CN106317170A - Antioxidant peptide in perilla seeds and application thereof - Google Patents
Antioxidant peptide in perilla seeds and application thereof Download PDFInfo
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Abstract
The invention provides an antioxidant peptide in perilla seeds and an application thereof. The sequence of the antioxidant peptide is Tyr-Leu(YL). In vitro experiments show that the polypeptide can effectively remove ABTS, superoxide anions and various reactive oxide species (ORAC). Meanwhile, the peptide can effectively inhibit linoleic acid and rat liver lipid peroxidation. Cell experiments prove that the peptide is safe to cells and has obvious inhibiting effects on oxidative damages of the cell HepG-2. The antioxidant peptide has the characteristics of simple structure, safety, strong antioxidation activity, and the like, can serve as an excellent substitute for existing synthetic antioxidants and has important value for development and application of novel antioxidant health care products and food additives.
Description
Technical field
The present invention relates to a kind of Fructus Perillae anti-oxidation peptide and application thereof, belong to technical field of food biotechnology.
Background technology
China is a large agricultural country, crops of a great variety, but due to being limited by process technology condition, crops
Processing be mostly in the junior stage, the residue after its processing still has bigger content nutrient substance, such as protein etc., and it
Be mostly mainly used in animal feed or fertilizer industry and naturally discharge, not only waste resource, also result in environmental pollution simultaneously.
The proposition of plant active peptides concept and the raising of separation detection technique, make people start gradually to pay attention to rich in proteins is agricultural and sideline
The deep processing of product also therefrom obtains various active peptide, such as phosphopeptide caseinate, blood pressure lowering peptide, antibacterial peptide and absorption peptide easy to digest
Deng, the research and development of plant origin bioactive peptide has important function to the added value improving agricultural products in China deep processing.
Antioxidant the most at home and abroad develops quickly, and purposes is more and more wider.Research thinks that people's vivo oxidation produces
Raw free radical is relevant with the aging of people and numerous disease, and therefore antioxidant is applied not only to the antioxidation of fat-containing food, and
And the exploitation of health food and cosmetics etc. it is used for as functional factor.The kind of antioxidant is a lot, but some chemosynthesis
Antioxidant such as BHA, BHT etc., due to itself agree can have toxic and side effects, national governments are one after another to its mandatory provision
ADI value controls its excess and adds.Then people progressively turn to sight and extract natural antioxygen from various plant and animal tissues
Agent.Antioxidant activity polypeptide is due to low toxicity, the feature such as efficiently, as the antioxidant of food and body it is considered to be artificial
The preferable replacer of synthetized oxidation preventive agent.
Reactive oxygen free radical superfluous in antioxidation Toplink purged body effectively, protection cell and mitochondrial normal configuration
And function, prevent the generation of lipid peroxidation, and aoxidize and the mankind and the numerous disease such as cancer of other animals, aging, dynamic
The pathogeny of arteries and veins hardening etc. is relevant.The material that suitable absorption has antioxidant activity can reduce interior free yl level, anti-
Only lipid peroxidation, helps body to resist disease.
Fructus Perillae, also known as Fructus Perillae, is done for Labiatae Perilla annual herb plant Folium Perillae (Perilla frutescens)
Dry mature fruit.Folium Perillae originates in China, all has wild species and cultivation in China North China, south China, Central China, southwest and Taiwan Province
Kind, it is one of 60 kinds of medicinal and edible plants permitting in the first batch of China's Ministry of Public Health.Folium Perillae begins to be loaded in as China's Chinese medicine medical material
The holy Li Shizhen (1518-1593 A.D.) Compendium of Material Medica of Ming Dynasty doctor, has the merit of " promoting the circulation of QI to alleviate the stagnation in middle-JIAO, clearing away phlegm profit lung, and blood, warming middle-JIAO, pain relieving, Dingchuan, antiabortive "
Effect.Fructus Perillae is rich in oils and fats, and owing to being subject to processing the restriction of technical conditions, current Fructus Perillae is main office in China's food industry
It is limited to refine oils and fats.
Research to Fructus Perillae the most both at home and abroad concentrates on refinement and the biological activity aspect of oils and fats in a large number, and to its by-product
In thing rich in protein development and utilization research less, to its active polypeptide or its pharmacological research both at home and abroad almost without report
Road.Therefore, it is necessary to make full use of China's the abundantest Folium Perillae (Fructus Perillae) plant resources, it is goed deep into system and grinds
Study carefully, for realizing the Fructus Perillae modernization of Chinese medicine and developing novel, efficient, safe health food offer scientific basis.
Summary of the invention
It is an object of the invention to provide one and prepare simply, the Fructus Perillae anti-oxidation peptide that antioxidant activity is strong, and permissible
This anti-oxidation peptide is applied to health food and the development of medicine and exploitation.
A kind of antioxidation oligopeptide of the present invention, its aminoacid sequence Tyr-Leu, it is expressed as YL with single-letter, i.e. by cheese ammonia
Acid and 2 amino acid residues of leucine are constituted.
This antioxidation oligopeptide can effectively remove ABTS, ultra-oxygen anion free radical and various reactive oxygen free radical
(ORAC), there is good lipid peroxidation inhibition simultaneously；
This antioxidation oligopeptide does not has toxic action to CHO and HepG-2 cell, and HepG-2 cell can be protected from peroxidating
The cell injury of hydrogen induction.
The enzymolysis preparation of Fructus Perillae anti-oxidation peptide: use alkali extraction and acid precipitation to obtain Fructus Perillae egg from the Fructus Perillae dregs of rice
In vain, use alkaline protease by it at enzymatic hydrolysis condition: pH 9.80, enzyme addition 3000U/g, concentration of substrate 3%(w/v), solve enzyme
Time 5.00h, boiling water bath 10min enzyme denaturing, it is centrifuged 10min through 12000rpm, takes supernatant lyophilization, be Fructus Perillae albumen
Matter enzymatic hydrolysate；
Described anti-oxidation peptide isolation and purification method: Fructus Perillae protein digestion product utilization Sephadex G-25 gel chromatography is entered
Row separates, with deionized water as eluent, flow velocity 0.3 mL/min, measures the light absorption value at elution fraction 214 nm wavelength；Receive
Collection has optimal antioxidant activity part, utilizes reversed-phase high-performance liquid chromatography (RP-HPLC) to separate further；RP-HPLC washes
De-gradient is 0 ~ 60 min, 5% ~ 40%(V/V) acetonitrile；Flow velocity is 2.0 mL/min, detects wavelength 214 nm, collects retention time
For the eluting peak of 34min, lyophilization i.e. obtains described anti-oxidation peptide.
It is an advantage of the current invention that:
Described anti-oxidation peptide has strong free radical scavenging activity, to superoxide anion and ABTS free radical and reactive oxygen free radical all
There is strong scavenging action, show that it has important value in terms of antioxidant activity development and application.Fructus Perillae Natural Antioxidant Peptides
The YL IC to ABTS free radical scavenging activity50Value is 3.71 μ g/mL;When concentration is 1.0mg/mL, it is to superoxide anion freely
The clearance rate of base is 42.63 ± 0.47%.The ORAC value of Natural Antioxidant Peptides YL is 3.90 ± 0.10 μm ol TE/mg polypeptide.
Additionally, external linoleic acid lipid peroxidation and Hepar Mus lipid peroxidation are had very well by this Natural Antioxidant Peptides YL
Inhibition.Concentration is under conditions of the anti-oxidation peptide YL of 1.0mg/mL mixes with linoleic acid and is placed on 40 DEG C, first four days
It close to 30%, illustrates that this Natural Antioxidant Peptides YL can alleviate lipid peroxidation effectively to the suppression ratio of linoleic acid peroxidation
Speed, it is possible to prevent containing fatty foods corrupt, Shelf-life.This Natural Antioxidant Peptides YL is to mouse liver extract lipid mistake
The IC of oxidizing and depressing rate50Value is 3.02 mg/mL, illustrates that certain density anti-oxidation peptide YL has well protection to hepatic tissue
Effect.
This Natural Antioxidant Peptides YL is to normal Chinese hamster ovary celI and the equal no cytotoxicity of HepG-2 cell, and has protection HepG-
2 from the effect of hydrogen peroxide induced injury.Chinese hamster ovary celI processes after 24 hours through 0.1mg/mL YL, normal cell ratio
96.3% does not has significant change relative to untreated fish group 96.7%, illustrates that YL does not has toxic action to Chinese hamster ovary celI.HepG-2 cell warp
After 0.1mg/mL YL processes 24 hours, normal cell ratio 77.0% is declined slightly relative to untreated fish group 79.1%, but the thinnest
Born of the same parents' ratio 9.8% is decreased obviously, and illustrates that YL can induce HepG-2 cell slightly to become feeble and die but the most lethal.HepG-2 cell is through 1mM H2O2
After induced damage, normal cell ratio drops to 44.3%, and dead cell reaches 39.9%, if at hydrogen peroxide-induced oxidative damage
Before with 0.1mg/mL YL pretreatment HepG-2 cell, then can reduce cell apoptosis rate (normal cell 61.7%, dead thin
Born of the same parents 29.6%), illustrate that this Natural Antioxidant Peptides YL can be effectively protected HepG-2 cell oxidative damage.
Accompanying drawing explanation
Fig. 1: Fructus Perillae dregs of rice crude protein enzymatic hydrolysate Sephedex G-25 gel filtration chromatography figure.
Fig. 2: gel chromatography eluting peak c peak reversed-phase high-performance liquid chromatography figure.
The mass spectrum of Fig. 3: anti-oxidation peptide.
Fig. 4: the anti-oxidation peptide YL scavenging action to free radical.A:ABTS free radical scavenging activity；B: superoxide anion is certainly
By base scavenging capacity；C: reactive oxygen free radical Scavenging activity (ORAC).
Fig. 5: anti-oxidation peptide YL to linoleic acid lipid peroxidation (A) and the inhibitory action of Hepar Mus lipid peroxidation (B).
Fig. 6: anti-oxidation peptide YL cytotoxicity experiment.A: normal Chinese hamster ovary celI；B:CHO+0.1mg/mL YL；C:HepG-2 is thin
Born of the same parents；D:HepG-2+0.1mg/mL YL.
Fig. 7: the anti-oxidation peptide YL protective effect to hydrogen peroxide-induced HepG-2 cell oxidative damage.A:HepG-2 is thin
Born of the same parents；B:HepG-2+1mM H2O2；C:HepG-2+1mM H2O2+0.1mg/mL YL。
Detailed description of the invention
The present embodiment is implemented under premised on technical solution of the present invention, gives detailed embodiment and mistake
Journey, but protection scope of the present invention is not limited only to following embodiment.
Embodiment 1
The isolated and purified of anti-oxidation peptide of the present invention includes Sephadex G-25 gel filtration chromatography and RP-HPLC color
Spectrum (RP-HPLC) two steps.
The preparation of Fructus Perillae crude protein enzymatic hydrolysate: first Fructus Perillae petroleum ether is carried out ungrease treatment, then use alkali
Put forward the heavy method of acid and obtain Fructus Perillae crude protein, with alkaline protease enzymolysis Fructus Perillae crude protein, use alkaline protease optimal at it
Enzymatic hydrolysis condition (pH 9.80, enzyme addition 3000U/g, concentration of substrate 3%(w/v)) under solve enzyme time 5.00h, boiling water bath
10min inactivates, and is centrifuged 10min through 12000rpm, takes supernatant lyophilization, is Fructus Perillae protease hydrolysis products；
Sephadex G-25 gel filtration chromatography: by Fructus Perillae enzymatic hydrolysate lyophilized powder, be dissolved in deionized water, 12000
Rpm is centrifuged 30 min.Take supernatant 0.22 μm aperture micro-filtrate membrane filtration.Sephadex G-25 gel column (1.6cm × 100
Cm) with deionized water balance, by filtered sample upper prop.Being washed with deionized water de-, flow velocity 0.3 mL/min, in 214 nm ripples
Strong point detection eluent light absorption value, draws elution curve, as shown in Figure 1.Collecting eluting c peak, lyophilization ,-80 DEG C of low temperature are protected
Deposit standby.
High performance liquid chromatography: deionized water dissolving above-mentioned c peak lyophilized powder, uses RP-HPLC to separate further.Liquid chromatograph
System is LC-20A, and assembling Gemini 5 μ C18 (250mm × 10mm) reversed-phase column (Phenomenex, UK), by water and second
Nitrile is (containing 0.05%(V/V) trifluoroacetic acid) elution system that constitutes carries out gradient elution.Gradient: 0 ~ 60 min, 5% ~ 40%
(V/V) acetonitrile；Elution flow rate 2.0 mL/min, detects wavelength 214 nm, and elution curve is as shown in Figure 2.Collect eluting and stay the time
For 34.17min, lyophilization is antioxidation oligopeptide of the present invention.
The antioxidant composition lyophilization that will collect, uses the compositional purity that high performance liquid chromatography inspection institute obtains.Warp
Detection, this anti-oxidation peptide compositional purity reaches 95%, can measure its aminoacid sequence.
It is used in conjunction (LC-MS/MS) method with liquid chromatograph and mass spectrum and measures aminoacid sequence (Fig. 3), obtain this anti-oxidation peptide
Aminoacid sequence be Tyr-Leu (YL).
Embodiment 2
The Natural Antioxidant Peptides activity obtained in embodiment 1 is studied:
ABTS free radical scavenging effect: prepare the ABTS of 7 mmol/L with distilled water+Solution and the potassium peroxydisulfate of 2.45 mmol/L
Solution (first must at room temperature place 16 h before using), respectively by ABTS+With potassium persulfate solution by 1: 1 volume ratio mixing,
By the phosphate buffered solution of pH 7.4,5 mmol/L, mixed liquor is diluted to light absorption value A before use734Be 0.70 ±
0.02.Take the ABTS free-atom aqueous solution mixing of the sample of 0.5 mL variable concentrations and 0.5 mL and stand after 10 min under 734 nm
Survey its light absorption value.Sample is replaced to make blank and positive control respectively with deionized water and reduced glutathion.ABTS is certainly
Removed vigor by base to calculate by following equation (1):
(1)
In formula, A0: blank group light absorption value；As: sample sets light absorption value.
Ultra-oxygen anion free radical scavenging action: take the Tris-HCl buffering that 0.4 mL sample solution adds 50 mmol/L
Solution (pH 8.3) 0.4 mL, replaces sample as blank pipe with distilled water.Concussion mixing, is incubated 10 in 25 DEG C of water-baths
The mL(25 DEG C of water-bath preheating of pyrogallol hydrochloric acid solution 0.1 of 1.5 mmol/L is added after min), mixing reaction 5 min rapidly,
At 320 nm, absorbance A is measured every 30 s320.Sample is replaced to make sky respectively with deionized water and reduced glutathion
White comparison and positive control.Making the time dependent regression equation of light absorption value, the slope of curve is mouse thymus cells speed, sample
The removing vigor of superoxide anion is calculated by product by following equation (2):
(2)
In formula, Δ A0/ min: the blank group light absorption value slope of curve；ΔAs/ min: the sample sets light absorption value slope of curve.
Reactive oxygen free radical Scavenging activity (ORAC): 50 μ L sample solution and 100 μ L 70nM Fluresses are mixed
It is combined in opaque 96 orifice plates, and at 37 DEG C, hatches 15min, then, add rapidly 50 μ L 200mM AAPH in each hole
Solution, puts into rapidly fluorescence microplate reader after vibration 30s and carries out fluorescence measurement.Excitation wavelength 485nm, launches wavelength 530nm, often
Measure once until fluorescence intensity no longer changes every 1min.In test, all reagent must be with 75 mM pH 7.0 phosphoric acid buffers
Liquid configures.With phosphate buffer and GSH solution respectively as blank and positive control.Configure 0.625 μM, 1.25 μMs, 2.5
μM, 5 μMs, 10 μMs, 20 μMs, 40 μMs of watermiscible vitamin E (Trolox) solution make antioxidation standard definition non-oxidizability.Fluorescence declines
Subtract area under the curve (AUC) to calculate by following equation (3):
(3)
In formula: f0Represent initial fluorescence intensity, fiFluorescence intensity when representing i-th minute.Clean fluorescence decay curve area
(net-AUC) calculate by following equation (4):
(4)
Determining Trolox standard curve according to oxidation resistance is the most relevant to net-AUC, final ORAC value is expressed as μM
Trolox equivalent (TE)/mg polypeptide.
Linoleic acid peroxidation inhibitory activity: by 1.0 mg/ml sample 1.0 mL, 2.5%(V/V) linoleic acid dehydrated alcohol is molten
Liquid 1.0 mL, 50 mmol/L phosphate (pH 7.0) 2.0 mL and deionized water 1.0 mL are placed in tool plug test tube, keep away at 40 DEG C
Light constant temperature is cultivated 7 days.Measure thiocyanation ferrum value every 24 h and represent the degree of linoleic acid peroxidation.Take the 0.1 above-mentioned reactant liquor of mL
With 4.7 mL 75%(V/V) ethanol, 0.1 mL 30%(W/V) ammonium thiocyanate solution, 0.1 mL 20 mmol/L protochloride iron salt
Acid (3.5%, W/V) solution mixing, measures absorbance value, light absorption value A500nm and linoleic acid oxygen under 500 nm after reacting 3 min
Change degree is proportionate.Make negative control with deionized water, make positive control with GSH and BHA.
The inhibitory activity of Hepar Mus lipid peroxidation: take murine liver tissue 1.0 g, after ice-cold normal saline rinsing,
Filter paper blots, and weighs.It is placed in 5 mL small beakers, measures the normal saline that 9 times of volume blocks are long-pending, by the physiology of 2/3
Saline is poured in beaker, shreds rapidly piece of tissue with eye scissors, then pours, together with normal saline, the tissue shredded into glass
In glass homogenizer, then with remaining normal saline flushing beaker and eye scissors, and pour homogenizer into, be homogenized 6～8
Min, makes tissue homogenate, whole process should be placed in ice bath and complete.By homogenate with 3000～4000 after homogenate completely
R/min is centrifuged 10～15 min, and supernatant is 10% liver tissue homogenate's liquid.
Quickly by 0.1ml anti-oxidation peptide sample (concentration is 0.5 respectively, 1,2,4,8mg/ml) even with 1ml 0.5% hepatic tissue
Serosity, 0.1ml 6mM FeSO4Solution, 0.1ml 60mM H2O2Solution mixes, and at 37 DEG C of water-bath 1h.It is subsequently added 1ml
15% trichloroacetic acid and 1ml 0.67% thiobarbituricacidα-solution, and at 95 DEG C, heat 15min.It is cooled to 25 DEG C, then
3000rpm is centrifuged 10min.Measure the absorbance under 532nm wavelength.As a control group, distilled water is as blank group for GSH.Mus
Liver lipid peroxidation suppression ratio is calculated by following equation (5):
(5)
In formula, A0: blank group light absorption value；As: sample sets light absorption value.
Measure through the present embodiment, the Fructus Perillae Natural Antioxidant Peptides YL IC to ABTS free radical scavenging activity50Value is 3.71 μ g/
ML(Fig. 4 A);When concentration is 1.0mg/mL, it is 42.63 ± 0.47%(figure to the clearance rate of ultra-oxygen anion free radical
4B).The ORAC value of Natural Antioxidant Peptides YL is 3.90 ± 0.10 μm ol TE/mg polypeptide (Fig. 4 C).
Concentration be 1.0mg/mL first four days of anti-oxidation peptide YL its suppression ratio of linoleic acid peroxidation is schemed close to 30%(
5A), illustrate that this Natural Antioxidant Peptides YL can alleviate lipid peroxidation speed effectively.This Natural Antioxidant Peptides YL is to Mouse Liver
The IC of dirty extract lipid peroxidation suppression ratio50Value is 3.02 mg/mL, illustrates that certain density anti-oxidation peptide YL is to hepatic tissue
There is good protective effect (Fig. 5 B).
Embodiment 3:
The Natural Antioxidant Peptides obtained in embodiment 1 is carried out cytotoxicity and Study of Antioxidation:
Cytotoxicity experiment: through recovery Chinese hamster ovary cell (CHO) and hepatoma carcinoma cell (HepG-2) with containing 10% tire
The modified form RPMI-1640 culture medium of Ox blood serum, 2.0 mmol/L glutamate, Glus and 100 U/mL Pen .-Strep is moistening
, containing 5% carbon dioxide, temperature be 37 DEG C in the environment of adhere-wall culture 24 ~ 48 h.Add after digesting at 37 DEG C with pancreatin
RPMI-1640 culture medium terminates digestion, adds appropriate RPMI-1640 culture medium and carries out piping and druming and make cell fully dispersed, inoculation
Continue to cultivate until cell attachment area reaches more than 80% in 6 orifice plates.Cell is processed with the YL of final concentration of 0.1mg/mL
With cellular through the every porocyte of flow cytomery of cell double transfection reagent Annexin V-FITC and PI dyeing after 24 h
State.
Protection to hydrogen peroxide-induced hepatoma carcinoma cell oxidative damage: the hepatoma carcinoma cell (HepG-2) through recovering is by upper
Method of stating is cultivated, and reaches more than 80% at the 6 adherent areas of orifice plate inner cell, and the YL of final concentration of 0.1mg/mL processes cell 18 h
After again with after 1mmol/L hydrogen peroxide treatment cell 6h, dye through streaming with double transfection reagent Annexin V-FITC and PI of cell
Cell instrument detects the cell state of every porocyte.
Measuring through this embodiment, Chinese hamster ovary celI processes after 24 hours through 0.1mg/mL YL, normal cell ratio 96.3%
(Fig. 6 B) does not has significant change relative to untreated fish group 96.7% (Fig. 6 A), illustrates that YL does not has toxic action to Chinese hamster ovary celI.
HepG-2 cell is after 0.1mg/mL YL processes 24 hours, and normal cell ratio 77.0% (Fig. 6 D) is relative to untreated fish group
79.1% (Fig. 6 C) is declined slightly, but dead cell ratio 9.8% is decreased obviously, and illustrates that YL can induce HepG-2 cell slightly to become feeble and die
But the most till death.HepG-2 cell is through 1mM H2O2After induced damage, normal cell ratio drops to 44.3%, and dead cell reaches
39.9% (Fig. 7 B), if with 0.1mg/mL YL pretreatment HepG-2 cell before hydrogen peroxide-induced oxidative damage, then can
Reduce the apoptosis rate (normal cell 61.7%, dead cell 29.6%) (Fig. 7 C) of cell, illustrate that this Natural Antioxidant Peptides YL can
It is effectively protected HepG-2 cell oxidative damage.
SEQUENCE LISTING
<110>University of Fuzhou
<120>a kind of Fructus Perillae anti-oxidation peptide and application thereof
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 2
<212> PRT
<213>artificial sequence
<400> 1
Tyr Leu
1
Claims (3)
1. a Fructus Perillae anti-oxidation peptide, it is characterised in that: described anti-oxidation peptide aminoacid sequence is Tyr-Leu.
2. the preparation method of a Fructus Perillae anti-oxidation peptide as claimed in claim 1, it is characterised in that: from natural Fructus Perillae
Isolated and purified or artificial synthesis obtains；Concrete steps include the following:
(1) preparation of Fructus Perillae protein hydrolysate: Fructus Perillae petroleum ether first carries out ungrease treatment, then uses alkali to propose acid
Heavy method obtains Fructus Perillae crude protein, with alkaline protease enzymolysis Fructus Perillae crude protein, its enzymatic hydrolysis condition: pH 9.80, enzyme addition
3000U/g, concentration of substrate 3%w/v, solution enzyme time 5.00h, boiling water bath 10min inactivation, be centrifuged 10min through 12000rpm, take
Clear liquid lyophilization, is Fructus Perillae protease hydrolysis products；
(2) enzymatic hydrolysate of Fructus Perillae albumen utilizes Sephadex G-25 gel chromatography to separate, with deionized water as eluting
Liquid, flow velocity 0.3 mL/min, measures the light absorption value at elution fraction 214 nm wavelength, draws elution curve, collect eluting peak；Profit
Separate further with reversed-phase high-performance liquid chromatography RP-HPLC；The gradient of RP-HPLC is 0 ~ 60 min, 5% ~ 40%V/VSecond
Nitrile；Flow velocity is 2.0 mL/min, detects wavelength 214 nm, and collecting retention time is the eluting peak of about 34min, and lyophilization is i.e.
Obtain described antioxidation oligopeptide.
3. Fructus Perillae anti-oxidation peptide as claimed in claim 1 answering in terms of antioxidant health-care product exploitation and food industry
With.
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|---|---|---|---|---|
| CN111647072A (en) * | 2020-06-18 | 2020-09-11 | 天津商业大学 | Antioxidant peptide and preparation method and application thereof |
| CN114409735A (en) * | 2022-01-28 | 2022-04-29 | 福州大学 | Clinacanthus nutans antioxidant undecapeptide and preparation method and application thereof |
| CN114573664A (en) * | 2022-01-28 | 2022-06-03 | 福州大学 | Clinacanthus nutans antioxidant tridecapeptide as well as preparation method and application thereof |
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| 马文领等主编: "《心脑血管疾病预警与干预 CWPAS健康管理系统概论》", 31 January 2014, 军事医学科学出版社 * |
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| CN111647072A (en) * | 2020-06-18 | 2020-09-11 | 天津商业大学 | Antioxidant peptide and preparation method and application thereof |
| CN114409735A (en) * | 2022-01-28 | 2022-04-29 | 福州大学 | Clinacanthus nutans antioxidant undecapeptide and preparation method and application thereof |
| CN114573664A (en) * | 2022-01-28 | 2022-06-03 | 福州大学 | Clinacanthus nutans antioxidant tridecapeptide as well as preparation method and application thereof |
| CN114573664B (en) * | 2022-01-28 | 2023-11-21 | 福州大学 | Anti-oxidation tridecape of clindamianum as well as preparation method and application thereof |
| CN114409735B (en) * | 2022-01-28 | 2023-11-21 | 福州大学 | Clinopodium polycephalum antioxidation undecapeptide and preparation method and application thereof |
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