Albumin peptide composition capable of promoting immune cell proliferation and preparation method and application thereof

Abstract

The invention relates to the technical field of food, in particular to an albumin peptide composition capable of promoting immune cell proliferation and a preparation method and application thereof. The albumin peptide composition comprises the following components in parts by weight: 10-50 parts of albumin peptide, 5-20 parts of bovine spleen peptide, 2-10 parts of wheat oligopeptide and 1-5 parts of sea cucumber peptide. The albumin peptide composition provided by the invention has the advantages of higher stability and better immune enhancing effect.

Description

Technical Field

[0001] The present invention relates to the field of food technology, and in particular to an albumin peptide composition capable of promoting the proliferation of immune cells, and a preparation method and application thereof. Background Art

[0002] With the accelerated pace of modern life, irregular diets and lifestyles, and weakened immunity, sub-health conditions are becoming increasingly common. Weak immunity is also a potential risk factor for disease, making the body vulnerable to pathogenic invasion and damage. As a crucial protective barrier, the immune system is directly linked to human health. Strengthening immune function is crucial for preventing various diseases and promoting patient recovery. Therefore, seeking scientifically sound measures to enhance immune function is both urgent and necessary.

[0003] Functional foods are foods with special health benefits, suitable for specific populations. They regulate physiological functions without causing any acute, subacute, or chronic harm to the body. Natural functional foods with immune-modulating properties are particularly popular due to their green and safe properties. Related technical research is discussed below.

[0004] Chinese patent publication number CN109430413A discloses a composition for enhancing immunity. The composition comprises, by weight, 40-70 parts of milk powder, 0.05-2 parts of lactoferrin, 1-10 parts of bovine colostrum, 0.1-5 parts of yeast β-glucan, 5-20 parts of concentrated whey protein and 0.1-4 parts of ferric pyrophosphate. There is no stability conflict between the components of the provided composition, and the components can achieve the relatively best effect at a specific ratio. The composition can be used to enhance immunity and solve the common problem of low immunity and poor resistance among the current population.

[0005] Chinese Patent Publication No. CN1920049A discloses a small-molecule bioactive peptide derived from a mixture of soy protein, whey protein, egg, and milk. Over 90% of the active peptides have a molecular weight of less than 5 kDa. This small-molecule bioactive peptide can enhance immunity, promote wound healing, and improve physical fitness.

[0006] While these existing technologies have achieved some improvements in boosting immunity and enhancing physical fitness, the relatively limited functional ingredients they employ make it difficult to fully activate the immune system, resulting in limited improvements. Some existing technologies suffer from low absorption rates, insufficient bioavailability, and even the risk of side effects. Furthermore, traditional Chinese herbal formulas (such as Ganoderma lucidum spore powder) are slow to take effect and lack the ability to precisely regulate immune factors.

[0007] Existing technologies (Chen Xuehua, Yang Wangen. Research progress on functional foods that enhance the body's immunity [J]. Grain and Oils, 2022, 35(8): 20-22) also point out that there are some problems with current immune-enhancing products. For example, the focus is currently on the development of functional foods that enhance nonspecific immunity, while there is less research on functional foods that enhance specific immunity; basic research is insufficient, and there are fewer categories of functional factors that enhance immunity.

[0008] Therefore, it is very necessary to develop an albumin peptide composition that can promote the proliferation of immune cells and a preparation method and application thereof that can solve the above technical problems. Summary of the Invention

[0009] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an albumin peptide composition with high stability, good immune-enhancing effect and the ability to promote immune cell proliferation, as well as a preparation method and application thereof.

[0010] The present invention is achieved through the following technical solutions: The first aspect of the present invention provides an albumin peptide composition that can promote the production of immune factors, which comprises the following components in parts by weight: 10-50 parts of albumin peptide, 5-20 parts of bovine spleen peptide, 2-10 parts of wheat oligopeptide and 1-5 parts of sea cucumber peptide.

[0011] As an embodiment of the present invention, the albumin peptide composition further comprises yeast β-glucan.

[0012] Preferably, the amount of yeast β-glucan used is 1-5 parts.

[0013] As an embodiment of the present invention, the mass ratio of the albumin peptide, bovine spleen peptide and wheat oligopeptide is preferably 5:2:1.

[0014] As an embodiment of the present invention, the average molecular weight of the albumin peptide is 500-600, wherein the mass percentage of polypeptides with a relative molecular mass between 180-500 is above 50%.

[0015] The raw materials in the composition of the present invention can be directly purchased from the market or prepared by the user.

[0016] As an embodiment of the present invention, the albumin peptide extraction process includes the following steps: adding water to egg white for homogenization, then adding biological enzymes for enzymatic hydrolysis, and solid-liquid separation to obtain albumin peptide.

[0017] As a preferred embodiment of the present invention, egg white is homogenized by adding water, and the egg white concentration is 5-10% w / v.

[0018] As a preferred embodiment of the present invention, the biological enzymes include but are not limited to trypsin, chymotrypsin, pepsin, alkaline protease, neutral protease, papain, flavor protease, etc., and can be used alone or in combination.

[0019] As a preferred embodiment of the present invention, before the enzymatic hydrolysis is performed, the pH is adjusted to 7.0-8.5.

[0020] As a preferred embodiment of the present invention, the enzymatic hydrolysis temperature is 40-50° C., and the enzymatic hydrolysis time is 4-5 h.

[0021] As a preferred embodiment of the present invention, enzyme inactivation treatment is performed after the enzymatic hydrolysis is completed.

[0022] As a preferred embodiment of the present invention, after the solid-liquid separation, the liquid can be subjected to at least one of decolorization, deodorization, concentration, sterilization, and drying. The drying method is preferably spray drying.

[0023] More preferably, the preparation method of the albumin peptide comprises the following steps: S1: adding egg white to water for homogenization to a concentration of 5-10% w / v, adjusting the pH, adding biological enzymes for enzymatic hydrolysis at a temperature of 40-50°C for 4-5 hours, heating at 80-100°C for 5-15 minutes to inactivate the enzyme, and performing solid-liquid separation to obtain an enzymatic hydrolyzate; S2: The enzymatic hydrolysate is decolorized and deodorized, filtered with activated carbon, concentrated, sterilized, and spray-dried to obtain albumin peptide.

[0024] In the above preparation process, the solid-liquid separation method in step S1 includes centrifugation or filtration (such as diatomaceous earth filtration) to remove the precipitate and obtain a clarified enzymatic hydrolysate.

[0025] In the above preparation process, the decolorization and deodorization in step S2 can be carried out by conventional treatment methods.

[0026] In the above preparation process, the sterilization in step S2 can be carried out by a general sterilization method, such as pasteurization (e.g., 72°C, 15s) or membrane filtration sterilization (0.22μm filter membrane).

[0027] A second aspect of the present invention provides a method for preparing the above-mentioned albumin peptide composition, comprising the following steps: (1) mixing albumin peptide, bovine spleen peptide and wheat oligopeptide to obtain mixture A; (2) mixing the sea cucumber peptide and yeast β-glucan to obtain a mixture B; (3) Mix mixture A and mixture B in equal amounts.

[0028] As an embodiment of the present invention, in step (1), the albumin peptide, bovine spleen peptide and wheat oligopeptide are mixed, passed through a 60-100 mesh sieve, and dried to obtain a mixture A.

[0029] Preferably, the drying method in step (1) is vacuum drying.

[0030] Preferably, the drying temperature in step (1) is 50-80°C.

[0031] Preferably, the moisture content of the mixture A in step (1) is ≤5%.

[0032] As an embodiment of the present invention, step (2) is performed by mixing under an inert atmosphere.

[0033] As an embodiment of the present invention, in step (3), the mixture is mixed in equal amounts, and the mixing times are preferably 3 times to avoid agglomeration.

[0034] As an embodiment of the present invention, when the composition does not contain yeast β-glucan, the preparation method is as follows: (1) mixing albumin peptide, bovine spleen peptide and wheat oligopeptide to obtain mixture A; (2) Mix the mixture A and the sea cucumber peptide in equal amounts.

[0035] Wherein, as an embodiment of the present invention, in step (1), the albumin peptide, bovine spleen peptide and wheat oligopeptide are mixed and then passed through a 60-100 mesh sieve and dried to obtain a mixture A.

[0036] Preferably, the drying method in step (1) is vacuum drying.

[0037] Preferably, the drying temperature in step (1) is 50-80°C.

[0038] Preferably, the moisture content of the mixture A in step (1) is ≤5%.

[0039] In step (2), the mixture is mixed in equal amounts, preferably three times, to avoid agglomeration.

[0040] A third aspect of the present invention provides a use of the above-mentioned albumin peptide composition or the albumin peptide composition prepared by the above-mentioned preparation method in the preparation of an immunity-enhancing product.

[0041] The beneficial effects of the present invention are: The present invention improves the immune-enhancing effect of the composition by optimizing the raw material composition and raw material dosage ratio. The present invention utilizes albumin peptide, bovine spleen peptide, wheat oligopeptide, and sea cucumber peptide to regulate immunity. These components exhibit significant synergistic effects in enhancing immunity. Furthermore, yeast β-glucan is added as an active polysaccharide to synergistically enhance immunity. The rational formulation results in a more pronounced immune-enhancing effect.

[0042] The present invention optimizes the preparation process of the composition, thereby enhancing the immune effect of the composition and improving the stability of the product. DETAILED DESCRIPTION

[0043] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are merely exemplary and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solutions of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, and such modifications and replacements fall within the scope of protection of the present invention.

[0044] The raw material information used in the following examples and comparative examples is shown in Table 1.

[0045] Table 1 Raw material information

[0046] The formulations of Examples 1 to 3 and Comparative Examples 1 to 8 are shown in Table 2.

[0047] Table 2 Formulations of Examples 1-3 and Comparative Examples 1-8

[0048] The preparation methods of the albumin peptide compositions of Examples 1 to 3 and Comparative Examples 1 to 4 above were prepared according to the following steps: (1) Mix the formulated amounts of albumin peptide (or whey protein peptide), bovine spleen peptide (0 parts in comparative example 2) and wheat oligopeptide (0 parts in comparative example 3), pass through a 100-mesh sieve, and vacuum dry at 60°C until the moisture content is ≤5% to obtain a mixture A; (2) Mixing the formulated amount of sea cucumber peptide and yeast β-glucan under nitrogen protection to obtain mixture B; (3) Mix mixture A and mixture B in equal amounts in three times to obtain the product.

[0049] The preparation method of the albumin peptide composition of the above comparative example 5 comprises the following steps: (1) Mix the formulated amount of albumin peptide, bovine spleen peptide and wheat oligopeptide, pass through a 100-mesh sieve, and vacuum dry at 60°C until the moisture content is ≤5% to obtain mixture A; (2) Mix the mixture A and yeast β-glucan in equal amounts for three times to obtain the product.

[0050] The preparation method of the albumin peptide composition of the above comparative example 6 comprises the following steps: mixing the formulated amount of sea cucumber peptide and yeast β-glucan under nitrogen protection to obtain the albumin peptide composition.

[0051] The preparation method of the albumin peptide composition of Comparative Example 7 above comprises the following steps: (1) Mix the formulated amount of albumin peptide, bovine spleen peptide and wheat oligopeptide, pass through a 100-mesh sieve, and vacuum dry at 60°C until the moisture content is ≤5% to obtain mixture A; (2) Mix the mixture A and the sea cucumber peptide in equal amounts for three times to obtain the product.

[0052] Comparative Example 9 The only difference from Example 1 is the preparation method of the composition, which is to directly mix the components evenly to obtain the composition.

[0053] Test Example 1 Stability Test Test Method: 40g of the albumin peptide composition powder was placed flat at 60°C and 30% relative humidity. Samples were taken on the first, seventh, and fourteenth days and their appearance was observed. The results are shown in Table 3.

[0054] Table 3 Stability results

[0055] Test Example 2: Immunity Enhancement Test First prepare the three stock solutions for preparing buffered water. The configuration method is as follows: 100×E2A stock solution: Dissolve 140 g NaCl, 6 g KCl, 19.2 g MgSO4, 3.3 g KH2PO4, and 1.1 g Na2HPO4 in 1600 mL sterile deionized water and store at 4°C for long-term storage.

[0056] 500×E2B storage solution: Dissolve 11 g of CaCl2 in 200 mL of sterile deionized water and store at -20°C for a long term.

[0057] 500×E2C storage solution: Dissolve 6 g of NaHCO3 in 200 mL of sterile deionized water and store at -20°C for a long time.

[0058] Buffered water formula: Take 100 mL of 100×E2A stock solution, 20 mL of 500×E2B stock solution, and 20 mL of 500×E2C stock solution and add them to 19 L of sterile deionized water. Adjust the pH to 7.0 and make up to 20 L to prepare buffered water. After high-temperature sterilization, store at room temperature for a short period of time for subsequent experiments.

[0059] 2.1 Zebrafish Immunity Enhancement Project - T Cell Detection Experimental instruments and reagents: Stereo microscope (SZX7, Olympus, Japan), berbamine hydrochloride, and the albumin peptide compositions of Examples 1-3 and Comparative Examples 1-9.

[0060] Detection system and sample size: Experimental system: Transgenic T cell fluorescent zebrafish; Zebrafish age: 3 days after fertilization; Experimental sample size per group: 10 fish per well, 1 well per group.

[0061] Experimental process: The zebrafish were divided into 14 groups: a blank control group, a positive control group, and groups treated with samples from Examples and Comparative Examples. The albumin peptide compositions from Examples 1-3 and Comparative Examples 1-9 were diluted with buffered water using the half-dilution method to prepare uniformly dispersed suspensions at concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, and 250 μg / mL, respectively, to serve as the test sample solutions. The blank control group received no treatment, while the positive control group was supplemented with berbamine hydrochloride (5 μg / mL). The sample solutions from Examples 1-3 and Comparative Examples 1-9 were added to the sample treatment groups, respectively. After 24 hours of treatment with the test sample solutions, the fluorescence intensity of zebrafish neutrophils was measured. The 500 μg / mL concentration of the sample treatment group showed no significant difference compared to the blank control group. Therefore, 500 μg / mL is the maximum safe concentration, and the 500 μg / mL concentration was used for testing in all Examples and Comparative Examples. The results are shown in Table 4.

[0062] Table 4 Fluorescence intensity of zebrafish T cells

[0063] T test method was used to compare with the blank control group, ***, P <0.001;**, P <0.01;*, P <0.05;ns, P >0.05. Compared with the group in Example 1, ##, P <0.01;#, P <0.05.

[0064] 2.2 Zebrafish Immunity Enhancement Project - Macrophage Detection Experimental instruments and reagents: Stereo microscope (SZX7, Olympus, Japan), berbamine hydrochloride, and the albumin peptide compositions of Examples 1-3 and Comparative Examples 1-9.

[0065] Detection system and sample size: Experimental system: Transgenic giant cell fluorescent zebrafish; Zebrafish age: 3 days after fertilization; The number of experimental samples per group: 10 per well, 1 well per group.

[0066] Experimental process: The zebrafish were divided into 14 groups, namely a blank control group, a positive control group, and a sample treatment group. The albumin peptide composition test substances of Examples 1-3 and Comparative Examples 1-9 were diluted with buffered water by the half-dilution method to prepare a group of uniformly dispersed suspensions with concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, and 250 μg / mL, respectively, as the test sample solutions. Among them, the blank control group was not treated in any way. The positive control group and the sample treatment group were added with berbamine hydrochloride (5 μg / mL) and the test sample solutions of the above concentrations, respectively. After 24 hours of incubation, the fluorescence intensity of the zebrafish macrophages was tested. There was no significant difference between the sample treatment group with a concentration of 500 μg / mL and the blank control group. Therefore, the concentration of 500 μg / mL is the maximum safe concentration. The concentration of 500 μg / mL was used for testing in each Example and Comparative Example. The results are shown in Table 5.

[0067] Table 5 Fluorescence intensity of zebrafish macrophages

[0068] T test method was used to compare with the blank control group, ***, P <0.001;**, P <0.01;*, P <0.05;ns, P >0.05. Compared with the group in Example 1, ##, P <0.01;#, P <0.05.

[0069] 2.3 Zebrafish Immunity Enhancement Project - Neutrophil Detection Experimental instruments and reagents: Stereo microscope (SZX7, Olympus, Japan), berbamine hydrochloride, and the albumin peptide compositions of Examples 1-3 and Comparative Examples 1-9.

[0070] Detection system and sample size: Experimental system: Transgenic neutrophil fluorescent zebrafish; Zebrafish age: 3 days after fertilization; The number of experimental samples per group: 10 per well, 1 well per group.

[0071] Experimental process: The zebrafish were divided into 14 groups, namely a blank control group, a positive control group, and a sample treatment group. The albumin peptide composition test substances of Examples 1-3 and Comparative Examples 1-9 were diluted with buffered water by the half-dilution method to prepare a group of uniformly dispersed suspensions with concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, and 250 μg / mL, respectively, as the test sample solutions. Among them, the blank control group did not receive any treatment, and the positive control group and the sample treatment group were added with berbamine hydrochloride (5 μg / mL) and the test sample solutions of the above concentrations, respectively. After 24 hours of incubation, the neutrophils of the zebrafish in each group were photographed using a fluorescence microscope, and the number of fluorescent neutrophils was counted. The results are expressed as mean ± standard deviation. There was no significant difference between the sample treatment group at a concentration of 500 μg / mL and the blank control group. Therefore, a concentration of 500 μg / mL was the maximum safe concentration. Each embodiment and comparative example was tested at a concentration of 500 μg / mL. The results are shown in Table 6.

[0072] Table 6 Number of neutrophils in zebrafish (units)

[0073] T test method was used to compare with the blank control group, ***, P <0.001;**, P <0.01;*, P <0.05;ns, P >0.05. Compared with the group in Example 1, ##, P <0.01;#, P <0.05.

[0074] The above detailed description is a specific description of one feasible embodiment of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not depart from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. An albumin peptide composition capable of promoting the proliferation of immune cells, characterized in that: The invention comprises the following components in parts by weight: 10-50 parts of albumin peptide, 5-20 parts of bovine spleen peptide, 2-10 parts of wheat oligopeptide and 1-5 parts of sea cucumber peptide.

2. The albumin peptide composition according to claim 1, characterized in that The albumin peptide composition also includes yeast beta-glucan.

3. The albumin peptide composition according to claim 2, characterized in that The amount of yeast beta-glucan used is 1-5 parts.

4. The albumin peptide composition according to claim 1, characterized in that The mass ratio of the albumin peptide, bovine spleen peptide and wheat oligopeptide is 5:2:

1.

5. The albumin peptide composition according to claim 1, characterized in that The average molecular weight of the albumin peptide is 500-600, and the mass percentage of polypeptides with relative molecular masses between 180-500 in the albumin peptide is above 50%.

6. A method for preparing the albumin peptide composition according to claim 1 or any one of claims 4-5, characterized in that: The steps include: Step 1: mixing albumin peptide, bovine spleen peptide and wheat oligopeptide to obtain mixture A; Step 2: Mix the mixture A and the sea cucumber peptide in equal amounts.

7. A method for preparing the albumin peptide composition according to any one of claims 2-3, characterized in that: The steps include: (1) mixing albumin peptide, bovine spleen peptide and wheat oligopeptide to obtain mixture A; (2) mixing the sea cucumber peptide and yeast β-glucan to obtain a mixture B; (3) Mix mixture A and mixture B in equal amounts.

8. The preparation method according to claim 7, characterized in that In step (1), the albumin peptide, bovine spleen peptide and wheat oligopeptide are mixed, passed through a 60-100 mesh sieve, and dried to obtain a mixture A; and in step (2), the mixture is mixed under an inert atmosphere.

9. The preparation method according to claim 8, characterized in that The drying method in step (1) is vacuum drying, the drying temperature is 50-80°C, and the moisture content of the mixture A is ≤5%.

10. Use of the albumin peptide composition according to any one of claims 1 to 5 or the albumin peptide composition prepared by the preparation method according to any one of claims 6 to 9 in preparing a product that enhances immunity.

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