CN103352065A - Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof - Google Patents
Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof Download PDFInfo
- Publication number
- CN103352065A CN103352065A CN2013102941683A CN201310294168A CN103352065A CN 103352065 A CN103352065 A CN 103352065A CN 2013102941683 A CN2013102941683 A CN 2013102941683A CN 201310294168 A CN201310294168 A CN 201310294168A CN 103352065 A CN103352065 A CN 103352065A
- Authority
- CN
- China
- Prior art keywords
- henan
- china
- polypeptide
- rhiizoma dioscoreae
- antioxidation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses Huai rhizoma dioscoreae antioxidation polypeptide and a preparation method thereof, and belongs to the technical field of biology. The preparation method comprises the following steps: taking Huai rhizoma dioscoreae as raw materials, enzymatically hydrolyzing the Huai rhizoma dioscoreae by neutral protease to obtain a crude Huai rhizoma dioscoreae polypeptide solution, and performing separation and purification processes such as absolute ethanol desugaring, dialysis and dextran gel chromatography on the crude Huai rhizoma dioscoreae polypeptide solution to obtain the Huai rhizoma dioscoreae antioxidation polypeptide, wherein the molecular weight of the Huai rhizoma dioscoreae antioxidation polypeptide is 600-5000 da. The Huai rhizoma dioscoreae antioxidation polypeptide prepared by the method is higher in purity and oxidation resistance; and if the Huai rhizoma dioscoreae antioxidation polypeptide is developed into a natural antioxidant, the safety worry of people on an artificially synthesized oxidant is eliminated, the development and the comprehensive utilization of the Huai rhizoma dioscoreae are promoted, and a basis is laid for an application of the antioxidation polypeptide to foods and medicines.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide and preparation method thereof.
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.Therefore, the antioxidant of seeking safety with suppress polyphenoils effect be the study hotspot that biochemistry nutrition is learned always.Polypeptide relates to the active substance of various kinds of cell function in the organism.Along with the development of modern protein engineering, bio-enzyme engineering technology, the biologically active peptides that has in a large number characteristic functions is found, develops, and is applied to the fields such as food formulations, makeup.The research of the antioxygenation mechanism of polypeptide and preparation method's research of antioxidation polypeptide are also day by day ripe.Although the antioxidant low price of chemosynthesis, effective, but Many researchers finds that the chemosynthesis antioxidant can cause the property accumulated carcinogenesis to organs such as human liver, lungs at present, thereby cause people to the worry of chemosynthesis antioxidant, and beginning limits its Applications in food gradually.So people have turned to natural antioxidants to sight, therefore, it is particularly important that the preparation research of natural anti-oxidation polypeptide seems.
It is domestic that the RHIIZOMA DIOSCOREAE from Henan of China that have a title in " bosom ginseng " originates from Jiaozhuo, Henan, and have high medicinal and health care and be worth, be one of main raw material of a lot of health care medicines and functional foodstuff.Contain abundant albumen in the RHIIZOMA DIOSCOREAE from Henan of China, its protein extraction is out become polypeptide liquid by protease hydrolyzed, again polypeptide liquid is carried out purifying, can obtain having the RHIIZOMA DIOSCOREAE from Henan of China polypeptide than the strong anti-oxidation activity.Biologically active peptides is extremely important active substance in the human body, and it has participated in, and human body grows, metabolism, disease and aging, dead whole process, and plays vital effect.Research finds that peptide is compared with protein, has good solubility, low viscosity, anti-gel formative, and low molecular peptide can not produce the advantages such as anaphylaxis.In addition, also has numerous physiologically actives, such as anti-oxidant, hypotensive, decreasing cholesterol, antifatigue, promotion alcohol metabolism isoreactivity.
Gel permeation chromatography also claims sieve chromatography, exclusion chromatography, is to utilize the molecular sieve effect with cancellated gel, separates according to the molecular size difference of separated material.Filler in the chromatography column is the porous network structure material of some inertia, mostly be crosslinked glycan (such as dextran or agarose) class material, small-molecule substance can enter its inside, distance is longer when flowing down, the section and macromolecular substance is left out, the distance of getting off is short, and when a mixing solutions passed through the gel permeation chromatography post, the material in the solution had just been opened by the different molecular weight screening.It is one of the most widely used separation method of present polypeptide, also is the method the simplest in all chromatographic techniques, that condition is the gentleest.
Summary of the invention
Primary and foremost purpose of the present invention is to provide a kind of preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide, the method good separating effect, and the polypeptide purity of acquisition reaches more than 90%.
Another object of the present invention is to provide the RHIIZOMA DIOSCOREAE from Henan of China for preparing by above-mentioned preparation method antioxidation polypeptide.
Purpose of the present invention is achieved through the following technical solutions:
A kind of preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide comprises the steps: to adopt neutral protease that RHIIZOMA DIOSCOREAE from Henan of China is carried out enzymolysis and obtains the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China, obtains the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide through separation and purification, lyophilize; The means of described separation and purification comprise desugar, dialysis and gel chromatography.
The preparation method of described RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide specifically comprises the steps:
(1) preparation of the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China: take by weighing RHIIZOMA DIOSCOREAE from Henan of China, add the entry making beating; Elder generation's thermal treatment, be cooled to again the enzyme digestion reaction temperature, transfer to the required pH value of enzyme reaction with NaOH solution, add neutral protease, constantly drip NaOH solution in the reaction process, make the pH value stabilization in the required pH value of enzyme reaction, after reaction finishes, the enzyme that goes out, cooling, the centrifuging and taking supernatant liquor namely obtains the thick polypeptide liquid YP of filemot RHIIZOMA DIOSCOREAE from Henan of China.
(2) desugar: thick polypeptide liquid YP is carried out polypeptide liquid YP after desugar obtains desugar with dehydrated alcohol
(3) dialysis: with polypeptide liquid YP
Process with trichoroacetic acid(TCA), can remove remaining most of albumen in the polypeptide liquid, behind the deproteination, by the dialysis of 1000da dialysis tubing, all fallen by dialysis less than the small molecules amino acid of 1000da and ion etc. in the polypeptide liquid, obtain purer mixed peptide component YPP.
(4) gel chromatography: separate mixed peptide component YPP with Sephadex G-50 sephadex chromatography, collect the chromatography elutriant of best anti-oxidant activity component, again through 1000da dialysis tubing dialysis desalting.
(5) freezing: the polypeptide liquid after the collection desalination places the above ice cube that forms of the freezing 24h of Ultralow Temperature Freezer below-50 ℃.
(6) drying: place Vacuumdrier dry freezing ice cube, obtain the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide.
The preparation of the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China described in the step (1) is preferably: the concentration of substrate by 20% mass concentration takes by weighing RHIIZOMA DIOSCOREAE from Henan of China, adds the distilled water making beating; Then at 90 ℃ of lower thermal treatment 30min, after being cooled to 45 ℃, transferring to pH with the NaOH solution of 0.1mol/L is 6, adding enzyme substrates according to used enzyme activity unit is the neutral protease of 1.2-6 ten thousand U/g than [E]/[S], constantly drip the NaOH solution of 0.1mol/L in the reaction process, make pH value stabilization 6; Behind the reaction 6h, the boiling water bath enzyme 10min that goes out, centrifugal 15 min of 10000r/min get supernatant liquor after the cooling, namely obtain the thick polypeptide liquid YP of filemot RHIIZOMA DIOSCOREAE from Henan of China.
Desugar described in the step (2) is preferably: mix with the ratio of thick polypeptide liquid YP volume ratio 1:1 according to dehydrated alcohol, 170rpm/min shakes 1h in shaking table, and the centrifugal 15min of 8000rpm/min gets the polypeptide liquid YP after supernatant liquor is desugar
Trichoroacetic acid(TCA) described in the step (3) is preferably the trichoroacetic acid(TCA) of volumetric concentration 15%, polypeptide liquid YP
Be preferably 1:1 with the volume ratio of 15% trichoroacetic acid(TCA).
The condition optimization of the gel chromatography described in the step (4) is: the chromatography column post of employing directly is 2cm, dress post height is 45-70cm, sample size is 0.5-1mL, elutriant is Tutofusin tris/HCI of pH 8.0 final concentration 50mmol/L and the mixed solution of final concentration 0.lmol/L sodium-chlor, flow velocity 3-6mL/10min measures under 280nm; Collect the elutriant of different absorption peaks, by the anti-oxidant activity of each elution fraction of hydroxy radical qiao kit measurement, and then collect the chromatography elutriant of best anti-oxidant activity component.
The preparation method of described RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide also comprises further crushing screening and can, and is specific as follows: crushing screening: the RHIIZOMA DIOSCOREAE from Henan of China polypeptide freezing, that drying obtains is pulverized through pulverizer, crossed 100 order vibratory screening apparatuss; Can: the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide powder after sieving carries out sterile filling, obtains RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide powder.
A kind of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide prepares by aforesaid method, and its molecular weight is 600-5000da.
The present invention has following advantage and effect with respect to prior art:
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.This anti-oxidation peptide also can be used as main active ingredient and be developed to various healthcare products; such as protective foods, makeup etc.; this not only has very important meaning to the health level that improves people; and can promote large-scale production and the deep processing of Chinese yam, for anti-oxidation peptide application is in food, medicine laid a good foundation.
Description of drawings
Fig. 1 is that Sephadex G-50 dextrane gel is to the separating effect figure of RHIIZOMA DIOSCOREAE from Henan of China mixed peptide;
Time: 1 hour 46 minutes 29 seconds, A=0.014;
Time: 2 hours 13 minutes 47 seconds, A=0.018;
Time: 4 hours 55 minutes 33 seconds, A=0.035;
Time: 14 hours 31 minutes 06 second, A=0.032.
Fig. 2 is the elutriant canonical plotting; Bovine serum albumin (M=68000) appearance time: 0 hour 51 minutes 07 second, A=0.109; Sigma I8405 (M=5700) appearance time: 2 hours 03 minute 19 seconds, A=0.031.
Fig. 3 is molecular weight logarithm and elution volume graph of a relation, and the molecular weight distribution curve equation is: y=-0.0498x+5.5951.
Fig. 4 is different elution fraction reducing power comparison diagrams, and YPP III component has stronger reducing power.
Fig. 5 is different elution fraction Scavenging activity on hydroxyl free radical figure, and YPP III component has stronger Scavenging activity on hydroxyl free radical, and is suitable with vitamins C.
Embodiment
Below in conjunction with embodiment and accompanying drawing the present invention is done further detailed description, but embodiments of the present invention are not limited to this.If do not specialize, the conventional means that used technique means is well known to those skilled in the art among the embodiment.
Embodiment 1
(1) preparation of the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China: the concentration of substrate by 20% accurately takes by weighing the fresh Chinese yam of 100g, adds the making beating of 400mL distilled water; Then at 90 ℃ of lower thermal treatment 30min, be cooled to the enzyme digestion reaction temperature 45 C after, transfer to the required pH value of enzyme reaction pH 6 with the NaOH solution of 0.1 mol/L; Add 3g neutral protease (enzyme work is 1,200,000 U/g for the prosperous industrial chemicals in Zhengzhou poly-hundred million company limited, food grade), constantly drip the NaOH solution of 0.1mol/L in the reaction process, make the pH value stabilization 6; Behind the reaction 6h, the boiling water bath enzyme 10min that goes out, centrifugal 15 min of 10000r/min get supernatant liquor after the cooling, namely obtain the thick polypeptide liquid YP of filemot RHIIZOMA DIOSCOREAE from Henan of China.Measure to such an extent that thick polypeptide liquid YP content of peptides is 0.98g/L through biuret method, the yield of polypeptide is the 0.26g/100g Chinese yam.
(2) desugar: with dehydrated alcohol thick polypeptide liquid YP is carried out desugar, mix with the ratio of thick enzymolysis solution volume ratio 1:1 according to dehydrated alcohol, 170rpm/min shakes 1h in shaking table, and the centrifugal 15min of 8000rpm/min gets the polypeptide liquid YP after supernatant liquor is desugar
Measure to such an extent that the polysaccharide clearance reaches 58.07% through phenol sulfuric acid process and biuret method, the polypeptide retention rate reaches 75.17%.
(3) dialysis: polypeptide liquid YP
By volume 1:1 ratio processing of trichoroacetic acid(TCA) through volumetric concentration 15%, can remove remaining most of albumen in the polypeptide liquid, behind the deproteination, dialyse by the 1000da dialysis tubing, all fallen by dialysis less than the small molecules amino acid of 1000da and ion etc. in the polypeptide liquid, obtain purer mixed peptide component YPP.
(4) gel chromatography: with Sephadex G-50(pharmacia company) sephadex chromatography separates mixed peptide component YPP, the chromatography column post that adopts directly is 2cm, dress post height is 70cm, sample size is 1mL, elutriant is the mixed solution of pH 8.0 final concentration 50mmol/L Tutofusin tris/HCI and final concentration 0.lmol/L sodium-chlor, flow velocity 3mL/10min, under 280nm, measure, collect the elutriant of different absorption peaks, by the anti-oxidant activity of each elution fraction of hydroxy radical qiao kit measurement.
The instrument, the detection means that adopt are as follows:
Utilize automatic collector (BSZ-100, Industrial Co., Ltd. of upper Nereid section) and computer Ultraviolet Detector (HD-9707, Industrial Co., Ltd. of upper Nereid section) to detect and collect different elution fractions.
Molecular weight and the Antioxidative Activity Determination of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide are as follows:
1) molecular weight determination of antioxidation polypeptide
Measure best anti-oxidant activity component molecular weight, quantitative sample injection bovine serum albumin and Sigma I8405 standard substance mixed solution, drawing standard protein adsorption curve (see figure 2), the corresponding elutriant time of the protein peak value that settles the standard, again take this elutriant time as X-coordinate, the corresponding molecular weight logarithmic value of protein standard substance is that logM-V typical curve (see figure 3) is drawn in the ordinate zou mapping, adopt the one-variable linear regression method to try to achieve the relational expression of molecular weight logarithmic value and elutriant time, thereby obtain the standard substance molecular weight distribution curve.Can be obtained the molecular weight ranges of best antioxidation polypeptide component by the elution time of best antioxidation polypeptide component according to this distribution curve.
2) test of anti-oxidant activity
Anti-oxidant activity comprehensively compares by reducing power and Scavenging activity on hydroxyl free radical.
3) test of reducing power
Get certain density sample (0.1mg/mL-500mg/mL) 2.5mL, add the phosphate buffered saline buffer (0.2mol/L) of 2.5mL pH6.6 and the potassium ferricyanide solution of 2.5mL mass concentration l%, mixing, the trichoroacetic acid(TCA) that adds 2.5mL volumetric concentration 10% behind 50 ℃ of insulation 20min mixes rear with the centrifugal 10min of 3000r/min.Get supernatant 2.5mL, add the FeC1 of 2.5mL distilled water and 0.50mL mass concentration 0.1%
3, then the 5min that develops the color under the room temperature measures absorbancy at 700nm wavelength place.Blank tube namely changes sample into 2.5mL distilled water, and all the other operation stepss are identical with sample hose.The absorbancy that records is larger, and then the reducing power of sample is stronger.
4) test of Scavenging activity on hydroxyl free radical
Utilize hydroxy radical qiao test kit (bio-engineering research institute is built up in Nanjing) to measure and wash Scavenging activity on hydroxyl free radical, each solution mixing in will showing according to the described method of table 1,37 ℃ of reactions 1 minute (accurately with manual time-keeping), begin to finish by one minute from adding reagent three, add immediately the developer termination reaction, the amount of every colorimetric cylinder adding developer is 2mL, once can only make a colorimetric cylinder.After adding developer, mixing, room temperature was placed after 20 minutes, the 1cm optical path, 550nm, the distilled water zeroing is surveyed and is respectively managed absorbance.
Table 1 Scavenging activity on hydroxyl free radical measuring method table
| ? | Blank tube | Standard pipe | Control tube | Measure pipe |
| Distilled water (mL) | 0.4 | 0.2 | 0.2 | ? |
| 0.03％H 2O 2Standard Applying Solution (mL) | ? | 0.2 | ? | ? |
| Substrate is used liquid (mL) | ? | ? | 0.2 | 0.2 |
| Sample * (mL) | ? | ? | ? | 0.2 |
| Reagent three is used liquid (mL) | 0.4 | 0.4 | 0.4 | 0.4 |
5) remove the active calculation formula of hydroxy radical qiao
Remove hydroxyl radicals (U/mL)=(contrast OD value-measure OD value)/front extension rate of (standard OD value-blank OD value) * standard substance concentration (8.824mmol/L) * (1mL/ sampling amount) * test sample.
The result is as follows: the design sketch of Sephadex G-50 sephadex chromatography separation mixed peptide component YPP is collected absorption peak as shown in Figure 1
,
,
,
Elutriant, respectively called after YPP I, YPP II, YPP III, YPP IV.YPP I, YPP II, YPP III, four kinds of components of YPP IV reducing power and Scavenging activity on hydroxyl free radical result such as Fig. 4 and Fig. 5 (Chinese yam protain is that RHIIZOMA DIOSCOREAE from Henan of China is through the standby RHIIZOMA DIOSCOREAE from Henan of China crude protein that obtains of the heavy legal system of the molten acid of salt, vitamins C is commercially available vitamins C medicine) shown in, YPP III component is good anti-oxidant activity component, and its molecular weight is 600-5000da.
(5) desalination: collect the chromatography elutriant of best anti-oxidant activity component YPP III, through 1000da dialysis tubing dialysis desalting.
(6) freezing: polypeptide liquid is put the above ice cube that forms of the freezing 24h of Ultralow Temperature Freezer below-50 ℃ after collecting desalination.
(7) drying: freezing ice cube zymolyte is dry to Vacuumdrier.
(8) crushing screening: the RHIIZOMA DIOSCOREAE from Henan of China polypeptide that lyophilize obtains is pulverized through pulverizer, crosses 100 order vibratory screening apparatuss.
(9) can: the antioxidation polypeptide powder after sieving carries out sterile filling, obtains RHIIZOMA DIOSCOREAE from Henan of China polypeptide antioxidant.It is 90.8% that the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide that obtains is measured the mass content that can get antioxidation polypeptide through biuret method.
The present invention uses the multiple separation and purification means such as ethanol desugar, dialysis tubing dialysis and Sephadex G-50 sephadex chromatography, and realization has the high efficiency separation purifying of the antioxidation polypeptide of remarkable activity.
Above-described embodiment is the better embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. the preparation method of a RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide is characterized in that comprising the steps: to adopt neutral protease that RHIIZOMA DIOSCOREAE from Henan of China is carried out enzymolysis obtains the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China, obtains the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide through separation and purification, lyophilize.
2. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 1, it is characterized in that: the means of described separation and purification comprise desugar, dialysis and gel chromatography.
3. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 1 is characterized in that specifically comprising the steps:
(1) preparation of the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China: take by weighing RHIIZOMA DIOSCOREAE from Henan of China, add the entry making beating; Elder generation's thermal treatment, be cooled to again the enzyme digestion reaction temperature, transfer to the required pH value of enzyme reaction with NaOH solution, add neutral protease, constantly drip NaOH solution in the reaction process, make the required pH value of pH value stabilization enzyme reaction, after reaction finishes, the enzyme that goes out, cooling, the centrifuging and taking supernatant liquor namely obtains the thick polypeptide liquid YP of RHIIZOMA DIOSCOREAE from Henan of China;
(2) desugar: thick polypeptide liquid YP is carried out polypeptide liquid YP after desugar obtains desugar with dehydrated alcohol
(3) dialysis: with polypeptide liquid YP
Process with trichoroacetic acid(TCA), behind the deproteination, by the dialysis of 1000da dialysis tubing, obtain mixed peptide component YPP;
(4) gel chromatography: separate mixed peptide component YPP with Sephadex G-50 sephadex chromatography, collect the chromatography elutriant of best anti-oxidant activity component, again through 1000da dialysis tubing dialysis desalting;
(5) freezing: the polypeptide liquid after the collection desalination places the above ice cube that forms of the freezing 24h of Ultralow Temperature Freezer below-50 ℃;
(6) drying: place Vacuumdrier dry freezing ice cube, obtain the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide.
4. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 3 is characterized in that: being prepared as of the thick polypeptide liquid of RHIIZOMA DIOSCOREAE from Henan of China described in the step (1): the concentration of substrate by 20% mass concentration takes by weighing RHIIZOMA DIOSCOREAE from Henan of China, adds the distilled water making beating; Then at 90 ℃ of lower thermal treatment 30min, after being cooled to 45 ℃, transferring to pH with the NaOH solution of 0.1 mol/L is 6, adds enzyme substrates than being the neutral protease of 1.2-6 ten thousand U/g according to used enzyme activity unit, constantly drip the NaOH solution of 0.1 mol/L in the reaction process, make pH value stabilization 6; Behind the reaction 6h, the boiling water bath enzyme 10min that goes out, centrifugal 15 min of 10000r/min get supernatant liquor after the cooling, namely obtain the thick polypeptide liquid YP of RHIIZOMA DIOSCOREAE from Henan of China.
5. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 3, it is characterized in that: the desugar described in the step (2) is: mix with the ratio of thick polypeptide liquid YP volume ratio 1:1 according to dehydrated alcohol, 170rpm/min shakes 1h in shaking table, the centrifugal 15min of 8000rpm/min gets supernatant liquor and is polypeptide liquid YP after the desugar
6. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 3, it is characterized in that: the trichoroacetic acid(TCA) described in the step (3) is the trichoroacetic acid(TCA) of volumetric concentration 15%, polypeptide liquid YP
With the volume ratio of the trichoroacetic acid(TCA) of volumetric concentration 15% be 1:1.
7. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 3, it is characterized in that: the condition of the gel chromatography described in the step (4) is: the chromatography column post of employing directly is 2cm, dress post height is 45-70cm, sample size is 0.5-1mL, elutriant is Tutofusin tris/HCI of pH 8.0 final concentration 50mmol/L and the mixed solution of final concentration 0.lmol/L sodium-chlor, flow velocity 3-6mL/10min measures under 280nm.
8. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 1, it is characterized in that: freezing, drying obtains also comprising crushing screening and can behind the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide.
9. the preparation method of RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide according to claim 8 is characterized in that: described crushing screening is crossed 100 order vibratory screening apparatuss for the RHIIZOMA DIOSCOREAE from Henan of China polypeptide freezing, that drying obtains is pulverized through pulverizer; Described can obtains RHIIZOMA DIOSCOREAE from Henan of China polypeptide antioxidant for the RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide powder after will sieving carries out sterile filling.
10. a RHIIZOMA DIOSCOREAE from Henan of China antioxidation polypeptide is characterized in that: prepare by each described method of claim 1-9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013102941683A CN103352065A (en) | 2013-07-12 | 2013-07-12 | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013102941683A CN103352065A (en) | 2013-07-12 | 2013-07-12 | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103352065A true CN103352065A (en) | 2013-10-16 |
Family
ID=49308476
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2013102941683A Pending CN103352065A (en) | 2013-07-12 | 2013-07-12 | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103352065A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105111294A (en) * | 2015-09-08 | 2015-12-02 | 苏州普罗达生物科技有限公司 | Chinese yam peptide and application thereof |
| CN107467192A (en) * | 2017-08-11 | 2017-12-15 | 烟台市华昕生物医药科技有限公司 | A kind of apple peptide milk tea of the active matter containing oligopeptides |
| CN109315584A (en) * | 2017-08-01 | 2019-02-12 | 天津天肽生物科技有限公司 | A kind of preparation method of Chinese yam polypeptide |
| CN113527411A (en) * | 2021-07-24 | 2021-10-22 | 中国科学院新疆理化技术研究所 | Separation and purification method of dioscorea opposita glycopeptide |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009091305A (en) * | 2007-10-10 | 2009-04-30 | Yaizu Suisankagaku Industry Co Ltd | Novel peptide and method of manufacturing the same, and antioxidant |
-
2013
- 2013-07-12 CN CN2013102941683A patent/CN103352065A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009091305A (en) * | 2007-10-10 | 2009-04-30 | Yaizu Suisankagaku Industry Co Ltd | Novel peptide and method of manufacturing the same, and antioxidant |
Non-Patent Citations (2)
| Title |
|---|
| 李小强: "山药蛋白酶解多肽液及多肽酒制备工艺研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
| 董怀灿等: "怀山药肽的开发及应用", 《科技风》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105111294A (en) * | 2015-09-08 | 2015-12-02 | 苏州普罗达生物科技有限公司 | Chinese yam peptide and application thereof |
| CN109315584A (en) * | 2017-08-01 | 2019-02-12 | 天津天肽生物科技有限公司 | A kind of preparation method of Chinese yam polypeptide |
| CN107467192A (en) * | 2017-08-11 | 2017-12-15 | 烟台市华昕生物医药科技有限公司 | A kind of apple peptide milk tea of the active matter containing oligopeptides |
| CN113527411A (en) * | 2021-07-24 | 2021-10-22 | 中国科学院新疆理化技术研究所 | Separation and purification method of dioscorea opposita glycopeptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102336840B (en) | Triple-helical Tremellan, preparation method and application thereof | |
| CN103919712B (en) | Cordyceps militaris extract, and preparation method and application thereof | |
| CN102584963B (en) | Active Ganoderma lucidum polysaccharide peptide reference substance, and preparation method and application thereof | |
| CN103352065A (en) | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof | |
| CN105695545A (en) | Method for preparing sea cucumber fucoidan and sea cucumber glycoprotein | |
| WO2022016644A1 (en) | Acanthopanax senticosus harms homogeneous polysaccharide, preparation method therefor and use thereof | |
| CN102676613A (en) | Preparation method for disaccharide, tetrasccharide and hexaose of chondroitin sulfuric acid | |
| CN105037577B (en) | Procoagulant blackberry seed polysaccharide, and extraction separation method and application thereof | |
| CN109400753A (en) | Giant salamander cartilage preparation | |
| CN110916198A (en) | Method for simultaneously preparing pectic polysaccharide and viscous glycoprotein by using okra fermented wine lees | |
| CN103172724A (en) | Method for extracting ovomacroglobulin from egg white | |
| CN113717296B (en) | Eucommia acidic polysaccharide, extraction method and application of eucommia acidic polysaccharide in preparation of anti-colon cancer drugs | |
| CN106520877A (en) | Method for preparing pig cerebral protein antioxidative peptide | |
| CN104610446A (en) | Jerk filefish skin collagen antioxidative peptide and preparation method thereof | |
| CN102000133B (en) | Preparation method of traditional Chinese medicine anti-oxidation preparation | |
| CN103382229A (en) | Preparation method and structural identification for novel strongylocentrotus nudus egg polysaccharide having immunoregulation effect | |
| CN104865333B (en) | A kind of detection method of Arillus Longan proteoglycan | |
| CN117281885A (en) | Application of selenium-enriched yam glycoprotein as immunomodulating drug | |
| CN104817646B (en) | Polysaccharide and its extracting method and application in ganoderma capense | |
| CN101962402A (en) | Astragalus membranaceus glycoprotein as well as preparation method and usage thereof | |
| CN107177650A (en) | A kind of preparation method of the anti-oxidant enzymolysis oligopeptide of North Pacific squid spawn tangled gland | |
| CN104861080A (en) | Polysaccharide in guava and preparation method and application thereof | |
| CN101857645B (en) | Water-soluble neutral Ganoderma lucidum polysaccharide GLP1 and preparation method thereof | |
| CN109081864A (en) | A kind of method for extraction and purification of Chinese potato glucoprotein | |
| CN101643515A (en) | Method for separating and purifying lentinan for injection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20131016 |