Yolk immunoglobulin-proteoglycan compound for improving intestinal biological activity of yolk immunoglobulin and preparation method of yolk immunoglobulin-proteoglycan compound

Abstract

The invention relates to a yolk immunoglobulin-proteoglycan compound for improving the intestinal biological activity of yolk immunoglobulin and a preparation method of the yolk immunoglobulin-proteoglycan compound, and relates to the technical field of yolk immunoglobulin. The yolk immunoglobulin-proteoglycan compound comprises the following components: 5-20 parts of yolk immunoglobulin, 1-5 parts of tremella polysaccharide and 75-90 parts of formula protein, the formula protein comprises at least one of mung bean protein, whey protein and egg white protein. The yolk immunoglobulin-proteoglycan compound provided by the invention can effectively improve the antigen binding capacity of the yolk immunoglobulin in the intestinal tract, can be used in combination with the existing gastrointestinal dissolution technology, adopts additive components allowed to be used by the state or food-grade materials in the prescription, can be applied to food, and has the advantages of simple preparation process and low cost. And the blank of domestic food-grade enteric yolk immunoglobulin is filled.

Description

Technical Field

[0001] The present invention relates to the technical field of yolk immunoglobulin, and in particular to a yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of yolk immunoglobulin and a preparation method thereof. Background Art

[0002] Chicken yolk immunoglobulin (abbreviated as IgY, also known as yolk antibody, yolk immunoglobulin or yolk immunoglobulin) is a special immunoglobulin with important biological functions, which is widely used in medicine, food and other fields. Chicken yolk immunoglobulin is mainly composed of multiple antibody molecules, each of which is composed of two heavy chains and two light chains. It is characterized by rich natural antibody components and good stability and solubility. In addition, it also has high affinity and selective binding ability, and can react with a variety of specific antigens. Its therapeutic effects include immunomodulation, anti-oxidation, antibacterial and anti-inflammatory, and it can be used as a drug-assisted treatment for infectious diseases, tumors, etc.

[0003] However, IgY is sensitive to environmental factors such as pH, high temperature, and enzymes. Strong acidic environments and proteases can reduce the activity of IgY, affecting its actual physiological effects. Existing technologies currently focus only on enteric-coated technology based on active protein protection through the stomach, and cannot solve the stability of yolk immunoglobulin after release in the intestine. Summary of the invention

[0004] In order to solve the above problems, the present invention provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof.

[0005] In a first aspect, the present invention provides an egg yolk immunoglobulin-proteoglycan complex, wherein the egg yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight:

[0006] 5-20 parts of egg yolk immunoglobulin, 1-5 parts of Tremella polysaccharide and 75-90 parts of formula egg white;

[0007] The formula protein includes at least one of mung bean protein, whey protein and egg white protein.

[0008] Furthermore, the yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight:

[0009] 9 parts of egg yolk immunoglobulin, 1 part of Tremella polysaccharide and 90 parts of formula egg white.

[0010] Furthermore, the property parameters of the Tremella polysaccharide include: mannose as the skeleton, a molecular weight between 200000D and 800000D, and a glucuronic acid weight content between 10wt% and 25wt%.

[0011] Furthermore, the property parameters of the formula protein include: the protein weight content is above 80wt%.

[0012] In a second aspect, the present invention provides a method for preparing an egg yolk immunoglobulin-proteoglycan complex, the preparation method comprising the following steps:

[0013] The egg yolk immunoglobulin and the tremella polysaccharide are first mixed and stirred evenly to obtain an egg yolk immunoglobulin-tremella polysaccharide complex;

[0014] The egg yolk immunoglobulin-tremella polysaccharide complex and the formula protein are mixed and stirred evenly for a second time, and then freeze-dried at low temperature to obtain the egg yolk immunoglobulin-proteoglycan complex.

[0015] Furthermore, the working condition parameters for the first mixing and stirring uniformly include: a temperature of 25° C., a stirring speed of 1400-2000 rpm, and a stirring time of 15 minutes.

[0016] Furthermore, the working condition parameters for the second mixing and stirring uniformly include: temperature of 25° C., stirring speed of 1400-2000 rpm, and stirring time of 15 minutes.

[0017] Furthermore, the working condition parameters of the low-temperature freeze drying include: pre-freezing temperature -25°C--50°C, cold trap temperature -50°C, pre-freezing time 2 hours, vacuuming time 24 hours, heating, drying temperature 20°C-30°C.

[0018] The above technical solution provided by the embodiment of the present invention has at least the following advantages compared with the prior art:

[0019] The embodiment of the present invention provides a yolk immunoglobulin-proteoglycan complex and a preparation method thereof for improving the intestinal biological activity of yolk immunoglobulin. The present invention patent finds that the preparation of yolk protein microparticles coated with mung bean protein and tremella polysaccharide can increase the structural stability of yolk immunoglobulin in the intestine and improve the binding efficiency of yolk immunoglobulin with mucosal receptors. The mung bean protein used in this patent has a protein content of more than 80%, and the tremella polysaccharide has a molecular weight between 200000D-800000D. The yolk immunoglobulin microparticles designed in this patent can effectively improve the antigen binding ability of yolk immunoglobulin in the intestine, and can be used in combination with existing gastric enteric technology. The prescription uses additive ingredients allowed by the state or food-grade materials, which can be used in food, filling the gap in domestic food-grade enteric yolk immunoglobulin. DETAILED DESCRIPTION

[0020] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0022] In a first aspect, the present invention provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin. The egg yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight:

[0023] 5-20 parts of egg yolk immunoglobulin, 1-5 parts of Tremella polysaccharide and 75-90 parts of formula egg white;

[0024] The formula protein includes at least one of mung bean protein, whey protein and egg white protein.

[0025] The embodiment of the present invention provides a yolk immunoglobulin-proteoglycan complex that improves the intestinal biological activity of yolk immunoglobulin. The present invention patent finds that the preparation of yolk protein microparticles coated with mung bean protein and tremella polysaccharide can increase the structural stability of yolk immunoglobulin in the intestine and improve the binding efficiency of yolk immunoglobulin with mucosal receptors. The mung bean protein used in this patent has a protein content of more than 80%, and the tremella polysaccharide has a molecular weight between 200000D-800000D. The yolk immunoglobulin microparticles designed in this patent can effectively improve the antigen binding ability of yolk immunoglobulin in the intestine, and can be used in combination with existing gastric enteric technology. The prescription uses additive ingredients allowed by the state or food-grade materials, which can be used in food, filling the gap in domestic food-grade enteric yolk immunoglobulin.

[0026] In some specific embodiments, the yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight:

[0027] 9 parts of egg yolk immunoglobulin, 1 part of Tremella polysaccharide and 90 parts of formula egg white.

[0028] In some specific embodiments, the property parameters of the Tremella polysaccharide include: mannose as the skeleton, a molecular weight between 200000D-800000D, and a glucuronic acid weight content between 10wt%-25wt%; the property parameters of the formula protein include: a protein weight content of more than 80wt%.

[0029] In some specific embodiments, Tremella polysaccharide can be prepared by existing processes (Tremella is prepared by soaking, extracting, separating and drying) or commercial products with a Tremella polysaccharide content of not less than 80% can be directly purchased.

[0030] In some specific embodiments, the egg yolk immunoglobulin can be prepared by existing processes (egg liquid is extracted at low temperature, filtered, concentrated, and freeze-dried) or purchased from the market as a product of the same process.

[0031] In some specific embodiments, mung bean protein can be prepared by existing processes (mung bean is crushed, extracted, refined, separated, and dried), or commercial mung bean protein products with a protein content of not less than 65% can be directly purchased. In some specific embodiments, whey protein can be prepared by existing processes (cow's milk is separated and dried), or commercial products with a whey protein content of not less than 75% can be directly purchased.

[0032] In some specific embodiments, the egg white protein can be prepared by itself using existing processes (egg liquid is dissolved, separated, and dried), or a product with an egg white protein content of not less than 65% can be directly purchased from the market.

[0033] In a second aspect, the present invention provides a method for preparing an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin, the preparation method comprising the following steps:

[0034] The egg yolk immunoglobulin and the tremella polysaccharide are first mixed and stirred evenly to obtain an egg yolk immunoglobulin-tremella polysaccharide complex;

[0035] The egg yolk immunoglobulin-tremella polysaccharide complex and the formula protein are mixed and stirred evenly for a second time, and then freeze-dried at low temperature to obtain the egg yolk immunoglobulin-proteoglycan complex.

[0036] In some specific embodiments, the working condition parameters for the first mixing and stirring uniformly include: a temperature of 25° C., a stirring speed of 1400-2000 rpm, and a stirring time of 15 minutes.

[0037] In some specific embodiments, the working condition parameters for the second mixing and stirring uniformly include: a temperature of 25° C., a stirring speed of 1400-2000 rpm, and a stirring time of 15 minutes.

[0038] In some specific embodiments, the working condition parameters of the low-temperature freeze drying include: pre-freezing temperature -25°C--50°C, cold trap temperature -50°C, pre-freezing time 2 hours, vacuum time 24 hours, heating, drying temperature 20°C-30°C. It should be noted that the components and raw materials involved in the yolk immunoglobulin-proteoglycan complex and the preparation method thereof provided in the embodiment of the present invention, if there is no special limitation or specific description, can be directly commercially available products or self-made using existing public preparation methods; at the same time, the steps and parameters involved, if there is no special limitation or specific description, can be carried out according to the existing processing technology or directly using existing equipment, and the present invention document will not repeat them one by one.

[0039] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually measured according to national standards. If there is no corresponding national standard, then the conditions recommended by the manufacturer are followed.

[0040] Example 1

[0041] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 1.

[0042] Table 1

[0043]

[0044] The method for preparing the above-mentioned yolk immunoglobulin-proteoglycan complex comprises the following steps:

[0045] The egg yolk immunoglobulin and the tremella polysaccharide are first mixed and stirred evenly to obtain an egg yolk immunoglobulin-tremella polysaccharide complex;

[0046] The yolk immunoglobulin-tremella polysaccharide complex and the formula protein are mixed and stirred evenly for a second time, and then freeze-dried at low temperature to obtain the yolk immunoglobulin-proteoglycan complex;

[0047] Among them, the working condition parameters for the first mixing and stirring include: temperature of 25°C, stirring speed of 1400-2000rpm, and stirring time of 15 minutes; the working condition parameters for the second mixing and stirring include: temperature of 25°C, stirring speed of 1400-2000rpm, and stirring time of 15 minutes; the working condition parameters for low-temperature freeze-drying include: pre-freezing temperature of -25°C--50°C, cold trap temperature of -50°C, pre-freezing time of 2 hours, vacuuming time of 24 hours, heating, and drying temperature of 20°C-30°C.

[0048] Example 2

[0049] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 2.

[0050] Table 2

[0051]

[0052] The preparation method of the above-mentioned yolk immunoglobulin-proteoglycan complex is the same as that in Example 1.

[0053] Example 3

[0054] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 3.

[0055] Table 3

[0056]

[0057] The preparation method of the above-mentioned yolk immunoglobulin-proteoglycan complex is the same as that in Example 1.

[0058] Example 4

[0059] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 4.

[0060] Table 4

[0061]

[0062]

[0063] The preparation method of the above-mentioned yolk immunoglobulin-proteoglycan complex is the same as that in Example 1.

[0064] Example 5

[0065] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 5.

[0066] Table 5

[0067]

[0068] The preparation method of the above-mentioned yolk immunoglobulin-proteoglycan complex is the same as that in Example 1.

[0069] Example 6

[0070] This example provides an egg yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of egg yolk immunoglobulin and a preparation method thereof. The components and raw materials involved are shown in Table 6.

[0071] Table 6

[0072]

[0073] The preparation method of the above-mentioned yolk immunoglobulin-proteoglycan complex is the same as that in Example 1.

[0074] Comparative Example 1

[0075] This example provides an egg yolk immunoglobulin complex and a preparation method thereof, which is different from Example 1 in that no Tremella polysaccharide is added.

[0076] The components and raw materials involved in this example are shown in Table 7.

[0077] Table 7

[0078]

[0079] Comparative Example 2

[0080] This example provides an egg yolk immunoglobulin complex and a preparation method thereof, which is different from Example 1 in that no mung bean protein is added.

[0081] The components and raw materials involved in this example are shown in Table 8.

[0082] Table 8

[0083]

[0084] Test Case

[0085] Bioactivity experiment of egg yolk immunoglobulin: Take Example 1: 3 mg, Example 2: 3.75 mg, Example 3: 1.67 mg, Example 4: 1.875 mg, Example 5: 3 mg, Example 6: 3 mg, and Comparative Example 1: 3 mg, Comparative Example 2: 0.3 mg, take 3 mg of egg yolk immunoglobulin sample, add it to a beaker containing 10 ml of artificial intestinal fluid, bathe at 37°C, and treat for 15 min, 30 min, 1 h, and 2 h respectively; the obtained samples were tested for urease (Ure) antibody activity according to the requirements of enzyme-linked immunosorbent assay kit ELISA (Helicobacter pylori typing detection); at the same time, 3 mg of egg yolk immunoglobulin sample was used as a control, and urease (Ure) antibody activity was tested according to the requirements of enzyme-linked immunosorbent assay kit ELISA (Helicobacter pylori typing detection), and the results are shown in Tables 9 and 10.

[0086] Table 9 Urease (Ure) antibody activity detection value table

[0087] project 0min 15min 30min 1h 2h Yolk immunoglobulin sample 29.51 19.62 9.47 2.51 0.79 Example 1 29.13 24.32 17.36 8.07 4.63 Example 2 30.02 24.77 18.16 8.59 4.83 Example 3 28.98 22.49 14.43 5.68 2.87 Example 4 28.32 21.98 14.73 6.34 3.17 Example 5 28.64 23.28 16.58 7.10 4.18 Example 6 29.17 23.36 16.98 7.26 4.40 Comparative Example 1 28.26 22.69 15.51 6.19 2.52 Comparative Example 2 29.5 21.09 11.68 3.69 1.48

[0088] Table 10 Percentage Conversion Table

[0089]

[0090]

[0091] The results of the experiment show that the solutions of Demonstration Examples 1-4 can achieve good results, among which the formula of Demonstration Example 2 (9 yolk immunoglobulin: 1 tremella polysaccharide: 90 mung bean protein) has the best effect. The results of mung bean protein, whey protein and egg white protein are similar, with no significant difference. Using mung bean protein or tremella polysaccharide alone also has a protective effect on yolk immunoglobulin, but the effect is not as good as the combination.

[0092] In summary, the present invention provides a yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of yolk immunoglobulin and a preparation method thereof, which can effectively enhance the antigen binding ability of yolk immunoglobulin in the intestine and can be used in combination with the existing gastrointestinal enteric technology. The prescription uses additive ingredients permitted by the state or uses food-grade materials, and can be used in food, filling the gap in domestic food-grade oral yolk immunoglobulin.

[0093] Various embodiments of the present invention may be presented in the form of a range; it should be understood that the description in the form of a range is only for convenience and brevity, and should not be understood as a rigid limitation on the scope of the present invention; therefore, the range description should be considered to have specifically disclosed all possible sub-ranges and single numerical values ​​within the range. For example, the range description from 1 to 6 should be considered to have specifically disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5 and 6, which apply regardless of the range. In addition, whenever a numerical range is indicated herein, it is meant to include any cited number (fractional or integer) within the indicated range.

[0094] The foregoing is merely a specific embodiment of the present invention, which enables those skilled in the art to understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of yolk immunoglobulin, characterized in that: The yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight: 5-20 parts of egg yolk immunoglobulin, 1-5 parts of Tremella polysaccharide and 75-90 parts of formula egg white; The formula protein includes at least one of mung bean protein, whey protein and egg white protein.

2. The yolk immunoglobulin-proteoglycan complex according to claim 1, characterized in that: The yolk immunoglobulin-proteoglycan complex comprises the following components in parts by weight: 9 parts of egg yolk immunoglobulin, 1 part of Tremella polysaccharide and 90 parts of formula egg white.

3. The yolk immunoglobulin-proteoglycan complex according to claim 1, characterized in that: The property parameters of the Tremella polysaccharide include: mannose as the skeleton, a molecular weight between 200000D and 800000D, and a glucuronic acid weight content between 10wt% and 25wt%.

4. The yolk immunoglobulin-proteoglycan complex according to claim 1, characterized in that: The property parameters of the formula protein include: the protein weight content is above 80wt%.

5. A method for preparing the yolk immunoglobulin-proteoglycan complex for improving the intestinal biological activity of yolk immunoglobulin according to any one of claims 1 to 4, characterized in that: The preparation method comprises the following steps: The egg yolk immunoglobulin and the tremella polysaccharide are first mixed and stirred evenly to obtain an egg yolk immunoglobulin-tremella polysaccharide complex; The egg yolk immunoglobulin-tremella polysaccharide complex and the formula protein are mixed and stirred evenly for a second time, and then freeze-dried at low temperature to obtain the egg yolk immunoglobulin-proteoglycan complex.

6. The preparation method according to claim 5, characterized in that: The working condition parameters for the first mixing and stirring uniformly include: temperature of 25° C., stirring speed of 1400-2000 rpm, and stirring time of 15 minutes.

7. The preparation method according to claim 5, characterized in that: The working condition parameters for the second mixing and stirring uniformly include: temperature of 25° C., stirring speed of 1400-2000 rpm, and stirring time of 15 minutes.

8. The preparation method according to claim 5, characterized in that: The working condition parameters of the low-temperature freeze drying include: pre-freezing temperature -25°C--50°C, cold trap temperature -50°C, pre-freezing time 2 hours, vacuuming time 24 hours, heating, and drying temperature 20°C-30°C.

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