A complex functional protein peptide derived from sea cucumber, its formulation and its application

Abstract

This invention discloses a complex functional protein peptide derived from sea cucumber, its formulation, and its applications, belonging to the field of bioactive peptide technology. The amino acid sequence of the complex functional protein peptide derived from sea cucumber in this invention is at least one of FFGG, FGAF, and FDGF. Using sea cucumber blanching solution as raw material, this invention employs methods such as protease hydrolysis, LC-MS / MS, and bioinformatics to separate and purify three bioactive peptides with antioxidant and anti-allergic effects. Based on the identified peptide sequences, chemical synthesis is performed, and the antioxidant and anti-allergic effects of the synthesized single peptides and complex peptides are measured. These bioactive peptides all exhibit high DPPH scavenging rates, ABTS free radical scavenging rates, and hyaluronic acid inhibition rates. Therefore, these bioactive peptides have significant application prospects in the development of products with antioxidant and anti-allergic effects.

Description

Technical Field

[0001] This invention belongs to the field of bioactive peptide technology, specifically relating to a complex functional protein peptide derived from sea cucumber, its formulation, and its application. Background Technology

[0002] Oxidative stress (OS) is a core factor inducing aging, inflammation, and various chronic diseases. This is due to an imbalance between the production and clearance of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the body. Excessive accumulation of oxidative substances not only damages cells, tissues, and biomolecules (lipids, proteins, DNA), but also exacerbates the progression of cardiovascular diseases, neurodegenerative diseases, and inflammatory lesions. Current antioxidant interventions have significant limitations. Traditional chemical antioxidants (such as BHT and TBHQ) are controversial in terms of safety, while natural antioxidants, although capable of scavenging free radicals, often suffer from poor stability and low bioavailability, making it difficult to meet the development needs of functional foods.

[0003] Meanwhile, allergic diseases also face the challenge of insufficient intervention methods. Allergic reactions, as a prevalent immune system disorder worldwide, are characterized by an imbalance in the Th2 immune response, excessive IgE production, and the release of allergic mediators such as histamine. Clinical manifestations include skin redness and swelling, itching, rhinitis, asthma, and even anaphylactic shock, severely impacting patients' quality of life. Currently, anti-allergy treatments largely rely on antihistamines or glucocorticoids. These drugs only relieve symptoms and cannot fundamentally regulate the immune imbalance; long-term use may also lead to side effects such as drowsiness and metabolic disorders. Commercially available anti-allergy functional foods mostly rely on traditional ingredients such as probiotics and vitamins, with unclear active ingredients and mechanisms of action. Research on immunomodulatory anti-allergy peptides is still in its early stages.

[0004] Bioactive peptides are a class of small molecule polypeptides derived from natural products with specific physiological regulatory functions. Due to their diverse structures, good biocompatibility, and low toxicity, they have received widespread attention in the fields of functional foods, pharmaceuticals, and health products in recent years. Among them, peptides with antioxidant and anti-allergic activities show promising application prospects in the prevention and relief of oxidative stress-related diseases and the regulation of immune responses.

[0005] Sea cucumber blanching solution, a major byproduct of sea cucumber processing, is typically discharged directly, wasting valuable biological resources such as proteins and polysaccharides and creating environmental pollution. Studies have shown that sea cucumber blanching solution is rich in various bioactive components, including sea cucumber polysaccharides, polypeptides, and saponins, exhibiting significant potential in antioxidant, anti-inflammatory, and immunomodulatory effects. Currently, research on extracting functional bioactive peptides from sea cucumber blanching solution is insufficient, particularly lacking systematic screening, structural identification, and application studies of small molecule bioactive peptides with both antioxidant and anti-allergic properties.

[0006] Therefore, developing a small molecule bioactive peptide with a clear source, well-defined structure, and both high antioxidant and anti-allergic activity, and applying it to food, medicine, or health products, has significant scientific research value and practical application significance. Summary of the Invention

[0007] In view of the problems existing in the prior art, the purpose of this invention is to provide a composite functional protein peptide, preparation and application of sea cucumber.

[0008] To achieve the above objectives, the present invention adopts the following technical solution: A complex functional protein peptide derived from sea cucumber, wherein the amino acid sequence of the protein peptide is at least one of FFGG, FGAF, and FDGF.

[0009] The above-mentioned sea cucumber-derived complex functional protein peptides are used in the preparation of products with antioxidant and anti-allergic effects.

[0010] Based on the above scheme, the product is food, medicine or health product.

[0011] Based on the above scheme, the product further includes excipients acceptable for food, pharmaceutical or health products.

[0012] Based on the above scheme, the antioxidant mentioned has a free radical scavenging effect.

[0013] Based on the above scheme, the free radical is at least one of DPPH free radical and ABTS free radical.

[0014] Based on the above scheme, the anti-allergy effect is described as having a hyaluronic acid inhibitory effect.

[0015] A complex functional protein peptide preparation derived from sea cucumber, wherein the active ingredient is at least one of FFGG, FGAF, and FDGF.

[0016] Based on the above scheme, the concentration of the active ingredient is 1 mg / mL.

[0017] Advantages of the technical solution of this invention This invention uses sea cucumber blanching liquid as raw material and employs methods such as protease hydrolysis, LC-MS / MS, and bioinformatics to separate and purify three bioactive peptides with antioxidant and anti-allergic effects. Based on the identified peptide sequences, these peptides were chemically synthesized, and the antioxidant and anti-allergic effects of the synthesized single and compound peptides were measured. These bioactive peptides all exhibited high DPPH scavenging rates, ABTS free radical scavenging rates, and hyaluronic acid inhibition rates. Therefore, these bioactive peptides can not only be used as natural antioxidants in food, pharmaceuticals, and / or cosmetics, but also as natural anti-allergens in pharmaceuticals; they have significant application prospects in the development of products with antioxidant and anti-allergic effects.

[0018] This invention uses sea cucumber processing by-product blanching liquid as raw material, which not only provides a product with natural and safe antioxidant and anti-allergy effects, but also promotes the high-value utilization of sea cucumber processing by-product blanching liquid, responds to the strategies of "green development of marine economy" and "circular economy", and provides a new direction for the research and development of high-purity, high-activity marine source functional ingredients and anti-allergy preparations. Attached Figure Description

[0019] Figure 1 Scavenging rate of DPPH free radicals of active peptides with different molecular weights; Figure 2 Scavenging rate of ABTS free radicals by active peptides of different molecular weights; Figure 3 Hyaluronic acid inhibition rate of active peptides with different molecular weights; Figure 4 Elution peaks after chromatography; Figure 5 The scavenging rate of DPPH radicals at the three peaks; Figure 6 The scavenging rate of ABTS radicals in the three peaks; Figure 7 Hyaluronic acid inhibition rate of the three peaks; Figure 8 This is a second-order mass spectrum of FFGG. Figure 9 This is a secondary mass spectrum of FGAF; Figure 10 This is a secondary mass spectrum of FDGF; Figure 11 Results of hydrolysis degree determination by different proteases; Figure 12 DPPH free radical scavenging rate after enzymatic hydrolysis by different enzymes; Figure 13 ABTS free radical scavenging rate after enzymatic hydrolysis with different enzymes; Figure 14 Inhibition rate of hyaluronic acid after enzymatic hydrolysis with different enzymes; Figure 15 DPPH radical scavenging rate and ABTS radical scavenging rate of a single peptide; Figure 16 DPPH radical scavenging rate and ABTS radical scavenging rate of the complex peptides; Figure 17 Hyaluronic acid inhibition rate of a single peptide segment; Figure 18 Hyaluronic acid inhibition rate of compound peptides. Detailed Implementation

[0020] The terminology used in this invention, unless otherwise specified, generally has the meanings commonly understood by those skilled in the art. The invention is further described in detail below with reference to specific embodiments and data. The following embodiments are merely illustrative and are not intended to limit the scope of the invention in any way.

[0021] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the experimental materials, reagents, and chemicals used in the following embodiments can be obtained through general channels.

[0022] In the following embodiments, 1. Determination of degree of hydrolysis The degree of protein hydrolysis in the lyophilized sea cucumber blanching solution was determined using the ophthaldialdehyde (OPA) method. The specific method is as follows: 80 mg of OPA was dissolved in 2 mL of anhydrous ethanol, 200 μL of β-mercaptoethanol, 5 mL of 10% SDS (w / v), and 92.8 mL of 0.1 mol / L sodium tetraborate to prepare a 100 mL OPA reagent solution. 40 μL of the enzymatic hydrolysis product from the sea cucumber blanching solution was mixed with 4 mL of OPA reagent and incubated at room temperature for 2 min, and the absorbance was measured at 340 nm. The lyophilized sea cucumber blanching solution powder was placed in 6 mol / L HCl and reacted at 115 °C for 24 h. The number of free amino groups was determined using a serine standard curve, which was taken as the number of free amino acids produced by the complete hydrolysis of the sea cucumber blanching solution lyophilized protein.

[0023] The degree of protein hydrolysis in freeze-dried sea cucumber blanching solution powder is calculated using the following formula: In the formula: (NH2) t Indicates the amount of free amino acids released; (NH2) t0 Indicates the amount of free amino acids that have not undergone enzymatic hydrolysis; (NH2) T This indicates the number of free amino acids after complete hydrolysis.

[0024] 2. Determination of DPPH free radical scavenging rate Prepare a 0.1 mmol / L DPPH ethanol solution using 2,2-biphenyl-1-picrylhydrazine and anhydrous ethanol. Mix the protein peptide solution with an equal volume of the 0.1 mmol / L DPPH ethanol solution and react at room temperature in the dark for 30 min. Measure the absorbance at 517 nm and record this as sample A. Replace the DPPH ethanol solution with an equal volume of ethanol and mix with an equal volume of protein peptide solution, then measure the corresponding absorbance and record this as blank A. Replace the protein peptide solution with an equal volume of distilled water and mix with an equal volume of DPPH ethanol solution, then measure the corresponding absorbance and record this as control A. Calculate the DPPH free radical scavenging rate using the following formula: 3. Determination of ABTS free radical scavenging rate Preparation of ABTS radical solution: Mix 7 mM ABTS solution and 2.45 mM potassium persulfate in equal volumes, place in the dark, and react for 12-16 hours. Then dilute with anhydrous ethanol to obtain an absorbance of 0.700 ± 0.02 at 734 nm. Let stand at room temperature for 30 minutes before use. Use anhydrous ethanol as a reference solution.

[0025] A certain amount of protein peptide solution was weighed and prepared into a sample solution with a concentration of 2 mg / mL using PBS buffer. 30 μL of the sample was mixed thoroughly with 570 μL of ABTS free radical solution, and reacted at room temperature in the dark for 10 min. The absorbance value A1 of the sample was then measured at 734 nm. 30 μL of distilled water was used instead of the sample solution and mixed thoroughly with 570 μL of ABTS free radical solution to serve as a blank control. The absorbance value A0 of the blank control was measured at 734 nm. The ABTS free radical scavenging rate was calculated using the following formula: In the formula, A1 is the absorbance value of the experimental group and A0 is the absorbance value of the blank group.

[0026] 4. Determination of hyaluronic acid inhibition rate 100 μL of 0.2 M sodium acetate buffer (pH 4.6, containing 0.15 M NaCl) was added to the positive control group (OD1) and negative control group (OD2), respectively. 100 μL of sample solution was added to the sample test group (OD3) and sample background group (OD4). The positive control group (OD1) and sample test group (OD3) tubes contained 50 μL of hyaluronidase (500 U / mL, prepared with buffer), and the negative control group (OD2) and sample background group (OD4) tubes contained 50 μL of buffer. After incubation at 37 °C for 20 min, 20 μL of 2.5 mol / L CaCl2 solution was added, and incubation was continued at 37 °C for 20 min. Then, 50 μL of hyaluronic acid (3 mg / mL, prepared with buffer), 100 μL of buffer, and 250 μL of deionized water were added to each tube, and incubation was continued at 37 °C for 40 min. The enzymatic reaction was then terminated by adding 110 μL of alkaline borate solution followed by heating in a boiling water bath for 5 min. The test tube was then placed in ice water for 20 min, followed by the addition of 1.5 mL of p-dimethylaminobenzaldehyde solution. To maximize the coloring of the reaction mixture, the test tube was incubated at 37 °C for 20 min. The absorbance of the colored product was measured at 585 nm. The hyaluronic acid inhibition rate was calculated using the following formula: Table 1. Determination of hyaluronic acid inhibition rate Sea cucumber scalding solution is a byproduct of sea cucumber processing. The sea cucumber scalding solution used in the following examples comes from Haozhenfa Trading Company (a sole proprietorship) in Laoshan District. Example 1

[0027] The method for extracting protein peptides with antioxidant and anti-allergic complex functions from sea cucumber blanching solution includes the following steps: (1) Take 600mL of sea cucumber blanching liquid, thaw it, centrifuge it at 6000g for 20 minutes, remove the precipitate, and freeze-dry the supernatant at -80℃. (2) Take 2g of sea cucumber blanching liquid freeze-dried powder and add it to 20 mL of PBS solution (solid-to-liquid ratio 1:10); then add 5% (relative to substrate mass) of papain and enzymatically hydrolyze at 50℃ and pH 7 for 4 h. The entire process is carried out in a water bath incubator. After the enzymatic hydrolysis reaction is completed, heat the hydrolysate at 100℃ for 15 min and then quickly cool it to 4℃ with ice water. Centrifuge at 8000 r / min for 20 min in a refrigerated centrifuge at 4℃ and retain the supernatant.

[0028] (3) After passing the supernatant obtained from the above enzymatic hydrolysis through 0.45 and 0.22 micrometer filter membranes, it was fractionated into different molecular weights using an ultrafiltration centrifuge tube. The ultrafiltration separation conditions were 4000 g / min and centrifugation at 4℃ for 20 min, with molecular weight cutoffs of 10 kDa and 3 kDa. Fractions >10 kDa were collected, where the peptides were retained but did not pass through the 10 kDa membrane; 310 kDa, where the peptides permeated into the 10 kDa membrane but not into the 3 kDa membrane; and <3 kDa, where the peptides permeated into the 3 kDa membrane. Thus, proteolytic hydrolysates with molecular weights ranging from <3 kDa, 3-10 kDa, to 10 kDa were obtained. The scavenging rates of ABTS and DPPH free radicals and the inhibition rate of hyaluronic acid of active peptides with different molecular weights were measured. The results are as follows: Figure 1 , Figure 2 and Figure 3 As shown, protein peptides with smaller molecular weights (<3 kDa) exhibited better performance in scavenging ABTS and DPPH free radicals and inhibiting hyaluronic acid in vitro. This may be because small peptides have advantages such as simple structure, strong biological activity, low toxicity, and good stability, thus potentially offering advantages in in vivo efficacy.

[0029] (4) Protein peptides with a molecular weight <3 kDa were further purified by filtration on a Superdex™ Increase 10 / 300 GL gel permeation column (10 × 300 mm). The column was eluted with deionized water, and the fractions were collected at a flow rate of 1 mL / min. The fractions were detected at 280 nm. The peptide fractions were collected in the order of peak elution.

[0030] Gel filtration chromatography was used to separate and purify the enzymatic hydrolysates of sea cucumber blanching solution with molecular weights <3 kDa. The elution curve is shown below. Figure 4 As shown in the figure, three main elution peaks were obtained, named F1, F2, and F3 in ascending order of elution volume. The components of each peak were collected, and their antioxidant and anti-allergic activities were determined. The results are as follows. Figure 5 , Figure 6 and Figure 7 As shown, the F3 component performed best in all three indicators of ABTS scavenging, DPPH scavenging, and hyaluronidase inhibition, indicating that this component is enriched with active ingredients that have both antioxidant and anti-allergic activities. In addition, its elution volume is the largest, suggesting that its molecular weight is small. Further structural identification and structure-activity relationship studies can be conducted on it.

[0031] (5) Peptide sequences in components with high free radical scavenging rate and anti-allergic potential were identified by LC-MS / MS. The LC conditions were: C18 analytical column (75 μm × 150 mm, 3 μm), flow rate 300 nL / min. Mobile phase A was 0.1% formic acid, 2% ACN; mobile phase B was 0.1% formic acid, 80% ACN; elution was performed with 6–9% B for 8 min, 9–14% B for 14 min, 14–30% B for 36 min, 30–40% B for 15 min, 40–95% B for 3 min, and 95% B hold for 5 min. The mass spectrometry conditions were: MSAS scan range (m / z) 100–1500, AGC target: 3e6; resolution: 70,000, etc.

[0032] (6) The potential bioactivity of peptides in sea cucumber blanching liquid protein hydrolysate was predicted by the Peptide Ranker program (0-1). Peptides with a score >0.97 were selected, and peptides with a toxicity threshold prediction <0.4 were selected by the ToxinPred3.0 website.

[0033] After screening, three peptides, FFGG, FGAF, and FDGF, were finally obtained. The screened peptide sequences were then chemically synthesized. The secondary mass spectra of the three peptides are shown below. Figure 8 , Figure 9 and Figure 10 As shown. Example 2

[0034] The effect of protease type on the proteolytic effect of sea cucumber blanching solution Single enzymatic hydrolysis: After thawing 600 mL of sea cucumber blanching solution, centrifuge at 6000 g for 20 minutes to remove the precipitate. The supernatant was then freeze-dried at -80℃. 2 g of the freeze-dried sea cucumber blanching solution powder was added to 20 mL of PBS solution (solid-to-liquid ratio 1:10). Then, 5% (relative to substrate mass) of alkaline protease, papain, and trypsin were added separately, and enzymatic hydrolysis was performed for 4 hours at their respective optimal temperatures and pH values ​​(Table 2). The entire process was carried out in a water bath incubator. After the enzymatic hydrolysis reaction, the hydrolysate was heated at 100℃ for 15 minutes and then rapidly cooled to 4℃ with ice water. It was then centrifuged at 8000 r / min for 20 minutes in a refrigerated centrifuge at 4℃, and the supernatant was retained.

[0035] Complex enzymatic hydrolysis: After thawing 600 mL of sea cucumber blanching solution, centrifuge at 6000 g for 20 minutes to remove the precipitate. The resulting supernatant is then freeze-dried at -80℃. 18 g of the freeze-dried sea cucumber blanching solution powder is added to 180 mL of PBS solution (solid-to-liquid ratio 1:10). The solution is divided into three groups, and subjected to compound enzymatic hydrolysis using alkaline protease and papain, papain and trypsin, and alkaline protease and trypsin, respectively. The specific method is as follows: first, add the first protease and hydrolyze for 4 hours, then inactivate the enzyme in a boiling water bath for 15 minutes, cool to room temperature, adjust the pH to the optimal pH for the second protease, and then add the second protease for 4 hours of hydrolysis. After hydrolysis, heat the hydrolysate at 100℃ for 15 minutes and quickly cool it to 4℃ with ice water. Centrifuge at 8000 r / min for 20 minutes in a refrigerated centrifuge at 4℃, and retain the supernatant. The amounts of the first and second proteases added are 5% of the substrate mass, respectively.

[0036] Table 2 Optimal reaction conditions for different proteases The degree of protein hydrolysis in the sea cucumber blanching solution was determined after single and combined enzymatic hydrolysis, and the results are as follows: Figure 11 As shown, in the single-enzyme hydrolysis system, papain exhibited the highest degree of hydrolysis (58%), significantly higher than alkaline protease (44%) and trypsin (45%), indicating that papain has a stronger cleavage ability for this substrate protein, possibly related to its broad substrate specificity and high catalytic efficiency. In the complex enzymatic hydrolysis system, pairwise enzymatic hydrolysis did not significantly improve the degree of hydrolysis; in fact, some combinations were slightly lower than that of single papain, suggesting that there may be substrate competition or overlapping cleavage sites between the enzymes, failing to form a synergistic effect. In summary, from the perspective of hydrolysis efficiency, single papain can achieve highly efficient hydrolysis of sea cucumber blanching liquid protein powder, with the advantages of simple operation and low cost, making it suitable as the preferred enzyme for subsequent functional peptide preparation.

[0037] The ABTS free radical scavenging rate, DPPH free radical scavenging rate, and anti-allergic activity of the sea cucumber blanching extract peptides after single and combined enzymatic hydrolysis were determined, and the results are as follows: Figure 12 , Figure 13 and Figure 14 As shown in the figure, papain performed exceptionally well across all indicators in single enzymatic hydrolysis. While some combinations showed slightly higher performance than papain alone in certain indicators in combined enzymatic hydrolysis, papain maintained high levels across all three indicators, demonstrating the most balanced overall performance. Considering both antioxidant activity (ABTS and DPPH scavenging rates) and anti-allergic activity, as well as the economics and simplicity of the enzymatic hydrolysis process, papain-based single enzymatic hydrolysis is the most suitable method. Therefore, papain can be used for further enzymatic hydrolysis to obtain higher content small molecule peptides. Example 3

[0038] A complex functional protein peptide derived from sea cucumber, wherein the protein peptide is at least one of the peptide segments of the amino acid sequences shown in SEQ ID NO:1 to SEQ ID NO:3.

[0039] SEQ ID NO:1: FFGG; SEQ ID NO:2: FGAF; SEQ ID NO:3: FDGF. Example 4

[0040] A complex functional protein peptide preparation derived from sea cucumber, wherein the active ingredient is at least one of FFGG, FGAF, and FDGF active peptides.

[0041] Using pure water as a solvent, single peptides FFGG, FGAF, and FDGF, and complex peptides FFGG+FGAF, FFGG+FDGF, FGAF+FDGF, and FFGG+FGAF+FDGF were prepared into solutions with a concentration of 1 mg / mL. In the complex peptides, the peptides were combined at volume ratios of 1:1 and 1:1:1. The concentration of both single and complex peptides was 1 mg / mL. The antioxidant and anti-allergic effects of the single peptides FFGG, FGAF, and FDGF, and the complex peptides FFGG+FGAF, FFGG+FDGF, FGAF+FDGF, and FFGG+FGAF+FDGF were determined.

[0042] The results of single peptide solutions on the DPPH radical scavenging rate and ABTS radical scavenging rate are as follows: Figure 15 As shown, FFGG exhibits the most significant scavenging effect on DPPH free radicals, with a scavenging rate of 88.05%, followed by FGAF at approximately 40.29%, and FDGF at the lowest, with a scavenging rate of approximately 31.34%. For ABTS free radicals, FFGG also demonstrates the highest scavenging rate at approximately 91.77%, followed by FDGF at approximately 62.41%, and FGAF at the lowest at approximately 72.1%. In conclusion, FFGG shows the best antioxidant effect among single peptides.

[0043] The results of the DPPH radical scavenging rate and ABTS radical scavenging rate of the complex peptide solution are as follows: Figure 16 As shown, the free radical scavenging rate of the complex peptides was not as significant as that of the single peptide FFGG.

[0044] In summary, the FFGG single peptide exhibits the best scavenging rate for both DPPH and ABTS free radicals.

[0045] Results of hyaluronic acid inhibition rate of single peptide solution are as follows: Figure 17As shown, FFGG showed the most significant effect with an inhibition rate of 67.67%, followed by FGAF with an inhibition rate of 49.33%, while FDGF had the lowest inhibition rate of only 43.33%.

[0046] The results of the hyaluronic acid inhibition rate of the complex peptide solution are as follows: Figure 18 As shown, the combination of FFGG+FDGF has the best hyaluronic acid inhibition rate, followed by the combination of FFGG+FGAF, then FGAF+FDGF, and the combination of all three has the lowest effect.

[0047] The hyaluronic acid inhibition rate of single peptides was generally higher than that of mixed samples, especially FFGG, which was higher than that of other single peptides and combined peptides. After compounding, there may be antagonistic effects between different peptides, especially when three peptides are mixed, the antagonistic effect is the most obvious. Furthermore, after compounding, the effective concentration of highly active peptides is reduced, and there is no synergistic effect.

[0048] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A complex functional protein peptide derived from sea cucumber, characterized in that, The amino acid sequence of the protein peptide is at least one of FFGG, FGAF, and FDGF.

2. The application of the sea cucumber-derived complex functional protein peptides of claim 1 in the preparation of products with antioxidant and anti-allergic effects.

3. The application according to claim 2, characterized in that, The products mentioned are food, medicine, or health products.

4. The application according to claim 3, characterized in that, The product further includes food, pharmaceutical or health food-grade excipients.

5. The application according to claim 2, characterized in that, The antioxidant described herein has a free radical scavenging effect.

6. The application according to claim 5, characterized in that, The free radical is at least one of DPPH free radical and ABTS free radical.

7. The application according to claim 2, characterized in that, The anti-allergic effect is due to the inhibition of hyaluronic acid.

8. A complex functional protein peptide preparation derived from sea cucumber, characterized in that, The active ingredient is at least one of FFGG, FGAF, and FDGF.

9. The sea cucumber-derived compound functional protein peptide preparation according to claim 8, characterized in that, The concentration of the active ingredient is 1 mg / mL.

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