Anti-inflammatory peptide derived from western-style dry-cured ham, preparation method and application thereof

Abstract

The present invention belongs to the field of food biotechnology, and discloses an anti-inflammatory peptide derived from western-style dry-cured ham, and a preparation method and application thereof. The present invention uses western-style dry-cured ham as a raw material, obtains a natural anti-inflammatory peptide complex through direct extraction, separation and purification, and obtains a natural small molecule anti-inflammatory peptide by screening in combination with bioinformatics technology. The bioactive peptide can effectively inhibit the secretion of NO and inflammatory cytokines (IL-1 or IL-6 or TNF-α) by RAW264.7 cells stimulated by lipopolysaccharide, and has anti-inflammatory properties. The present invention expands the application scope of western-style dry-cured ham polypeptides, which can not only promote their high-value utilization, but also can be used as a potential functional factor in anti-inflammatory products.

Description

Technical Field

[0001] The invention belongs to the field of food biotechnology, and in particular relates to an anti-inflammatory peptide derived from western-style dry-cured ham and a preparation method and application thereof. Background Art

[0002] Inflammation is a biological process that maintains homeostasis by protecting the host from pathogens and tissue damage, and involves tissue repair, regeneration and remodeling. It plays a key role in the host's immune defense system. Excessive inflammatory response can have a significant impact on the body, and even lead to the occurrence and deterioration of diseases such as cardiovascular and cerebrovascular diseases, tumors, cancer, diabetes, atherosclerosis and asthma. At present, inhibiting or intervening in inflammation-related signaling pathways and reducing the secretion of inflammatory cytokines (IL-6, TNF-α, IL-1β) have become the mainstream of research on the prevention and treatment of inflammation. Commonly used anti-inflammatory drugs (steroids, non-steroidal anti-inflammatory drugs and antibiotics, etc.) have certain side effects. Long-term use can reduce the repairability of tissues and cause gastrointestinal mucosal damage. Therefore, the research and development of natural active substances that have the function of treating or assisting the treatment of inflammation and are safe and without side effects has become the focus of researchers.

[0003] Bioactive peptides are one of the research hotspots in the field of life and food science. They are usually composed of 2-20 amino acid residues. Depending on the amino acid composition, sequence and structure, they can play a variety of physiological functions such as anti-hypertension, anti-diabetes, antioxidant, anti-bacterial, anti-inflammatory, and anti-cancer. Currently, 5135 bioactive peptides are listed in the BIOPEP database. Compared with traditional chemical drugs, bioactive peptides have higher tissue affinity and specificity, and fewer side effects, and have broad prospects in the prevention and treatment of diseases.

[0004] The unique processing technology and long processing process of Western dry-cured ham make its muscle protein highly degraded under the combined action of endogenous enzymes and exogenous microorganisms, becoming an important carrier rich in bioactive peptides and a direct source of active peptides. At present, bioactive peptides from various sources of dry-cured ham have been successfully separated, purified and identified. The research mainly focuses on angiotensin converting enzyme inhibitory activity, antioxidant activity, antibacterial activity, etc. However, there are relatively few studies and reports on the preparation and application of anti-inflammatory peptides from Western dry-cured ham at home and abroad. Therefore, the preparation and identification of ham peptides and the discovery of new active functions are of great significance for the full exploration and high-value utilization of bioactive peptide resources from Western dry-cured ham.

[0005] At present, the main methods for preparing anti-inflammatory peptides include microbial fermentation, enzymatic hydrolysis and chemical synthesis. Microbial fermentation is a process that uses proteases in microbial metabolic activities to degrade proteins into active peptides during the fermentation process. It has a simple process and low cost, but the product is affected by the raw materials, strains and processes. The fermentation process is difficult to control, which can easily lead to unstable product quality and low product standardization. Enzymatic hydrolysis is the most commonly used method for preparing active peptides, which is to break the peptide bonds in proteins through protease hydrolysis to release bioactive peptides. However, due to the randomness and uncertainty of protease cleavage sites, the workload of screening proteases is heavy and has a certain degree of blindness. The chemical synthesis method is based on the premise of a known active peptide amino acid sequence, and a condensation reaction is carried out from the C-terminus to the N-terminus of the amino acid. Several methods have their own advantages and disadvantages, and in practical applications, they need to be selected according to actual conditions.

[0006] The separation, purification and identification of anti-inflammatory peptides are also important links. At present, most studies combine traditional separation technology with modern biotechnology to carry out targeted separation of target products, but lack the preparation, identification and verification of components related to the anti-inflammatory activity of Western dry-cured ham. In addition, different preparation and separation methods will result in different active products. At present, there are only 50 peptides with anti-inflammatory activity and amino acid sequence data in the BIOPEP-UWM database. Therefore, the development of anti-inflammatory peptides from Western dry-cured ham and the discovery of new sequence peptides are of great significance for their high-value utilization and in the field of anti-inflammatory product research. Summary of the invention

[0007] The purpose of the present invention is to provide an anti-inflammatory peptide derived from western-style dry-cured ham and a preparation method and application thereof.

[0008] In order to achieve the purpose of the present invention, in a first aspect, the present invention provides an anti-inflammatory peptide (anti-inflammatory active peptide), the amino acid sequence of the anti-inflammatory active peptide is as shown in any one of LPFP (SEQ ID NO: 1), PYPPP (SEQ ID NO: 2) and IPPPL (SEQ ID NO: 3), preferably LPFP (SEQ ID NO: 1).

[0009] In a second aspect, the present invention provides an anti-inflammatory peptide complex comprising two or three peptides of any one of the sequences shown in SEQ ID NOs: 1-3, or a complex with the three active peptides as main components.

[0010] In a third aspect, the present invention also provides a method for preparing the anti-inflammatory peptide or the anti-inflammatory peptide complex, wherein the anti-inflammatory peptide or the anti-inflammatory peptide complex is extracted using western-style dry-cured ham as a raw material, and the fermentation and maturation period of the western-style dry-cured ham is 1 year or more.

[0011] Furthermore, the preparation method of the western-style dry-cured ham comprises: trimming and shaping the raw pork leg, removing blood, and ripening it through the following processing procedures: salting (temperature of 0-4°C, about 20 days), pickling (temperature of 3-7°C, relative humidity of 60-80%, about 60 days), and aging (temperature of 12-18°C, relative humidity of 60-85%, not less than 280 days);

[0012] The specific preparation method comprises the following steps:

[0013] (1) Preparation of crude peptide: Take mature Western dry-cured ham lean meat, remove the fascia tissue, cut into pieces and mince, then add hydrochloric acid solution to homogenize, centrifuge the homogenate, take the supernatant and add anhydrous ethanol to precipitate protein, then take the supernatant and filter it with a 0.45 μm filter membrane, concentrate it by rotary evaporation, and freeze-dry it to obtain crude peptide powder I.

[0014] (2) Ultrafiltration and gel filtration chromatography separation: The crude peptide powder I was re-dissolved into a crude peptide solution with a mass concentration of 25 mg / mL, and the peptides with a molecular weight cutoff of less than 3 kDa were ultrafiltered and freeze-dried to obtain peptide powder II. Peptide powder II was prepared into a peptide solution with a mass concentration of 10-50 mg / mL, and separated using a Sephadex g-15 dextran gel column with a sample volume of 2.0-4.0 mL. The eluent was ultrapure water after ultrasonic degassing, the flow rate was 1-1.5 mL / min, and the detection wavelength was 280 nm. Two main components (F1 and F2) were collected.

[0015] (3) Identification and screening: The components with the best anti-inflammatory effects were selected, analyzed and screened by sequencing combined with bioinformatics, and verified by in vitro cell experiments to obtain an anti-inflammatory peptide complex and three new anti-inflammatory peptides (SEQ ID NO: 1-3).

[0016] Preferably, in step (2), the peptide powder II is configured with a mass concentration of 50 mg / mL, and is separated by a Sephadex g-15 dextran gel column with a sample volume of 4.0 mL and an eluent flow rate of 1.5 mL / min.

[0017] Preferably, the specific operation for determining the anti-inflammatory effect in step (3) is as follows: a lipopolysaccharide-induced RAW264.7 macrophage inflammation model is established; when the NO release amount and the inflammatory cytokine (IL-6 or TNF-α or IL-1β) secretion amount of the peptide incubation group are lower than those of the model group (LPS group), the peptide is considered to have anti-inflammatory potential; otherwise, the peptide is considered to have no anti-inflammatory potential.

[0018] To achieve the above object and other related objects, the present invention provides a method for screening an anti-inflammatory peptide complex, comprising the following steps:

[0019] (1) Use the PeptideRanker tool to score the potential biological activity of peptides;

[0020] (2) The biological activity of peptides with a score of 0.8 or above in (1) was predicted using the BIOPEP-UWM database. Peptides containing anti-inflammatory amino acid sequences such as IPP, PY, and LPF were screened based on the anti-inflammatory amino acid and its constituent sequence characteristics.

[0021] (3) The novelty of the peptide sequences in (2) was checked using the BIOPEP-UWM database, and three new anti-inflammatory peptides were obtained.

[0022] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0023] (I) The present invention uses Western dry-cured ham as raw material, and obtains a natural small molecule anti-inflammatory peptide complex through direct extraction, separation, purification and screening combined with bioinformatics technology. The preparation cost is low, the conditions are mild, the anti-inflammatory activity of the peptide is effectively maintained, and the preparation process is less polluting and green and environmentally friendly;

[0024] (ii) The present invention expands the application scope of Western-style dry-cured ham polypeptide, which can not only promote its high-value utilization, but also be used as a potential functional factor in anti-inflammatory products. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 These are two components (F1 and F2) separated and purified from the western-style dry-cured ham polypeptide (<3 kDa) by dextran gel filtration chromatography in a preferred embodiment of the present invention.

[0026] Figure 2 The figure shows the effect of F1 (A) and F2 (B) on the proliferation activity of RAW264.7 macrophages in the preferred embodiment of the present invention.

[0027] Figure 3 The effect of F1 and F2 on NO production by RAW264.7 macrophages in the preferred embodiment of the present invention. The ordinate is the NO content (μM), and the abscissa is NC (negative control group), LPS (lipopolysaccharide stimulation group) and polypeptide group (100μg / mL, 250μg / mL, 500μg / mL, 1000μg / mL).

[0028] Figure 4 This is the effect of F1 on cytokine production by RAW264.7 macrophages in a preferred embodiment of the present invention. The ordinate is the concentration of cytokines, and the abscissa is NC (negative control group), LPS (lipopolysaccharide stimulation group) and polypeptide group (100 μg / mL, 250 μg / mL, 500 μg / mL, 1000 μg / mL).

[0029] Figure 5 The inhibition rate of LPFP, PYPPP and IPPPL on the proliferation activity of RAW264.7 macrophages (A) and the NO production of RAW264.7 macrophages stimulated by lipopolysaccharide (B) in the preferred embodiments of the present invention.

[0030] Figure 6 This is the effect of LPFP, PYPPP and IPPPL on cytokine production by RAW264.7 macrophages in a preferred embodiment of the present invention. DETAILED DESCRIPTION

[0031] The present invention aims to provide a natural anti-inflammatory peptide complex derived from western dry-cured ham, wherein the molecular weight of all peptide segments is less than 2.5 kDa, the peptide segments are composed of 3 to 20 amino acids, and the complex comprises three new peptide segments with anti-inflammatory activity, and the amino acid sequences are LPFP (SEQ ID NO: 1), PYPPP (SEQ ID NO: 2) and IPPPL (SEQ ID NO: 3).

[0032] The present invention also provides a method for preparing western-style dry-cured ham and its anti-inflammatory peptide complex, and the specific preparation steps are as follows:

[0033] (1) Processing of Western-style dry-cured ham and preparation of crude peptides: The raw pig legs are trimmed and shaped, and the blood is removed by massage using a massage machine. The ham is matured through salting (temperature 0-4°C, about 20 days), pickling (temperature 3-7°C, relative humidity 60-80%, about 60 days), and aging (temperature 12-18°C, relative humidity 60-85%, not less than 280 days). The mature lean meat of Western-style dry-cured ham is taken, the fascia tissue is removed, the ham is cut into pieces and minced, and then a hydrochloric acid solution is added to homogenize the slurry. After the homogenate is centrifuged, the supernatant is taken and anhydrous ethanol is added to precipitate the protein. Then, the supernatant is filtered with a 0.45 μm filter membrane, concentrated by rotary evaporation, and freeze-dried to obtain crude peptide powder I.

[0034] (2) Ultrafiltration and gel filtration chromatography separation: The crude peptide powder I was re-dissolved into a crude peptide solution with a mass concentration of 25 mg / mL, and the peptides with a molecular weight cutoff of less than 3 kDa were ultrafiltered and freeze-dried to obtain peptide powder II. Peptide powder II was prepared into a peptide solution with a mass concentration of 10-50 mg / mL, and separated using a Sephadex g-15 dextran gel column with a sample volume of 2.0-4.0 mL. The eluent was ultrapure water after ultrasonic degassing, the flow rate was 1-1.5 mL / min, and the detection wavelength was 280 nm. Two main components (F1 and F2) were collected.

[0035] (3) Identification and screening: The components with the best anti-inflammatory effect were selected through peptide sequence identification combined with bioinformatics analysis and screening and in vitro cell experiment verification to obtain anti-inflammatory peptide complexes and 3 new peptide sequences. The peptide sequences of the components are shown in SEQ ID NO: 1-3.

[0036] The present invention also provides a method for screening an anti-inflammatory peptide complex, comprising the following steps:

[0037] (1) Use the PeptideRanker tool to score the potential biological activity of the peptides in the complex;

[0038] (2) The biological activity of peptides with a score of 0.8 or above in (1) was predicted using the BIOPEP-UWM database, and peptides with potential anti-inflammatory activity were screened based on the anti-inflammatory active amino acids and their constituent sequence characteristics;

[0039] (3) The peptide sequences in (2) were checked using the BIOPEP-UWM database and three new peptides were obtained.

[0040] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0041] Example 1 Processing of Western-style dry-cured ham and preparation, separation and purification of active peptides

[0042] 1. Processing of Western-style dry-cured ham and preparation of crude peptide: The raw pig leg (taken from Beijing black pig) is trimmed and shaped, and the blood is removed by massage machine. After salting (temperature 0~4℃, about 20d), pickling (temperature 3~7℃, relative humidity 60~80%, about 60d), aging (temperature 12~18℃, relative humidity 60~85%, not less than 280d) and other processing procedures, the lean meat of Western-style dry-cured ham is taken, the fascia tissue is removed, and 30g is weighed after cutting and mincing. 120mL hydrochloric acid solution (0.01mol / L) is added, and the mixture is homogenized at 22000r / min for 3 times, each time for 10s, with a rest of 10s in between. The ham homogenate is left to stand at 4℃ for 2h, and then centrifuged at 4℃ and 12,000×g for 15min. Take the supernatant, add 3 times the volume of anhydrous ethanol solution, stand at 4°C for 20 hours to remove protein, and then centrifuge at 4°C and 12,000×g for 15 minutes. Take the supernatant, filter it with a 0.45μm filter membrane, concentrate it by rotary evaporation, and freeze-dry it to obtain crude peptide powder I.

[0043] 2. Ultrafiltration and gel filtration chromatography separation: The crude peptide powder I was re-dissolved into a crude peptide solution with a mass concentration of 25 mg / mL, and centrifuged using an ultrafiltration centrifuge tube with a molecular weight cutoff of 3 kDa. The centrifugal speed was 4000 rpm and the centrifugal volume was 15 mL. The liquid below the retention membrane was collected and freeze-dried to obtain peptide powder II. Peptide powder II was configured into a peptide solution with a mass concentration of 50 mg / mL, and separated using a Sephadexg-15 dextran gel column. The sample volume was 4.0 mL, the eluent was ultrapure water after ultrasonic degassing, the flow rate was 1.5 mL / min, and the detection wavelength was 280 nm. After the sample peak appears, 5 mL of eluent is collected per tube with a protein collector, and the eluents belonging to the same peak segment are mixed into the same sample, and then freeze-dried to obtain polypeptide powder.

[0044] 3. Results Analysis

[0045] The crude peptides from western dry-cured ham were separated by ultrafiltration and gel filtration chromatography to obtain two main components (F1 and F2) ( Figure 1 ), the molecular weights are in the order of F1>F2, and the activities of F1 and F2 are subsequently determined.

[0046] Example 2 Determination of anti-inflammatory activity of active peptide complex

[0047] 1. Recovery and culture of RAW264.7 macrophages

[0048] Take the cell cryopreservation tube out of the -80℃ ultra-low temperature refrigerator, place it in a 37℃ warm water bath for the shortest time, shake it gently to melt it quickly, wipe the outer wall of the cryopreservation tube with 75% alcohol and put it on the workbench. Open the cryopreservation tube, aspirate the cell suspension into a 15mL centrifuge tube, add 10mL of complete culture medium (DMEM high glucose culture medium + 10% fetal bovine serum + 1% double antibody), and blow it evenly (the general number of operations is 20-30 times). Centrifuge with a small centrifuge, centrifuge at 1000rpm for 5min, aspirate the supernatant, add 10mL of new complete culture medium, and gently blow it evenly to make a cell suspension. Aspirate the appropriate cell suspension and add it to the cell culture dish, move it into a cell culture incubator at 37℃ with 5% CO2 for static culture. Replace the cell culture medium every 24 hours and observe the cell adhesion and growth. If the cells are observed to be 70%-80% full of the bottom of the culture dish under a microscope, they should be subcultured in time. After cell subculture, replace the culture medium every 48h.

[0049] 2. Detection of cell proliferation activity (CCK-8 method)

[0050] The concentration of RAW264.7 macrophage suspension was adjusted to 3 × 10 5 / mL, add 100uL cell suspension to each well of a 96-well plate, and pre-culture the culture plate in an incubator for 24h. Add 10uL of peptide solution to the culture plate and continue incubation for 24h, add 10uL of CCK-8 solution to each well, continue incubation for 4h, and measure the absorbance at 450nm with an enzyme marker. Cell viability is calculated according to the following formula:

[0051] Cell viability (%) = [A(drug added) - A(blank)] / [A(0 drug added) - A(blank)] × 100

[0052] A (drug added): absorbance of wells with cells, CCK-8 solution, and peptide solution

[0053] A (blank): absorbance of wells with culture medium and CCK-8 solution but no cells

[0054] A(0 drug addition): absorbance of the well with cells and CCK-8 solution but no peptide solution

[0055] 3. NO content and cytokine determination

[0056] The macrophage cell concentration was adjusted to 5 × 10 5 Each well was added with 100 μL of cell suspension. After 4 hours of pre-culture, the supernatant was carefully aspirated and discarded, taking care not to touch the adherent cells. 100 μL of peptide samples of different mass concentrations were added to each well and cultured for 20 hours. After that, the supernatant was discarded. 100 μL of lipopolysaccharide (LPS) with a concentration of 2 μg / mL was added to each well for stimulation and culture for 20 hours. The supernatant was taken and the NO content was determined using a nitric oxide detection kit. The ELISA kit was used to determine the effect of LPS-induced cell inflammatory factors (IL-1β, IL-6, TNF-α) production. The inhibition rate of NO or cytokines = (LPS group-experimental group) / LPS group × 100%. The experimental groups were set up as follows:

[0057] Blank (NC) group: normal culture 24h+20h;

[0058] LPS group: normal culture for 24 h + LPS (2 μg / mL) stimulation culture for 20 h;

[0059] Experimental group: normal culture for 4 h + normal culture with sample for 20 h + LPS (2 μg / mL) stimulation culture for 24 h.

[0060] 4. Results Analysis

[0061] like Figure 2 As shown in the figure, F1 and F2 have no toxic effect on the proliferation activity of RAW264.7 cells in the mass concentration range of 100~1000μg / mL, and can promote cell proliferation. Figure 3It can be seen that compared with the blank control group, LPS induction significantly increased the secretion of NO in macrophages, and after pre-incubation with different mass concentrations of F1 and F2, and then induced by LPS, the secretion of NO in the cell supernatant of the treatment group was significantly reduced. At the same time, the NO inhibition rate was calculated (Table 1). Comprehensive analysis showed that F1 had a better inhibitory effect, and then its effect on cytokine secretion was determined, such as Figure 4 As shown in the figure, compared with the negative control group, the secretion levels of inflammatory cytokines IL-1β, IL-6, and TNF-α increased significantly after LPS induction, while the secretion of inflammatory cytokines was effectively inhibited after F1 pre-incubation and then induced with LPS, with the most obvious inhibitory effect on IL-6, with the highest inhibition rate of 76%, followed by the inhibitory effect on IL-1β, with the highest inhibition rate of 49%, and the highest inhibition rate on TNF-α of 9%. This shows that F1 can effectively reduce the secretion of inflammatory factors in RAW264.7 cells stimulated by LPS.

[0062] Table 1 Nitric oxide inhibition rate under F1 and F2 treatments

[0063]

[0064] Example 3 Preliminary screening of anti-inflammatory active peptides

[0065] 1. After preliminary peptide identification of F1 using Nano-HPLC-MS / MS, the potential biological activity of the peptides was scored using the PeptideRanker tool.

[0066] 2. The anti-inflammatory activity of peptides with a score of 0.8 or above in (1) was predicted using the BIOPEP-UWM database. Combined with the abundance values ​​of the peptides, peptides with anti-inflammatory active amino acid sequences such as IPP, PY, and LPF were screened.

[0067] 3. The novelty of the five peptide sequences was checked using the BIOPEP-UWM database.

[0068] 4. Results Analysis

[0069] Through peptide identification, the molecular weight of all peptides of the anti-inflammatory peptide complex F1 was less than 2.5 kDa, and the peptides were composed of 3 to 20 amino acids. Five peptides containing amino acid sequences with anti-inflammatory activity were obtained by bioinformatics screening. The sequences, molecular weights and scores of the five peptides are shown in Table 2. Three new peptides were obtained by querying the active peptide database, namely LPFP, PYPPP and IPPPL.

[0070] Table 2 Peptides with potential anti-inflammatory activity

[0071]

[0072] Example 4 Determination of anti-inflammatory activity of new peptides LPFP, PYPPP and IPPPL

[0073] 1. Determination of cell proliferation activity of the three peptides and their effects on NO production and cytokine production in RAW264.7 macrophages

[0074] The concentration of RAW264.7 macrophage suspension was adjusted to 3 × 10 5 / mL, add 100uL cell suspension to each well of a 96-well plate, and pre-culture the culture plate in an incubator for 24h. Add 10uL of peptide solution to the culture plate and continue incubation for 24h, add 10uL of CCK-8 solution to each well, continue incubation for 4h, and measure the absorbance at 450nm with an enzyme marker. Cell viability is calculated according to the following formula:

[0075] Cell viability (%) = [A(drug added) - A(blank)] / [A(0 drug added) - A(blank)] × 100

[0076] A (drug added): absorbance of wells with cells, CCK-8 solution, and peptide solution

[0077] A (blank): absorbance of wells with culture medium and CCK-8 solution but no cells

[0078] A(0 drug addition): absorbance of the well with cells and CCK-8 solution but no peptide solution

[0079] The macrophage cell concentration was adjusted to 5 × 10 5 Each well was added with 100 μL of cell suspension, and the supernatant was carefully discarded after 4 hours of pre-culture. 100 μL of peptide sample was added to each well and cultured for 20 hours. After that, the supernatant was discarded. 100 μL of lipopolysaccharide (LPS) with a concentration of 2 μg / mL was added to each well to stimulate the culture for 20 hours. The supernatant was taken and the NO content was determined using a nitric oxide detection kit. The ELISA kit was used to determine the effect of LPS-induced cell inflammatory factors (IL-1β, IL-6, TNF-α) production. The experimental groups were set up as follows:

[0080] Blank (NC) group: normal culture 24h+20h;

[0081] LPS group: normal culture for 24 h + LPS (2 μg / mL) stimulation culture for 20 h;

[0082] Experimental group: normal culture for 4 h + normal culture with sample for 20 h + LPS (2 μg / mL) stimulation culture for 24 h.

[0083] 2. Results Analysis

[0084] Figure 5A in the middle shows the effects of LPFP, PYPPP and IPPPL on the proliferation activity of macrophages. The results show that all three peptides can promote the activity of macrophages without toxic effects. Figure 5 Figure B shows the inhibition rate of LPFP, PYPPP and IPPPL on NO production by RAW264.7 macrophages stimulated by lipopolysaccharide, indicating that all three peptides can inhibit NO production by macrophages, and LPFP has the best inhibitory effect. Figure 6 The results showed that the three peptides could inhibit the secretion of three inflammatory cytokines, and the inhibitory effect on IL-6 was the best.

[0085] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. An anti-inflammatory peptide derived from western-style dry-cured ham, characterized in that: The amino acid sequence of the anti-inflammatory peptide is shown in SEQ ID NO:

2.

2. Use of the anti-inflammatory peptide according to claim 1 in the preparation of anti-inflammatory preparations.

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