Albumin peptide compositions that can promote immune cell proliferation, methods of making and using the same

Abstract

The present application relates to the field of food technology, in particular to a kind of albumin peptide composition for promoting immune cell proliferation and its preparation method and application.The albumin peptide composition includes the following components by weight fraction: albumin peptide 10-50 parts, bovine spleen peptide 5-20 parts, wheat oligopeptide 2-10 parts and sea cucumber peptide 1-5 parts.The albumin peptide composition provided by the present application has the advantages of higher stability and better immune-enhancing effect.

Description

TECHNICAL FIELD

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

[0002] With the acceleration of modern social life pace, irregular diet and living schedule, low immunity has become a common sub-health state of modern people, and weak immunity is also a potential factor for disease occurrence. The human body is vulnerable to pathogenic bacteria invasion in this state, causing damage to the body. The immune system, as an important protective barrier of the human body, is directly related to the health of the human body. Enhancing the immune function of the body is the key to preventing various diseases and patient recovery. Therefore, it is urgent and necessary to seek scientific and reasonable measures to enhance the immune function of the body.

[0003] Functional food is a kind of food with special health care function, which is suitable for specific population to eat, has the function of regulating the body's physiological function, and does not produce any acute, subacute or chronic harm to the human body. Natural functional food with immune regulation function, green and safe, is more favored by people, and the related technical research is as follows.

[0004] Chinese patent publication No. CN109430413A discloses a composition for enhancing immunity, which comprises milk powder 40-70 parts, lactoferrin 0.05-2 parts, bovine colostrum 1-10 parts, yeast beta-glucan 0.1-5 parts, concentrated whey protein 5-20 parts and pyrophosphate iron 0.1-4 parts by weight. There is no stability conflict between the components of the provided composition, and the composition can play the best effect under a specific ratio, and can be used for enhancing immunity to solve the common problem of low immunity and poor resistance of the present population.

[0005] Chinese patent publication No. CN1920049A discloses a small molecule bioactive peptide, which is derived from a mixture containing soybean protein, whey protein, chicken egg and milk. More than 90% of the active peptides in the small molecule bioactive peptide have a molecular weight less than 5KDa. The small molecule bioactive peptide can improve immunity, promote wound healing of body tissues and enhance physical fitness.

[0006] Although the above prior art has obtained some improvement in improving immunity and enhancing physical fitness, the functional components used are relatively single, which is difficult to activate the immune system comprehensively, and the improvement degree is still limited. The absorption rate of some prior art is low, the bioavailability is insufficient, and there may be side effects. In addition, traditional Chinese medicine compounds (such as ganoderma lucidum spore powder) have slow effect and lack precise immune factor regulation ability.

[0007] The prior art (Chen X H, Yang W G. Research progress of functional foods for enhancing immune function [J]. Cereals and oils, 2022, 35 (8): 20-22) also points out that there are some problems in the current immune-enhancing products, such as the current main focus on the development of functional foods for enhancing non-specific immunity, and less research on functional foods for enhancing specific immunity; the basic research is not enough, and the category of functional factors for enhancing immunity is less.

[0008] Therefore, it is necessary to develop an albumin peptide composition capable of promoting immune cell proliferation, a preparation method and application thereof, which can solve the above technical problems. SUMMARY

[0009] The purpose of the present application is to overcome the shortcomings of the prior art and provide an albumin peptide composition capable of promoting immune cell proliferation, which has high stability and good immune-enhancing effect, and a preparation method and application thereof.

[0010] The present application is realized by the following technical solutions:

[0011] The present application provides an albumin peptide composition capable of promoting immune factor generation, comprising the following components by weight fraction: 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.

[0012] As an embodiment of the present application, the albumin peptide composition further comprises yeast beta-glucan.

[0013] Preferably, the amount of yeast beta-glucan is 1-5 parts.

[0014] As an embodiment of the present application, the mass ratio of albumin peptide, bovine spleen peptide and wheat oligopeptide is preferably 5:2:1.

[0015] As an embodiment of the present application, the average molecular weight of the albumin peptide is 500-600, and the mass percentage of polypeptides with a relative molecular mass of 180-500 is more than 50%.

[0016] Each raw material in the composition of the present application can be directly purchased from commercially available products or can be prepared by itself.

[0017] As an embodiment of the present application, the extraction process of the albumin peptide comprises the following steps: homogenizing chicken egg white with water, then adding biological enzyme for enzymolysis, solid-liquid separation, and obtaining albumin peptide.

[0018] As a preferred embodiment of the present application, the chicken egg white is homogenized with water, and the concentration of chicken egg white is 5-10% w / v.

[0019] As a preferred embodiment of the present application, the biological enzyme includes but is not limited to trypsin, chymotrypsin, pepsin, alkaline protease, neutral protease, papain, flavor protease, etc., and can be used alone or in combination.

[0020] As a preferred embodiment of the present application, the pH is adjusted to 7.0-8.5 before the enzymolysis.

[0021] As a preferred embodiment of the present application, the temperature of the enzymolysis is 40-50℃, and the time of the enzymolysis is 4-5h.

[0022] As a preferred embodiment of the present application, the enzyme inactivation treatment is performed after the enzymolysis.

[0023] As a preferred embodiment of the present application, at least one of decolorization, deodorization, concentration, sterilization and drying can be performed on the liquid after the solid-liquid separation. The drying mode is preferably spray drying.

[0024] More preferably, the preparation method of the albumin peptide comprises the following steps:

[0025] S1: homogenate the egg white in water, the concentration of the egg white is 5-10% w / v, adjust the pH, add biological enzyme for enzymolysis, the temperature of the enzymolysis is 40-50℃, the time of the enzymolysis is 4-5h, after the enzymolysis, heat at 80-100℃ for 5-15min for enzyme inactivation, solid-liquid separation, and obtain the enzymolysis liquid;

[0026] S2: decolorize and deodorize the enzymolysis liquid, filter with activated carbon, concentrate, sterilize, and spray dry to obtain the albumin peptide.

[0027] In the above preparation process, the solid-liquid separation mode in step S1 includes centrifugation or filtration (such as diatomite filtration), and the precipitate is removed to obtain the clear enzymolysis liquid.

[0028] In the above preparation process, the decolorization and deodorization in step S2 can be performed by using a conventional treatment method.

[0029] In the above preparation process, the sterilization in step S2 can be performed by using a general sterilization method, such as commonly used pasteurization (such as 72℃, 15s) or membrane filtration sterilization (0.22μm filter membrane).

[0030] The second aspect of the present application provides a preparation method of the above-mentioned albumin peptide composition, comprising the following steps:

[0031] (1) mix the albumin peptide, the bovine spleen peptide and the wheat oligopeptide to obtain a mixture A;

[0032] (2) mixing the sea cucumber peptide with the yeast β-glucan to obtain a mixture B;

[0033] (3) mixing the mixture A and the mixture B in an equal amount and adding manner, to obtain the albumin peptide composition.

[0034] As an embodiment of the present application, the albumin peptide, the bovine spleen peptide and the wheat oligopeptide are mixed and then sieved through a 60-100 mesh sieve in step (1), and then dried to obtain the mixture A.

[0035] Preferably, the drying method in step (1) is vacuum drying.

[0036] Preferably, the drying temperature in step (1) is 50-80℃.

[0037] Preferably, the moisture content of the mixture A in step (1) is ≤5%.

[0038] As an embodiment of the present application, the mixing in step (2) is carried out in an inert atmosphere.

[0039] As an embodiment of the present application, the mixing in step (3) is carried out in an equal amount and adding manner, and the mixing number is preferably 3 times to avoid caking.

[0040] As an embodiment of the present application, when the composition does not contain the yeast β-glucan, the preparation method is as follows:

[0041] (1) mixing the albumin peptide, the bovine spleen peptide and the wheat oligopeptide to obtain a mixture A;

[0042] (2) mixing the mixture A and the sea cucumber peptide in an equal amount and adding manner, to obtain the albumin peptide composition.

[0043] As an embodiment of the present application, the albumin peptide, the bovine spleen peptide and the wheat oligopeptide are mixed and then sieved through a 60-100 mesh sieve in step (1), and then dried to obtain the mixture A.

[0044] Preferably, the drying method in step (1) is vacuum drying.

[0045] Preferably, the drying temperature in step (1) is 50-80℃.

[0046] Preferably, the moisture content of the mixture A in step (1) is ≤5%.

[0047] The mixing in step (2) is carried out in an equal amount and adding manner, and the mixing number is preferably 3 times to avoid caking.

[0048] The present application provides an application of the albumin peptide composition or the albumin peptide composition prepared by the preparation method in the preparation of an immune-enhancing product.

[0049] The beneficial effects of the present application are:

[0050] The present application improves the immune-enhancing effect of the composition by optimizing the composition of raw materials and the ratio of raw material usage. The present application uses albumin peptide, bovine spleen peptide, wheat oligopeptide and sea cucumber peptide to regulate immunity, and the synergistic effect of each component in enhancing immunity is significant. On this basis, yeast beta-glucan is added as an active polysaccharide to synergistically enhance immunity. The formula is reasonable, and the immune-enhancing effect is more significant.

[0051] The present application optimizes the preparation process of the composition, thereby improving the stability of the product while enhancing the immune effect of the composition. DETAILED DESCRIPTION

[0052] The advantages and characteristics of the present application will become clearer with the description. However, these examples are only exemplary and do not constitute any limitation on the scope of the present application. Those skilled in the art should understand that the details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and such modifications and replacements all fall within the protection scope of the present application.

[0053] The raw material information used in the following examples and comparative examples is shown in Table 1.

[0054] Table 1 Raw material information

[0055]

[0056] The formulations of Examples 1-3 and Comparative Examples 1-8 are shown in Table 2.

[0057] Table 2 Formulations of Examples 1-3 and Comparative Examples 1-8

[0058]

[0059] The preparation method of the albumin peptide composition of Examples 1-3 and Comparative Examples 1-4 above is prepared according to the following steps:

[0060] (1) Mix the formula amount of albumin peptide (or whey protein peptide), bovine spleen peptide (0 parts for Comparative Example 2) and wheat oligopeptide (0 parts for 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;

[0061] (2) Mix the formula amount of sea cucumber peptide and yeast beta-glucan under nitrogen protection to obtain a mixture B;

[0062] (3) Mix mixture A and mixture B in an equal amount of additive manner in three times.

[0063] The preparation method of the above Comparative Example 5 albumin peptide composition comprises the following steps:

[0064] (1) Mix the formula 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;

[0065] (2) Mix mixture A with yeast β-glucan in an equal incremental manner for three times, and obtain.

[0066] The preparation method of the above Comparative Example 6 albumin peptide composition comprises the following steps: mix the formula amount of sea cucumber peptide with yeast β-glucan under nitrogen protection, and obtain.

[0067] The preparation method of the above Comparative Example 7 albumin peptide composition comprises the following steps:

[0068] (1) Mix the formula 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;

[0069] (2) Mix mixture A with sea cucumber peptide in an equal incremental manner for three times, and obtain.

[0070] Comparative Example 9

[0071] The difference from Example 1 is only in the preparation method of the composition, directly mix the components uniformly, and obtain.

[0072] Test Example 1 Stability Test

[0073] Test method: Take the same gram weight (40g) of the albumin peptide composition powder and place it flat at 60°C and a relative humidity of 30%, take samples on the 1st day, 7th day and 14th day, and observe the appearance. The results are shown in Table 3.

[0074] Table 3 Stability Results

[0075]

[0076] Test Example 2 Immunity Enhancement Test

[0077] First prepare 3 kinds of buffer water solution mother liquor, and the configuration method is as follows:

[0078] 100×E2A storage solution: take 140g NaCl, 6g KCl, 19.2g MgSO4, 3.3g KH2PO4, 1.1g Na2HPO4, and dissolve in 1600mL sterile deionized water, store at 4°C for a long time.

[0079] 500x E2B stock solution: 11 g CaCl2 was dissolved in 200 mL sterile deionized water, and stored at -20 °C for long-term storage.

[0080] 500x E2C stock solution: 6 g NaHCO3 was dissolved in 200 mL sterile deionized water, and stored at -20 °C for long-term storage.

[0081] Buffer water formula:

[0082] 100 mL 100x E2A stock solution, 20 mL 500x E2B stock solution, and 20 mL 500x E2C stock solution were added to 19 L sterile deionized water, and the pH was adjusted to 7.0 and then diluted to 20 L to prepare buffer water. After high-temperature sterilization, the buffer water was stored at room temperature for short-term storage and used for subsequent experiments.

[0083] 2.1 Zebrafish enhanced immunity project-T cell detection

[0084] Experimental instruments and reagents:

[0085] Olympus SZX7 body microscope (Olympus, Japan), berberine hydrochloride, and albumin peptide compositions of Examples 1-3 and Comparative Examples 1-9.

[0086] Detection system and sample size:

[0087] Experimental system: transgenic T cell fluorescent strain zebrafish; zebrafish age: 3 days after fertilization; sample size per group: 10 fish per well, 1 well per group.

[0088] Experimental procedure:

[0089] The test zebrafish were divided into 14 groups, which were blank control group, positive control group, and sample treatment groups of Examples and Comparative Examples. The albumin peptide compositions of Examples 1-3 and Comparative Examples 1-9 were diluted with buffer water by half-dilution method to prepare a uniformly dispersed suspension with concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, and 250 μg / mL, respectively, as the test sample solution. Among them, the blank control group was not treated, the positive control group was added with berberine hydrochloride (5 μg / mL), and the sample treatment groups of Examples and Comparative Examples were added with the test sample solution of Examples 1-3 and Comparative Examples 1-9, respectively. After 24 hours of treatment with the test sample solution, the fluorescence intensity of neutrophils of zebrafish was detected. The sample treatment group with a concentration of 500 μg / mL had no significant difference compared with the blank control group, so the concentration of 500 μg / mL was the maximum safe concentration, and each Example and Comparative Example was tested at a concentration of 500 μg / mL. The results are shown in Table 4.

[0090] Table 4 Zebrafish T cell fluorescence intensity

[0091]

[0092] ***, P <0.001; **, P <0.01; *, P <0.05; ns, P >0.05. Compared with Example 1 group, ## P <0.01; #, P <0.05.

[0093] 2.2 Zebrafish enhanced immunity project - macrophage detection

[0094] Experimental instruments and reagents:

[0095] Olympus SZX7 body microscope (Olympus, Japan), berberine hydrochloride, Example 1-3 and Comparative Example 1-9 albumin peptide composition.

[0096] Detection system and sample size:

[0097] Experimental system: transgenic macrophage fluorescence strain zebrafish;

[0098] Zebrafish age: 3 days after fertilization;

[0099] Sample size of each group: 10 per well, 1 hole per group.

[0100] Experimental procedure:

[0101] The tested zebrafish were divided into 14 groups, namely blank control group, positive control group and sample treatment group. The Example 1-3 and Comparative Example 1-9 albumin peptide composition test substances were diluted with buffer water by half dilution method to prepare a uniformly dispersed suspension with concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, 250 μg / mL as the sample solution to be tested. Among them, the blank control group was not treated, the positive control group and the sample treatment group were added with berberine hydrochloride (5 μg / mL) and the above concentration of sample solution to be tested. After 24 hours of culture, the fluorescence intensity of macrophages in zebrafish was detected. The sample treatment group with a concentration of 500 μg / mL had no significant difference compared with the blank control group, so the maximum safe concentration was 500 μg / mL, and each example and comparative example was tested at a concentration of 500 μg / mL, and the results are shown in Table 5.

[0102] Table 5 Zebrafish macrophage fluorescence intensity

[0103]

[0104] T-test, compared with the blank control group, ***, P <0.001; **, P <0.01; *, P <0.05; ns, P >0.05. Compared with Example 1 group, ##, P <0.01; #, P <0.05.

[0105] 2.3 Zebrafish enhanced immunity project - neutrophil detection

[0106] Experimental instruments and reagents:

[0107] Olympus, Japan), berberine hydrochloride, Example 1-3 and Comparative Example 1-9 albumin peptide composition.

[0108] Detection system and sample size:

[0109] Experimental system: transgenic neutrophil fluorescence strain zebrafish;

[0110] Zebrafish age: 3 days after fertilization;

[0111] Sample size of each group: 10 per well, 1 hole per group.

[0112] Experimental procedure:

[0113] The tested zebrafish were divided into 14 groups, namely blank control group, positive control group and sample treatment group. The Example 1-3 and Comparative Example 1-9 albumin peptide composition test substances were diluted with buffer water by half dilution method to prepare a uniformly dispersed suspension with concentrations of 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, 250 μg / mL as the sample solution to be tested. Among them, the blank control group was not treated, the positive control group and the sample treatment group were added with berberine hydrochloride (5 μg / mL) and the above concentration of sample solution to be tested. After 24 hours of culture, the neutrophils of zebrafish were photographed by fluorescence microscope, and the number of neutrophils with fluorescence was counted, and the results were expressed as mean ± standard deviation. The sample treatment group with a concentration of 500 μg / mL had no significant difference compared with the blank control group, so the maximum safe concentration was 500 μg / mL, and each example and comparative example was tested at a concentration of 500 μg / mL, and the results are shown in Table 6.

[0114] Table 6 Number of zebrafish neutrophils (pieces)

[0115]

[0116] ​​​​​​​​***, P <0.001; **, P <0.01; *, P <0.05; ns, P >0.05. Compared with Example 1 group, ##, P <0.01; #, P <0.05.

[0117] The above detailed description is a specific description of one of the possible embodiments of the present application, which is not intended to limit the patent scope of the present application, and any equivalent implementation or change that does not deviate from the present application shall be included in the scope of the technical solutions of the present application.

Claims

1. An albumin peptide composition capable of promoting proliferation of immune cells, characterized in that, The albumin peptide composition comprises, 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 of claim 1, wherein, The albumin peptide composition further comprises yeast beta-glucan.

3. The albumin peptide composition of claim 2, wherein, The amount of the yeast beta-glucan is 1-5 parts.

4. The albumin peptide composition of claim 1, wherein, The mass ratio of the albumin peptide, the bovine spleen peptide, and the wheat oligopeptide is 5:2:

1.

5. The albumin peptide composition of claim 1, wherein, The average molecular weight of the albumin peptide is 500-600, and the mass percentage of polypeptides with a relative molecular weight of 180-500 in the albumin peptide is more than 50%.

6. A method of preparing the albumin peptide composition according to any one of claims 1 or 4 to 5, characterized in that, The method comprises the following steps: Step 1: mixing the albumin peptide, the bovine spleen peptide, and the wheat oligopeptide to obtain a mixture A; Step 2: mixing the mixture A and the sea cucumber peptide in an equal amount in a stepwise manner, and obtaining the albumin peptide composition.

7. A method of preparing the albumin peptide composition according to any one of claims 2 to 3, characterized by, The method comprises the following steps: (1) mixing the albumin peptide, the bovine spleen peptide, and the wheat oligopeptide to obtain a mixture A; (2) mixing the sea cucumber peptide and the yeast beta-glucan to obtain a mixture B; (3) mixing the mixture A and the mixture B in an equal amount in a stepwise manner, and obtaining the albumin peptide composition.

8. The preparation method according to claim 7, characterized in that, In step (1), the mixture A is obtained by mixing the albumin peptide, the bovine spleen peptide, and the wheat oligopeptide, passing through a 60-100 mesh sieve, and drying.

9. The production method according to claim 8, characterized by, In step (1), the drying is performed in a vacuum drying manner at a temperature of 50-80℃, and the moisture content of the mixture A is ≤5%.

10. Use of the albumin peptide composition of any one of claims 1-5 or the albumin peptide composition prepared by the method of any one of claims 6-9 in the preparation of an immune-enhancing product.

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