-allergy peptide, which mainly includes the following steps: preparation of Atlantic salmon viscera enzymatic hydrolysate: 12% substrate is mixed with distilled water (w / v) and pepsin, then 1M HCl is used to adjust the pH to 2.0, 37 ℃ is incubated for 8h to release the anti-allergy peptide, 100℃ is heated for 15 min to inactivate the pepsin, after cooling, 0.45μm filter membrane is used for suction filtration to remove unhydrolyzed proteins. The Chinese patent with the publication number CN110638681A discloses a preparation method of mussel anti-allergy peptide and its application in cosmetics, which mainly includes the following steps: pretreatment, homogenization, ultrahigh pressure extraction, subcritical extraction, enzymatic hydrolysis, ultrafiltration, vacuum concentration and drying, etc.

[0005] At present, there is no report on the preparation of anti-allergic peptides from ovalbumin. Therefore, it is of great significance to prepare a kind of albumin peptide with anti-allergic activity and immune-enhancing effect by enzymatic hydrolysis, which is of great significance for the further application of ovalbumin in food and medicine fields. SUMMARY

[0006] The purpose of the present application is to provide an albumin peptide with immune-enhancing and anti-allergic effects and a preparation method thereof, so as to fill the blank of the preparation of anti-allergic peptides from ovalbumin.

[0007] The technical scheme adopted by the present application to solve the technical problems is as follows: The present application provides a preparation method of an albumin peptide with immune-enhancing and anti-allergic effects, which specifically comprises the following steps: Step S1: extraction of ovalbumin; The egg white powder is added to pure water and stirred to dissolve, and the pH is adjusted and then the ovalbumin is precipitated by standing. Step S2: enzymatic hydrolysis; The ovalbumin precipitate is added to pure water and stirred to dissolve, and the pH and temperature are adjusted before adding the complex protease for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the material liquid is heated to inactivate the enzyme. Step S3: desalination; The inactivated enzyme material liquid is centrifuged, and the clear liquid is subjected to desalination treatment. Step S4: separation; The desalted material liquid is separated by membrane equipment, and the permeate is collected. Step S5: drying; The separated permeate is concentrated and dried to obtain small molecule albumin peptides.

[0008] Preferably, in step S1, the egg white powder is added to 5-10 times the mass of pure water for stirring and dissolving, the pH of the material liquid is adjusted to 4.3-4.7, and then the ovalbumin is precipitated by standing for 0.5-2h.

[0009] Preferably, in step S2, the ovalbumin precipitate is added to 5-20 times the mass of pure water for stirring and dissolving, the pH of the material liquid is adjusted to 6-10, the temperature is adjusted to 40-60℃, and 0.3-1% of complex protease is added for enzymatic hydrolysis for 3-6h.

[0010] More preferably, the complex protease is composed of alkaline protease, trypsin and ficin, and the mass ratio of the alkaline protease, trypsin and ficin is (3-5):(1-2):(0-2).

[0011] Preferably, in step S2, after the enzymatic hydrolysis is completed, the material liquid is heated to 80-90℃ for 10-15min to inactivate the enzyme.

[0012] The present application provides an albumin peptide with immunity and anti-allergy, which is prepared by the preparation method.

[0013] Preferably, the albumin peptide contains an anti-allergy active peptide segment with an amino acid sequence of Ala-Leu-Ala-Met.

[0014] The present application also provides an anti-allergy active peptide segment prepared by the albumin peptide with immunity and anti-allergy.

[0015] Preferably, the anti-allergy active peptide segment has an amino acid sequence of Ala-Leu-Ala-Met.

[0016] The present application has the following advantages: 1. The present application provides an albumin peptide with immunity and anti-allergy. The albumin peptide has significant physiological activity, and has good immunity and anti-allergy effects.

[0017] 2. The present application provides a preparation method of the albumin peptide with immunity and anti-allergy. The preparation method has the advantages of simple operation and mild reaction conditions, and is suitable for large-scale production of the albumin peptide.

[0018] 3. The present application provides an anti-allergy active peptide segment prepared by the albumin peptide. The anti-allergy active peptide segment has an amino acid sequence of Ala-Leu-Ala-Met. The anti-allergy active peptide segment is a new small molecule peptide segment with high anti-allergy activity from egg albumin.

[0019] 4. The albumin peptide with immunity and anti-allergy, the preparation method thereof, and the anti-allergy active peptide segment Ala-Leu-Ala-Met provided by the present application not only eliminate the sensitization of egg albumin, but also have anti-allergy activity, and have good application prospects. The albumin peptide with immunity and anti-allergy, the preparation method thereof, and the anti-allergy active peptide segment Ala-Leu-Ala-Met provided by the present application have important significance for further application of egg albumin related products in the fields of food and medicine. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 Dose-effect relationship curve of the albumin peptide prepared in Example 1 on the inhibition of hyaluronidase.

[0021] Figure 2 Dose-effect relationship curve of the albumin peptide prepared in Example 2 on the inhibition of hyaluronidase.

[0022] Figure 3 Dose-effect relationship curve of the anti-allergy active peptide segment Ala-Leu-Ala-Met on the inhibition of hyaluronidase. DETAILED DESCRIPTION

[0023] The present application provides an albumin peptide for increasing immunity and resisting allergy.

[0024] The present application provides an albumin peptide for increasing immunity and resisting allergy, which uses small molecule albumin peptide as active ingredient.

[0025] The present application provides an albumin peptide for increasing immunity and resisting allergy, which has immunity enhancing activity.

[0026] The present application provides an albumin peptide for increasing immunity and resisting allergy, which contains an anti-allergy peptide segment and thus has anti-allergy activity.

[0027] Specifically, the amino acid sequence of the anti-allergy peptide segment is Ala-Leu-Ala-Met.

[0028] The present application provides a preparation method of an albumin peptide for increasing immunity and resisting allergy.

[0029] The present application provides a preparation method of an albumin peptide for increasing immunity and resisting allergy, which has the following specific implementation process: Step S1: extraction of egg albumin; The egg white powder is added with 5-10 times of pure water for slow stirring and dissolution, and the pH of the material liquid is adjusted to 4.3-4.7 (preferably 4.5), and then the egg albumin is precipitated after standing for 0.5-2 h.

[0030] Preferably, the egg white powder is a commercial egg white powder product, and any product with a protein content of ≥80% can be used.

[0031] Step S2: enzymolysis; The obtained egg albumin precipitate is added with 5-20 times of pure water for stirring and dissolution, and the pH of the material liquid is adjusted to 6-10 and the temperature is adjusted to 40-60℃, and then 0.3-1% of complex protease is added for enzymolysis for 3-6 h; after the enzymolysis is completed, the material liquid is heated to 80-90℃ (preferably 90℃) for 10-15 min (preferably 15 min) for enzyme inactivation.

[0032] Preferably, the complex protease is mainly composed of alkaline protease, trypsin, and ficin, and the mass ratio of alkaline protease, trypsin, and ficin is (3-5):(1-2):(0-2).

[0033] Step S3: desalination; The enzyme-inactivated material liquid is centrifuged (6000-10000 rpm, 10 min), and the supernatant is taken and subjected to desalination treatment by an electrodialysis device (10-20 mA / cm², conductivity 200-300 μS / cm).

[0034] Step S4: separation; The desalted solution is separated by a membrane device with a molecular weight cut-off of 1000-2000 Da (preferably 1000 Da), and the permeate is collected.

[0035] Step S5: drying; The separated permeate is concentrated, spray-dried or freeze-dried to obtain small-molecule albumin peptides.

[0036] In a third aspect, the present application provides an anti-allergy active peptide segment.

[0037] The anti-allergy active peptide segment provided by the present application has an amino acid sequence of Ala-Leu-Ala-Met.

[0038] The anti-allergy active peptide segment provided by the present application can be synthesized by biotechnology. It has been determined that the anti-allergy active peptide segment Ala-Leu-Ala-Met has extremely strong anti-allergy activity and is superior to albumin peptides of the same concentration.

[0039] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application. Example 1: Preparation of an albumin peptide for increasing immunity and resisting allergy

[0040] Take 500 g of egg white powder and add 10 times the mass of pure water to slowly stir and dissolve. After adjusting the pH of the solution to 4.5, let it stand for 2 h to precipitate the ovalbumin. Take the obtained ovalbumin precipitate, add 15 times the mass of pure water to stir and dissolve again, adjust the pH of the solution to 9.0 and the temperature to 50℃, and add 0.5% of a compound protease (alkaline protease, chymotrypsin, and ficin in a mass ratio of 4:2:1) for enzymatic hydrolysis for 5 h. After the enzymatic hydrolysis is completed, raise the temperature of the solution to 90℃ for 15 min to inactivate the enzyme. Centrifuge the inactivated solution (10000 rpm, 10 min), take the supernatant, and desalt it by an electrodialysis device (20 mA / cm², conductivity 250 μS / cm). Separate the desalted solution by a membrane device with a molecular weight cut-off of 1000 Da, and collect the permeate. Concentrate, spray-dry or freeze-dry the separated permeate to obtain small-molecule albumin peptides. Example 2: Preparation of an albumin peptide for increasing immunity and resisting allergy

[0041] Take 500 g of egg white powder and add 10 times the mass of pure water to slowly stir and dissolve. Adjust the pH of the material liquid to 4.5 and stand for 1.5 h to precipitate ovalbumin. Take the ovalbumin precipitate obtained above, add 20 times the mass of pure water to stir and dissolve again, adjust the pH of the material liquid to 8.0 and the temperature to 45℃, and add 1% of a compound protease (alkaline protease, chymotrypsin, and ficin in a mass ratio of 3:3:2) for enzymatic hydrolysis for 4 h. After the enzymatic hydrolysis is complete, the material liquid is warmed to 90℃ for 15 min to inactivate the enzyme. The material liquid after enzyme inactivation is centrifuged (8000 rpm, 10 min), and the supernatant is taken and subjected to desalting treatment by an electrodialysis device (15 mA / cm2, conductivity 300 μS / cm). The desalted material liquid is separated by a membrane device with a molecular weight cut-off of 1000 Da, and the permeate is collected. The separated permeate is concentrated, spray dried, or freeze dried to obtain small-molecule albumin peptides. Test Example 1: Test the amino acid composition of the albumin peptides prepared in Example 1 as the test sample

[0042] After testing the amino acid composition of the albumin peptides prepared in Example 1, the amino acid composition of the albumin peptides is shown in Table 1. The albumin peptides prepared in Example 1 contain 18 hydrolyzed amino acids, with a total content of 86.64 g / 100 g, of which 8 adult essential amino acids have a content of 30.11 g / 100 g, indicating that they have an important material basis for promoting the recovery or enhancement of immune function.

[0043] Table 1: Test results of the amino acid composition of the albumin peptides prepared in Example 1 Test Example 2: Test the molecular weight distribution of the albumin peptides prepared in Example 1 as the test sample

[0044] After testing the molecular weight distribution of the albumin peptides prepared in Example 1, the molecular weight distribution of the albumin peptides is shown in Table 2. The albumin peptides prepared in Example 1 have a molecular weight of 93.58% below 1000 Da, of which 58.18% is between 189-500 Da, indicating that the preparation process of the present application has good hydrolysis effect, and the large molecular proteins are hydrolyzed into oligopeptides, providing a structural basis for the enhancement of immunity and anti-allergy activity. Because compared with proteins, oligopeptides have unique structure and function, and show stronger advantages in absorption efficiency, biological potency, and physiological activity functions.

[0045] Table 2: Molecular weight distribution results of the albumin peptides prepared in Example 1 Test Example 3: Test the immunity-enhancing activity of the albumin peptides prepared in Example 1 as the test sample

[0046] Experimental animals: female mice, body weight 18-22 g, 10 in each group.

[0047] Dose grouping and administration time of test sample: The recommended human oral dose of albumin peptide is 5 g / day, and the equivalent dose is 0.0833 g / kg·bw according to the standard body weight of adults 60 kg. Three dose groups (low dose group, medium dose group, and high dose group) and one negative control group (normal saline) were set up. The mice in the three dose groups were given test samples by gavage for 30 days, and the mice in the negative control group were given normal saline by gavage for 30 days. The mice in the low dose group were fed according to the standard of 0.416 g / kg·bw, the mice in the medium dose group were fed according to the standard of 0.833 g / kg·bw, and the mice in the high dose group were fed according to the standard of 2.499 g / kg·bw, which were about 5 times, 10 times, and 30 times of the human oral dose (0.0833 g / kg·bw), respectively.

[0048] Test items and methods are as follows: 1. Body weight and organ / body weight ratio determination: After 30 days of test, the mice were sacrificed, and the thymus and spleen were weighed to determine the thymus / body weight ratio and the spleen / body weight ratio.

[0049] 2. Cell immune function determination: ConA-induced mouse spleen lymphocyte transformation test (MTT method); The spleen of each group of mice was placed in a culture dish containing 5 mL of sterile PBS, and the fat and connective tissue was removed and washed twice with PBS. The single cell suspension was prepared by grinding, centrifuged at 1500 r / min for 5 min, and the supernatant was discarded. 5 mL of RPMI1640 medium was added to resuspend the cells, centrifuged at 1500 r / min for 5 min, and washed twice. The cell concentration was adjusted to 2×10 6 cells / mL with RPMI 1640 medium; 100 μL of cell suspension (2×10 5 μL of ConA solution (final concentration 5 μg / mL), 37℃, 5% CO2 incubator for 48 h, 4 h before the end of culture, 20 μL of MTT solution (5 mg / mL) was added to each well, and the culture was continued for 4 h. The supernatant in the well was carefully aspirated, 150 μL of DMSO was added to each well, and the culture was placed on a shaker for 10 min to fully dissolve the MTT crystals. The absorbance (OD value) of each well was measured at 570 nm wavelength by a microplate reader.

[0050] 3. Humoral immune function determination: serum hemolysin determination (half hemolysis value); Take each group of mice serum, respectively, with diluent (PBS) according to 1:5, 1:10, 1:20 dilution, guinea pig serum with PBS according to 1:10 dilution after as complement ready for use; Take 96 well plate, according to experimental group (low dose group, middle dose group, high dose group): 100 μL dilution serum + 50 μL 5% sheep red blood cell (SRBC) suspension + 50 μL complement; Positive control group (total hemolysis): 100 μL diluent + 50 μL 5% SRBC suspension + 50 μL distilled water (instead of complement, make SRBC complete hemolysis); Negative control group 1 (no complement): 100 μL dilution serum + 50 μL 5% SRBC suspension + 50 μL PBS; Negative control group 2 (inactivated complement): 100 μL dilution serum + 50 μL 5% SRBC suspension + 50 μL 56 ℃ inactivated complement; Blank control group: 100 μL diluent + 50 μL 5% SRBC suspension + 50 μL complement; The well plate is placed in a 37 ℃ CO2incubator for 30 minutes, and then immediately placed in ice bath to terminate the reaction, 20 μL 0.1% ice acetic acid (fixing non hemolytic cells) is added to each well, and it is left to stand for 5 minutes; centrifuged at 3000 r / min for 10 minutes, 100 μL supernatant was taken to a new 96 well plate, and the absorbance (OD value) was measured at 540 nm wavelength by enzyme label instrument. The hemolysis rate of each group = (experimental group OD value-blank control OD value) / (total hemolysis OD value-blank control group OD value) * 100%, and the half hemolysis value (HC 50 ) of each group was calculated by drawing the hemolysis curve when the hemolysis rate was 50%.

[0051] The test results are as follows: 1. The effect of albumin peptide on the body weight and organ / body weight ratio of mice; The effect of albumin peptide on the body weight and organ / body weight ratio of mice is shown in Table 3. There was no significant difference between the body weight and organ / body weight ratio of mice in each dose group and the negative control group, indicating that different doses of albumin peptide had no effect on the body weight and immune organ / body weight ratio of mice.

[0052] Table 3 Effect of albumin peptide prepared in Example 1 on the body weight and organ / body weight ratio of mice ±S)

[0053] 2. The effect of albumin peptide on the cellular immune function of mice; The effect of albumin peptide on the cellular immune function of mice is shown in Table 4. Each dose group can significantly improve the proliferation ability of mouse spleen lymphocytes, and with the increase of dose, the proliferation ability of spleen lymphocytes shows an upward trend, indicating that albumin peptide has obvious enhancing effect on the cellular immune function of mice.

[0054] Table 4 Effect of the albumin peptide prepared in Example 1 on ConA-induced spleen lymphocyte proliferation in mice ±S)

[0055] 3. Effect of the albumin peptide on the humoral immune function of mice The effect of the albumin peptide on the humoral immune function of mice is shown in Table 5. Each dose group can significantly increase the serum hemolysin of mice, indicating that the albumin peptide has a significant enhancing effect on the humoral immune function of mice.

[0056] Table 5 Effect of the albumin peptide prepared in Example 1 on the half hemolysis value of mice ±S)

[0057] According to the "Health Food Function Test and Evaluation Method (2023 Edition)", it is determined that the test sample has the effect of helping to enhance immunity if the results of any two of the cellular immune function, humoral immune function, monocyte-macrophage function, and NK cell activity are positive. Therefore, it can be determined that the albumin peptide prepared in the present application has the activity of enhancing immunity. Test Example 4: The albumin peptide prepared in Example 2 was used as a test sample to test the activity of enhancing immunity The test method was the same as that in Test Example 3, and the results are shown in the following table. The albumin peptide prepared in Example 2 showed the same effect of enhancing immunity as the albumin peptide prepared in Example 1.

[0058] Table 6 Effect of the albumin peptide prepared in Example 2 on the body weight and organ / body weight ratio of mice ±S)

[0059] Table 7 Effect of the albumin peptide prepared in Example 2 on ConA-induced spleen lymphocyte proliferation in mice ±S)

[0060] Table 8 Effect of the albumin peptide prepared in Example 2 on the half hemolysis value of mice ±S) Test Example 5: The albumin peptides prepared in Examples 1 and 2 were used as test samples to test the anti-allergy activity

[0061] Hyaluronidase inhibition experiment is one of the main in vitro methods for evaluating the anti-allergic activity of a substance. The anti-allergic activity of a substance can be evaluated by determining the inhibition rate of the substance on the activity of hyaluronidase. The greater the inhibition rate, the stronger the anti-allergic activity of the substance.

[0062] The experimental method is as follows: 0.25 mmol / L CaCl2 solution 0.1 mL, hyaluronidase solution (50 mg hyaluronidase is dissolved in 20 mL 0.1 mol / L acetic acid buffer solution, pH 4.01) is added, 37°C incubation for 20 min; different concentrations (20, 40, 60, 80, 100 mg / mL) of albumin peptide solution 0.5 mL is added, 37°C incubation for 20 min; then 0.6 mg / mL sodium hyaluronate solution 0.6 mL is added, 37°C incubation for 20 min; 0.4 mol / L NaOH solution 1 mL is added, rapidly cooled on ice water for 15 min; boric acid 0.2 mL is added, boiling water bath for 3 min, immediately ice water cooling for 5 min, P-DAB reagent 3 mL is added, 37°C incubation for 20 min, color development, OD value is determined at 585 nm.

[0063] The specific calculation formula of the anti-allergic activity is as follows:

[0064] In the formula, A is the OD value of the control solution (enzyme + buffer + substrate); B is the OD value of the control blank (buffer + buffer + substrate); C is the OD value of the sample (enzyme + albumin peptide solution + substrate); D is the OD value of the sample blank control (buffer + albumin peptide solution + substrate).

[0065] The experimental results are shown in Figure 1 and Figure 2 In the test concentration range, the inhibition rate of the albumin peptide on hyaluronidase increases with the increase of the concentration, indicating that the prepared albumin peptide has good anti-allergic activity. Preparation of anti-allergic activity peptide segment Ala-Leu-Ala-Met and anti-allergic activity test

[0066] 1. Preparation of anti-allergic activity peptide segment Ala-Leu-Ala-Met; The albumin peptide prepared in Example 1 was subjected to peptide sequence identification and relative abundance analysis using LC-MS / MS technology. A peptide with high anti-allergic activity was screened out, with the amino acid sequence Ala-Leu-Ala-Met. This amino acid sequence was found to be a new active peptide by searching the online databases BIOPEP and EROP-Moscow.

[0067] 2. Anti-allergic activity test of the anti-allergic active peptide Ala-Leu-Ala-Met; A peptide with the amino acid sequence Ala-Leu-Ala-Met was synthesized using biotechnology, with a purity of 99.31%. Different concentrations (20 mg / mL, 40 mg / mL, 60 mg / mL, 80 mg / mL, and 100 mg / mL) of the synthesized peptide were prepared, and its anti-allergic activity was determined.

[0068] Experimental results are as follows Figure 3 As shown, within the experimental concentration range, the inhibition rate of the Ala-Leu-Ala-Met active peptide against hyaluronidase increased with increasing concentration, and was related to... Figure 1 Compared with the results shown, the Ala-Leu-Ala-Met active peptide fragment showed a significantly higher inhibition rate of hyaluronidase than albumin peptide of the same concentration, indicating that the active Ala-Leu-Ala-Met peptide fragment has extremely strong anti-allergic activity and is superior to albumin peptide of the same concentration.

[0069] This invention discloses an albumin peptide that enhances the body's immunity and has anti-allergic properties, as well as a method for its preparation. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It is particularly important to note that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The product of this invention has been described through preferred embodiments, and those skilled in the art can clearly modify or appropriately change and combine the product described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

Claims

1. A method for preparing albumin peptides that enhance immunity and have anti-allergic properties, characterized in that, The method comprises the following steps: Step S1: extraction of egg white protein; The egg white protein is extracted by adding egg white powder into pure water and stirring to dissolve the powder, adjusting the pH value and then allowing the mixture to stand for 0.5-2 hours to precipitate the egg white protein. Step S2: enzymatic hydrolysis; The precipitated egg white protein is dissolved by adding pure water and stirring, the pH value and temperature of the solution are adjusted, and then complex protease is added for enzymatic hydrolysis; after the enzymatic hydrolysis is completed, the temperature of the solution is raised to inactivate the enzyme. Step S3: desalination; The solution after inactivation of the enzyme is centrifuged, and the supernatant is subjected to desalination treatment. Step S4: separation; The solution after desalination is subjected to separation by a membrane device, and the permeate is collected. Step S5: drying; The permeate after separation is concentrated and dried to obtain small-molecule albumin peptides.

2. The method for preparing albumin peptides that enhance immunity and have anti-allergic properties according to claim 1, characterized in that, In step S1, egg white powder is added into 5-10 times the mass of pure water to dissolve the powder by stirring, the pH value of the solution is adjusted to 4.3-4.7, and then the mixture is allowed to stand for 0.5-2 hours to precipitate the egg white protein.

3. The method for preparing albumin peptides that enhance immunity and have anti-allergic properties according to claim 1, characterized in that, In step S2, the precipitated egg white protein is dissolved by adding 5-20 times the mass of pure water and stirring, the pH value of the solution is adjusted to 6-10, the temperature is adjusted to 40-60℃, and then 0.3-1% of complex protease is added for enzymatic hydrolysis for 3-6 hours.

4. The method for preparing albumin peptides that enhance immunity and have anti-allergic properties as described in claim 3, characterized in that, The complex protease is composed of alkaline protease, chymotrypsin and ficin, and the mass ratio of the alkaline protease, chymotrypsin and ficin is (3-5):(1-2):(0-2).

5. The method for preparing albumin peptides that enhance immunity and have anti-allergic properties according to claim 1, characterized in that, In step S2, after the enzymatic hydrolysis is completed, the temperature of the solution is raised to 80-90℃ for 10-15 minutes to inactivate the enzyme.

6. A white albumin peptide with increased immunity and anti-allergy prepared by the method of any one of claims 1-5.

7. The albumin peptide for increasing immunity and resisting allergy according to claim 6, wherein the albumin peptide is a peptide of SEQ ID NO:

1. The white albumin peptide contains an anti-allergy active peptide segment, and the amino acid sequence of the anti-allergy active peptide segment is Ala-Leu-Ala-Met.

8. An anti-allergy active peptide segment prepared by the white albumin peptide with increased immunity and anti-allergy of claim 6.

9. The anti-allergic active peptide segment according to claim 8, characterized by The anti-allergy active peptide segment has an amino acid sequence of Ala-Leu-Ala-Met.

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