Preparation method of ovalbumin peptide powder
By using a combination of enzymatic hydrolysis and secondary dry yeast fermentation, short-chain peptides were generated and glycosylated, which solved the problems of egg white's easy denaturation and poor solubility at high temperatures, improved its antioxidant and heat resistance, and expanded its application range.
Patent Information
- Application Number
- CN202511601611.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-04
- Publication Date
- 2026-01-27
AI Technical Summary
Traditional egg white products are prone to denaturation, are difficult to dissolve, and lack functionality at high temperatures, which limits their application in food and cosmetics and leads to a decline in product quality.
The preparation method employs a combination of enzymatic hydrolysis and secondary dry yeast fermentation. By introducing aminopeptidase to cleave terminal amino acid residues of proteins to generate short-chain peptides, and by glycosylation modification to improve antioxidant properties and heat resistance.
An ovalbumin peptide powder with excellent antioxidant and heat resistance was prepared, expanding its application range in food, cosmetics and other fields.
Smart Images
Figure CN121406740A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of protein peptide powder, and particularly relates to a preparation method of egg white protein peptide powder. BACKGROUND
[0002] For a long time, the egg industry in China mainly sells fresh eggs and carries out primary processing, and the products have low added value and short industrial chains, and the industry development is faced with the dilemma of serious homogenization competition and limited profit space. Functional characteristics such as heat sensitivity are easily denatured at high temperatures, lose emulsifying and foaming properties, and limit their application in high-temperature processed foods such as baking and beverages. Soluble protein powder is prone to caking when reconstituted, and additional stabilizers need to be added to improve performance, increasing the complexity of the formula. Egg white is prone to egg odor after processing, and high-temperature treatment may cause Maillard reaction, making the product color darker and affecting the sensory acceptance of the end food. With the continuous development of social economy and the steady increase of residents' consumption ability, people's living standards are improving, which is not only reflected in the satisfaction of food "quantity", but also in the pursuit of "quality", the subdivision of "function" and the diversified demand of "application scene". Under this trend, egg white as a natural and high-quality source of protein, its application value in the fields of food industry, nutrition and health, biotechnology, etc. is increasingly concerned. However, the traditional egg white products gradually show limitations in utilization, functional diversity and applicability - functional heat stability, poor solubility and other problems, leading to the loss of some nutrients or the decline of product quality; applicability cannot meet the individual needs of different industries such as health products and cosmetics. Therefore, in order to break through these bottlenecks and fully tap the potential value of egg white, the industry has begun to focus on systematic research to improve its utilization, strengthen its functionality and expand its applicability, aiming to make egg white better adapt to the needs of modern industrial development and consumer upgrading through technological innovation.
[0003] Therefore, the current research focuses on the development of complex enzymatic hydrolysis and yeast fermentation system to obtain egg protein peptide powder with more excellent functionality and more extensive application range. SUMMARY
[0004] To solve the problems in the background art, the application provides a preparation method of egg white protein peptide powder, which simultaneously improves the antioxidant property and heat resistance, to obtain egg white protein peptide powder with more excellent functionality and more extensive application range.
[0005] To achieve the above-mentioned purpose, the application provides the following technical solutions. A preparation method of egg white protein peptide powder, comprising the following steps: S1, using a full-automatic egg beater to separate the egg yolk and the egg white of the egg, then transferring the egg white liquid to a fermentation tank, adding citric acid to adjust the pH to 6.0-6.9, then adding dry yeast, stirring for 30-40 min, then fermenting to obtain a fermentation liquid; S2, transferring the fermentation liquid obtained in S1 to an enzymolysis tank, heating to 50-55 DEG C, adding a compound enzyme preparation, naturally enzymolyzing for 7-10 h, stirring for 10 min every hour to obtain an enzymolysis liquid; the compound enzyme preparation comprises papain, subtilisin, trypsin and aminopeptidase; S3, transferring the enzymolysis liquid obtained in S2 back to the fermentation tank, cooling to 30-37 DEG C, adding dry yeast, stirring for 30-40 min, then fermenting, using a spray drying tower to spray dry, then placing in a hot chamber for 3-7 days to obtain the egg white peptide powder.
[0006] Further, in the compound enzyme preparation, the mass ratio of papain, subtilisin, trypsin and aminopeptidase is (5-6):(3-4):2:1.
[0007] Further, in S2, the addition amount of the compound enzyme preparation is 0.4-0.6% of the mass of the fermentation liquid.
[0008] Further, in S2, the compound enzyme preparation is diluted with normal temperature water at a ratio of 1:2 before being transferred to the enzymolysis tank.
[0009] Further, in S1, the addition amount of dry yeast is 0.7-1% of the mass of the egg white liquid; and in S3, the addition amount of dry yeast is 0.5-0.6% of the mass of the enzymolysis liquid.
[0010] Further, in S1, the fermentation temperature is 30-37 DEG C, and the fermentation time is 8-10 h.
[0011] Further, in S3, the fermentation temperature is 30-37 DEG C, and the fermentation time is 4-5 h.
[0012] Further, in S3, the specific operation of spray drying is as follows: using a spray drying tower to spray dry, the inlet air temperature of the spray drying tower is 160-170 DEG C, and the discharge temperature is 60-80 DEG C.
[0013] Further, in S3, the temperature of the hot chamber is 80-90 DEG C.
[0014] The application has the following beneficial effects: The amino peptidase introduced by the application removes the shielding of the anti-oxidation active groups (such as cysteine thiol and tyrosine phenolic hydroxyl) by specifically cutting the hydrophobic amino acid residues (such as leucine and phenylalanine) at the end of the protein to generate short-chain polypeptides; compared with the random cutting mode of the traditional protease, the amino peptidase is more inclined to retain the short peptides containing the anti-oxidation active sequence (such as the histidine-proline-glutamic acid motif) to provide a high-activity carrier for subsequent glycosylation.
[0015] The application also adds secondary dry yeast fermentation to improve the heat resistance and anti-oxidation of the product by modifying the dimensions through glycosylation modification, cell wall wrapping and metabolite deposition; and the glycosyltransferase secreted by the dry yeast in the secondary fermentation stage can form a glycopeptide complex with the protein peptides (especially short peptides) obtained by the amino peptidase to synergistically improve the heat resistance and anti-oxidation of the prepared egg white peptide powder. BRIEF DESCRIPTION OF DRAWINGS
[0016] Fig. 1 The figure is a comparison trend chart of the anti-oxidation-DPPH clearance rate data of the egg white peptide powder prepared in Examples 1-3 and Comparative Examples 1-3 in the test examples of the application. Fig. 2 The figure is a comparison trend chart of the heat resistance-DPPH clearance rate loss rate data of the egg white peptide powder prepared in Examples 1-3 and Comparative Examples 1-3 in the test examples of the application. DETAILED DESCRIPTION
[0017] The application will be further described in detail below in combination with the examples.
[0018] The raw materials of the examples and comparative examples of the application are all ordinary commercially available materials unless otherwise specified.
[0019] Example 1: A preparation method of egg white peptide powder, comprising the following steps: S1, using a full-automatic egg beater to separate the yolk and albumen of the egg, and then transferring the albumen liquid to a fermentation tank, adding citric acid to adjust the pH to 6.5, and then adding dry yeast, the addition amount of the dry yeast being 0.8% of the mass of the albumen liquid, stirring for 35 min, and then performing fermentation, the fermentation temperature being 32℃ and the fermentation time being 9h to obtain a fermentation liquid.
[0020] S2, transferring the fermentation liquid obtained in S1 to an enzymolysis tank, heating to 52℃, adding a compound enzyme preparation, and naturally enzymolysing for 8h, stirring for 10 min every hour to obtain an enzymolysis liquid.
[0021] The complex enzyme preparation includes papain, subtilisin, trypsin and aminopeptidase, and the mass ratio of papain, subtilisin, trypsin and aminopeptidase is 5.5:3.5:2:1. The adding amount of the complex enzyme preparation is 0.5% of the mass of the fermentation liquor. The adding method of the complex enzyme preparation is that the complex enzyme preparation is diluted with normal temperature water at a ratio of 1:2, and then is transferred to the enzymolysis tank.
[0022] Among them, papain (YT00018), subtilisin (YT1793), trypsin (YT2194) and aminopeptidase (YT6416) are purchased from Beijing Yita Biological Technology Co., Ltd.
[0023] S3, the enzyme solution obtained in S2 is transferred back to the fermentation tank, and the temperature is lowered to 32℃, dry yeast is added, the adding amount of dry yeast is 0.55% of the mass of the enzyme solution, after stirring for 35min, fermentation is carried out, the fermentation temperature is 32℃, and the fermentation time is 4.5h.
[0024] Spray drying is carried out by using a spray drying tower, the inlet air temperature of the spray drying tower is 165℃, and the discharge temperature is 70℃. Then, the spray drying tower is placed in a hot room at 85℃ for 5 days, and egg white peptide powder is obtained.
[0025] Example 2: The difference between this example and example 1 is that a preparation method of egg white peptide powder includes the following steps: S1, using a full-automatic egg beater, the yolk and the albumen of the egg are separated, then the albumen liquid is transferred to a fermentation tank, citric acid is added to adjust the pH to 6.0, then dry yeast is added, the adding amount of dry yeast is 0.7% of the mass of the albumen liquid, after stirring for 30min, fermentation is carried out, the fermentation temperature is 30℃, and the fermentation time is 8h, and a fermentation liquor is obtained.
[0026] S2, the fermentation liquor obtained in S1 is transferred to an enzyme solution tank, the temperature is raised to 50℃, a complex enzyme preparation is added, and natural enzymolysis is carried out for 7h, and stirring is carried out for 10min every hour, and an enzyme solution is obtained.
[0027] The complex enzyme preparation includes papain, subtilisin, trypsin and aminopeptidase, and the mass ratio of papain, subtilisin, trypsin and aminopeptidase is 5:3:2:1. The adding amount of the complex enzyme preparation is 0.4% of the mass of the fermentation liquor. The adding method of the complex enzyme preparation is that the complex enzyme preparation is diluted with normal temperature water at a ratio of 1:2, and then is transferred to the enzymolysis tank.
[0028] S3, the enzyme solution obtained in S2 is transferred back to the fermentation tank, and the temperature is lowered to 32℃, dry yeast is added, the adding amount of dry yeast is 0.55% of the mass of the enzyme solution, after stirring for 35min, fermentation is carried out, the fermentation temperature is 32℃, and the fermentation time is 4.5h.
[0029] Spray drying was performed using a spray drying tower, the inlet temperature of the spray drying tower was 165℃, and the discharge temperature was 70℃. Then, the obtained egg white peptide powder was placed in a hot chamber at 80℃ for 7 days.
[0030] Example 3: The difference between this example and Example 1 is that a method for preparing an egg white peptide powder comprises the following steps: S1, using a full-automatic egg beater to separate the yolk and the albumen of the egg, then transferring the albumen liquid to a fermentation tank, adding citric acid to adjust the pH to 6.9, then adding dry yeast, the addition amount of the dry yeast is 1% of the mass of the albumen liquid, stirring for 40 min, then performing fermentation, the fermentation temperature is 37℃, and the fermentation time is 10 h, to obtain a fermentation liquid.
[0031] S2, transferring the fermentation liquid obtained in S1 to an enzymolysis tank, heating to 55℃, adding a compound enzyme preparation, and naturally enzymolyzing for 10 h, stirring for 10 min every hour, to obtain an enzymolysis liquid.
[0032] The compound enzyme preparation comprises papain, subtilisin, trypsin and aminopeptidase, and the mass ratio of the papain, the subtilisin, the trypsin and the aminopeptidase is 6:4:2:1. The addition amount of the compound enzyme preparation is 0.6% of the mass of the fermentation liquid. The compound enzyme preparation is diluted with normal temperature water at a ratio of 1:2, and then transferred to the enzymolysis tank.
[0033] S3, transferring the enzymolysis liquid obtained in S2 back to the fermentation tank, cooling to 37℃, adding dry yeast, the addition amount of the dry yeast is 0.6% of the mass of the enzymolysis liquid, stirring for 40 min, then performing fermentation, the fermentation temperature is 37℃, and the fermentation time is 5 h.
[0034] Spray drying was performed using a spray drying tower, the inlet temperature of the spray drying tower was 165℃, and the discharge temperature was 70℃. Then, the obtained egg white peptide powder was placed in a hot chamber at 90℃ for 3 days.
[0035] Comparative Example 1: The difference between this comparative example and Example 1 is that the compound enzyme preparation does not contain aminopeptidase; and no secondary dry yeast fermentation is performed.
[0036] Specifically, a method for preparing an egg white peptide powder comprises the following steps: S1, using a full-automatic egg beater to separate the yolk and the albumen of the egg, then transferring the albumen liquid to a fermentation tank, adding citric acid to adjust the pH to 6.5, then adding dry yeast, the addition amount of the dry yeast is 0.8% of the mass of the albumen liquid, stirring for 35 min, then performing fermentation, the fermentation temperature is 32℃, and the fermentation time is 9 h, to obtain a fermentation liquid.
[0037] S2, transfer the fermentation broth obtained in S1 to an enzymolysis tank, warm to 52℃, add the compound enzyme preparation, and naturally enzymolyze for 8h, with stirring for 10min every hour, to obtain an enzymolysis liquid.
[0038] The compound enzyme preparation comprises papain, subtilisin and trypsin, and the mass ratio of papain, subtilisin and trypsin is 5.5:3.5:2. The addition amount of the compound enzyme preparation is 0.5% of the mass of the fermentation broth. The compound enzyme preparation is diluted with normal temperature water at a ratio of 1:2, and then transferred to the enzymolysis tank.
[0039] S3, spray dry the enzymolysis liquid obtained in S2 by using a spray drying tower, with the inlet air temperature of the spray drying tower being 165℃ and the discharge temperature being 70℃. Then, place in a hot chamber at 85℃ for 5 days to obtain egg white protein peptide powder.
[0040] Comparative Example 2: The difference between this comparative example and Example 1 is that no secondary dry yeast fermentation is performed.
[0041] Comparative Example 3: The difference between this comparative example and Example 1 is that no aminopeptidase is added in the compound enzyme preparation.
[0042] Test Example: Test objects: egg white protein peptide powders prepared in Examples 1-3 and Comparative Examples 1-3.
[0043] Test Items and Methods: 1. Antioxidant property-DPPH clearance rate: dissolve DPPH in anhydrous ethanol to prepare a DPPH solution with a concentration of 0.1mM, store in the dark and use within 3.5 hours; dissolve each test object in distilled water to prepare a sample solution with a concentration of 0.2mg / mL; mix 2mL of the DPPH solution with 2mL of the sample solution (A1 absorbance value); mix 2mL of the sample solution with 2mL of ethanol (A2 absorbance value); mix 2mL of the DPPH solution with 2mL of distilled water (A3 absorbance value). The DPPH clearance rate is [1-(A1-A2) / A3]x100%. The larger the DPPH clearance rate value, the better the antioxidant property of the corresponding test object.
[0044] Heat resistance-DPPH clearance rate loss rate: the DPPH clearance rate measured in Test Item 1 above is denoted as M0; after storing each test object at 65℃ for 4h, test the DPPH clearance rate again, denoted as M1, and the test method is the same as in Test Item 1 above; calculate the DPPH clearance rate loss rate, denoted as X; the calculation formula is X=(M0-M1) / M0x100%. The smaller the DPPH clearance rate loss rate value, the better the heat resistance of the corresponding test object.
[0045] Test Results: see Table 1.
[0046] Table 1. Test data of Test Example
[0047] Results analysis: combined with the data in Table 1 and Figs. 1-2 By analyzing Example 1-Example 3, it can be seen that the DPPH scavenging rate of the egg white peptide powder prepared by the present application (Example 1-Example 3) is above 93.8%, and the DPPH scavenging rate loss rate is as low as 5.1% or less. It shows that the egg white peptide powder prepared by the present application has excellent antioxidant and heat resistance.
[0048] Combined with the data in Table 1 and Figs. 1-2 By analyzing Example 1 and Comparative Example 1-Comparative Example 3, and specifically comparing Comparative Example 1 and Comparative Example 2, it can be seen that compared with Comparative Example 1, the amino peptidase is added to the complex enzyme preparation in Comparative Example 2. As a result, the DPPH scavenging rate of the egg white peptide powder prepared is increased from 88.7% (Comparative Example 1) to 92.1% (Comparative Example 2), that is, the antioxidant property is improved; however, the DPPH scavenging rate loss rate is increased from 6.1% (Comparative Example 1) to 6.6% (Comparative Example 2), that is, the heat resistance is actually reduced.
[0049] This is mainly because when the amino peptidase is added, it can cut off the amino acid residues from the amino or carboxyl end of the peptide chain one by one, tend to generate short-chain polypeptides and free amino acids, contain sequences with antioxidant activity, and the activity is higher than that of the original long-chain protein; and the active site originally wrapped inside the protein is destroyed during the enzymatic hydrolysis, which helps to expose the amino acid residues with antioxidant activity, thereby improving the antioxidant capacity. However, at the same time, excessive hydrolysis of the amino peptidase will cause the protein peptide fragments to become smaller, which is more prone to denaturation or degradation under high temperature conditions, resulting in a decrease in heat resistance.
[0050] Specifically, by comparing Comparative Example 1 and Comparative Example 3, it can be seen that compared with Comparative Example 1, the secondary dry yeast fermentation is added in Comparative Example 3. As a result, the DPPH scavenging rate of the egg white peptide powder prepared is increased from 88.7% (Comparative Example 1) to 89.8% (Comparative Example 3), and the DPPH scavenging rate loss rate is reduced from 6.1% (Comparative Example 1) to 5.3% (Comparative Example 3), that is, both the antioxidant property and the heat resistance are improved.
[0051] In combination with Example 1, it can be seen that the addition of the amino peptidase in the complex enzyme preparation, and at the same time, the addition of the secondary dry yeast fermentation, can produce a synergistic effect, and synergistically improve the antioxidant property and heat resistance of the egg white peptide powder prepared.
[0052] In addition, it should be noted that each specific technical feature described in the above specific embodiments can be combined in any appropriate manner without contradiction, and in order to avoid unnecessary repetition, various possible combinations are not described again in the present application.
[0053] Furthermore, the various embodiments can also be combined, if not in contradiction, as long as they do not deviate from the spirit of the present application, which should be considered as disclosed.
Claims
1. A method for preparing egg white protein peptide powder, characterized in that, Includes the following steps: S1. Transfer the protein solution to a fermenter, adjust the pH to 6.0-6.9, add dry yeast, stir, and ferment to obtain fermentation liquid; S2. Transfer the fermentation broth obtained in S1 to an enzymatic hydrolysis tank, heat it to 50-55℃, add the compound enzyme preparation, and enzymatically hydrolyze for 7-10 hours, stirring for 10 minutes every hour to obtain the enzymatic hydrolysate; the compound enzyme preparation includes papain, subtilisin, trypsin and aminopeptidase. S3. Transfer the enzymatic hydrolysate obtained in S2 back to the fermenter, cool it down, add dry yeast, stir, ferment, spray dry, and place it in a hot room for 3-7 days to obtain the ovalbumin peptide powder.
2. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In the compound enzyme preparation, the mass ratio of papain, subtilisin, trypsin and aminopeptidase is (5-6):(3-4):2:
1.
3. The method for preparing ovalbumin peptide powder according to claim 1 or 2, characterized in that, In S2, the amount of the compound enzyme preparation added is 0.4-0.6% of the mass of the fermentation broth.
4. The method for preparing egg white protein peptide powder according to claim 3, characterized in that, In S2, the compound enzyme preparation is added by diluting it with room temperature water at a ratio of 1:2 and then transferring it to the enzymatic hydrolysis tank.
5. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In S1, the amount of dry yeast added is 0.7-1% of the protein solution mass; in S3, the amount of dry yeast added is 0.5-0.6% of the enzyme hydrolysate mass.
6. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In S1, the fermentation temperature is 30-37℃ and the fermentation time is 8-10h.
7. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In S3, the fermentation temperature is 30-37℃ and the fermentation time is 4-5 hours.
8. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In S3, the specific operation of spray drying is as follows: spray drying is carried out using a spray drying tower, with an inlet air temperature of 160-170℃ and an outlet air temperature of 60-80℃.
9. The method for preparing egg white protein peptide powder according to claim 1, characterized in that, In S3, the temperature of the hot chamber is 80-90℃.