CN109369778B - Tripeptide and preparation method and application thereof - Google Patents
Tripeptide and preparation method and application thereof Download PDFInfo
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- CN109369778B CN109369778B CN201811365385.6A CN201811365385A CN109369778B CN 109369778 B CN109369778 B CN 109369778B CN 201811365385 A CN201811365385 A CN 201811365385A CN 109369778 B CN109369778 B CN 109369778B
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- China
- Prior art keywords
- tripeptide
- fqf
- polypeptide
- preparation
- antioxidant
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0802—Tripeptides with the first amino acid being neutral
- C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention relates to the technical field of polypeptides, and particularly relates to tripeptide and a preparation method and application thereof. Experiments show that FITC fluorescence intensity of high-dose and low-dose groups of FQF has significant difference, and the FQF can significantly reduce ROS level in cells and has an antioxidation effect. Can be widely applied to preparing medicaments or food for inhibiting CD38 enzyme activity or medicaments or food with the effects of resisting oxidation and delaying senility.
Description
Technical Field
The invention relates to the technical field of polypeptides, and particularly relates to tripeptide and a preparation method and application thereof.
Background
The CD38 enzyme catalyzes substrates, namely cyclic adenosine diphosphate ribose (cADPR), adenosine diphosphate ribose (ADPR) and Nicotinic Acid Adenosine Dinucleotide Phosphate (NAADP), to be combined with different receptors or channels so as to participate in regulating calcium signals in cells, and Ca is acted on a human body related receptor system2+The messenger small molecule can control the release of calcium ions in an intracellular calcium reservoir and the internal flow of extracellular calcium ions under physiological conditions, thereby regulating the functions and life activities of cells. Nicotinamide Adenine Dinucleotide (NAD)+) As substrates for Sir2-related enzymes (Sir2-related enzymes) and as molecules linking nicotinamide phosphoribosyltransferase (Nampt) and SIRT1, extensive studies have recently been conducted, since the level of NAD + directly determines the activity of the sirtuins family, which in turn has a strong importance on cell lifeThe desired effect. In the body NAD+Is associated with the combination of its synthetases and degradative enzymes, or both, NAD in the body+The synthetases include nicotinamide phosphoribosyltransferase (NMNPT1-3), nicotinic acid mononucleoyltransferase (NMNAT), and nicotinoyl phosphoribosyltransferase (NAPRT), NAD+The degrading enzymes comprise CD38, poly (ADP-ribose) polymerase (PARP), sirtuins and the like, and the research shows that the CD38 protein level in the liver, adipose tissue, spleen and skeletal muscle of the body is compared with other NAD protein levels in the aging process+The content of synthetase or degrading enzyme is obviously increased, the mRNA expression of CD38 is also obviously increased, and NAD+The level decreased, indicating that CD38 is compared to other NADs+Synthetase or degradative enzyme on NAD+The effect of the level is more pronounced, i.e. CD38 is NAD+The key factor of the lower level drop. CD38 is the predominant NAD in mammalian tissues+Degrading enzymes, more than 90% of CD38 exert NAD+Action of hydrolytic enzymes to convert NAD+Molecular hydrolysis to nicotinamide and adenosine diphosphate ribose (ADPR), less than 10% of CD38 acting as ADP-ribosyl cyclase cyclizing NAD+cADPR and nicotinamide. In addition, CD38 degrades NAD+The production of nicotinamide is also an endogenous inhibitor of sirtuins. Based on the maintenance of NAD+Level of importance in aging and age-related disorders by inhibiting the activity of the CD38 enzyme to increase NAD in aging populations+Level, is a potentially important means of delaying aging.
Aging (Aging) is an irreversible degenerative change of biological individuals with Aging, and oxidation is one of the main causes of Aging. At present, the aging of the population has become a global social problem. In order to delay aging, some synthetic antioxidants are usually used clinically to scavenge free radicals and metabolic wastes in vivo so as to prevent excessive oxidation, but long-term administration can generate toxic and side effects on the organism. Therefore, the exploration and development of natural antioxidants are of great significance for delaying aging. Free radical theory states that degenerative changes in the aging process are caused by toxic effects resulting from the accumulation of free radicals. The normal metabolic process of the body can generate a certain amount of free radicals, and the free radicals have physiological functions of signal transduction, gene transcription and expression regulation, cell division and differentiation regulation and the like in vivo. Meanwhile, a set of endogenous free radical scavenging system is arranged in the organism and consists of enzymatic and non-enzymatic antioxidant systems, so that the free radicals are maintained in a low-amount harmless dynamic equilibrium state. With age, the free radical scavenging system in the body gradually degrades and the dynamic equilibrium of free radical levels is disrupted. Excessive free radicals are accumulated in the body, and unsaturated fatty acids in the biological membrane are promoted to generate lipid peroxidation, so that structural damage and functional disorder of the cell membrane are caused. At the same time, the lipid peroxides produced promote cross-linking of protein molecules, destroy the enzyme structure and reduce the enzyme activity, thereby damaging the DNA double helix structure and affecting the correct expression of genetic information. The accumulation of these free radical damaging effects, which cause structural destruction, functional decline and even apoptosis or necrosis, is one of the important sources of aging and various senile diseases. Studies have confirmed that Reactive Oxygen Species (ROS) are normal metabolites of many redox reactions in cells, and the production and elimination in the body are in a state of homeostasis. When the body is stimulated by endogenous or exogenous harmful factors, increased ROS production, i.e., Oxidative Stress (OS), can be caused. Oxidative stress can cause DNA oxidative damage and abnormal expression of proteins, leaving the body vulnerable. Atherosclerosis, heart disease, parkinsonism, alzheimer's disease, tumor, senile diabetes and the like, most of which are diseases occurring on the basis of oxidative stress, and the older the age, the higher the risk of developing them. When the capacity of self-scavenging free radicals is insufficient and the balance between oxidation and antioxidation is broken, the supplement of external antioxidants can inhibit oxidative stress and relieve body injury, and has important significance for delaying body aging and preventing and treating a plurality of senile diseases.
Bioactive peptides are a class of peptide molecules with biological activity, and have various physiological functions on human bodies. Compared with amino acids and proteins, the polypeptide is easier to digest and absorb by human bodies, is safer and more specific than other small compound molecules, and is not easy to cause serious immunological rejection. At present, the research on the anti-aging peptide at home and abroad is still in the initial stage.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide a tripeptide, which has good antioxidant activity, and a preparation method and use thereof.
The invention provides tripeptide with an amino acid sequence shown as SEQ ID NO. 1.
The amino acid sequence of the tripeptide is Phe-Gln-Phe, which is called FQF for short. Wherein Phe is the amino acid corresponding residue of phenylalanine and Gln is the amino acid corresponding residue of glutamine.
The chemical formula is shown as formula I:
the invention also provides a DNA molecule for encoding the tripeptide with the amino acid sequence shown as SEQ ID NO. 1.
In the present invention, the nucleotide sequence of the DNA molecule encoding the tripeptide is shown in SEQ ID NO. 2, specifically AAAGTTAAA.
The tripeptide of the present invention may be artificially synthesized by a solid-phase synthesis method, a liquid-phase synthesis method or a solid-liquid combination method, or may be synthesized by a genetic engineering method.
The preparation method of the tripeptide comprises the following steps: according to the amino acid sequence shown in SEQ ID NO. 1, amino acids are coupled one by one on a solid phase carrier to obtain peptide resin, and tripeptide is obtained after cracking.
Specifically, the solid-phase synthesis method specifically comprises the following steps: coupling Fmoc protected amino acid and resin one by one according to the amino acid sequence of the polypeptide shown in SEQ ID NO. 1 from the C end to the N end of the amino acid sequence, then removing the resin and protecting the side chain protecting group of the amino acid by lysate to obtain a crude product, and purifying the crude product to obtain the tripeptide.
In the method for synthesizing tripeptide by solid phase synthesis, the solid phase carrier is dichloro resin.
In the method for synthesizing tripeptide by solid-phase synthesis, the coupling agent is DIEA. The coupling agent is preferably used in excess. Specifically, the molar amount of the resin was 10 times. The coupling conditions were 30min at room temperature.
In the solid phase synthesis method for synthesizing tripeptide, the cracking agent for cracking consists of TFA, water, EDT and TIS. Preferably, the volume ratio of TFA, water, EDT and TIS is 94.5:2:2.5: 1. The cracking condition is 30 ℃ and 2 h.
Further, the coupling mode one by one also comprises a step of removing Fmoc before each coupling step. In some embodiments, the Fmoc removal reagent is 20% piperidine DMF solution, i.e. piperidine: DMF 1:4 (volume ratio), reaction time 15 min.
The preparation method of the invention also comprises a step of purifying the prepared polypeptide. Preferably, the polypeptide is purified by high performance liquid chromatography.
The mobile phase was water and acetonitrile, and the initial gradient of preparative hplc was: water 95%, acetonitrile 5%, end gradient: 25% of water, 75% of acetonitrile and 40min of gradient time. The purity reaches more than 99 percent.
The tripeptide preparation method by genetic engineering comprises the following steps: transforming a host cell with an expression vector comprising a DNA molecule encoding said tripeptide, inducing expression of said tripeptide.
The host cell of the genetic engineering is prokaryote and eukaryote. In particular Escherichia coli. Yeast or insect cells.
Use of tripeptides for the preparation of a product inhibiting the activity of CD 38.
And (3) carrying out butt joint treatment on the FQF peptide ligand and the CD38 crystal structure by adopting a molecular butt joint technology. The results show that FQF can interact with the active site Lys129 and Asp156 of CD38 and can enter the hydrophobic region of CD38, thereby inhibiting its activity.
Use of a tripeptide in the preparation of an agent for reducing ROS levels.
The survival rate of Hek293 cells induced by different concentrations of hydrogen peroxide indicates that hydrogen peroxide-induced damage can increase intracellular ROS levels, ultimately leading to cell death. And the FITC fluorescence intensity of the FQF high-dose group and the FQF low-dose group have significant difference, which shows that the FQF can significantly reduce the ROS level in cells and has an antioxidant effect. Can be used for preparing antioxidant drugs or foods, or drugs or foods for preventing or treating diseases caused by peroxidation, and improving medical conditions of related diseases.
Application of tripeptide in preparing antioxidant and anti-aging products.
The invention also provides an antioxidant product which comprises the tripeptide.
The product is a medicament or a food.
The medicine provided by the invention also comprises pharmaceutically acceptable auxiliary materials.
The medicament of the invention also comprises other therapeutic agents which are selected from other substances with antioxidant activity.
The food is health food, and the dosage form comprises granule, capsule, syrup, tablet, powder, soft candy, emulsion or oral liquid.
The dosage form of the medicine comprises paste, granules, pills, powder, tablets, capsules, oral liquid or syrup.
The invention also provides an anti-oxidation treatment method, which is to administer the medicine.
Compared with the prior art, the invention has the following advantages and beneficial effects:
experiments show that FITC fluorescence intensity of high-dose and low-dose groups of FQF has significant difference, and the FQF can significantly reduce ROS level in cells and has an antioxidation effect. Can be widely applied to preparing medicaments or food for inhibiting CD38 enzyme activity or medicaments or food with the effects of resisting oxidation and delaying senility.
Drawings
FIG. 1a is a High Performance Liquid Chromatography (HPLC) chart of the polypeptide FQF synthesized in example 1;
FIG. 1b is a liquid chromatography-mass spectrometry (LC-MS) graph of the polypeptide FQF synthesized in example 1;
FIG. 2a is a diagram of the structure formula of FQF as a small molecule ligand in example 2;
FIG. 2b is a combination of FQF and CD38 hydrophobic cavities as shown in example 2;
FIG. 3a is a graph showing the viability of Hek293 cells induced by different concentrations of hydrogen peroxide in the cell model of example 3;
FIG. 3b is a graph of the change in ROS levels of Hek cells from the FQF-treated group and the model group in example 3;
fig. 3c is a flow histogram of FQF processing and model set in example 3.
Detailed Description
The invention provides tripeptide and a preparation method and application thereof, and a person skilled in the art can realize the tripeptide by properly improving process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The test materials adopted by the invention are all common commercial products and can be purchased in the market.
The invention is further illustrated by the following examples:
EXAMPLE 1 Synthesis of polypeptide FQF by solid phase Synthesis of polypeptide
Adopting a standard Fomc scheme, selecting dichloro resin, extending peptide chains from the C end to the N end one by one according to the sequence characteristics of an amino acid sequence Phe-Gln-Phe, treating each step of condensation by using 20% piperidine/N, N-Dimethylformamide (DMF) solution (15mL/g) for 15min, and removing Fmoc protecting groups; and then, detecting, pumping out the piperidine solution, taking dozens of resins, washing with ethanol for three times, adding ninhydrin, pyridine and phenol, dissolving and heating for 5min at 105-110 ℃, wherein the color turning to dark blue is positive reaction, the next amino acid can be continuously connected, and if the color is not changed, the next amino acid is negative and needs to be deprotected again. Each washing was performed twice with 15mL of methanol, and 15mL of DMMF, respectively. After the first cleaning, condensation is carried out, Fmoc-L-Gln (Trt) -OH with 3 times of resin molar weight and HBTU with 3 times of resin molar weight are added, and are dissolved by a small amount of DMF, DIEA with 10 times of resin molar weight is added immediately, and reaction is carried out for 30 min. And (3) detecting after the second cleaning, pumping out the solvent, taking dozens of resin, washing with ethanol for three times, adding ninhydrin, pyridine and phenol one drop by one drop, heating at 105-110 ℃ for 5min, wherein the colorless state is a positive reaction, and the condensation is needed again if the color is blue. Repeating the above steps to sequentially connect the rest amino acids. The synthesis of the whole peptide is completed after the last amino acid is ligated. And (3) entering the final contraction stage, washing and reacting for 3 times by using DMF (dimethyl formamide), washing and reacting for 3 times by using DCM (DCM), washing and reacting for 3 times by using methanol, and finally pumping out the peptide resin. After peptide side chain synthesis, a peptide chain containing a resin was added to a mixture of TFA (94.5%), water (2%), EDT (2.5%), and TIS (1%) in a volume ratio, and the peptide chain was cleaved from the resin. The resin was charged into a flask and shaken at a constant temperature (30 ℃ C.) for 2 hours. The lysate is blown dry as much as possible with nitrogen, then poured into a centrifuge tube, and slowly poured into ether. Sealing and placing in a centrifuge for 5min, pouring out the supernatant, and collecting the white solid below. Washing with ether for 6 times, volatilizing at normal temperature, dissolving with 50% acetonitrile solution, and performing chromatography with initial gradient: water 95%, acetonitrile 5%, end gradient: 25% of water, 75% of acetonitrile and 40min of gradient time. The sample from the detector is collected.
The high performance liquid chromatogram and the liquid chromatography-mass spectrometry (LC-MS) of the synthesized polypeptide are respectively shown in fig. 1a and fig. 1b, and the analysis of fig. 1a and fig. 1b shows that the mass-to-charge ratio of the synthesized polypeptide is 441.4, the primary amino acid sequence of the synthesized polypeptide is Phe-Gln-Phe, and the target polypeptide and the synthesized polypeptide with the effects of inhibiting CD38 enzyme activity and resisting oxidation are obtained.
Example 2 molecular docking evaluation of the polypeptide for CD38 inhibitory Activity
1. Target receptor structure determination
Depending on the mode of capture or the small molecule species bound, the stability of the key residue Glu226 and the complex towards NAD was considered by comparison of crystal structures in the protein database PDB+Simulation of binding of molecules to CD38 shows that CD38 and tobacco were selectedThe amide mononucleotide (NMN) cocrystallized structure was subjected to molecular docking, and PDBID was 3 DZK.
FQF ligand establishment
And (3) adopting Chem3D software to draw the polypeptide FQF to generate a three-dimensional structure, and carrying out force field optimization on the structure, and storing for later use.
3. Molecular docking
Downloading crystal junction 3DZK of CD38 from a PDB database, and performing energy lattice point operation on the A chain to determine the original ligand position, namely the position of the molecular docking ligand-receptor binding site. After the ligand-receptor binding site was determined, the original ligand NMN was deleted, the original water molecules were removed, polar hydrogen atoms were added, and the AD4 type atoms were redistributed after the gastiger charge was calculated. And (3) scoring the binding condition between the macromolecular receptor and the micromolecular ligand, and setting the docking center coordinate and the docking box size according to the position of the original ligand NMN in 3 DZK. The FQF peptide ligand was interfaced with the CD38 crystal structure. FIG. 2a is a diagram of the structure of a ligand of FQF polypeptide. From fig. 2b it can be seen that FQF can interact with the active sites Lys129 and Asp156 of CD38 and can enter the hydrophobic region of CD38, thereby inhibiting its activity.
EXAMPLE 3 in vitro antioxidant assay of the synthetic polypeptide FQF
1. Preparation of the solution
0.1mM and 0.5mM synthetic peptide (FQF) solutions: 4.405mg of the synthetic peptide was weighed out accurately, dissolved in 10mL of the medium, filtered through a 0.22 μm filter to give a 1mM stock solution, and the stock solution was diluted with the medium to the concentration required for the experiment.
H2O2Preparing a solution: preparing mother liquor with concentration of 2mmol/L with 30% hydrogen peroxide and DMEM, and diluting with culture medium to obtain solutions of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 and 2.0
2. Construction of hydrogen peroxide damage model
Hek293 cells were seeded in 96-well plates at a density of 10000/well, and after 24h adherence, hydrogen peroxide media were added to the respective fractions at concentrations of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 and 2.0mmol/L, respectively, at 37 ℃ and 5% CO2The culture box is used for culturing for 2 hours. The cells were washed 2 times with PBS to remove the residual hydrogen peroxide sufficiently. Then 20. mu.L of MTT (0.5mg/mL) was added to each well, incubated in an incubator for 4h, the medium was aspirated, and 150. mu.L of DMSO was added to each well; the Optical Density (OD) at 570nm was measured using a microplate reader. Each group was set with 5 replicate wells, and the experiment was repeated 3 times to calculate the cell viability.
Cell viability ═ (OD dosing-OD blank)/(OD control-OD blank).
The viability of Hek293 cells induced by different concentrations of hydrogen peroxide can be seen in FIG. 3a as H2O2Increased concentrations and decreased cell viability indicate that hydrogen peroxide-induced damage can raise intracellular ROS levels, ultimately leading to cell death. The concentration at which the cell viability is 60% was selected for molding, i.e. H2O2The concentration was 600. mu.M.
3. Protection of polypeptide-hydroperoxide-induced oxidative stress injury model
The Hek293 cells are plated in a 12-well plate at a density of 10w/mL, and a culture medium solution without polypeptide is added to a control group after complete adherence; the administration group is added with FQF polypeptide culture medium of 0.1mmol/L and 0.5 mmol/L. After 48h, 600. mu. M H was added2O2And the medium containing the same concentration of polypeptide was incubated for 2 h. The cells were washed 2 times with PBS, 0.5mL of DCFH-DA dye diluted 1:1000 was added and incubation continued at 37 ℃ for 30min, the DCFH-DA dye was rapidly aspirated away, and the cells were washed twice with PBS. Cells were digested with 0.25% EDTA, collected in 1.5mL Ep tubes, and placed on ice. The fluorescence intensity of DCF in cells was detected by Cyto Fle flow cytometer, and FITC channel was selected as the channel. Statistics were performed with the mean fluorescence intensity of FITC.
As can be seen from flow histogram 3b, the polypeptide administration group was left-shifted relative to the FITC histogram of the model group, indicating a decrease in intracellular ROS levels following polypeptide treatment. Statistics were performed based on the FITC mean fluorescence intensity of the histogram, as shown from FIG. 3 c. The fluorescence intensity of the FITC of the model group is significantly different from that of the FQF high-dose and low-dose groups, which shows that the FQF can significantly reduce the intracellular ROS level and has the function of oxidation resistance.
The results show that the polypeptide FQF has remarkable antioxidant effect (p is less than 0.05), and can be applied to preparation of antioxidant drugs or foods, or drugs or foods for preventing or treating diseases caused by peroxidation, and improvement of medical conditions of related diseases.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Sequence listing
<110> Infinite Limited (China) Co., Ltd
<120> tripeptide, preparation method and application thereof
<130>MP1826325
<160>2
<170>SIPOSequenceListing 1.0
<210>1
<211>3
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>1
Phe Gln Phe
1
<210>2
<211>9
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>2
Claims (9)
1. Tripeptide shown in SEQ ID NO 1.
2. A DNA molecule encoding the tripeptide of claim 1.
3. A process for preparing the tripeptide of claim 1, comprising: according to the amino acid sequence shown in SEQ ID NO. 1, amino acids are coupled one by one on a solid phase carrier to obtain peptide resin, and tripeptide is obtained after cracking.
4. The method of claim 3, wherein the coupling agent is DIEA.
5. The method of claim 3, wherein the cleaved cleaving agent consists of TFA, water, EDT and TIS.
6. A process for preparing the tripeptide of claim 1, comprising: transforming a host cell with an expression vector comprising the DNA molecule of claim 2, inducing expression of the tripeptide of claim 1.
7. Use of the tripeptide of claim 1 in the manufacture of a formulation for reducing the level of ROS.
8. Use of the tripeptide of claim 1 for the preparation of an antioxidant and/or anti-ageing product.
9. An antioxidant and/or anti-aging product comprising the tripeptide of claim 1.
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| CN103864891A (en) * | 2014-03-25 | 2014-06-18 | 福州大学 | Chinese chive seed antioxidative tripeptide, and preparation and application thereof |
| CN104558105A (en) * | 2015-01-04 | 2015-04-29 | 华南理工大学 | Antioxidative peptide of metapenaeus affinis as well as separation and extraction method and application of antioxidative peptide |
| CN105254708A (en) * | 2015-11-24 | 2016-01-20 | 福州大学 | Carrot seed antioxidant tripeptide as well as preparation method and application thereof |
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| CN103864891A (en) * | 2014-03-25 | 2014-06-18 | 福州大学 | Chinese chive seed antioxidative tripeptide, and preparation and application thereof |
| CN104558105A (en) * | 2015-01-04 | 2015-04-29 | 华南理工大学 | Antioxidative peptide of metapenaeus affinis as well as separation and extraction method and application of antioxidative peptide |
| CN105254708A (en) * | 2015-11-24 | 2016-01-20 | 福州大学 | Carrot seed antioxidant tripeptide as well as preparation method and application thereof |
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