CN102219828B - Antioxidation polypeptide prepared from sharkskin collagen - Google Patents
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Abstract
The invention provides an antioxidation polypeptide prepared from sharkskin collagen. In the method, the sharkskin collagen is adopted as a raw material; and acid protease is broken down by enzymes and separated and purified to obtain specific antioxidation polypeptide, wherein the molecular weight of the specific antioxidation polypeptide is 866 Da; and the complete sequence of the amino acid of the specific antioxidation polypeptide is Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala. According to the invention, the defects of a natural antioxidant are eliminated, the worries of the public about artificial antioxidant are eliminated, and theoretical basis is laid for developing the antioxidation polypeptide based on a food source and exploring wide application of the antioxidation polypeptide in foods and medical science.
Description
Technical field
The invention provides a kind of antioxidation polypeptide and preparation method thereof, related more specifically to a kind of antioxidation polypeptide of shark skin collagen protein preparation, belong to biological technical field.
Background technology
The oxidation of biomolecules is the process of a free radical mediated, and it can cause many adverse influences to food and biosystem.In aerobic organ, many with arteriosclerosis, cancer etc. in the free free radical of disease-relateds can produce along with the process of oxygen metabolism inevitably.In food, the oxidation meeting of food nutrient composition produces superoxide, and it not only can affect Nutritive value of food, causes that Food Quality descends serious even health diseases that also can cause absorption person.Therefore, seek safe antioxidant is the study hotspot that biochemistry nutrition is learned to suppress the superoxide generation always.Because the chemosynthesis antioxidants such as BHT, TBHQ have better effect and more cheap price than natural antioxidants, so it is widely used in the food service industry.But, has at present research to find that synthetized oxidation preventive agent has the property of accumulating carcinogenesis to organs such as human liver, spleen, lungs, thereby caused the worry of people to its security, and beginning limits its use in food gradually.So people turn to natural antioxidants to sight.Alpha-tocopherol is a kind of natural antioxidants by the most generally using, and it can effectively keep the stability of grease in the food, but but is unfavorable for Food preservation.Therefore, we are necessary to seek a kind of natural antioxidants of safety of other source.
Existing research is found, the protolysate of a lot of different sourcess all has certain resistance of oxidation, all has certain oxidation-resistance such as the hydrolyzate of casein, soybean protein, bovine serum albumin, Protalbinic acid, oil seed protein, wheat gliadin and zein etc.Although the mechanism of polypeptide performance anti-oxidant activity also is not clear especially, reports that some die aromatischen Aminosaeuren and Histidine have played vital role.Research both domestic and external finds that the peptide in collagen protein source has more high efficiency oxidation-resistance than other protein source peptide.And collagen protein accounts for more than 80% of gross protein in the fish-skin, and from obtaining the efficient of collagen protein, fish-skin is the important sources that obtains collagen protein.Be rich in hydrophobic amino acid and other and lipid in the gelatin and have than high-affinity and can form the amino acid of stable emulsion system, so be considered to prepare the very good material of antioxidation polypeptide.Therefore, the high-efficiency antioxidant polypeptide that how obtains to have the specific amino acid sequence from gelatin just becomes urgent research direction.
Summary of the invention
In order to address the above problem, the invention provides a kind of antioxidation polypeptide of shark skin collagen protein preparation, anti-oxidant activity is realized efficiently.
A kind of antioxidation polypeptide of the present invention, aminoacid sequence are Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala.
The present invention also provides a kind of preparation method of antioxidation polypeptide, extracts collagen protein take shark skin as raw material, then adopts aspartic protease that it is carried out enzymolysis, and separation and purification, lyophilize obtain antioxidation polypeptide; Described enzymatic hydrolysis condition is: pH is 3.0, temperature 50 C, enzymolysis time are that 5h, enzyme-substrate proportioning are 3000U/g; Described enzyme is aspartic protease.The means of described separation and purification comprise ultrafiltration, SP-Sephadex C-25 ion-exchange chromatography, Sephadex G-50 molecular sieve and RP-HPLC RPLC.
The concrete steps of described separation and purification are:
(1) enzymolysis product at first utilizes molecular weight to hold back scope for the ultra-filtration membrane of 8000Da and 3500Da enzymolysis product to be carried out ultra-filtration and separation, it is divided into 3 components that molecular weight ranges is different, be respectively molecular weight greater than the component of 8000Da, molecular weight between the component of 3500Da and 8000Da, the molecular weight component less than 3500Da; (2) collection has the component of best anti-oxidant activity, separate with SP-Sephadex C-25 cation-exchange chromatography again, flush away absorbed component not after the loading, carry out linear gradient elution with 0.02mol/L, the pH4.0 acetic acid-sodium acetate buffer that contains 0～0.35mol/L NaCl again, flow velocity is 0.5ml/min, under 225nm, measure, measure the anti-oxidant activity of elution fraction corresponding to each absorption peak; (3) collect the peak with best anti-oxidant activity, separate with Sephadex G-50 gel chromatography, elutriant is deionized water, and flow velocity is 0.5ml/min, measures under 225nm, measures the anti-oxidant activity of elution fraction corresponding to each absorption peak; (4) collect the peak with best anti-oxidant activity, sharp RP-HPLC RPLC further separates again, the separation of reversed-phase HPLC is to contain the 0.1%(volume) concentration gradient of trifluoroacetic acid carries out linear elution as elutriant as 5%～40% acetonitrile solution, used chromatographic column is Gemini 5 μ C18, applied sample amount is 100 μ L, flow velocity is 1mL/min, the detection wavelength is 215nm, measure the anti-oxidant activity of elutriant corresponding to each absorption peak, collect and obtain three the highest components of anti-oxidant activity; (5) utilize the RP-HPLC RPLC that three components are identified, carry out linear elution take the concentration gradient that contains the 10mM ammonium acetate acetonitrile solution as 5%～27.5% as elutriant, used chromatographic column is Gemini 5 μ C18, applied sample amount is 20 μ L, flow velocity is 1ml/min, the detection wavelength is 215nm, and three components are separated respectively and obtained single peak, namely obtain three pure active ingredients; Utilize proteinaceous solid facies sequence analysis instrument to identify the aminoacid sequence of three peptides, the aminoacid sequence that obtains antioxidation polypeptide of the present invention is: Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala.
In order to realize such scheme, the concrete steps that the present invention takes are as follows:
(1) extraction of collagen protein
Shark skin of the present invention is commercially available.
Then shark skin smashs (length＜1cm), use again the NaOH solution (solid-liquid ratio w/v=1:6) of 0.1mol/L at 4 ℃ of lower 24h of immersion to pieces through washing 3 times with removal of impurities.Drain away the water after being washed to neutrality, add again HCl solution (the solid-liquid ratio w/v=1:6), behind 4 ℃ of lower immersion 4h, be washed to again neutrality of 0.03w%.At last, add distilled water (solid-liquid ratio w/v=1:6), 60 ℃ of water-bath 6h, after filtration, obtain collagen protein after the concentrated and lyophilize.
(2) enzymolysis of collagen protein
Enzyme is available from Foochow good fortune large hundred special biological reagent companies (Fuzhou of China).
Adopt aspartic protease enzymolysis shark skin collagen protein, protein concentration is 30mg/ml, enzymatic hydrolysis condition gets that pH is 3.0, temperature 50 C, enzymolysis time are that 5h, enzyme-to-substrate are than being (3000U/g), it is stable to regulate pH with 2M HCl, and behind the hydrolysis 5h, enzyme 15min goes out in the boiling water bath, then be cooled to rapidly room temperature, place whizzer, with the centrifugal 15min of 4000r/min, it is for subsequent use to get supernatant liquor.
(3) separation of enzymolysis product, purifying
Utilize molecular weight to hold back scope the enzymolysis product that obtains and for the ultra-filtration membrane of 8000Da and 3500Da enzymolysis product is carried out ultra-filtration and separation, it is divided into 3 components that molecular weight ranges is different, be respectively molecular weight greater than the component of 8000Da, molecular weight between the component of 3500Da and 8000Da, the molecular weight component less than 3500Da; Collection has the component of best anti-oxidant activity, pass through again SP-Sephadex C-25 cation-exchange chromatography (long 20cm, diameter 1.6cm) separates, carry out linear gradient elution with 0.02mol/L, the pH4.0 acetic acid-sodium acetate buffer that contains 0～0.35mol/L NaCl, flow velocity is 0.5ml/min; Collection has the peak of best anti-oxidant activity, again with Sephadex G-50(long 100cm, diameter 2.6cm) gel chromatography separates, and elutriant is deionized water, and flow velocity is 0.5ml/min, and elution peak is measured under 225nm; Collection has the peak of best anti-oxidant activity, utilize the RP-HPLC RPLC further to separate, the separation of reversed-phase HPLC is carried out gradient elution take the concentration gradient that contains 0.1% trifluoroacetic acid (volume) acetonitrile solution as 5%～40%, used chromatographic column is Gemini 5 μ C18, applied sample amount is 100 μ L, flow velocity is 1ml/min, and the detection wavelength is 215nm, collects to obtain three the highest components of anti-oxidant activity.
(4) evaluation of antioxidation polypeptide
Utilize the RP-HPLC RPLC that three components are identified, carry out linear elution take the concentration gradient that contains the 10mM ammonium acetate acetonitrile solution as 5%～27.5% as elutriant, used chromatographic column is Gemini 5 μ C18, applied sample amount is 20 μ L, flow velocity is 1ml/min, and the detection wavelength is 215nm, and three components are separated respectively and obtained single peak, illustrate to obtain three pure active ingredients, further identify the aminoacid sequence of peptide.
(5) test of anti-oxidant activity
Utilize DPPH(1,1-Diphenyl-2-picryl-hydrazyl) free radical scavenging activity assay method research antioxidation polypeptide.Compound concentration is 1 * 10
-5The DPPH ethanolic soln of mol/L keeps in Dark Place.With 2ml, the DPPH ethanol solution of 0.1mM joins in the clean tube that contains the different enzymolysis samples of 2mL, mixing.After placing 30min under the room temperature, measure absorbancy in 517 nm places, light absorption value is less, shows that radical scavenging activity is stronger.
Clearance rate (%)=(1-(A
i-A
j)/A
0) * 100%
In the formula, A
0Be i.e. 2 mL of the light absorption value of the DPPH solution that do not add sample, the sample solvent of DPPH ethanol solution+2ml of 0.1mM, the light absorption value of blank; A
iFor the light absorption value that adds the DPPH solution behind the sample is 2ml, the light absorption value of the sample of DPPH ethanol solution+2mL of 0.1mM; A
jThe light absorption value of the sample of the dehydrated alcohol that is 2ml for solvent and the light absorption value after the sample mix of DPPH solution+2 ml.
(6) determined amino acid sequence
Utilize proteinaceous solid facies sequence analysis instrument (Applied Biosystems Model 476A, Perkin Elmer Co. MA, U.S.A) the amino acid total order of measuring antioxidation polypeptide of the present invention is classified Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala as, and molecular weight is 866 Da.
The present invention sees through the defective of present natural antioxidants and the public to the worry of synthetic antioxidant, be based on seeking a kind of antioxidant of efficiency natural, take the collagen protein that comes from shark skin as starting point, be conceived to the cutting condition control by aspartic protease, be cut into the active polypeptide with specific peptide chain length and structural domain composition, and anti-oxidant activity is realized efficiently.
The present invention has changed thinking and the method for extraction and the utilization of existing antioxidant, has eliminated the side effect that the synthetic antioxidant may cause, is a kind of natural antioxidants, can replace traditional synthetic antioxidants.And the present invention has also improved China to collagen protein utilization ratio situation on the low side in the aquatic animal skin, both can solve the recycling problem of a large amount of aquatic resources, can remove again the human consumer to antioxidant in the misgivings aspect the food safety, will have profound significance to the development of science and technology, economy and grocery trade.
Description of drawings
Fig. 1 is the removing effect of the DPPH free radical of collagen protein enzymolysis thing after ultra-filtration and separation;
Fig. 2 is the SP-Sephadex C-25 elution profile of collagen protein enzymolysis thing;
Fig. 3 is that four chromatographic separation components of SP-Sephadex C-25 are to the removing effect of DPPH free radical;
Fig. 4 is the Sepadex G-50 elution profile of B component;
Fig. 5 is that Sepadex G-50 elution fraction is to the removing effect of DPPH free radical;
Fig. 6 is the RP-HPLC elution profile of B component-III;
Fig. 7 is the DPPH free radical scavenging activity of the RP-HPLC purified components of B-III;
Fig. 8 is the RP-HPLC evaluation figure of antioxidation polypeptide B-III a;
Fig. 9 is the RP-HPLC evaluation figure of antioxidation polypeptide B-III c;
Figure 10 is the RP-HPLC evaluation figure of antioxidation polypeptide B-III d;
Figure 11 is " amount-effect " relation curve that the antioxidation polypeptide B-III c of purifying removes the DPPH free radical.
Figure 12 is " amount-effect " relation curve that the antioxidation polypeptide B-III c of purifying removes the ABTS free radical.
Embodiment
The aminoacid sequence of antioxidation polypeptide of the present invention is Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala.
The preparation method is as follows:
Extract collagen protein take shark skin as raw material, then use aspartic protease that it is carried out enzymolysis, protein concentration is 30mg/ml, and enzymatic hydrolysis condition is: pH is 3.0, temperature is that 50 ℃, enzymolysis time are that 5 hours, enzyme-substrate proportioning are 3000U/g; Utilize molecular weight to hold back scope the enzymolysis product that obtains and for the ultra-filtration membrane of 8000Da and 3500Da enzymolysis product is carried out ultra-filtration and separation, it is divided into 3 components that molecular weight ranges is different, be respectively molecular weight greater than the component of 8000Da, molecular weight between the component of 3500Da and 8000Da, the molecular weight component less than 3500Da; Collection has the component of best anti-oxidant activity, pass through again SP-Sephadex C-25 cation-exchange chromatography (long 20cm, diameter 1.6cm) separates, carry out linear gradient elution with 0.02mol/L, the pH4.0 acetic acid-sodium acetate buffer that contains 0～0.35mol/L NaCl, flow velocity is 0.5ml/min, measures under 225nm; Collection has the peak of best anti-oxidant activity, again with Sephadex G-50(long 100cm, diameter 2.6cm) gel chromatography separates, and elutriant is deionized water, and flow velocity is 0.5ml/min, and elution peak is measured under 225nm; Collection has the peak of best anti-oxidant activity, utilize the RP-HPLC RPLC further to separate, the separation of reversed-phase HPLC is carried out gradient elution take the concentration gradient that contains 0.1% trifluoroacetic acid (volume) acetonitrile solution as 5%～40%, used chromatographic column is Gemini 5 μ C18, applied sample amount is 100 μ L, flow velocity is 1ml/min, and the detection wavelength is 215nm, collects to obtain three the highest components of anti-oxidant activity.Further identify through the RP-HPLC RPLC, obtain three pure active ingredients.
Instrument, detection means that the present invention adopts are as follows:
The present invention uses the multiple separation and purification means such as ultrafiltration, SP-Sephadex C-25 ion-exchange chromatography, Sephadex G-50 gel filtration chromatography, RP-HPLC RPLC, and realization has the high efficiency separation purifying of the antioxidation polypeptide of remarkable activity.
Utilize protein solid phase sequenator (Applied Biosystems Model 476A, Perkin Elmer Co. MA, U.S.A) to measure the aminoacid sequence of the antioxidation polypeptide behind the purifying.
In order further to understand content of the present invention, Characteristic, hereby exemplify following examples:
Embodiment 1
Take by weighing 7.5 gram shark skin collagen proteins with deionized water dissolving and be settled to 250ml, then use 2mol/L HCl with its pH regulator to 3.0.First this solution water-bath being heated to 50 ℃, is the enzyme of the ratio adding respective amount of 3000U/g according to the enzyme-substrate proportioning more then, and enzymolysis time is 5 hours.Then in boiling water bath, go out enzyme 15 minutes, centrifugal 15 minutes of 4000rpm again after the cooling.The collection supernatant liquor is for subsequent use.
It is that the ultra-filtration membrane of 8000Da and 3500Da carries out ultra-filtration and separation that supernatant liquor is held back scope with molecular weight, it is divided into 3 components that molecular weight ranges is different, be respectively molecular weight greater than the component of 8000Da, molecular weight between the component of 3500Da and 8000Da, the molecular weight component less than 3500Da, and measure its anti-oxidant activity (as shown in Figure 1).Can find out that molecular weight has best anti-oxidant activity less than the component of 3500Da.
Collect molecular weight less than the component of 3500Da, with SP-Sephadex C-25 cation-exchange chromatography (long 20cm, diameter 1.6cm) further separates, carry out linear gradient elution with 0.02mol/L, the pH4.0 acetic acid-sodium acetate buffer that contains 0～0.35mol/L NaCl, flow velocity is 0.5ml/min, elution peak is measured under 225nm, sees Fig. 2.Collect each peak and measure anti-oxidant activity (as shown in Figure 3).Can find out that B component has best anti-oxidant activity.
To separate B component and separate with Sepadex G-50 gel filtration chromatography (long 20cm, diameter 1.6cm) again, elutriant is deionized water, and flow velocity is 0.5ml/min, and elution peak is measured under 225nm, sees Fig. 4.Collect each peak and measure anti-oxidant activity (as shown in Figure 5).Can find out that B component-III has best anti-oxidant activity.
Utilize the RP-HPLC RPLC further to separate (as shown in Figure 6) B component-III of separating, elutriant is 5%～40% acetonitrile solution for the concentration gradient that contains 0.1% trifluoroacetic acid (volume), used chromatographic column is Gemini 5 μ C18, applied sample amount is 100 μ L, flow velocity is 1ml/min, the detection wavelength is 215nm, measure the anti-oxidant activity (as shown in Figure 7) of elutriant corresponding to each absorption peak, collection obtains three the highest components of anti-oxidant activity, is respectively B-III a, B-III c and B-III d.
Utilize the purity of three antioxidation polypeptides that the RP-HPLC RPLC obtains separation to identify, carry out linear gradient elution take the concentration gradient that contains the 10mM/L ammonium acetate acetonitrile solution as 5%～27.5% as elutriant, used chromatographic column is Gemini 5 μ C18, applied sample amount is 20 μ L, flow velocity is 1ml/min, and the detection wavelength is 215nm, and the result shows, it is single peak, illustrates that it has reached higher degree (such as Fig. 8, Fig. 9 and shown in Figure 10).
Antioxidation polypeptide B-III c behind the purifying has very strong resistance of oxidation, can be found out by Figure 11, Figure 12, and the 503nhibiting concentration that it removes DPPH free radical and ABTS free radical is respectively 98.70mg/ml and 90.97mg/ml.
Utilize protein solid phase sequenator (Applied Biosystems Model 476A, Perkin Elmer Co. MA, U.S.A) to measure the aminoacid sequence of the antioxidation polypeptide B-III c behind the purifying.Obtaining its amino acid total order classifies as: the Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala(molecular weight is 866 Da).
＜110〉University of Fuzhou
＜120〉antioxidation polypeptide of shark skin collagen protein preparation
<160> 1
<210> 1
<211> 10
<212> PRT
＜213〉artificial sequence
<220>
<400> 1
Gly Ala Ile Gly Pro Ala Gly Pro Arg Ala
1 5 10
Claims (2)
1. antioxidation polypeptide, it is characterized in that: the aminoacid sequence of described antioxidation polypeptide is Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala.
2. the preparation method of an antioxidation polypeptide is characterized in that: extract collagen protein take shark skin as raw material, then adopt aspartic protease that it is carried out enzymolysis, separation and purification, lyophilize obtain antioxidation polypeptide; Described enzymatic hydrolysis condition is: pH is 3.0, temperature 50 C, enzymolysis time are that 5h, enzyme-substrate proportioning are 3000U/g; Described enzyme is aspartic protease; The concrete steps of described separation and purification are: (1) enzymolysis product at first utilizes molecular weight to hold back scope for the ultra-filtration membrane of 8000Da and 3500Da enzymolysis product to be carried out ultra-filtration and separation, it is divided into 3 components that molecular weight ranges is different, be respectively molecular weight greater than the component of 8000Da, molecular weight between the component of 3500Da and 8000Da, the molecular weight component less than 3500Da; (2) collection has the component of best anti-oxidant activity, separate with SP-Sephadex C-25 cation-exchange chromatography again, flush away absorbed component not after the loading, carry out linear gradient elution with 0.02mol/L, the pH4.0 acetic acid-sodium acetate buffer that contains 0～0.35mol/L NaCl again, flow velocity is 0.5ml/min, under 225nm, measure, measure the anti-oxidant activity of elution fraction corresponding to each absorption peak; (3) collect the peak with best anti-oxidant activity, separate with Sephadex G-50 gel chromatography, elutriant is deionized water, and flow velocity is 0.5ml/min, measures under 225nm, measures the anti-oxidant activity of elution fraction corresponding to each absorption peak; (4) collect the peak with best anti-oxidant activity, recycling RP-HPLC RPLC further separates, the separation of reversed-phase HPLC is to contain the 0.1%(volume) concentration gradient of trifluoroacetic acid carries out linear elution as elutriant as 5%～40% acetonitrile solution, used chromatographic column is Gemini 5 μ C18, applied sample amount is 100 μ L, flow velocity is 1mL/min, the detection wavelength is 215nm, measure the anti-oxidant activity of elutriant corresponding to each absorption peak, collect and obtain three the highest components of anti-oxidant activity; (5) utilize the RP-HPLC RPLC that three components are identified, carry out linear elution take the concentration gradient that contains the 10mM ammonium acetate acetonitrile solution as 5%～27.5% as elutriant, used chromatographic column is Gemini 5 μ C18, applied sample amount is 20 μ L, flow velocity is 1ml/min, the detection wavelength is 215nm, and three components are separated respectively and obtained single peak, namely obtain three pure active ingredients; Utilize proteinaceous solid facies sequence analysis instrument to identify the aminoacid sequence of three peptides, obtain the aminoacid sequence of antioxidation polypeptide as claimed in claim 1: Gly-Ala-Ile-Gly-Pro-Ala-Gly-Pro-Arg-Ala.
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| CN103468776B (en) * | 2013-09-25 | 2015-04-15 | 福州大学 | Method for preparing fish-derived antifreeze polypeptide with acid protease and application of antifreeze polypeptide |
| CN104356200B (en) * | 2014-11-05 | 2018-03-16 | 福州大学 | A kind of anti-oxidation peptide and preparation method thereof |
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| CN108117597A (en) * | 2016-11-30 | 2018-06-05 | 中南大学 | A kind of preparation process of new dace fish collagen antioxidant peptide |
| CN107629113B (en) * | 2017-11-02 | 2023-09-15 | 中国海洋大学 | Method for identifying Pacific cod skin collagen |
| CN108191959A (en) * | 2018-04-02 | 2018-06-22 | 广州赛莱拉干细胞科技股份有限公司 | Anti-oxidation peptide and preparation method thereof and the cosmetics comprising the anti-oxidation peptide |
| CN108822208A (en) * | 2018-06-26 | 2018-11-16 | 福州大学 | The method of antioxidation polypeptide is prepared from black sharkskin |
| CN108794571A (en) * | 2018-06-26 | 2018-11-13 | 福州大学 | Antioxidation polypeptide is isolated and purified using black sharkskin |
| CN108794575A (en) * | 2018-06-26 | 2018-11-13 | 福州大学 | A kind of preparation method of black sharkskin antioxidation polypeptide |
| CN108484721B (en) * | 2018-06-26 | 2020-09-01 | 福州大学 | Method for preparing antioxidant polypeptide by enzymolysis of black sharkskin and separating and purifying antioxidant polypeptide |
| CN108794577A (en) * | 2018-06-26 | 2018-11-13 | 福州大学 | A kind of preparation method of antioxidation polypeptide |
| CN108484722B (en) * | 2018-06-26 | 2020-09-01 | 福州大学 | Preparation, separation and purification method of antioxidant polypeptide |
| CN108822207A (en) * | 2018-06-26 | 2018-11-16 | 福州大学 | The method for preparing antioxidation polypeptide using alkali protease |
| CN108794576B (en) * | 2018-06-26 | 2020-09-01 | 福州大学 | Method for preparing antioxidant polypeptide by taking black sharkskin as raw material |
| CN109078164B (en) * | 2018-08-22 | 2022-04-01 | 厦门肽王基因科技有限公司 | Application of shark skin mixed protein peptide of Astrocaryum platypomum as skeletal development promoter and microelement supplement |
| CN113425829B (en) * | 2021-07-27 | 2022-06-03 | 时垠(上海)生物科技有限公司 | Cosmetic or pharmaceutical product comprising active polypeptide and process for preparing same |
-
2011
- 2011-05-18 CN CN 201110128241 patent/CN102219828B/en active Active
Non-Patent Citations (6)
| Title |
|---|
| 三文鱼皮胶原肽的制备及其抗氧化活性的研究;贾建萍 等;《中国食品学报》;20100831;第10卷(第4期);94-99 * |
| 江勇 等.鲨鱼皮明胶水解肽的抗氧化活性与初步分离研究.《经济发展方式转变与自主创新——第十二届中国科学技术协会年会》.2010,1-9. |
| 王运改 等.鮰鱼皮明胶抗氧化肽的制备工艺研究.《食品科学》.2010,第31卷(第19期),254-258. |
| 贾建萍 等.三文鱼皮胶原肽的制备及其抗氧化活性的研究.《中国食品学报》.2010,第10卷(第4期),94-99. |
| 鮰鱼皮明胶抗氧化肽的制备工艺研究;王运改 等;《食品科学》;20101231;第31卷(第19期);254-258 * |
| 鲨鱼皮明胶水解肽的抗氧化活性与初步分离研究;江勇 等;《经济发展方式转变与自主创新——第十二届中国科学技术协会年会》;20101231;1-9 * |
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