CN101012272A - Halobios oxidation resistance active peptide - Google Patents
Halobios oxidation resistance active peptide Download PDFInfo
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- CN101012272A CN101012272A CNA2007100027127A CN200710002712A CN101012272A CN 101012272 A CN101012272 A CN 101012272A CN A2007100027127 A CNA2007100027127 A CN A2007100027127A CN 200710002712 A CN200710002712 A CN 200710002712A CN 101012272 A CN101012272 A CN 101012272A
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Abstract
The invention discloses a new-typed sea biological anti-oxidizing active peptide, whose amino acid sequence is Asp-Thr-Pro-Ala-Gly-Lys-Val-Pro. The invention possesses strong anti-oxidizing activity and solubility, which is fit for food, cosmetics and medicine.
Description
Technical field
The present invention relates to biological peptide field, particularly relate to a kind of novel sea biological antioxidant bioactive peptide and the preparation method that extract from the marine animal shellfish.
Background technology
As everyone knows, the marine organisms product is key protein source in people's daily life, illustrates and contains various rich in protein in the marine prods.It is reported, be the protein that a class contains acid heteroglycan from the albumen that has anti-oxidant activity in the marine prods shellfish, and thermotolerance is strong.Also contain a large amount of glycogen and other polysaccharide in the seashells internal organ, and abundant protein, the in the past employing saltoutd and step such as acetone precipitation is separated polysaccharide component from extract thing more.
The enzymatic activity polypeptide mostly is the polypeptide of small molecule as proteinic post-treatment product, has the normal amino acid structure identical with protein molecule.This class polypeptide majority all is to play the important function regulating effect in the vital movement process, so in a single day structure illustrate, by means of chemistry route, but analogues such as synthesis antagonist, active centre segment.Study the restriction enzyme site of active polypeptide in catabolic pathway of metabolism simultaneously, keep the main position of the prerequisite of its conformation and they and special receptors bind and start or the active centre of conducts information, thereby can consciously whole molecule be changed looks, prepare and tire higher and the stronger medicine of selectivity than native peptides.
The separation of relevant polypeptide proteinoid both at home and abroad at present, the existing extensive studies of purifying generally generally adopt HPLC to carry out the purifying of peptide matters, and its activity recovery can reach more than 80%.People such as domestic Ma Jiannong adopt CM-52, Bio-Gel P-6, DEAE-52 and R-HPLC respectively to go on foot chromatography, and purifying has obtained pure insulin antagonistic peptide from the acid alcohol extracting solution of Pancreas Sus domestica; People such as Dong Wenyu adopt high performance liquid chromatography that combinatorial chemistry composite structure micromolecule polypeptide is analyzed, and the result successfully separates 10 kinds of target polypeptides with other impurity, and a kind of analysis means of routine is provided.But from the marine animal shellfish, separate, purifying antioxidation active peptides and resolve its structure, still belong to blank at present.
Summary of the invention
The objective of the invention is to overcome above-mentioned prior art exists not enough, be engaged in for a long time in the development research of marine organisms, microorganism and following process processing thereof through the contriver, a kind of a kind of new oxidation resistant bioactive peptide of halobios that makes from the seashells visceral mass is provided.
A kind of novel sea biological antioxidant bioactive peptide provided by the invention, the aminoacid sequence that it is characterized by this oxidation resistant bioactive peptide of halobios is aspartic acid (Asp)-Threonine (Thr)-proline(Pro) (Pro)-L-Ala (Ala)-glycine (Gly)-Methionin (Lys)-Xie Ansuan (Val)-proline(Pro) (Pro), i.e. Asp-Thr-Pro-Ala-Gly-Lys-Val-Pro in proper order; Be a small molecular weight polypeptides matter, molecular weight is 794Dal; Form by 7 seed amino acids, be respectively aspartic acid (Asp), glycine (Gly), Threonine (Thr), L-Ala (Ala), proline(Pro) (Pro), Xie Ansuan (Val) and Methionin (Lys), the ratio of each amino acid amount of substance is Asp: Gly: Thr: Ala: Pro: Val: Lys=1: 1: 1: 1: 2: 1: 1, contain 8 amino-acid residues, wherein the relative content of acidic amino acid is 9.22%, the relative content of basic aminoacids is 12.62%, the relative content of polare Aminosaeren is 44.8%, each amino acid relative content such as table 1
Table 1.
| Amino acid | Relative content (mol%) |
| Asp | 9.2 |
| Gly | 10.3 |
| Thr | 12.6 |
| Ala | 14.9 |
| Pro | 27.6 |
| Val | 12.6 |
| Lys | 12.6 |
This polypeptide acidic amino acid and basic aminoacids content relative equilibrium, so acid-basicity is not strong, polare Aminosaeren and nonpolar amino acid relative content are also than balance, so do not demonstrate stronger or more weak hydrophobic interaction.
Ninhydrin reaction is positive, and shows that this peptide N end is not closed and is the chain polypeptide.
The absorption spectrum scanning result shows that the maximum absorption of this peptide is 280nm, and is identical with typical protein absorption peak.
Purity is identified purity greater than 96.42%, and it is pure to reach high performance liquid phase.
The antioxidation biology activity
Adopt the ferrous ion catalyzing hydrogen peroxide to produce hydroxy radical qiao (Fenton reaction), the hydroxy radical qiao that this reaction produces can make the sarranine premium fade.
Get 0.025M, the phosphoric acid buffer 1ml of pH7.4, the sarranine premium 1ml of 40ug/ml, for reagent product 0.5ml, 3% hydrogen peroxide 1ml (fresh preparation), 0.945mM EDTA-Fe (II) 1ml (fresh preparation), mix the back in 37 ℃ of water-baths behind the reaction 30min in 520nm place mensuration optical density.Blank group replaces for test agent with 0.5ml distilled water, and control group is with 1.5ml distilled water replacement EDTA-Fe (II) with for test agent.And be calculated as follows clearance rate:
Ferrous ion and hydroperoxidation produce hydroxy radical qiao in the Fenton reaction, and hydroxy radical qiao can make the sarranine premium fade.After adding this antioxidation active peptides,, fade thereby suppress the sarranine premium by eliminating hydroxy radical qiao.According to the inhibition degree of fading is calculated the power of removing the hydroxy radical qiao ability.
The result shows, oxidation resistant bioactive peptide of halobios (in 0.1～1.0mg/mL) scope hydroxy radical qiao is had extremely strong scavenging(action), and along with the increase of concentration, clearance rate continues to increase in experimental concentration.And when lower concentration, anti-oxidant activity increases rapidly, when concentration increases, activity gather way reduce not obvious.Carry out logarithm according to the gained curve and return, calculate IC
50Be 0.39mg/mL.
Find that by the specificity chemical experiment method of a series of anti-oxidant activities this peptide has restraining effect and other anti-oxidant reaction is not had tangible effect sarranine premium color fading reaction, also can't determine but whether this polypeptide really has anti-hydroxy radical qiao activity.Because have following several may be that this polypeptide has restraining effect to color fading reaction: 1. polypeptide can be caught hydroxy radical qiao, thereby has suppressed fading of sarranine premium; 2. polypeptide and H
2O
2Or EDTA-Fe (II) combination, destroyed the generation of Fenton hydroxy radical qiao reaction, cause the sarranine premium to fade; 3. polypeptide has intensive to absorb at the 513nm place, has offset the degree that the sarranine premium fades when reaction; 4. polypeptide and sarranine premium are combined into the stable material of chemical property, thereby have shielded the reaction of hydroxy radical qiao and sarranine premium.In view of above four kinds of possibilities, we have done some experiments targetedly, the results are shown in following table 2:
The table 2 sarranine premium mechanism measurement result of fading
| The application of sample amount | A 513nm | A 513nm mean value | ||||
| 1ml EDTA-Fe(II) | 1mlH 2O | 0.1ml peptide | 0.709； | 0.710； | 0.711 | 0.710 |
| 1ml 1 EDTA-Fe(II) | 1mlH 2O | 0.1mlH 2O | 0.700； | 0.704； | 0.697 | 0.700 |
| 1mlH 2O | 1mlH 2O 2 | 0.1ml peptide | 0.680； | 0.683； | 0.684 | 0.682 |
| 1mlH 2O | 1mlH 2O 2 | 0.1mlH 2O | 0.620； | 0.624； | 0.626 | 0.623 |
Can find from last table 2 data: 1. fading of sarranine premium is because the hydroxy radical qiao that the Fenton reaction produces causes, because EDTA-Fe (II) or H really
2O
2The sarranine premium is faded doing the time spent separately; Can not induce reaction the basically bigger increase of body absorbancy of the adding of polypeptide.Therefore, the sarranine premium color fading reaction that causes by the Fenton reaction can be used for detecting the activity of anti-hydroxy radical qiao polypeptide fully.
Simultaneously, adopt ESR spin trapping technique and chemoluminescence method that the anti-oxidant activity of this antioxidation polypeptide is studied.It is very strong to found that this little peptide hydroxy radical qiao that reaction produces to Fenton is removed ability, further confirms the catch effect of this polypeptide to hydroxy radical qiao.
The preparation method of a kind of novel sea biological antioxidant bioactive peptide provided by the invention comprises:
(1) the proteic preparation of oxidation activity:
1. shellfish visceral mass such as fresh scallop visceral mass are rubbed, the NaCl solution by weight in wet base (w/w) adding 1.0% in 1: 1 (includes 0.2%Triton-100 then; 0.1mmol/L EDTA) put and make homogenate in the tissue homogenizer;
2. transferring pH is 5.6,70 ℃ of heating in water bath 5min, rapidly cooling.Cooling fluid is collected supernatant liquor through as the centrifugal 10min of 15000rpm, collects (the NH of 60～90% saturation ratios behind the accent pH to 6.7
4)
2SO
4Precipitation, precipitation is dissolved, is dialysed with Tutofusin tris (Tris)-HCl (pH7.5) damping fluid of 5mmol/L.
3. spin dialysis liquid is got supernatant liquor, add 1.5 times of pre-cold acetones.The acetone precipitation thing is dissolved in again in Tris-HCl (pH7.5) damping fluid of 5mmol/L and fully dialyse crude protein liquid;
4. crude protein liquid is got DEAE-Sephadex A-50 post (20 * 400mm) on the supernatant liquor through Macrogol 2000 0 concentrated, the centrifugal insolubles that removes.Earlier with the abundant balance of the Tris-HCl damping fluid (pH7.5) of 5mmol/L, again with the above-mentioned buffer solution for gradient elution that contains 0～0.2M sodium-chlor, flow velocity 20ml/h.Being collected in 280nm has the fraction of absorption, merges the fraction of tool anti-oxidant activity, concentrate and with distilled water fully dialyse, freeze-drying preserves (above test temperature condition is 4 ℃ except that indicating especially).
(2) anti-oxidant activity polypeptide preparation
1. choosing the top condition that enzymolysis shellfish visceral mass such as fresh scallop visceral mass make anti-oxidation peptide according to orthogonal experiment is: to anti-oxidant albumen, add the 0.016g low-temperature alkaline protease, protein and enzyme weight ratio are that 33: 1.6 accent pH are 9.0, and enzymolysis is 24 hours in 30 ℃ of waters bath with thermostatic control; Place the boiling water bath heating to make enzyme deactivation in 10 minutes, cooling enzymolysis solution.The centrifugal high molecular weight protein of removing sex change is got the supernatant lyophilize.
2. crude product deionized water dissolving behind the enzymolysis, further remove high molecular weight protein through Sephadex G-25 gel filtration chromatography: with adorning post (18 * 1000mm) after Tris-HCl damping fluid (pH7.2) balance, use the level pad wash-out behind the last sample, flow velocity is 3ml/min, and the 280nm place is detected.
(3) purify
1. CM Sepharose cation exchange column chromatography:
Cationite CM Sepharose Fast Flow, through adorning post (30 * 160mm) after with 2mM Sodium phosphate dibasic-citrate buffer solution (pH3.0) balance after the pre-treatment, behind the last sample with buffer solution elution to baseline occurring, again with containing the above-mentioned buffer solution for gradient elution of 0～0.5M sodium-chlor.Collect active peak.
2. RPLC (RP-HPLC) is analyzed:
Waters 2690 highly effective liquid phase chromatographic systems, Symmetry C
18The analysis mode chromatographic column (5um, 3.9 * 150mm), last sample solution was the film deionized water, elutriant is 30% acetonitrile-aqueous solution.0%～100% elutriant wash-out 15ml.The lyophilize of sample collection final vacuum gets pure oxidation resistant bioactive peptide of halobios.
In oxidation resistant bioactive peptide of halobios provided by the invention and preparation method thereof, described shellfish visceral mass comprises various shellfish visceral masses, for example clam, ball mother, cutter freshwater mussel, clam, scallop etc., particularly scallop such as chlamys farreri, usually peel off remaining whole visceral masses behind the scallop post, be beneficial to economical and practically, preferably adopt fresh shellfish such as new fresh scallops to peel off remaining whole visceral masses behind the scallop post.Also can adopt the remaining tankage in various fishing class processing backs in addition.
The animal physiological activity of described a kind of novel sea biological antioxidant bioactive peptide provided by the invention:
1. active dexamethasone (DEX) and spleen and the thymic lymphocytes co-cultivation of adopting of polyphenoils set up lymphocyte immunity inhibition test model.Experimental result; DEX can significantly reduce the activity of spleen and thymic lymphocytes; and this oxidation resistant bioactive peptide of halobios can not only alleviate its restraining effect to immunocyte significantly; simultaneously can also promote activity of immune cells, show that this oxidation resistant bioactive peptide of halobios suppresses to have provide protection to the lymphocyte that DEX causes.
2. adopt the thiazole salt colorimetry to inquire into
60This oxidation resistant bioactive peptide of halobios is to the provide protection of thymocyte and right under the Co radiation injury condition
60The influence of the thymocyte repair ability of Co radiation injury.Results suggest: point out this oxidation resistant bioactive peptide of halobios to have opposing
60The Co radiation is to the damaging action of thymocyte, and is dose-dependently; And in the regular hour scope, the thymocyte after being subjected to radiation injury had the effect that promotes its reparation.
3. probed into this oxidation resistant bioactive peptide of halobios to intraluminal middle cerebral artery occlusion in rats ischemia-reperfusion injury model (MCAO) the damage neuronic provide protection in back; conclusion: the neurone of this oxidation resistant bioactive peptide of halobios after to the intraluminal middle cerebral artery occlusion in rats ischemical reperfusion injury has provide protection; its mechanism can improve antioxidase content with this oxidation resistant bioactive peptide of halobios, and it is relevant to suppress lipid peroxidation.
4. described this oxidation resistant bioactive peptide of halobios can obviously increase old and feeble skin epidermis mean thickness and inoblast number.Improve intradermal spandex fiber content, have significant resisting age of skin effect.And the dermatoxicology experiment shows that this marine peptide does not cause the skin acute toxic reaction, and nonirritant is slight sensitizer.
5. this oxidation resistant bioactive peptide of halobios has the effect of uvioresistant UVA to the hairless mouse skin oxidative damage.Its mechanism raises the Bcl-2 protein expression with this oxidation resistant bioactive peptide of halobios, and the proteic expression of downward modulation NOS improves antioxidase content, and it is relevant to suppress lipid peroxidation.
Determine that by biology and animal physiological determination of activity resulting polypeptide is the anti-oxidant activity polypeptide.
In the anti-oxidant activity protein Preparation, commercially available refiner such as tissue refiner's (as sea-gull YQ-3 type refiner, the new northern five metals skin in Jiangyin City is moulded factory and sold) are adopted in homogenate usually.
The albumen that collection has anti-oxidant activity is the enzymolysis target, and the preparation antioxidation polypeptide has improved the targeted and the feasibility of each later step operation like this.
The step of long period heating is adopted in this experiment, makes unwanted foreign protein sex change, precipitation, and then has improved degree of purity of production, helps subsequent operations step (Fig. 1).
Contain a large amount of glycogen and other polysaccharide in the marine shellfish internal organ, the existence of polysaccharide increases the water absorbability of composition, viscosity, is difficult for freeze-drying and preserves, and adopts to saltout and step such as acetone precipitation is separated polysaccharide component from extract.
The ocean alkaline low-temperature protease because of its from the marine low temperature environment, so compare, have singularity in the properties of enzyme with land proteolytic enzyme.When enzymolysis prepared polypeptide class active substance, the reaction conditions of various enzymes was different, influenced the variation of substrate protein conformation, and various enzyme specificity impels N-end, the C-end amino acid composition of product polypeptide and arranges different.Choose the marine low temperature Sumizyme MP anti-oxidant activity albumen is carried out enzymolysis, targeted is stronger.
The anti-oxidation peptide crude product that enzymolysis obtains is analyzed through Sephadex G-25, and record shows two peaks (Fig. 2).
According to the action principle of molecular sieve, material appearance time and molecular weight size are negative correlation.Comparison shows that with the anti-oxidant albumen without enzymolysis: first peak of first peak of albumen absorption curve and peptide absorption curve is a same substance among the figure, and second peak of peptide absorption curve is for to have the more material of small molecular weight than first peak.Therefore second peak of albumen absorption curve is the product of anti-oxidant albumen behind the Sumizyme MP enzymolysis among the figure, is the small peptide material.Polypeptide peak is collected freeze-drying preserves.
This experiment mainly is for the foreign protein behind the enzymolysis is separated with polypeptide in step, and the polypeptide molecular weight difference behind albumen and the enzymolysis is bigger, so when conducting molecule sieves purifying, chosen bigger flow velocity, albumen is flowed out faster.Through this step purifying, realized that albumen separates with polypeptide fast.Target component is concentrated, be convenient to further operation.
To the results are shown in Figure 3 behind the CM Sepharose FF cation-exchange chromatography post on the marine peptide.
Low pH value can guarantee that most of peptide molecule and Zeo-karb produce adsorption preferably, but the tolerance pH value scope of considering resin reaches to avoiding polypeptide to be hydrolyzed, therefore, the pH value of determining sample-loading buffer is 3.0, as elutriant, should be the damping fluid of high salt concentration, all elute to guarantee the peptide molecule that to be adsorbed.
Second peak that cationic exchange obtains is analyzed through RP-HPLC, result such as Fig. 4, and retention time is that the peak of 20.919min is active peak.Because of closing on Za Feng and have part to intersect in this peak, so only collect the peak nose part and freeze-drying is preserved.
The reversed phase high efficiency liquid phase process is to be used to separate one of the most frequently used method of small-molecule peptide material at present.
Ocean anti-oxidation peptide raw product is through steps such as CM Sepharose cation seperation column chromatography, RP-HPLC column chromatographies, and this bioactive peptide has been purified 35.88 times, the results are shown in Table 3.
Table 3 purification result
| Step | Total protein content (mg) | EC 50 (mg/mL) | The purifying multiple |
| Enzymolysis solution | 1.702 | 13.99 | 1.00 |
| CM Sepharose cation seperation column chromatography | 0.736 | 1.794 | 7.8 |
| RP-HPLC | 0.016 | 0.39 | 35.88 |
The invention provides oxidation resistant bioactive peptide of halobios and preparation method thereof is a kind of quick, convenient and reliable method, obtains having the hydroxy radical qiao that reaction produces to Fenton and removes the EC of very strong its removing of ability
50Has significant antioxidant role about=0.39Mg/ml.Oxyradical inductive peroxidatic reaction of lipid and numerous disease close getting in touch arranged.Wherein hydroxy radical qiao is a strongest known oxygenant, and it almost can react with all cellular constituents, and is very harmful to body.Therefore, the ocean anti-oxidation peptide may be used as natural food antioxidant and is used to suppress the lipid autoxidation.Hydroxy radical qiao can make the hyaluronic acid depolymerization, contains waterpower decline and causes xerosis cutis wrinkle to occur, and the ocean anti-oxidation peptide also can be thought a kind of marine active substance of effective resisting age of skin.In addition, studies show that the peptide molecular weight that has than strong antioxidant action is generally less, so they are worked at body directly by intestinal absorption easily, safe again, nontoxic, have no side effect.At present, free radical theory and relevant index determining thereof have become the universal method of setting forth anti-ageing material mechanism of action.In addition, it is good generally to present solvability behind the marine protein resource enzymolysis, and emulsifying property is strong, advantages such as mobile increase.Therefore, this anti-oxidation peptide all has wide development and utilization prospects as natural antioxidants at aspects such as medicine, daily-use chemical industry, food.
Description of drawings
Fig. 1 is anti-oxidant proteic DEAE-Sephadex A-50 column chromatography curve.
Fig. 2 is the gel chromatography curve of anti-oxidant albumen and peptide.
Fig. 3 is an anti-oxidation peptide CM Sepharose cation seperation column chromatography curve.
Fig. 4 is an anti-oxidation peptide RP-HPLC analytical results.
Embodiment
The present invention further specifies the present invention with the following example, but protection scope of the present invention is not limited to embodiment.
Embodiment 1
The proteic preparation of anti-oxidant activity
(1) the fresh scallop visceral mass of 600 grams is rubbed, the NaCl solution by weight in wet base (w/w) adding 1.0% in 1: 1 (includes 0.2%Triton-100 then; 0.1mmol/L EDTA) put and make homogenate in the tissue homogenizer.Transferring pH is 5.6,70 ℃ of heating in water bath 5min, rapidly cooling.Cooling fluid is collected supernatant liquor through the centrifugal 10min of 15000rpm, collects (the NH of 60～90% saturation ratios behind the accent pH to 6.7
4)
2SO
4Precipitation, precipitation is dissolved, is dialysed with Tris-HCl (pH 7.5) damping fluid of 5mmol/L.Spin dialysis liquid is got supernatant liquor, slowly add 1.5 times of pre-cold acetones.The acetone precipitation thing is dissolved in again in Tris-HCl (pH 7.5) damping fluid of 5mmol/L and fully dialyse crude protein liquid, (sample I) preserved in freeze-drying.
(2) crude protein liquid is got DEAE-Sephadex A-50 post (20 * 400mm) on the supernatant liquor through Macrogol 2000 0 concentrated, the centrifugal insolubles that removes.Earlier with the abundant balance of the Tris-HCl damping fluid (pH 7.5) of 5mmol/L, again with the above-mentioned buffer solution for gradient elution that contains 0～0.2M sodium-chlor, flow velocity 20ml/h.Being collected in 280nm has the fraction of absorption, merges the fraction of tool anti-oxidant activity, concentrate and with distilled water fully dialyse, freeze-drying preserves (sample II).
Enzymolysis
Sample thief II90ml (containing protein is 0.33g) adds 0.016g marine low temperature Sumizyme MP (Huanghai Sea aquatic products institute product), regulates pH=9.0, and enzymolysis is 24 hours in 30 ℃ of waters bath with thermostatic control.Enzymolysis finishes, and places the boiling water bath heating to make enzyme deactivation in 10 minutes enzymolysis solution.Cooling, the centrifugal high molecular weight protein of removing sex change.Get supernatant lyophilize, Vacuum Package (sample III).
Separate
Crude product deionized water dissolving behind the enzymolysis, further remove high molecular weight protein through the SephadexG-25 gel filtration chromatography: with adorning post (18 * 1000mm) after Tris-HCl damping fluid (pH7.2) balance, use the level pad wash-out behind the last sample, collect polypeptide peak, (sample IV) preserved in freeze-drying.
Purify
(1) cationite CM Sepharose Fast Flow, through adorning post (30 * 160mm) after with 2mM Sodium phosphate dibasic-citrate buffer solution (pH3.0) balance after the pre-treatment, behind the last sample with buffer solution elution to baseline occurring, again with containing the above-mentioned buffer solution for gradient elution of 0～0.5M sodium-chlor.Collect active peak (sample V).
(2) Waters 2690 highly effective liquid phase chromatographic systems, Symmetry C
18The analysis mode chromatographic column (5um, 3.9 * 150mm), last sample solution was the film deionized water, elutriant is 30% acetonitrile-aqueous solution.0%～100% elutriant wash-out 15ml.Sample collection final vacuum lyophilize (sample VI) gets pure oxidation resistant bioactive peptide of halobios.
The aminoacid sequence of this oxidation resistant bioactive peptide of halobios is Asp-Thr-Pro-Ala-Gly-Lys-Val-Pro in proper order.
Claims (2)
1, a kind of novel sea biological antioxidant bioactive peptide is characterized in that: the aminoacid sequence of this oxidation resistant bioactive peptide of halobios is Asp-Thr-Pro-Ala-Gly-Lys-Val-Pro in proper order.
2, according to the oxidation resistant bioactive peptide of halobios of claim 1, it is characterized in that being a small molecular weight polypeptides matter, molecular weight is 794Dal; The relative mol% content of each amino acid is respectively aspartic acid 9.2, glycine 10.3, Threonine 12.6, L-Ala 14.9, proline(Pro) 27.6, Xie Ansuan 12.6, Methionin 12.6, the ratio of its amount of substance is an aspartic acid: glycine: Threonine: L-Ala: proline(Pro): Xie Ansuan: Methionin=1: 1: 1: 1: 2: 1: 1, contain 8 amino-acid residues, wherein the relative content of acidic amino acid is 9.22%, the relative content of basic aminoacids is 12.62%, and the relative content of polare Aminosaeren is 44.8%;
This polypeptide acidic amino acid and basic aminoacids content relative equilibrium, so acid-basicity is not strong, polare Aminosaeren and nonpolar amino acid relative content are also than balance, so do not demonstrate stronger or more weak hydrophobic interaction;
Ninhydrin reaction: be positive, show that this material N end is not closed and be the chain polypeptide;
Absorption spectrum: scanning result shows that the maximum absorption of this material is 280nm, and is identical with typical protein absorption peak;
Purity is identified: purity is greater than 96.42%, and it is pure to reach high performance liquid phase;
The antioxidation biology activity: this antioxidation active peptides hydroxy radical qiao that reaction produces to Fenton is removed very competent, the EC of its removing
50=0.39mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100027127A CN101012272A (en) | 2007-01-23 | 2007-01-23 | Halobios oxidation resistance active peptide |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100027127A CN101012272A (en) | 2007-01-23 | 2007-01-23 | Halobios oxidation resistance active peptide |
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ID=38700012
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103524597A (en) * | 2012-07-03 | 2014-01-22 | 浙江海洋学院 | Antioxidant peptide prepared from shark prolamin and preparation method and application thereof |
| CN106916868A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | A kind of white clam polypeptide with anti-oxidant in vivo and neuroprotective function and preparation method and application |
| CN106916206A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | Hard clam polypeptide and preparation method and application |
| CN114671961A (en) * | 2022-03-28 | 2022-06-28 | 内蒙古大漠魂生物科技有限公司 | Cistanche deserticola composition with antioxidant effect |
-
2007
- 2007-01-23 CN CNA2007100027127A patent/CN101012272A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103524597A (en) * | 2012-07-03 | 2014-01-22 | 浙江海洋学院 | Antioxidant peptide prepared from shark prolamin and preparation method and application thereof |
| CN106916868A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | A kind of white clam polypeptide with anti-oxidant in vivo and neuroprotective function and preparation method and application |
| CN106916206A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | Hard clam polypeptide and preparation method and application |
| CN114671961A (en) * | 2022-03-28 | 2022-06-28 | 内蒙古大漠魂生物科技有限公司 | Cistanche deserticola composition with antioxidant effect |
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