Method for preparing low-sensitization protein peptide through microwave and enzymolysis combined composite modification and application of low-sensitization protein peptide

Abstract

The invention discloses a method for preparing low-sensitization protein peptide through microwave combined enzymolysis composite modification and application, and relates to the technical field of food processing. The treatment method comprises the following steps: preparing protein powder into a solution, adding tea polyphenol into the solution, and uniformly mixing to prepare a protein solution; carrying out microwave modification treatment on the protein solution; adding compound protease into the protein solution subjected to microwave treatment for enzymolysis and enzyme deactivation; and filtering the enzymolysis product, and drying the filtrate to obtain the low-sensitization protein peptide. According to the method, microwave and enzymolysis combined composite modification is adopted, so that the sensitization of protein can be reduced, the reaction time is shortened, and the method is simple in process, free of pollution and high in safety. In the enzymolysis process, active peptide substances which are easy to digest and absorb, can enhance immunity and delay senescence are synchronously generated, and tea polyphenol added in the formula can synergistically relieve anaphylactic reaction and play an anti-oxidation role, so that multi-element synergy of desensitization, function enhancement and flavor optimization is realized, and the added value and market application potential of the product are improved.

Description

Technical Field

[0001] This invention relates to the field of food processing technology, specifically to a method and application for preparing low-allergenic protein peptides through microwave-enzymatic hydrolysis and composite modification. Background Technology

[0002] Egg white protein is characterized by its high nutritional value, strong foaming ability, low price, and high bioactivity, making it an important source of animal protein in the food industry. It is rich in carbohydrates, protein, methionine, vitamins, minerals, and other nutrients, containing all nine essential amino acids required by the human body. Compared to other food proteins, its amino acid pattern is closest to that of human proteins. It has benefits such as supplementing nutrition, enhancing immunity, and delaying aging, thus contributing to overall health.

[0003] Although ovalbumin is a high-quality source of nutrition, it is also one of the eight most common allergens. Its allergenic potential manifests in several ways, with key characteristics including its abundance in food, the presence of multiple linear IgE binding sites, and resistance to protease hydrolysis and denaturation. Protease resistance is considered an important characteristic of food allergens. The more intact the protein polypeptide sequence is, the more likely its potential allergenic fragments—the digested peptides—are to retain intact IgE binding sites. This increases the likelihood that they will be recognized and bound by specific IgE antibodies in the body's immune system, thereby increasing the risk of inducing allergic reactions. Surveys show that up to 54% of subjects exhibit allergic reactions to ovalbumin, with a food allergy prevalence of approximately 5% in adults and 8% in children. It can cause symptoms such as rashes, papules, abdominal pain, and bronchial asthma, posing potential health risks to consumers. Therefore, the development of egg products with high nutritional value and low allergenicity is urgently needed.

[0004] Enzymatic hydrolysis is an effective method for modifying food protein allergens. Chinese patent CN 117603331 A discloses a low-allergenic whey protein peptide and its preparation method, using endopeptidase and exopeptidase for one-step and two-step enzymatic hydrolysis respectively to obtain the whey protein peptide. However, this method is complex, and the two-step hydrolysis is time-consuming, which is not conducive to industrial production. Chinese patent CN 117694402 A discloses a low-allergenic buffalo milk dairy product and its preparation method, using fig protease to hydrolyze buffalo milk whey protein. However, due to the strong protease resistance of whey protein, the degree of hydrolysis is not high, and the enzymatic hydrolysis product still retains a large number of IgE allergic epitopes, resulting in an unsatisfactory desensitization effect. Summary of the Invention

[0005] The purpose of this invention is to at least solve one of the technical problems existing in the prior art. This invention provides a method and application for preparing low-allergenic protein peptides by microwave combined with enzymatic hydrolysis and compound modification. The method utilizes microwave technology to pretreat protein solutions such as ovalbumin solution, egg white protein solution, casein solution, and soy protein isolate solution to reduce their protease resistance. Then, it uses compound protease hydrolysis and modification. The prepared protein peptides, such as ovalbumin peptide, egg white protein peptide, casein peptide, and soy protein isolate peptide, have the characteristics of low allergenicity and strong antioxidant properties, and effectively reduce allergic epitopes. The technical solution is simple to operate, has high yield, and good effect, making it suitable for large-scale industrial production.

[0006] The technical solution of the present invention is as follows: In a first aspect, the present invention provides a method for preparing low-allergenic protein peptides by microwave-combined enzymatic hydrolysis and composite modification, comprising the following steps: S1. Prepare a protein powder solution, add tea polyphenols to the solution, mix well to obtain a protein solution; S2. The protein solution is subjected to microwave modification treatment to obtain a microwave-treated protein solution; S3. Add a complex protease to the protein solution after microwave treatment for enzymatic hydrolysis. After enzymatic hydrolysis, inactivate the enzyme to obtain the enzymatic hydrolysis product. S4. Filter the enzymatic hydrolysis product to remove protein molecules, collect the filtrate, and dry the filtrate to obtain low-allergenic protein peptides.

[0007] This invention employs a combination of microwave and enzymatic hydrolysis for compound modification, which not only reduces the allergenicity of proteins but also significantly shortens the reaction time. The process is simple, pollution-free, and highly safe. Furthermore, tea polyphenols can synergistically alleviate allergic reactions and exert antioxidant effects.

[0008] In a preferred embodiment of the present invention, S1 further includes the addition of calcium citrate and erythritol to the solution.

[0009] In a preferred embodiment of the present invention, in S1, the mass ratio of protein powder, tea polyphenols, calcium citrate and erythritol is 10:0.1~0.2:0.01~0.02:0.25~0.5.

[0010] In a preferred embodiment of the present invention, in step S1, the protein powder is prepared into a solution with a solute mass fraction of 10-50%, wherein the protein powder is at least one selected from egg white protein powder, ovalbumin powder, casein powder, soy protein isolate powder, α-lactalbumin powder, and β-lactoglobulin powder.

[0011] In a preferred embodiment of the present invention, in S2, the power of the microwave modification treatment is 100~800 W and the time is 30~90s.

[0012] In a preferred embodiment of the present invention, in step S3, the pH of the microwave-treated protein solution is adjusted to 6-8 using hydrochloric acid solution, and the mass ratio of the complex protease to the microwave-treated protein solution is 1:50-100.

[0013] In a preferred embodiment of the present invention, in S3, the complex protease is bromelain and alkaline protease.

[0014] In a preferred embodiment of the present invention, in step S3, the mass ratio of the two enzymes in the complex protease is 1:1 to 2. More preferably, the mass ratio of the two enzymes in the complex protease is 1:1.5.

[0015] In a preferred embodiment of the present invention, a method for preparing low-allergenic protein peptides by microwave-combined enzymatic hydrolysis and compound modification includes the following steps: (1) Preparation of protein solution: First, use distilled water to prepare a solution with a solute mass fraction of 10-50% for the protein powder. Then, add tea polyphenols, calcium citrate and erythritol to the solution, stir evenly and prevent the generation of a large number of bubbles to obtain the protein solution. The ratio of each raw material is determined according to the mass fraction of solute as ovalbumin: tea polyphenols: calcium citrate: erythritol = 10: 0.1-0.2: 0.01-0.02: 0.25-0.5. (2) Microwave treatment: The protein solution obtained in step (1) is microwave-modified for 30 to 90 seconds at a power of 100 to 800 W to obtain the microwave-treated sample; (3) Enzymatic hydrolysis: Adjust the pH of the sample in step (2) to 6-8 with hydrochloric acid solution, add the complex protease at a ratio of 1:50-100 (w / w) between enzyme and sample, and hydrolyze by shaking at 40-60 ℃ for 1-2 h; (4) Enzyme inactivation: Under conditions of 80 ~ 100℃, enzyme inactivation is performed for 5 ~ 10 min to obtain the enzymatic hydrolysis product of the microwave-treated protein solution; (5) Filtration: The enzymatic hydrolysis product obtained in step (4) is filtered through a small molecular weight filter membrane to separate protein molecules; (6) Spray drying: The filtrate of each component is spray dried to obtain the low-allergenic protein peptide.

[0016] Secondly, the present invention provides a low-allergenic protein peptide, which is prepared by the method described above.

[0017] The protein peptides prepared by this invention have low antigenicity, which can reduce or eliminate allergic reactions to proteins such as eggs.

[0018] Thirdly, the present invention provides the application of the aforementioned low-allergenic protein peptide in the preparation of low-allergenic food, health products, or pharmaceuticals.

[0019] This invention has at least one of the following beneficial effects: 1. This invention utilizes a combination of microwave and enzymatic hydrolysis to reduce sensitization. On one hand, microwave technology is used to pretreat the ovalbumin solution to reduce protease resistance. Microwave treatment unfolds the ovalbumin, exposing its hydrophobic regions and disrupting its stable α-helix structure and disulfide bonds. This increases the accessibility of the complex protease to the ovalbumin and reduces its resistance to the complex protease, thereby promoting the generation of smaller polypeptide fragments with significantly reduced sensitization during ovalbumin hydrolysis. On the other hand, enzymatic hydrolysis has been proven to be an effective method for reducing protein sensitization. Proteases selectively cleave conformational and linear epitopes in protein molecules that determine their allergenicity, thereby significantly regulating and reducing their ability to induce allergic reactions. Furthermore, this invention uses two proteases to form a complex protease. Compared to a single protease, the complex protease exhibits better hydrolysis performance, more thoroughly hydrolyzing large molecular weight antigen proteins, significantly reducing the number of allergenic epitopes in the hydrolysis products, and synergistically reducing the sensitization risk of ovalbumin peptides.

[0020] 2. This invention also incorporates tea polyphenols, calcium citrate, and erythritol into the protein peptides. Firstly, tea polyphenols strongly inhibit histamine release, exhibiting 2-10 times stronger anti-allergic and anti-skin allergy effects than commonly used anti-allergy drugs, thus alleviating allergies. According to a survey by the Chinese Medical Association, up to 900 million people in my country suffer from calcium deficiency, with 200 million experiencing severe deficiency, which can lead to fatigue, memory loss, and even osteoporosis and rickets. Secondly, calcium citrate has a low risk of kidney stones and is easily absorbed, effectively supplementing the body's calcium needs. Finally, erythritol, as a filler sweetener, has the characteristic mild flavor of sugar alcohols, is extremely low in calories, and does not cause fluctuations in blood sugar levels, making it suitable for obese and diabetic patients.

[0021] In summary, this invention, through the above-described technical solution, can prepare a low-allergenic protein peptide. Egg protein products prepared using this method exhibit low antigenicity and can reduce or eliminate egg allergic reactions. The use of microwave combined with enzymatic hydrolysis significantly shortens the reaction time and is characterized by a simple, pollution-free, and highly safe process. Furthermore, the enzymatic hydrolysis process simultaneously generates easily digestible and absorbable active peptides that enhance immunity and delay aging. The added tea polyphenols in the formula synergistically alleviate allergic reactions and exert antioxidant effects, while sodium citrate supplements the body's calcium needs and regulates system stability. Erythritol improves product flavor and reduces calories, achieving a multi-faceted synergistic effect of "desensitization-functional enhancement-flavor optimization," significantly increasing the product's added value and market application potential. Attached Figure Description

[0022] Figure 1 This is the preparation process for low-sensitivity ovalbumin peptides.

[0023] Figure 2 This is a comparison chart of the degree of hydrolysis of ovalbumin peptides prepared in various embodiments and comparative examples.

[0024] Figure 3 This is a comparison chart of the sensitization properties of ovalbumin peptides prepared in various examples and comparative examples.

[0025] Figure 4 This is a comparison chart of the β-HEX release rates of ovalbumin peptides prepared in various examples and comparative examples. Detailed Implementation

[0026] To make the technical problems solved, the technical solutions, and the beneficial effects of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0027] Example 1 A method for preparing low-allergenic ovalbumin peptides by microwave combined with enzymatic hydrolysis and compound modification, the preparation process is as follows: Figure 1 As shown, it includes the following steps: (1) Preparation of protein solution: First, prepare a solution with a solute mass fraction of 25% by distilled water using egg white protein powder. Then, add tea polyphenols, calcium citrate and erythritol to the solution, stir evenly and prevent the generation of a large number of bubbles to obtain the protein solution. The ratio of each raw material is determined according to the mass fraction of solute as egg white protein powder: tea polyphenols: calcium citrate: erythritol = 10: 0.2: 0.02: 0.5. (2) Microwave treatment: The protein solution obtained in step (1) was microwave-modified for 60 s at a power of 400 W to obtain the microwave-treated sample; (3) Enzymatic hydrolysis: Adjust the pH of the sample in step (2) to 6.8 with hydrochloric acid solution, add the complex protease at a ratio of 1:50 (w / w) between enzyme and sample, and hydrolyze at 50 °C for 2 h with shaking; wherein, the complex protease is composed of bromelain and alkaline protease at a mass ratio of 1:1.5.

[0028] (4) Enzyme inactivation: The enzyme was inactivated at 100 °C for 10 min to obtain the enzymatic hydrolysis product of the microwave-treated protein solution; (5) Filtration: The enzymatic hydrolysis product obtained in step (4) is filtered through a small molecular weight filter membrane to separate protein molecules; (6) Spray drying: The filtrate of each component is spray dried to obtain the low-sensitivity ovalbumin peptide.

[0029] Example 2 A method for preparing low-allergenic ovalbumin peptides by microwave combined with enzymatic hydrolysis and compound modification, the preparation process is as follows: Figure 1 As shown, it includes the following steps: (1) Preparation of protein solution: First, prepare a solution with a solute mass fraction of 25% by distilled water using egg white protein powder. Then, add tea polyphenols, calcium citrate and erythritol to the solution, stir evenly and prevent the generation of a large number of bubbles to obtain the protein solution. The ratio of each raw material is determined according to the mass fraction of solute as egg white protein powder: tea polyphenols: calcium citrate: erythritol = 10: 0.2: 0.02: 0.5. (2) Microwave treatment: The protein solution obtained in step (1) was microwave-modified for 90 s at a power of 800 W to obtain the microwave-treated sample; (3) Enzymatic hydrolysis: Adjust the pH of the sample in step (2) to 8 with hydrochloric acid solution, add the complex protease at a ratio of 1:50 (w / w) between enzyme and sample, and hydrolyze under shaking at 50 °C for 1.5 h; wherein, the complex protease is composed of bromelain and alkaline protease at a mass ratio of 1:1.5.

[0030] (4) Enzyme inactivation: The enzyme was inactivated at 100 °C for 10 min to obtain the enzymatic hydrolysis product of the microwave-treated protein solution; (5) Filtration: The enzymatic hydrolysis product obtained in step (4) is filtered through a small molecular weight filter membrane to separate protein molecules; (6) Spray drying: The filtrate of each component is spray dried to obtain the low-sensitivity ovalbumin peptide.

[0031] Comparative Example 1 The preparation method of ovalbumin peptide in Example 1 is followed, except that after the protein solution is prepared in step (1), it is not microwaved, that is, step (2) is not performed, but enzymatic hydrolysis in step (3) is performed directly, and other processing steps remain unchanged to obtain protein peptide.

[0032] Comparative Example 2 The preparation method of ovalbumin peptide in Example 2 was followed, except that only ovalbumin, calcium citrate and erythritol were added in step (1), and the ratio of ovalbumin:calcium citrate:erythritol was 10:0.01:0.25. That is, tea polyphenols were not added in step (1), and the other processing steps remained unchanged to obtain the protein peptide.

[0033] Comparative Example 3 The preparation method of ovalbumin peptide in Example 1 was followed, except that step (3) used a single enzyme (bromelain) for enzymatic hydrolysis, while other processing steps remained unchanged to obtain the protein peptide.

[0034] The protein peptides prepared in Examples 1 and 2, as well as Comparative Examples 1 to 3, were measured using the following methods: 1. The degree of hydrolysis of the protein peptides prepared in the examples and comparative examples was measured using the OPA method.

[0035] The results are as follows Figure 2 Therefore, the degrees of hydrolysis of ovalbumin peptides obtained in Examples 1 and 2 were 85.33% and 94.15%, respectively, while the degrees of hydrolysis of ovalbumin peptides obtained in Comparative Examples 1, 2, and 3 were 75.75%, 81.87%, and 57.06%, respectively. The degree of hydrolysis is a core indicator reflecting the extent of protein hydrolysis; a higher degree of hydrolysis usually means that the protein molecules have been broken down into shorter peptide segments, reducing the residue of large protein molecules. Comparing Example 1 with Comparative Example 1 (without microwave treatment), it can be seen that the degree of hydrolysis in Comparative Example 1 (without microwave treatment) is significantly lower than that in Example 1. This indicates that microwave treatment can increase the accessibility and enzymatic hydrolysis effect of the complex protease on ovalbumin. This may be because microwave treatment causes ovalbumin to unfold, expose the hydrophobic region, and disrupt the stable α-helix structure and disulfide bonds. Comparing Example 2 with Comparative Example 2 (without tea polyphenols), it can be seen that the degree of hydrolysis in Comparative Example 2 (without tea polyphenols) is significantly lower than that in Example 2. This indicates that adding tea polyphenols in step (1) helps to improve the hydrolysis effect of the complex protease on ovalbumin. This may be because tea polyphenols, as a natural active polyphenol, can interact with proteins, change the spatial structure of ovalbumin, partially unfold it, and expose more enzyme cleavage sites, making it easier for the complex protease to hydrolyze it. Comparing Example 1 with Comparative Example 3 (using a single enzyme for hydrolysis), it can be seen that the degree of hydrolysis in Comparative Example 3 (using a single enzyme for hydrolysis) is much lower than that in Example 1. This indicates that using a complex enzyme hydrolysis process can significantly improve the hydrolysis efficiency of ovalbumin, obtain more low molecular weight peptides, and thus reduce the residue of potentially allergenic large molecular weight proteins. This may lead to a reduction in the number of intact IgE epitopes retained in the ovalbumin of Examples 1 and 2, which may reduce their allergenic risk.

[0036] 2. Allergenicity assessment: The sensitization of ovalbumin peptides prepared in Examples 1-2 and Comparative Examples 1-3 was evaluated using enzyme-linked immunosorbent assay (ELISA). The analytical methods are as follows: The concentration of ovalbumin peptide was prepared to 1 μg / mL, and 50 μL was added to an ELISA plate and coated at 37 °C for 2 h. The coated ELISA plate wells were then washed three times with PBST and patted dry. 250 μL of 1% fish gelatin solution was added to each well, and the plate was blocked at 37 °C for 2 h, followed by washing five times with PBST and patting dry. 50 μL of rabbit serum / human serum and 50 μL of protein sample were mixed and incubated at 37 °C for 2 h. The wells were washed five times with PBST, patted dry, and 100 μL of goat anti-rabbit IgG ~ HRP was added, and the plate was incubated at 37 °C for 1 h. The wells were washed five times with PBST, patted dry, and 100 μL of TMB chromogenic solution was added, and the plate was incubated at 37 °C for 15 min. The reaction was then terminated with 100 μL of concentrated sulfuric acid, and the absorbance was measured at 500 nm using an ELISA reader.

[0037] IgG / IgE binding rate (%) = In the formula, B represents the absorbance of the positive control (CBS instead of the sample); Bs represents the absorbance of the experimental group (OVA enzymatic digest product). This indicates the absorbance value of the negative control (using PBST instead of the sample).

[0038] The lower the IgG / IgE binding rate, the lower the sensitizing effect of ovalbumin peptides. For example... Figure 3 As shown, comparing Example 1 and Comparative Example 1 (without microwave treatment), it can be seen that the microwave treatment step described in this invention results in lower allergenicity of the ovalbumin peptide obtained after enzymatic hydrolysis. Comparing Example 2 and Comparative Example 2 (without tea polyphenols), it can be seen that the addition of tea polyphenols has an inhibitory effect on the allergenicity of the product. Comparing Example 1, Example 2 and Comparative Example 3 (enzymatic hydrolysis using a single enzyme), it can be seen that the combined enzymatic hydrolysis treatment can more thoroughly hydrolyze large molecular antigen proteins, thereby synergistically reducing the allergenicity risk of ovalbumin peptides.

[0039] 3. β HEX release rate determination: Serum pool: A mixture of serum from 30 patients with general egg allergy (specific IgE or IgG > 0.8 KUA / L); KU812 cells (Wuhan Shangen) were cultured and passaged in 1640 complete medium; cells were stimulated with serum from egg-allergic patients for 24 h. KU812 cells were then stimulated with 2% v / v sample (1 mg / ml) for 4 h. Positive controls (PBS instead of the sample) and negative controls (PBS instead of serum and sample) were set up. After cell lysis, the supernatant was collected by centrifugation. A 96-well microplate was used, and β-labeled... HEX ELISA kit (Quanzhou Ruixin) for detecting beta HEX, and calculate β HEX release rate.

[0040] β HEX release rate = (OD) 样品 - OD 空白 ) / (OD 对照 - OD 空白 )×100% The results are as follows Figure 4 As shown, compared to Comparative Example 1, which only underwent combined enzymatic hydrolysis without microwave treatment, the microwave-assisted enzymatic hydrolysis treatment in Example 1 significantly reduced β-cell degradation. The HEX release rate indicates that microwave pretreatment may alter protein conformation, making antigenic epitopes more susceptible to enzymatic cleavage. Comparing Example 2 with Comparative Example 2 (without tea polyphenols), the addition of tea polyphenols further inhibited β-cell cleavage. The release of HEX may originate from the binding of tea polyphenols to proteins, which may mask or alter some sensitizing epitopes. Furthermore, the combined results of Examples 1, 2, and Comparative Example 3 (using a single enzyme for hydrolysis) show that combined enzymatic hydrolysis has a significant effect on β-sensitization. The inhibitory effect on HEX release was the most significant, indicating that multiple treatments have a synergistic effect in reducing sensitization.

[0041] 4. Detection of allergic epitopes: Using LC MS (Orbitrap Fusion) analysis of the peptide sequences of egg white protein hydrolysates.

[0042] The components were identified by HPLC-MS / MS. A Pep Map RSLC column (50 μm × 150 mm, C18, 2 μm, 100 Å) was used. A and B were prepared using 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile solution, respectively. The samples were eluted at a flow rate of 220 nL / min under the following gradient conditions: 0–2 min, 4–12% B; 2–25 min, 12–22% B; 25–32 min, 22–32% B; 32–37 min, 32–75% B; 37–40 min, 75% B. Positive ion mode, mass range m / z 250 ~ 1250, HCD cleavage mode, cleavage energy 27%, the first 20 peptides were selected for cleavage based on the MS signal intensity, and PEAKS ~ DB functional module software (Bioinformatics Solution Inc., Waterloo, ON, Canada) was used to process MS / MS data.

[0043] References: Matsuo H, Yokooji T, Taogoshi T. Common food allergens and their IgE ~ binding epitopes[J]. Allergology International, 2015, 64(4):332 ~ 343; Mao Jihua, Study on the sensitization of glycosylated ovalbumin and development of hypoallergenic products[D], Nanchang University, focusing on the 11 ~ 19, 20 ~ 33, 34 ~ 46, 38 ~ 49, 47 ~ 55, 53 ~ 60, 55 ~ 58, 55 ~ 60, 56 ~ 70, 77 ~ 84, 95 ~ 102, 103 ~ 108, 125 ~ 134, 127 ~ 136, 141 ~ 154, 159 ~ 172, 164 ~ 176, 188 ~ 198, 191 ~ Tablets 200, 243 ~ 248, 251 ~ 260, 275 ~ 280, 277 ~ 282, 279 ~ 282, 301 ~ 306, 303 ~ 308, 323 ~ 332, 370 ~ 385, 375 ~ 384.

[0044] The results are shown in Table 1. Comparing Example 1 and Comparative Example 1 (without microwave treatment), it can be seen that microwave modification pretreatment can significantly reduce the types and numbers of allergic epitopes in the hydrolysis products, especially epitopes 20-33, 47-55, 77-84, and 279-282. Comparing Example 2 and Comparative Example 2 (without tea polyphenols), it can be seen that the addition of tea polyphenols can reduce the number of allergic epitopes in the hydrolysis products, indicating that it may interfere with the spatial accessibility of epitopes by binding to or masking proteins. Comparing Example 1, Example 2, and Comparative Example 3 (using a single enzyme for enzymatic hydrolysis), it can be seen that the combined enzymatic hydrolysis treatment significantly reduces the number of allergic epitopes in the hydrolysis products.

[0045] Table 1. Results of allergic epitope detection for different hydrolysis products Note: √ indicates detected, × indicates not detected; the number represents the number of corresponding table positions detected.

[0046] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing low-allergenic protein peptides by microwave combined with enzymatic hydrolysis and compound modification, characterized in that, Includes the following steps: S1. Prepare a protein powder solution, add tea polyphenols to the solution, mix well to obtain a protein solution; S2. The protein solution is subjected to microwave modification treatment to obtain a microwave-treated protein solution; S3. Add a complex protease to the microwave-treated protein solution for enzymatic hydrolysis. After enzymatic hydrolysis, inactivate the enzyme to obtain the enzymatic hydrolysis product. S4. Filter the enzymatic hydrolysis product to remove protein molecules, collect the filtrate, and dry the filtrate to obtain low-allergenic protein peptides.

2. The method according to claim 1, characterized in that, S1 also includes the addition of calcium citrate and erythritol to the solution.

3. The method according to claim 2, characterized in that, In S1, the mass ratio of added protein powder, tea polyphenols, calcium citrate and erythritol is 10:0.1 ~ 0.2:0.01 ~ 0.02:0.25 ~ 0.

5.

4. The method according to claim 1, characterized in that, In step S1, the protein powder is prepared into a solution with a solute mass fraction of 10-50%, wherein the protein powder is at least one of egg white protein powder, ovalbumin powder, casein powder, soy protein isolate powder, α-lactalbumin powder, and β-lactoglobulin powder.

5. The method according to claim 1, characterized in that, In S2, the power of the microwave modification treatment is 100 ~ 800W, and the time is 30 ~ 90s.

6. The method according to claim 1, characterized in that, In step S3, the pH of the microwave-treated protein solution is adjusted to 6-8 using hydrochloric acid solution, and the mass ratio of the complex protease to the microwave-treated protein solution is 1:50-100.

7. The method according to claim 1, characterized in that, In S3, the complex protease is any two of bromelain, alkaline protease, flavor protease, neutral protease, papain, and pepsin.

8. The method according to claim 7, characterized in that, In S3, the mass ratio of the two enzymes in the complex protease is 1:1~2.

9. A low-allergenic protein peptide, characterized in that, It is prepared by the method described in any one of claims 1 to 8.

10. The use of the hypoallergenic protein peptide according to claim 9 in the preparation of hypoallergenic food, health product or pharmaceutical.

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