CN114805518A

CN114805518A - Protein WG-SP01 with immunoregulation activity

Info

Publication number: CN114805518A
Application number: CN202210609841.7A
Authority: CN
Inventors: 侯银臣; 黄继红; 廖爱美; 潘龙; 魏兆军; 董得平; 杨盛茹; 连伟帅; 李琰; 袁晓晴
Original assignee: Henan University of Animal Husbandry and Economy
Current assignee: Henan Jianjiu Industrial Co.,Ltd.
Priority date: 2022-05-31
Filing date: 2022-05-31
Publication date: 2022-07-29
Anticipated expiration: 2042-05-31
Also published as: CN114805518B

Abstract

The invention discloses an immunoregulation active protein WG-SP01, wherein the protein WG-SP01 can activate the activities of LDH and ACP, so that macrophages of an organism are in an activated state; can obviously improve the ATP activity of the lymphocyte; the proliferation capacity of spleen T lymphocytes and B lymphocytes is obviously improved; the protein WG-SP01 has remarkable immunoregulation effect. Therefore, the protein WG-SP01 can be used as an active ingredient of an immunomodulator or an immunopotentiator to prepare medicines or functional foods for treating immune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism and the like, and can also be used for other medicines or functional foods with immunoregulatory efficacy.

Description

Protein WG-SP01 with immunoregulation activity

Technical Field

The invention relates to the field of protein engineering, in particular to a protein WG-SP01 with immunoregulation activity.

Background

Immune diseases (immunological diseases) refer to diseases caused by the imbalance of immune regulation affecting the immune response of the body. Autoimmune diseases are mainly caused by the accumulation of a large number of immune cells and immunoglobulins produced by an excessive immune response, thereby damaging normal tissues and causing pathological changes and functional damage. Allergic dermatitis and various skin lesions occur in the skin, arthritis occurs in joints, and nephritis … … occurs in kidneys

Common autoimmune diseases are: systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism, juvenile diabetes, idiopathic platelet purpura, autoimmune hemolytic anemia, ulcerative colitis, and many skin diseases, chronic liver disease … …, and the like.

There is a common point in the treatment of these diseases, which requires the use of immunosuppressive agents to suppress the immune response against the body. The most commonly used are corticoid-like agents such as: prednisone, hydrocortisone, dexamethasone and the like, which are often referred to as "hormones" for short. If the treatment is poor, cytotoxic drugs such as cyclophosphamide and methotrexate may be used, which are used for both suppression of immunity and treatment of cancer. All immunosuppressive agents have a major common adverse effect, and they affect the body's anti-infective, anti-tumor immune function to varying degrees. Thus, numerous experts have been studying other therapeutic approaches and have found that certain biologies and natural drugs suppress the autoimmune response, but the immune function against infections, tumors has little or no adverse effect. At present, the food or the medicine for treating the immune diseases is needed to be lacked.

Disclosure of Invention

The invention aims to provide a protein WG-SP01 with immunoregulatory activity and application thereof, so as to solve the problems.

According to a first aspect of the present invention, there is provided a protein WG-SP01 having immunomodulatory activity, the amino acid sequence of the protein WG-SP01 being shown in SEQ ID NO: 1. Therefore, the protein WG-SP01 with the amino acid sequence can activate LDH and ACP activity, so that macrophages of the organism are in an activated state; can obviously improve the ATP activity of the lymphocyte; the proliferation capacity of spleen T lymphocytes and B lymphocytes is obviously improved; WG-SP01 has significant immunomodulatory efficacy.

In certain embodiments, the secondary structure of the protein WG-SP01 comprises an alpha helix 17%, a beta sheet 27%, a random coil 17%, and a beta turn 21%. Therefore, the protein WG-SP01 with the secondary structure can activate LDH and ACP activity, so that macrophages of the organism are in an activated state; can obviously improve the ATP activity of the lymphocyte; the proliferation capacity of spleen T lymphocytes and B lymphocytes is obviously improved; WG-SP01 has significant immunomodulatory efficacy.

According to a second aspect of the present invention, there is provided the use of an immunomodulatory protein WG-SP01, the amino acid sequence of said protein WG-SP01 is shown in SEQ ID NO 1; further, the secondary structure of the protein WG-SP01 comprises 17% of alpha helix, 27% of beta sheet, 17% of random coil and 21% of beta turn. Therefore, the food or medicine containing the protein WG-SP01 has high-efficiency immune regulation function, and can be used for treating immune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism and the like.

According to a third aspect of the present invention, there is provided an immunomodulator comprising a protein WG-SP01 having immunomodulatory activity, the amino acid sequence of said protein WG-SP01 being set forth in SEQ ID NO 1; further, the secondary structure of the protein WG-SP01 comprises 17% of alpha helix, 27% of beta sheet, 17% of random coil and 21% of beta turn. Therefore, the immunomodulator has remarkable immunoregulation effect, and can be used for preparing food or medicines with immunoregulation effect.

In certain embodiments, the immunomodulator comprises protein WG-SP01 as an active ingredient.

According to a fourth aspect of the present invention, there is provided a functional food or pharmaceutical product comprising an immunomodulator comprising a protein WG-SP01 having immunomodulatory activity, the amino acid sequence of said protein WG-SP01 being represented in SEQ ID NO: 1. Further, the secondary structure of the protein WG-SP01 comprises 17% of alpha helix, 27% of beta sheet, 17% of random coil and 21% of beta turn. Therefore, the functional food or medicine has high-efficiency immunoregulation function, and can be used for treating immune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism and the like.

According to a fifth aspect of the present invention there is provided an immunopotentiator comprising a protein WG-SP01 having immunomodulatory activity, the amino acid sequence of the protein WG-SP01 being shown in SEQ ID NO. 1. Further, the secondary structure of the protein WG-SP01 comprises 17% of alpha helix, 27% of beta sheet, 17% of random coil and 21% of beta turn. Therefore, the immunopotentiator has a remarkable immunopotentiating effect, can enhance the immune function of organisms, and can also be used for preparing foods or medicines with immunoregulation effect.

In certain embodiments, the immunopotentiator has protein WG-SP01 as an active ingredient.

According to a sixth aspect of the present invention, there is provided a functional food or pharmaceutical product comprising an immunopotentiator comprising a protein WG-SP01 having an immunomodulatory activity, the amino acid sequence of the protein WG-SP01 being shown in SEQ ID NO: 1. Further, the secondary structure of the protein WG-SP01 comprises 17% of alpha helix, 27% of beta sheet, 17% of random coil and 21% of beta turn. Therefore, the functional food or medicine has high-efficiency immune enhancing function, and can be used for treating immune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism and the like.

In conclusion, the invention has the beneficial effects that:

1. the immunoregulation active protein WG-SP01 can activate LDH and ACP activity, so that macrophages of an organism are in an activated state; can obviously improve the ATP activity of the lymphocyte; obviously improves the proliferation capacity of spleen T lymphocytes and B lymphocytes, and WG-SP01 has obvious immunoregulation effect.

2. The immunoregulatory active protein WG-SP01 can be used as an active ingredient of an immunomodulator or an immunopotentiator to prepare medicines or functional foods for treating immune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism and the like, and can also be used for other medicines or functional foods with immunoregulatory efficacy.

3. The protein WG-SP01 is derived from wheat germ, is used as an effective component of food or medicines for treating immune diseases, is used for treating the immune diseases, and is safe without side effects.

Drawings

FIG. 1 is a three-dimensional map of the secondary and tertiary structures of immunomodulatory wheat germ protein WG-SP01, and the distribution of hydrophilic and hydrophobic sites and charges: FIG. A, B, C is a three-level structural view of WG-SP01 (A front view, B left view, C top view). D is a secondary structure diagram of WG-SP01, as above; FIG. E is a graph of the hydrophilicity/hydrophobicity profile of WG-SP01, with hydrophobicity/non-polarity being in the light gray areas and hydrophilicity/polarity being in the dark gray area portions; panel F shows the charge distribution of WG-SP01, with white short arrows indicating positively charged color regions (primarily arginine and lysine), white long arrows indicating negatively charged color regions (primarily aspartic acid and glutamic acid), and white boxes indicating no charge;

FIG. 2 is a graph showing the effect of WG-SP01 on mouse spleen acid phosphatase and lactate dehydrogenase activity;

FIG. 3 is a two-dimensional dot-matrix diagram of peripheral blood lymphocyte subpopulation cells of normal control mice;

FIG. 4 is a two-dimensional dot-matrix diagram of peripheral blood lymphocyte subpopulation cells of mice in cyclophosphamide model group;

FIG. 5 is a two-dimensional dot-matrix of cells of peripheral blood lymphocytes subpopulation of WG-SP01 low dose group mice;

FIG. 6 is a two-dimensional dot-matrix of cells of a subpopulation of peripheral blood lymphocytes of mice of a dose group in WG-SP 01;

FIG. 7 is a two-dimensional dot-matrix diagram of the peripheral blood lymphocyte subpopulation of WG-SP01 high dose group mice.

Detailed Description

The invention is described in further detail below with reference to the accompanying drawings.

Example 1 preparation, screening and characterization of protein WG-SP01 having immunomodulatory activity.

Wheat germ is used as a raw material, and after enzymolysis by alkaline protease, protein WG-SP01 with immunoregulatory activity is obtained by centrifugation, ultrafiltration, separation and purification, screening and identification.

The specific method comprises the following steps: mixing wheat germ and 3% NaCl solution according to the proportion of 1: 10, adding alkaline protein (2000U/g), adding alkaline protease, performing enzymolysis for 0.5h at 55 ℃ and pH9.0, adjusting pH to 7.0 with NaOH to finish reaction, then centrifuging at 5000rpm for 10min, taking supernatant, separating protein by membrane filtration and column chromatography, screening through in vitro lymphocyte proliferation experiment to obtain immunoregulatory active protein WG-SP01, and obtaining data information such as amino acid sequence, tertiary structure and the like by means of mass spectrometry and the like.

The amino acid sequence of the immunoregulatory active wheat germ protein WG-SP01 is shown in SEQ ID NO: 1: MASKGGNKGEGPAIGIDLGTTYSCVGVWQHDRVEIVANDQGNRTTPSYVAFTDTERLIGDAAKNQVAMNPTNTVFDAKRLIGRRFSDASVQSDMKMWPFKVVPGAGDKPMIVVTYKGEEKTFSAEEISSMVLTKMREIAEAFLSTTINNAVVTVPAYFNDSQRQATKDAGVIAGLNVMRIINEPTAAAIAYGLDKKATSTGEKNVLIFDLGGGTFDVSILTIEEGIFEVKSTAGDTHLGGEDFDNRMVNHFVQEFKRKNKKDISGNPRALRRLRTACERAKRTLSSTAQTTIEIDSLYEGIDFYATITRARFEELNMDLFRKCMEPVEKCLRDAKMDKTQIHDIVLVGGSTRIPKVQQLLQDFFNGKELCKSINPDEAVAYGAAVQAAILSGEGNQKVQDLLLLDVTPLSLGLETAGGVMTTLIPRNTTIPTKKEQVFSTYSDNQPGVLIQVYEGERTRTKDNNLLGKFELSGIPPAPRGVPQITVTFDIDANGILNVSAEDKTTGQKNKITITNDKGRLSKEEIERMVQEAEKYKSEDEQVRHKVEARNALENYAYNMRNTLRDDKIASKLPADDKKKIEDSIEDAIKWLDGNQLAEADEFEDKMKELESICNPIISKMYQGAGPGGAAGMDEDMPGGGAGTGGGSGAGPKIEEVD are provided. The molecular weight is 71847MW, it contains 657 amino acids, the secondary structure includes alpha helix 17%, beta sheet 27%, random coil 17%, beta turn 21%, the tertiary structure is shown in figure 1.

Example 2 detection of the immunomodulatory Activity of protein WG-SP 01.

1. Determination of in vitro immune Activity-mouse spleen lymphocyte proliferation assay

(1) Preparation of spleen cell suspension

Taking 6-8 weeks old Balb/c mice to be killed after neck removal, placing the mice in an ultra-clean workbench after being soaked in 75% ethanol for aseptic laparotomy, taking out spleen, placing the spleen on a 200-mesh cell sieve above a culture dish containing Hank's solution, then cleaning the mice with Hank's solution, extruding and grinding the mice by using a disposable glass injector core, repeatedly cleaning the mice twice with Hank's solution, transferring the mice to a centrifuge tube to prepare suspension, centrifuging the mice at 1000rpm for 5-10 min, and discarding the supernatant. Taking 6mL of cell lysate which is sterilized under high pressure or filtered by a 0.22 mu m filter, adding a centrifuge tube to lyse erythrocytes, uniformly blowing, adding Hank's solution after acting for 1-2min, centrifuging at 1000rpm for 5-10 min, and removing supernatant. Adding Hank's solution, blowing uniformly, centrifuging at 1000rpm for 5min, and discarding supernatant. Adding appropriate amount of Hank's solution, blowing uniformly, taking a small amount of cell suspension for counting living cells, centrifuging at the rest 1000rpm for 5min, and discarding supernatant to obtain cell precipitate (at this time, if the precipitate is redder, it is indicated that the red blood cells are not completely lysed, the red blood cells can be lysed again according to the above steps).

Adding a proper amount of pre-cooled RPMI-1640 complete culture solution into the obtained cell sediment, blowing and beating to prepare cell suspension, and counting the living cells by a trypan blue staining method. The method comprises the steps of taking a certain volume of cell suspension, adding RPMI-1640 complete culture solution for dilution, and then adding trypan blue solution with the final concentration of 0.04%. The blood count plate was used for counting, and when observed under a low power microscope, live cells were not stained, and dead cells were stained dark blue by trypan blue. The cell counting formula is shown in (1):

(2) inoculation culture and MTT activity detection

According to the cell density of the obtained cell suspension, pre-cooled RPMI-1640 complete culture solution is added to prepare lymphocyte suspension with the cell density of 1.0-2.0X 106 cells/mL. To a 96-well plate, 100. mu.L of mouse spleen lymphocyte suspension was added. And respectively setting a blank control group and an experimental group, and detecting the capability of the experimental sample for promoting the spleen lymphocyte proliferation. Wherein, the blank control group is added with 100 mu L of complete culture solution, and the experimental groups are added with 100 mu L of samples to be tested respectively. After mixing gently, the mixture was cultured in an incubator at 37 ℃ with 5% CO2 for 48 hours.

10. mu.l of 5mg/mL MTT solution was added to each well 4 hours before the end of cell culture, and the culture was terminated after 4 hours of further culture. Centrifuging at 3000rpm for 10min, removing liquid in the wells, adding 150 μ l DMSO into each well, and shaking for 5min to dissolve the crystals completely. The wavelength of 570nm is selected, the light absorption value of each hole is measured on an enzyme linked immunosorbent assay instrument, and the result is recorded.

The greater the stimulation index, the greater its immunomodulatory activity. The stimulation index of protein WG-SP01 was measured according to the above method using different concentrations, and the stimulation index of protein WG-SP01 was 237% under the condition of 5. mu.g/mL, with EC50 being 1.2. mu.g/mL.

And (3) measuring the in vivo immunological activity, and carrying out animal experiments and evaluating according to the experimental scheme for enhancing the immunological function in the implementation manual of the health food inspection and evaluation technical specifications.

(3) Grouping and administration of laboratory animals

The mice were randomly divided into 6 groups of 12 mice each, and the specific groups are shown in table 1. Cyclophosphamide (CPA) is adopted and is injected into the abdominal cavity to establish an immunosuppression experimental animal model.

Table 1 groups of animal models (n ═ 12)

Note: NC, Normal control group (Normal control group); CPA, a group of Cyclophosphamide (Cyclophosphamide groups).

TABLE 2 influence of WG-SP01 on mouse organ index (n. 12)

Note: the difference was extremely significant compared to the normal group (P < 0.01); *. were significantly different compared to the normal group (P < 0.05); the difference is extremely significant compared with the cyclophosphamide group (P < 0.01); #. was very significantly different (P <0.01) compared to the cyclophosphamide group; the same applies below.

Table 2 it can be seen that CPA had no significant effect on the liver, kidney and heart indices of mice, with a significant decrease in spleen and thymus indices (P < 0.01). The spleen index of mice was affected to some extent by WG-SP01 at each dose compared to CPA group, with the difference in spleen index between the high, medium and low dose WG-SP01 and CPA groups being more pronounced (P <0.01), but the effect between doses was not as pronounced. The WG-SP01 of each dose has an ascending trend on the spleen index of the mice, and the ascending trend of low dose and high dose is more obvious compared with that of CPA group; the results show that the wheat blastoglobulin has certain weight increasing effect on immune organ indexes of the immunosuppressed mice, can improve the atrophy state of the immune organs of the immunosuppressed mice, and improves the immune function of the mice to a certain extent.

The detection results of the activity of the spleen lactate dehydrogenase and the acid phosphatase of the mice show that (as shown in figure 2), after the mice are injected with cyclophosphamide, the activity of the spleen acid phosphatase and the activity of the lactate dehydrogenase are both remarkably reduced (P is less than 0.01); each dose of WG-SP01 can antagonize immunosuppression caused by cyclophosphamide, has obvious influence (P is less than 0.01) on the activity of mouse spleen acid phosphatase and the activity of lactate dehydrogenase in an immunosuppression state, can obviously improve the reduction of the activity of mouse spleen acid phosphatase and the activity of lactate dehydrogenase caused by immunosuppression, and tends to recover to normal enzyme activity. Experiments show that the WG-SP01 can regulate the immune function of an organism in an immunosuppressive state and can improve the activation state of macrophages, so that the aim of improving the immune function of the organism is fulfilled.

TABLE 3 Effect of WG-SP01 on the phagocytosis of neutral Red and iNOS Activity by macrophages in the peritoneal cavity of immunosuppressed mice

As can be seen in Table 3, the phagocytic ability of macrophages in the abdominal cavity of mice in the cyclophosphamide group is significantly lower than that of the normal control group (P < 0.01); WG-SP01 can effectively and remarkably improve the phagocytosis neutral red ability of abdominal cavity macrophages of immunosuppressive mice (P is less than 0.01); except for the WG-SP01 low dose group, which is significantly lower than the normal group (P <0.01), the levels of the other treatment groups were not significantly different from the normal group (P > 0.05). Compared with a normal control group, the activity of iNOS (nitric oxide synthase) in the mouse abdominal cavity macrophage of the cyclophosphamide group is obviously lower than that of the normal control group (P < 0.01); after administration, iNOS activity of abdominal cavity macrophage of WG-SP01 in each dose group is obviously higher than that of cyclophosphamide group (P <0.01), and no significant difference exists in comparison with normal group (P > 0.05).

TABLE 4 Effect of WG-SP01 on mouse serum IL-2, IL-4, IFN-. gamma.and TNF-. alpha.

As can be seen from Table 4, compared with the normal control group, the contents of IL-2, IL-4, IFN-gamma and TNF-alpha in the serum of the mice in the model group are all significantly reduced (P is less than 0.01), which indicates that the immunosuppression model is successful and the immunity of the mice is at a low level; compared with the model group, each treatment group can obviously improve the concentration of the serum cytokine of the immunosuppressed mice (P is less than 0.01); experiments show that WG-SP01 has a remarkable recovery effect on immunosuppressive mouse serum cytokines (P < 0.01). The increase of the level of the related immune molecules indicates that the proportion or the activity of the lymphocyte subpopulation related to the immune molecules is obviously increased, thereby promoting the recovery of the immune function.

The flow cytometry double antibody labeling method is adopted to detect the lymphocyte CD3+, CD4+, CD8+, and CD25+ cell percentage in the peripheral blood of the mice. After the separated mouse blood mononuclear cells are marked by the specific monoclonal fluorescent antibody, lymphocyte groups with obvious areas can be obtained on a flow cytometer according to the size of the cells. And analyzing and processing by using CELLQuest software, and representing by adopting a two-dimensional lattice diagram form. The results are shown in FIGS. 3, 4, 5, 6 and 7. The number of cells in each subpopulation was calculated after two-dimensional dot-matrix analysis of the lymphocyte subpopulations as shown in table 5.

TABLE 5 Effect of WG-SP01 on immunosuppressed mouse lymphoid subpopulation

Lymphocytes, the main cell population that constitutes the immune system of the body, can be divided into a number of populations that differ in phenotype and function, and when infection occurs or immune function is altered, the change in each subset of T lymphocytes often reflects the state of the body with great sensitivity. As can be seen from table 5, the peripheral blood proportion of CD3+ and CD4+ cells in the CPA immunosuppression model mice was significantly decreased (P <0.01) compared to the normal group; this is consistent with the results of the previous studies and also suggests that the CPA immunosuppressive model is established. After the mouse is treated by WG-SP01, the proportion of CD3+ and CD4+ T cells in peripheral blood of an immunosuppressed mouse is obviously improved, is obviously higher than that of a CPA immunosuppression model group (P <0.01), and is obviously higher than that of a normal group mouse (P < 0.01).

The relative immune balance state of the body is mainly maintained by the mutual influence of CD4+ cells and CD8+ cells, and the maladjustment of the proportion of the two subgroups can cause the immune dysfunction, so that CD4+/CD8+ represents the balance of the overall immune function, and the reduction of the balance probably reflects the maladjustment of the immune function of the body. In the experiment, the ratio of CD4+/CD8+ of peripheral blood of mice in a CPA model group is found to be remarkably lower than the level of a normal group (p < 0.01); the ratio of CD4+/CD8+ of the WG-SP01 high, medium and low concentration treatment groups is significantly higher than that of the CPA model group (P <0.01), and has no significant difference compared with the normal group (P > 0.05).

The CD4+ CD25+ regulatory T cells play an important role in maintaining the immune tolerance and the immune response steady state of the organism, and Table 5 shows that the proportion of CD4+ CD25+ T cells in the peripheral blood of mice in the CPA immunosuppression model group is obviously reduced, and compared with a normal control group, the significant difference (P <0.01) is obtained, which indicates that the CPA immunosuppression model is successful. After WG-SP01 is drenched, the proportion of CD4+ CD25+ T cells in peripheral blood of an immunosuppressed mouse is obviously increased, and the immunosuppressed mouse has a significant difference (P is less than 0.01) compared with a CPA immunosuppression model group; slightly higher than normal mice, but no significant difference (P > 0.05). Experiments show that each dose of WG-SP01 has an effect of improving the proportion of CD4+ CD25+ T cells in the peripheral blood T cell subset of CPA immunosuppressive mice.

What has been described above are merely some embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made without departing from the inventive concept herein, and it is intended to cover all such modifications and variations as fall within the scope of the invention.

Sequence listing

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<120> a protein WG-SP01 having immunoregulatory activity

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Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly Val Trp Gln His Asp Arg

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Val Glu Ile Val Ala Asn Asp Gln Gly Asn Arg Thr Thr Pro Ser Tyr

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Val Ala Phe Thr Asp Thr Glu Arg Leu Ile Gly Asp Ala Ala Lys Asn

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Gln Val Ala Met Asn Pro Thr Asn Thr Val Phe Asp Ala Lys Arg Leu

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Ile Gly Arg Arg Phe Ser Asp Ala Ser Val Gln Ser Asp Met Lys Met

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Trp Pro Phe Lys Val Val Pro Gly Ala Gly Asp Lys Pro Met Ile Val

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Val Thr Tyr Lys Gly Glu Glu Lys Thr Phe Ser Ala Glu Glu Ile Ser

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Ser Met Val Leu Thr Lys Met Arg Glu Ile Ala Glu Ala Phe Leu Ser

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Thr Thr Ile Asn Asn Ala Val Val Thr Val Pro Ala Tyr Phe Asn Asp

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Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly Val Ile Ala Gly Leu Asn

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Val Met Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala Tyr Gly

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Leu Asp Lys Lys Ala Thr Ser Thr Gly Glu Lys Asn Val Leu Ile Phe

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Asp Leu Gly Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile Glu Glu

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Gly Ile Phe Glu Val Lys Ser Thr Ala Gly Asp Thr His Leu Gly Gly

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Glu Asp Phe Asp Asn Arg Met Val Asn His Phe Val Gln Glu Phe Lys

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Arg Lys Asn Lys Lys Asp Ile Ser Gly Asn Pro Arg Ala Leu Arg Arg

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Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg Thr Leu Ser Ser Thr Ala

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Gln Thr Thr Ile Glu Ile Asp Ser Leu Tyr Glu Gly Ile Asp Phe Tyr

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Ala Thr Ile Thr Arg Ala Arg Phe Glu Glu Leu Asn Met Asp Leu Phe

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Arg Lys Cys Met Glu Pro Val Glu Lys Cys Leu Arg Asp Ala Lys Met

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Asp Lys Thr Gln Ile His Asp Ile Val Leu Val Gly Gly Ser Thr Arg

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Ile Pro Lys Val Gln Gln Leu Leu Gln Asp Phe Phe Asn Gly Lys Glu

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Leu Cys Lys Ser Ile Asn Pro Asp Glu Ala Val Ala Tyr Gly Ala Ala

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Val Gln Ala Ala Ile Leu Ser Gly Glu Gly Asn Gln Lys Val Gln Asp

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Leu Leu Leu Leu Asp Val Thr Pro Leu Ser Leu Gly Leu Glu Thr Ala

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Gly Gly Val Met Thr Thr Leu Ile Pro Arg Asn Thr Thr Ile Pro Thr

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Lys Lys Glu Gln Val Phe Ser Thr Tyr Ser Asp Asn Gln Pro Gly Val

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Leu Leu Gly Lys Phe Glu Leu Ser Gly Ile Pro Pro Ala Pro Arg Gly

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Asn Thr Leu Arg Asp Asp Lys Ile Ala Ser Lys Leu Pro Ala Asp Asp

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Gly Asn Gln Leu Ala Glu Ala Asp Glu Phe Glu Asp Lys Met Lys Glu

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Leu Glu Ser Ile Cys Asn Pro Ile Ile Ser Lys Met Tyr Gln Gly Ala

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Gly Pro Gly Gly Ala Ala Gly Met Asp Glu Asp Met Pro Gly Gly Gly

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Ala Gly Thr Gly Gly Gly Ser Gly Ala Gly Pro Lys Ile Glu Glu Val

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Asp

Claims

1. A protein WG-SP01 with immunoregulatory activity, wherein the amino acid sequence of the protein WG-SP01 is shown as SEQ ID NO. 1.

2. The protein WG-SP01 having immunomodulatory activity of claim 1, wherein the secondary structure of the protein WG-SP01 comprises an alpha helix 17%, a beta sheet 27%, a random coil 17%, and a beta turn 21%.

3. Use of the protein WG-SP01 with immunomodulatory activity of claim 1 or 2 in the preparation of a food or pharmaceutical product with immunomodulatory efficacy.

4. An immunomodulator, wherein the immunomodulator comprises the protein WG-SP01 of claim 1 or 2.

5. The immunomodulator according to claim 4, wherein said protein WG-SP01 is used as an active ingredient in an immunomodulator.

6. A functional food or pharmaceutical product comprising the immunomodulator according to claim 4.

7. An immunopotentiator, wherein the immunopotentiator comprises the protein WG-SP01 according to claim 1 or 2.

8. The immunopotentiator according to claim 7, wherein the protein WG-SP01 is used as an active ingredient in an immunopotentiator.

9. A functional food or pharmaceutical product comprising the immunopotentiator according to claim 7.