Active peptides, active peptide compositions and use thereof in the manufacture of products having an anti-aging effect

Abstract

The application relates to the technical field of medicine, and particularly discloses an active peptide, an active peptide composition and application of the active peptide and the active peptide composition in preparation of products with anti-aging effects. The active peptide has an amino acid sequence shown in SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3; wherein the amino acid sequence of SEQ ID NO:1 is Gly-Phe-Glu-Tyr; the amino acid sequence of SEQ ID NO:2 is Ala-Glu-Phe-Gly; and the amino acid sequence of SEQ ID NO:3 is Val-Glu-Phe-Ala-His. The composition comprises a combination of any two or three of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3. The active peptide and the active peptide composition have anti-aging effects; therefore, the active peptide or the active peptide composition is used as an effective component to prepare products with anti-aging effects, which has important application value.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to an active peptide, an active peptide composition, and its application in the preparation of products with anti-aging effects. Background Technology

[0002] The factors inducing aging are complex and diverse, among which oxidative stress and oxidative damage are important contributing factors. Therefore, alleviating and reducing oxidative stress and oxidative damage are crucial measures to address aging. Numerous studies have found that probiotics and their metabolites colonizing the body possess excellent antioxidant activity. Further research has revealed that the regulation of the gut microbiota by probiotics and their metabolites can significantly enhance the gut's antioxidant capacity, thereby delaying aging.

[0003] Gamma-aminobutyric acid (GABA), a short-chain fatty acid secreted by probiotics, has activities such as lowering blood pressure, aiding sleep, and enhancing liver function. In skincare products, GABA exhibits anti-wrinkle and anti-aging activities. Numerous studies have shown that bioactive peptides, especially bioactive oligopeptides, also possess antioxidant and anti-aging physiological activities. Furthermore, their high water solubility and absorption make them widely used in food, pharmaceuticals, and skincare products.

[0004] Lactobacillus reuteri has a strong adhesion ability to the intestinal mucosa, which can improve the distribution of intestinal flora and antagonize the colonization of harmful bacteria, making it one of the common probiotics. However, the amount of γ-aminobutyric acid (GABA) secreted by the known Lactobacillus reuteri is relatively small, and its anti-aging effect needs to be further improved; there are also few reports on the preparation of anti-aging active peptides from Lactobacillus reuteri. Summary of the Invention

[0005] To overcome at least one of the technical problems existing in the prior art, the present invention first provides an active peptide. This active peptide was prepared for the first time from the lysate of *Limosilactobacillus reuteri* HCLR01 cells; it possesses anti-aging activity.

[0006] The technical solution of the present invention is as follows: The present invention first provides an active peptide having the amino acid sequence shown in SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3; The amino acid sequence of SEQ ID NO:1 is: Gly-Phe-Glu-Tyr; The amino acid sequence of SEQ ID NO:2 is: Ala-Glu-Phe-Gly; The amino acid sequence of SEQ ID NO:3 is: Val-Glu-Phe-Ala-His.

[0007] This invention provides a novel bioactive peptide; studies have shown that the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 all have anti-aging activity; in particular, the bioactive peptide with the amino acid sequence shown in SEQ ID NO:2 has the most significant anti-aging activity, which is significantly better than that of the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2.

[0008] The present invention also provides a method for preparing the above-mentioned active peptide, wherein the active peptide is isolated from the lysate of Limosilactobacillus reuteri HCLR01 cells.

[0009] The Lactobacillus reuteri HCLR01 mentioned above has the accession number GDMCC No: 63144.

[0010] The strain was deposited on January 13, 2023 at the Guangdong Provincial Center for Microbial Culture Collection (Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences Institute of Microbiology).

[0011] The *Limosilactobacillus reuteri* HCLR01 is rod-shaped with rounded ends. Its 16S rDNA sequence (as shown in SEQ ID NO:4) was analyzed and compared with the sequencing results in the NCBI database. Therefore, it was finally identified as *Limosilactobacillus reuteri*.

[0012] Preferably, the active peptide is an active peptide with anti-aging effects.

[0013] This invention provides a novel method for preparing bioactive peptides with anti-aging effects; this method is the first to prepare bioactive peptides with anti-aging effects from the lysate of Limosilactobacillus reuteri HCLR01 cells.

[0014] Preferably, the *Limosilactobacillus reuteri* HCLR01 cell lysis product is prepared by the following method: (1) Take Limosilactobacillus reuteri HCLR01 and mix it with water, then sonicate it to obtain a lysate; freeze-dry the lysate and then defatt it to obtain freeze-dried product A; (2) Mix the lyophilized product A with water, and then dialyze it with a dialysis membrane with a pore size of 0.8~1.2 kD. Take the lyophilized product B after lyophilizing the dialysate, and you will get the lyophilized product of Limosilactobacillus reuteri HCLR01 cells.

[0015] Preferably, the ultrasonic fragmentation time is 10-30 minutes.

[0016] Preferably, the ultrasonic power of the ultrasonic fragmentation is 500~1000W, and the ultrasonic frequency of the ultrasonic fragmentation is 20~40KHz.

[0017] Preferably, the Lactobacillus reuteri HCLR01 mentioned in step (1) is a lyophilized powder of Lactobacillus reuteri HCLR01.

[0018] Preferably, in step (1), the weight ratio of Limosilactobacillus reuteri HCLR01 lyophilized powder to water is 1:8~15.

[0019] Most preferably, in step (1), the weight ratio of Limosilactobacillus reuteri HCLR01 lyophilized powder to water is 1:10.

[0020] Preferably, in step (2), dialysis is performed using a dialysis membrane with a pore size of 1 kD.

[0021] Preferably, in step (2), the weight ratio of freeze-dried product A to water is 1:8~15.

[0022] Most preferably, in step (2), the weight ratio of freeze-dried product A to water is 1:10.

[0023] Preferably, the method for preparing the active peptide specifically includes the following steps: Dextran from the lysate of *Limosilactobacillus reuteri* HCLR01 cells Gel column; then elution, collection of eluent, concentration and drying to obtain dextran gel column eluent; The active peptide was obtained by separating the dextran gel column eluent using preparative HPLC.

[0024] Preferably, the dextran gel column is eluted with a (NH4)2SO4 buffer solution with a concentration of 1.0~1.5mM.

[0025] Most preferably, the dextran gel column is eluted with a 1.2 mM (NH4)2SO4 buffer solution.

[0026] Preferably, the specific elution conditions in the dextran gel column are as follows: Dextran from the lysate of *Limosilactobacillus reuteri* HCLR01 cells The gel column was prepared, and then eluted with 1.20 mM (NH4)2SO4 buffer. Each elution of 1 / 20 column volume was recorded as a fraction and numbered. Fractions 20-30 were combined, concentrated, and dried to obtain the dextran gel column eluate.

[0027] Preferably, the specific conditions for the preparative HPLC are as follows: gradient elution is performed using 0.05~0.2% trifluoroacetic acid-water solution as mobile phase A and 0.05~0.2% trifluoroacetic acid-acetonitrile solution as mobile phase B; the detection wavelength is 210~230 nm and the flow rate is 10 mL / min.

[0028] Most preferably, the gradient elution conditions for preparing the active peptide with the amino acid sequence shown in SEQ ID NO:1 are specified. Including: at 0 min, the volume fraction of mobile phase A is 18.0% and the volume fraction of mobile phase B is 82.0%; at 25 min, the volume fraction of mobile phase A is 43.0% and the volume fraction of mobile phase B is 57.0%; at 25.1 min, the volume fraction of mobile phase A is 100% and the volume fraction of mobile phase B is 0%. The eluent corresponding to the chromatographic peak at 10.276 min was collected, concentrated, and dried to obtain the active peptide with the amino acid sequence shown in SEQ ID NO:1.

[0029] Most preferably, the gradient elution conditions for preparing the active peptides with the amino acid sequences shown in SEQ ID NO:2 and SEQ ID NO:3 include: at 0 min, the volume fraction of mobile phase A is 10.0% and the volume fraction of mobile phase B is 90.0%; at 25 min, the volume fraction of mobile phase A is 35.0% and the volume fraction of mobile phase B is 65.0%; at 25.1 min, the volume fraction of mobile phase A is 100% and the volume fraction of mobile phase B is 0%. Collect the eluent corresponding to the chromatographic peak at 11.960 min, concentrate and dry it to obtain the active peptide with the amino acid sequence shown in SEQ ID NO:2; The eluent corresponding to the chromatographic peak at 12.484 min was collected, concentrated, and dried to obtain the active peptide with the amino acid sequence shown in SEQ ID NO:3.

[0030] The present invention also provides an active peptide composition comprising any two or three of the active peptides with amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3.

[0031] Preferably, the active peptide composition comprises active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3.

[0032] More preferably, the weight ratio of the active peptides of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 is 1~3:1~10:1~5.

[0033] Most preferably, the weight ratio of the active peptides of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 is 2:5:3.

[0034] In further research, the inventors surprisingly discovered that the composition obtained by combining the active peptides of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 exhibited a significantly enhanced anti-aging activity; its anti-aging activity was significantly higher than that of the active peptides of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 alone. This may be the result of a synergistic anti-aging effect among the three active peptides after combining them.

[0035] In some places below, for the sake of brevity, the active peptide of the amino acid sequence shown in SEQ ID NO:1 is referred to as oligopeptide-1 (abbreviated as HCOP-1); the active peptide of the amino acid sequence shown in SEQ ID NO:2 is referred to as oligopeptide-2 (abbreviated as HCOP-2); and the active peptide of the amino acid sequence shown in SEQ ID NO:3 is referred to as oligopeptide-3 (abbreviated as HCOP-3).

[0036] Beneficial effects: (1) This invention provides a novel bioactive peptide; studies have shown that the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 all possess anti-aging activity; in particular, the bioactive peptide with the amino acid sequence shown in SEQ ID NO:2 exhibits the most significant anti-aging activity, which is significantly superior to that of the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2. Furthermore, the composition obtained by combining the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 further significantly enhances its anti-aging activity; its anti-aging activity is significantly higher than that of the bioactive peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 alone.

[0037] (2) The present invention also provides a novel method for preparing an active peptide with anti-aging effects; the method is the first to prepare an active peptide with anti-aging effects from the lysate of Limosilactobacillus reuteri HCLR01 cells.

[0038] (3) Since the active peptides and active peptide compositions described in this invention have anti-aging effects, it is of great application value to use the active peptides or active peptide compositions described in this invention as effective ingredients to prepare products with anti-aging effects. Attached Figure Description

[0039] Figure 1 This is the mass spectrum of oligopeptide-1 (HCOP-1).

[0040] Figure 2 This is an HPLC chromatogram of oligopeptide-1 (HCOP-1).

[0041] Figure 3 This is the mass spectrum of oligopeptide-2 (HCOP-2).

[0042] Figure 4 This is an HPLC chromatogram of oligopeptide-2 (HCOP-2).

[0043] Figure 5 This is the mass spectrum of oligopeptide-3 (HCOP-3).

[0044] Figure 6 This is an HPLC chromatogram of oligopeptide-3 (HCOP-3). Detailed Implementation

[0045] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0046] Example 1: Isolation and Identification of Limosilactobacillus reuteri HCLR01 (1) Enrichment culture of microorganisms: Fecal treatment liquid from different healthy children was added to a culture medium containing glutamic acid (8.5 g / L peptone, 8.5 g / L yeast powder, 8.5 g / L glutamic acid, 9.5 g / L glucose, 1.2 mL / L Tween 80, 5.5 g / L sodium acetate, 2.2 g / L dipotassium hydrogen phosphate, 0.6 g / L magnesium sulfate, and 0.4 g / L manganese sulfate) and cultured statically at 33℃ for 6 h.

[0047] (2) Qualitative detection: The culture medium after step (1) was concentrated and qualitatively detected. Thin-layer chromatography was used. The concentrated culture medium was spotted onto a silica gel pre-prepared plate, and the chromatography solvent was developed. The chromatography solvent was chloroform: glacial acetic acid: water (65:25:10), with 0.5% ninhydrin added. After development, the plate was air-dried and developed at 98ºC for 2 min, and then compared with the standard.

[0048] (3) Isolation of single colonies: The culture medium with γ-aminobutyric acid production obtained in step (2) was examined under a microscope to determine the bacterial species. The culture medium was then diluted and spread on a plate containing the bacteria. The plate medium was a probiotic solid medium containing 12 g / L CaCO3 (8.8 g / L peptone, 5.5 g / L beef extract, 8.8 g / L yeast extract, 10.0 g / L glucose, 2.2 g / L dipotassium hydrogen phosphate, 2.0 g / L triammonium citrate, 5.5 g / L sodium acetate, 0.6 g / L magnesium sulfate, 0.4 g / L manganese sulfate, 1.2 g / L Tween-80, and 16.0 g / L agar). The culture was incubated at 33℃ for 6 h. Several colonies with calcium dissolution zones (smooth and with neat edges) were picked and inoculated into a medium containing glutamic acid. The culture was then kept still at 33℃ for 6 h. The γ-aminobutyric acid production was qualitatively detected and compared. The strain with the highest γ-aminobutyric acid (GABA) production, which is rod-shaped with rounded ends, was selected as Lactobacillus reuteri HCLR01.

[0049] The strain was identified as *Limosilactobacillus reuteri* by analyzing its 16S rDNA sequence (as shown in SEQ ID NO:4) and comparing the sequencing results with the NCBI database.

[0050] Example 2 Preparation of Limosilactobacillus reuteri HCLR01 lyophilized powder (1) Preparation of Limosilactobacillus reuteri HCLR01 seed culture: Limosilactobacillus reuteri HCLR01 was inoculated into a culture medium (the composition of the culture medium was: peptone 8.8 g / L, beef extract 5.5 g / L, yeast extract 8.8 g / L, glucose 10.0 g / L, dipotassium hydrogen phosphate 2.2 g / L, triammonium citrate 2.0 g / L, sodium acetate 5.5 g / L, magnesium sulfate 0.6 g / L, manganese sulfate 0.4 g / L, Tween-80 1.2 g / L, agar 16.0 g / L), and fermented in shake flasks for 12 h to obtain Limosilactobacillus reuteri HCLR01 seed culture with OD 600 > 1.5.

[0051] (2) Scale-up culture: The *Limosilactobacillus reuteri* HCLR01 seed culture obtained in step (1) was inoculated into a 20 L fermenter with liquid culture medium (the culture medium composition was: peptone 8.8 g / L, beef extract 5.5 g / L, yeast extract 8.8 g / L, glucose 10.0 g / L, dipotassium hydrogen phosphate 2.2 g / L, triammonium citrate 2.0 g / L, sodium acetate 5.5 g / L, magnesium sulfate 0.6 g / L, manganese sulfate 0.4 g / L, Tween-80 1.2 g / L, agar 16.0 g / L); the culture was stirred and cultured at pH 6 and 37℃ for 48 h; the fermentation was then stopped to obtain the fermentation broth. (3) Centrifuge the fermentation broth obtained in step (2) to collect the cells, then add water and skim milk powder as a freeze-drying protectant, mix evenly, and freeze-dry to obtain a freeze-dried powder of Lactobacillus reuteri HCLR01 with a viable count of 10 billion cfu / g.

[0052] Example 3 Preparation of Lactobacillus reuteri lysate (1) Take Limosilactobacillus reuteri HCLR01 lyophilized powder (10 billion CFU / g) and mix it with 10 times its weight of water; then, under the conditions of ultrasonic power of 900W and ultrasonic frequency of 25KHz, ultrasonically break it up for 15 minutes to obtain the broken liquid; freeze-dry the broken liquid and then defatt it with isopropanol to obtain lyophilized product A; (2) Mix the lyophilized product A with 10 times its weight of water, then dialyze it with a dialysis membrane with a pore size of 1 kD, and take the lyophilized product B after lyophilizing the dialysate to obtain the Lactobacillus reuteri cell lysis product.

[0053] Example 4 Preparation of active peptides (1) The lysate of Limosilactobacillus reuteri HCLR01 cells was loaded onto the plate. Dextran gel column (specifically dextran gel G-15 filler); then eluted with 1.20 mM (NH4)2SO4 buffer, each eluted 1 / 20 column volume of eluent is taken as one fraction and numbered; fractions 20-30 are combined, concentrated and dried to obtain dextran gel column eluent; (2) The dextran gel column eluent was separated by preparative HPLC to obtain the active peptide; The specific conditions for the preparative HPLC were as follows: gradient elution was performed using 0.1% trifluoroacetic acid-water solution as mobile phase A and 0.1% trifluoroacetic acid-acetonitrile solution as mobile phase B; the detection wavelength was 220 nm and the flow rate was 10 mL / min; the chromatographic column was an XBridge BEH C18 OBD Prep Column, 5 µm, 19 mm * 150 mm.

[0054] The gradient elution conditions for preparing the active peptide with the amino acid sequence shown in SEQ ID NO:1 are included. Including: at 0 min, the volume fraction of mobile phase A is 18.0% and the volume fraction of mobile phase B is 82.0%; at 25 min, the volume fraction of mobile phase A is 43.0% and the volume fraction of mobile phase B is 57.0%; at 25.1 min, the volume fraction of mobile phase A is 100% and the volume fraction of mobile phase B is 0%. The eluent corresponding to the chromatographic peak at 10.276 min was collected, concentrated, and dried to obtain oligopeptide-1 (HCOP-1), which is the active peptide with the amino acid sequence shown in SEQ ID NO:1.

[0055] The gradient elution conditions for preparing the active peptides with the amino acid sequences shown in SEQ ID NO:2 and SEQ ID NO:3 included: at 0 min, the volume fraction of mobile phase A was 10.0% and the volume fraction of mobile phase B was 90.0%; at 25 min, the volume fraction of mobile phase A was 35.0% and the volume fraction of mobile phase B was 65.0%; at 25.1 min, the volume fraction of mobile phase A was 100% and the volume fraction of mobile phase B was 0%. The eluent corresponding to the chromatographic peak at 11.960 min was collected, concentrated and dried to obtain oligopeptide-2 (HCOP-2), which is the active peptide with the amino acid sequence shown in SEQ ID NO:2. The eluent corresponding to the chromatographic peak at 12.484 min was collected, concentrated, and dried to obtain oligopeptide-3 (HCOP-3), which is the active peptide with the amino acid sequence shown in SEQ ID NO:3.

[0056] Furthermore, we used HPLC and mass spectrometry to determine and analyze the molecular weight and amino acid sequence of oligopeptide-1 (HCOP-1), oligopeptide-2 (HCOP-2), and oligopeptide-3 (HCOP-3), and the results are as follows: Oligopeptide-1 (HCOP-1): m / z 515.2122 is [M+H] + Ions; m / z 334.1394 is a Y3 ion; m / z 205.0970 is a Y2 ion; m / z 136.075 is [Tyr-COOH+H] + The ion with m / z 120.0804 is [Phe-COOH+H]. + Ion; m / z 84.0442 is [Glu-COOH-H2O+H] + Ions; 56.0498 is [Gly-H₂O+H] + Ions. Based on the results of the Edman degradation experiment, oligopeptide-1 (HCOP-1) was finally identified as an oligopeptide with the amino acid sequence Gly-Phe-Glu-Tyr; that is, the active peptide with the amino acid sequence shown in SEQ ID NO:1.

[0057] Oligopeptide-2 (HCOP-2): m / z 423.1870 is [M+H] + Ion; m / z 405.1763 is [M-H₂O+H] + Ion; m / z 388.1506 is [M-2H2O+H] + The ion with m / z 231.1122 is [y3-y1-COOH+H]. + Ions; m / z 201.0864 is γ2 ion; 120.0817 is [Phe-COOH+H] ion. +Ion; m / z 84.0443 is [Glu-COOH-H2O+H] + Ions. Based on the results of the Edman degradation experiment, oligopeptide-2 (HCOP-2) was finally identified as an oligopeptide with the amino acid sequence Ala-Glu-Phe-Gly; that is, the active peptide with the amino acid sequence shown in SEQ ID NO:2.

[0058] Oligopeptide-3 (HCOP-3): m / z 602.2942 is [M+H] + Ions; m / z 447.2242 is a Y4 ion; m / z 376.1864 is a Y3 ion; m / z 229.1184 is a Y2 ion; m / z 156.0766 is [His-H2O+H] + Ion; 110.0712 is [His-COOH+H] + Ions. Based on the results of the Edman degradation experiment, oligopeptide-3 (HCOP-3) was finally identified as an oligopeptide with the amino acid sequence Val-Glu-Phe-Ala-His; that is, the active peptide with the amino acid sequence shown in SEQ ID NO:3.

[0059] Example 5 Preparation of active peptide composition The active peptide composition is obtained by mixing the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 in a weight ratio of 2:5:3.

[0060] Example 6 Preparation of active peptide composition The active peptide composition is obtained by mixing the active peptides with the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2 at a weight ratio of 2:5.

[0061] Example 7 Preparation of active peptide composition The active peptide composition is obtained by mixing the active peptides with the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:3 at a weight ratio of 2:3.

[0062] Example 8 Preparation of active peptide composition The active peptide composition is obtained by mixing the active peptides with the amino acid sequences shown in SEQ ID NO:2 and SEQ ID NO:3 at a weight ratio of 5:3.

[0063] Experiment Example 1: Anti-aging Experiment Caenorhabditis elegans L4 larvae were randomly divided into 9 groups, with 100 larvae in each group; there were 8 experimental groups and 1 blank control group. The experimental group Caenorhabditis elegans L4 larvae were placed on NGM plates containing 1 mg / mL of the test sample and cultured. The blank control group Caenorhabditis elegans L4 larvae were placed on NGM plates without the test drug and cultured. Caenorhabditis elegans larvae that could not move were marked as dead. The number of larvae was counted daily, and each experiment was repeated 3 times. The average lifespan of Caenorhabditis elegans in each group was calculated. The test samples and experimental results of each experimental group are shown in Table 1.

[0064] Table 1. Results of anti-aging experimental tests Blank control group - 20 days Experimental group 1 The active peptide with the amino acid sequence shown in SEQ ID NO:1 23 days Experimental group 2 The active peptide with the amino acid sequence shown in SEQ ID NO:2 27 days Experimental group 3 The active peptide with the amino acid sequence shown in SEQ ID NO:3 22 days Experimental group 4 Example 5: Active Peptide Composition 32 days Experimental group 5 Example 6: Active Peptide Composition 25 days Experimental group 6 Example 7 Active peptide composition 22 days Experimental group 8 Example 8: Active Peptide Composition 24 days As can be seen from the experimental results in Table 1, the average lifespan of *C. elegans* in experimental groups 1-3 was significantly higher than that in the blank control group; among them, the average lifespan of *C. elegans* in experimental group 2 was the highest, significantly higher than that in the blank control group and significantly higher than that in experimental groups 1 and 3. This indicates that the active peptides of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 of this invention all have anti-aging activity; in particular, the active peptide of the amino acid sequence shown in SEQ ID NO:2 has the most significant anti-aging activity, which is significantly better than that of the active peptides of the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2.

[0065] As can be seen from the experimental results in Table 1, the average lifespan of Caenorhabditis elegans in experimental group 4 was significantly higher than that in experimental groups 1-3. This indicates that the composition obtained by combining the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 has a significantly enhanced anti-aging activity. Its anti-aging activity is significantly higher than that of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 alone. The combination of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 can produce a synergistic anti-aging effect.

[0066] As can be seen from the experimental results in Table 1, the average lifespan of Caenorhabditis elegans in experimental groups 6-7 was not significantly or substantially longer than that in experimental groups 1-3. This indicates that the combination of any two of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 does not significantly enhance the anti-aging activity; and the combination of any two of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 does not produce a synergistic anti-aging effect.

Claims

1. An active peptide, characterized in that, The amino acid sequence is shown in SEQ ID NO:3; The amino acid sequence of SEQ ID NO:3 is: Val-Glu-Phe-Ala-His.

2. An active peptide composition, characterized in that, An active peptide comprising the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3; The amino acid sequence of SEQ ID NO:1 is: Gly-Phe-Glu-Tyr; The amino acid sequence of SEQ ID NO:2 is: Ala-Glu-Phe-Gly; The amino acid sequence of SEQ ID NO:3 is: Val-Glu-Phe-Ala-His.

3. The active peptide composition according to claim 2, characterized in that, The weight ratio of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 is 1~3:1~10:1~5.

4. The active peptide composition according to claim 3, characterized in that, The weight ratio of the active peptides with the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 is 2:5:

3.

5. The use of the active peptide of claim 1 or the active peptide composition of any one of claims 2-4 in the preparation of products with anti-aging effects.

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