CN114958950A - Donkey milk antioxidant peptide and preparation method thereof - Google Patents

Donkey milk antioxidant peptide and preparation method thereof Download PDF

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CN114958950A
CN114958950A CN202210711742.XA CN202210711742A CN114958950A CN 114958950 A CN114958950 A CN 114958950A CN 202210711742 A CN202210711742 A CN 202210711742A CN 114958950 A CN114958950 A CN 114958950A
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donkey milk
peptide
donkey
whey protein
enzymolysis
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刘淼
马超越
梁星
吴晓彤
李超
刘志宇
刘端端
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Inner Mongolia University
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
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Abstract

The invention provides donkey milk antioxidant peptide and a preparation method thereof, and relates to the technical field of dairy products. The invention provides a preparation method of donkey milk antioxidant peptide, which can be used for preparing donkey milk antioxidant peptide by directly utilizing alpha-chymotrypsin to carry out enzymolysis on donkey milk whey protein. Proved by tests, the donkey milk antioxidant peptide prepared by the invention has high antioxidant activity, and has high activity on DPPH, OH, ABTS and O 2 ‑ Has good clearing effect, good reducing capability to iron ions, good anti-aging effect to yeast cells and important significance to the development and application of donkey milk.

Description

Donkey milk antioxidant peptide and preparation method thereof
Technical Field
The invention relates to the technical field of dairy products, and particularly relates to donkey milk antioxidant peptide and a preparation method thereof.
Background
Donkey milk is known as the best human milk substitute, has low fat content and high content of unsaturated fatty acid, vitamin and lactose, and has stronger health care function and the function of treating human body immunity related diseases. As whey protein milk, donkey whey protein contains high content of alpha-lactalbumin, beta-lactoglobulin and lysozyme. The alpha-lactalbumin can effectively inhibit harmful bacteria and is beneficial to the growth and development of infants; the beta-lactoglobulin can be specifically combined with the nutrient components in the digestion process to play a role in protecting the nutrient components; and the higher content of lysozyme enables the donkey milk to have stronger antibacterial function. In addition, donkey whey protein also contains various trace components such as immunoglobulin, serum albumin, epidermal growth factor and the like, and has important influence on the disease resistance mechanism of the organism. Therefore, the development and utilization of donkey milk have great social value and market prospect. However, at present, the donkey milk product in China is single, the utilization rate of donkey whey protein is greatly limited, and the economic benefit cannot be released to the maximum. The preparation of the bioactive peptide by using the protease enzymolysis donkey whey protein is beneficial to further research on the comprehensive value of donkey milk products.
The bioactive peptide refers to small peptide substances which are beneficial to the activity and health of a life organism and can optimize the metabolic environment of the organism, and has various bioactive functions of resisting bacteria, resisting oxidation, regulating immunity, reducing blood pressure and the like. Wherein, the antioxidant active peptide can remove redundant free radicals in vivo by providing protons, metal ion chelate, inhibiting lipid peroxidation and the like. Meanwhile, the proportion of hydrophobic amino acid and aromatic amino acid in the peptide chain is in positive correlation with the antioxidant activity of the polypeptide, and the higher the proportion is, the stronger the antioxidant activity is, and the more beneficial the oxidation defense of the organism is. At present, animal milk protein is a hot spot for researching antioxidant active peptide at home and abroad, but the milk protein is taken as a raw material mostly, and the research aiming at the donkey milk protein antioxidant active peptide is not common.
The research at home and abroad proves that the total antioxidant capacity of the donkey milk is higher than that of human milk, which indicates that the peptide released by enzymolysis of the donkey milk has certain antioxidant capacity, the donkey milk is a high-quality raw material for extracting antioxidant peptide, however, the research on the donkey milk source antioxidant peptide at home is less, a feasible process technical route for preparing the donkey whey protein antioxidant peptide is found, the antioxidant peptide has great significance in the application of the antioxidant peptide in the fields of medicine, food, health care and the like, and the development significance and market economic benefit of the donkey milk can be exerted.
Disclosure of Invention
In view of this, the present invention aims to provide a method for preparing donkey milk antioxidant peptide, which can simply and rapidly prepare donkey milk antioxidant peptide, and the prepared donkey milk antioxidant peptide has good scavenging effect on various free radicals.
In order to solve the technical problems, the invention provides the following technical scheme:
the invention provides a preparation method of donkey milk antioxidant peptide, which comprises the following steps:
carrying out enzymolysis on donkey milk whey protein by using alpha-chymotrypsin to obtain the donkey milk antioxidant peptide; the temperature of the enzymolysis is 30-60 ℃, the time of the enzymolysis is 2-8h, and the pH value of the enzymolysis is 6-10.
Preferably, the donkey milk whey protein is donkey milk whey protein solution, and the concentration of the whey protein in the donkey milk whey protein solution is 2-6%.
Preferably, the mass ratio of the alpha-chymotrypsin to the donkey milk whey protein is 2-6: 100.
Preferably, the donkey milk antioxidant peptide is obtained by adjusting the pH to be neutral after enzymolysis, centrifuging and drying.
Preferably, the solvent for adjusting the pH is hydrochloric acid or sodium hydroxide.
Preferably, the centrifugation is refrigerated centrifugation; the rotating speed of the centrifugation is 4000-4500rmp, and the time of the centrifugation is 5-15 min.
Preferably, the drying is freeze drying.
The invention also provides the donkey milk antioxidant peptide prepared by the preparation method.
The invention also provides application of the donkey milk antioxidant peptide in an antioxidant functional product.
The invention provides a preparation method of donkey milk antioxidant peptide, which can prepare donkey milk antioxidant peptide by utilizing alpha-chymotrypsin to carry out enzymolysis on donkey milk whey protein, wherein enzymolysis substrates are wide in source and easy to obtain, the preparation method is simple and quick, the preparation can be realized by only one-step enzymolysis, the preparation cost and time of the antioxidant peptide are greatly saved, and the expanded production is facilitated. Proved by tests, the donkey milk antioxidant peptide prepared by the invention has high antioxidant activity, and has high activity on DPPH, OH, ABTS and O 2 - Has good clearing effect, good reducing capability to iron ions, good anti-aging effect to yeast cells and important significance to the development and application of donkey milk.
Drawings
FIG. 1 shows the effect of donkey whey protein and zymolytic peptide on DPPH clearance, wherein DWP represents donkey whey protein, DWPP represents donkey whey protein zymolytic peptide, and VC represents positive control ascorbic acid.
FIG. 2 shows the effect of donkey whey protein and enzymolysis peptide on OH clearance, wherein DWP represents donkey whey protein, DWPP represents donkey whey protein enzymolysis peptide, and VC represents positive control ascorbic acid.
FIG. 3 shows the effect of donkey whey protein and zymolytic peptide on ABTS.clearance, where DWP represents donkey whey protein, DWPP represents donkey whey protein zymolytic peptide, and VC represents positive control ascorbic acid.
FIG. 4 shows donkey whey protein and polypeptide pair O 2 - Effect of clearance, where DWP represents donkey whey protein, DWPP represents donkey whey proteolytic peptide, and VC represents positive control ascorbic acid.
FIG. 5 shows the reducing power of donkey whey protein and enzymolysis peptide to iron ions, wherein DWP represents donkey whey protein, DWPP represents donkey whey proteolysis peptide, and VC represents positive control ascorbic acid.
FIG. 6 is a graph showing the effect of culture time on the survival rate of yeast cells.
FIG. 7 is a graph of Lv's basic methylene blue staining.
Detailed Description
The invention provides a preparation method of donkey milk antioxidant peptide, which comprises the following steps:
carrying out enzymolysis on donkey milk whey protein by using alpha-chymotrypsin to obtain the donkey milk antioxidant peptide; the temperature of the enzymolysis is 30-60 ℃, the time of the enzymolysis is 2-8h, and the pH value of the enzymolysis is 6-10.
In the invention, the donkey milk whey protein is preferably donkey milk whey protein solution, and the concentration of the whey protein in the donkey milk whey protein solution is preferably 2-6%, and more preferably 4%. In the invention, the enzymolysis temperature is preferably 38-45 ℃, and more preferably 40 ℃; the enzymolysis time is preferably 3-6h, more preferably 4-5 h; the pH of the enzymatic hydrolysis is preferably 7-9, more preferably 8. In the present invention, the mass ratio of the alpha-chymotrypsin to donkey milk whey protein (i.e. enzyme substrate ratio) is preferably 2-6:100, more preferably 4: 100.
The donkey milk antioxidant peptide is obtained by adjusting the pH to be neutral after enzymolysis, and centrifuging and drying. In the invention, the solvent for adjusting the pH is preferably hydrochloric acid or sodium hydroxide; the concentration of the hydrochloric acid is preferably 1mol/L, and the concentration of the sodium hydroxide is preferably 1 mol/L. In the present invention, the centrifugation is preferably refrigerated centrifugation, and the temperature of the refrigerated centrifugation is preferably 4 ℃; the rotation speed of the centrifugation is preferably 4000-4500rmp, more preferably 4200rmp, and the time of the centrifugation is preferably 5-15min, more preferably 10 min. In the present invention, the drying is preferably freeze-drying.
The invention also provides the donkey milk antioxidant peptide prepared by the preparation method.
The invention also provides application of the donkey milk antioxidant peptide in an antioxidant functional product.
The present invention will be described in detail with reference to examples for better understanding the objects, technical solutions and advantages of the present invention, but they should not be construed as limiting the scope of the present invention.
In the following examples, unless otherwise specified, all methods are conventional.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
1) Preparing the donkey milk source whey protein into a solution with the mass concentration of 4%, heating in a water bath at 85 ℃ for 10min, and adjusting the pH value of the solution to 8 after the solution is cooled;
2) adding an alpha-chymotrypsin solution into the solution obtained in the step 1), stirring the solution with a glass rod with the enzyme bottom being 4% of that of the solution, and then putting the stirred solution into a water bath kettle at 40 ℃ for enzymolysis for 4 hours; wherein, the pH value of the solution is measured at intervals of 30min, and the pH value is maintained to be 8 by 1mol/L sodium hydroxide solution;
3) and (3) after the enzymolysis is finished, inactivating enzyme in a water bath at 95 ℃ for 10min, then quickly cooling the solution to 25 ℃, adding 1mol/L hydrochloric acid solution to adjust the pH value to be neutral, centrifuging the solution at 4 ℃ for 10min at 4200rpm by using a high-speed refrigerated centrifuge, collecting supernatant, and freeze-drying the supernatant to obtain the enzymolysis donkey milk antioxidant peptide freeze-dried powder.
Example 2
1) Preparing the donkey milk source whey protein into a solution with the mass concentration of 5%, heating in a water bath at 85 ℃ for 10min, and adjusting the pH value of the solution to 7 after the solution is cooled;
2) adding an alpha-chymotrypsin solution into the solution obtained in the step 1), stirring by a glass rod, and then putting into a 35 ℃ water bath kettle for enzymolysis for 4 hours; wherein, the pH value of the solution is measured at intervals of 30min, and the pH value is maintained to be 8 by 1mol/L sodium hydroxide solution;
3) and (3) after the enzymolysis is finished, inactivating enzyme in a water bath at 95 ℃ for 10min, then quickly cooling the solution to 25 ℃, adding 1mol/L sodium hydroxide solution to adjust the pH value to be neutral, centrifuging the solution at 4 ℃ for 10min at 4200rpm by using a high-speed refrigerated centrifuge, collecting supernatant, and freeze-drying the supernatant to obtain the enzymolysis donkey milk antioxidant peptide freeze-dried powder.
Example 3
1) Preparing donkey milk source whey protein into a solution with the mass concentration of 3%, heating in a water bath at 85 ℃ for 10min, and adjusting the ph value of the solution to 9 after the solution is cooled;
2) adding an alpha-chymotrypsin solution into the solution obtained in the step 1), stirring by a glass rod, and then putting into a water bath kettle at 50 ℃ for enzymolysis for 4 hours; wherein, the pH value of the solution is measured at intervals of 30min, and the pH value is maintained to be 8 by 1mol/L sodium hydroxide solution;
3) and (3) after the enzymolysis is finished, inactivating enzyme in a water bath at 95 ℃ for 10min, then quickly cooling the solution to 25 ℃, adding 1mol/L hydrochloric acid solution to adjust the pH value to be neutral, centrifuging the solution at 4 ℃ for 10min at 4200rpm by using a high-speed refrigerated centrifuge, collecting supernatant, and freeze-drying the supernatant to obtain the enzymolysis donkey milk antioxidant peptide freeze-dried powder.
Example 4
Effect of donkey whey proteolytic peptide on DPPH clearance:
19.7mg of 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) is weighed and dissolved in ethanol, and the solution is added into a volumetric flask with the volume constant of 250mL to prepare 0.2mM of DPPH absolute ethanol solution.
Respectively mixing the enzymolysis peptide with the same volume with DPPH absolute ethanol solution, shaking uniformly, standing at room temperature in a dark place for 30min, setting the parameters of a high-speed refrigerated centrifuge at 4 ℃, 4200r/min, and centrifuging the mixed solution for 10 min.
Preheating an enzyme-linked immunosorbent assay (ELIAS) reader for 15min in advance, adding 200 mu L of centrifuged solution supernatant into a 96-pore plate, detecting a light absorption value Ai of a sample at 517nm by using the ELIAS reader, replacing a DPPH (dipeptidyl peptidase) absolute ethanol solution with an equal-volume absolute ethanol solution to form a blank group Aj, replacing an enzymolysis peptide sample with an equal-volume distilled water to form a control group A0, and utilizing the formula: DPPH-clearance was calculated as [1- (Ai-Aj)/a0] × 100%, and fig. 1 was obtained. Wherein A0 is light absorption value of control group (distilled water + DPPH anhydrous ethanol solution); ai is the light absorption value of a sample group (enzymolysis peptide + DPPH absolute ethanol solution); aj is the blank (zymolytic peptide + absolute ethyl alcohol) light absorption value.
As can be seen from figure 1, the DPPH and clearance of donkey whey protein increases with the increase of concentration, and at 10mg/mL, the clearance of whey protein enzymolysis peptide reaches 45.24%, and the clearance of whey protein reaches 36.90%.
Example 5
Effect of donkey whey proteolytic peptide on OH clearance:
0.1668g of ferrous sulfate crystal and 0.083g of salicylic acid are weighed and respectively dissolved in 100mL of distilled water to be prepared into a 6mM ferrous sulfate solution and a 6mM salicylic acid solution at constant volume, a plurality of test tubes are prepared, 2mL of the ferrous sulfate solution, the salicylic acid solution and a sample solution with a certain concentration are sequentially added into the test tubes, shaking and shaking are carried out, 2mL of hydrogen peroxide solution is added, and standing is carried out for 20min at room temperature after shaking and shaking are carried out. The light absorption value Ai of the reaction liquid is measured by using a microplate reader, isovolumetric distilled water replaces salicylic acid solution to be used as a blank group Aj, isovolumetric distilled water replaces enzymolysis peptide sample to be used as a control group A0, and the clearance rate of the sample to OH is determined by using a formula: OH clearance [% 1- (Ai-Aj)/a0] × 100%, and fig. 2 was obtained. Wherein, A0 is the light absorption value of a control group (distilled water + salicylic acid solution); ai is the light absorption value of a sample group (enzymolysis peptide + salicylic acid solution); aj is the blank (zymolytic peptide + distilled water) absorbance.
As can be seen from FIG. 2, both donkey whey protein and donkey whey proteolytic peptide can eliminate OH and have a dose-dependent relationship with concentration. The clearance rate of the whey protein enzymolysis peptide to OH is higher than that of whey protein, and at 10mg/mL, the clearance rate of the whey protein enzymolysis peptide to OH is 72.43%, and the clearance rate of the whey protein to OH is 62.2%.
Example 6
Influence of donkey whey protein enzymolysis peptide on ABTS clearance
Taking 3mg of 2,2' -dinitrobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and adding 0.735mL of distilled water to prepare 7.4mmol/L ABTS stock solution, then taking 1mg of potassium persulfate and adding 1.43mL of distilled water to prepare 2.6mmol/L potassium persulfate stock solution, mixing the ABTS diammonium salt stock solution and the potassium persulfate stock solution in equal volume, standing for 12h under the dark environment condition, and diluting the mixed solution by using absolute ethyl alcohol until the absorbance is 0.7 +/-0.02 when in use. 9mL of diluted ABTS solution is taken and transferred into a test tube, 1mL of sample is added, the mixture is immediately shaken and shaken up, then the mixture is kept stand for 6min, the absorbance of the reaction solution at the wavelength of 734nm is measured, meanwhile, the equal volume of absolute ethyl alcohol is used for replacing the ABTS solution to be a blank group A2, the equal volume of absolute ethyl alcohol is used for replacing an enzymolysis peptide sample to be a control group A0, and the clearance rate of the sample to ABTS free radicals can be calculated according to the formula of ABTS & clearance rate% [1- (A1-A2)/A0 ]. times.100%, and the result is shown in figure 3. Wherein, A0 is the light absorption value of a control group (distilled water + ABTS solution), A1 is the light absorption value of a sample group (enzymolysis peptide + ABTS solution), and A2 is the light absorption value of a blank group (enzymolysis peptide + absolute ethyl alcohol).
As can be seen from fig. 3, the ABTS clearance of both whey protein and whey proteolytic peptide increases with increasing concentration, but none of them has high ABTS clearance of ascorbic acid, and when the sample concentration is the same, the ascorbic acid clearance almost reaches 100%, and then whey proteolytic peptide has the lowest whey protein clearance.
Example 7
Donkey whey protein enzymolysis peptide pair O 2 - Effect of clearance
Respectively preparing 50mM Tris-HCl buffer solution with pH 8.2 and 25mM pyrogallol solution, preheating the Tris-HCl buffer solution in a water bath at 25 ℃ for 20min, sucking 4.5mL into a test tube, adding 0.5mL pyrogallol, uniformly mixing, measuring the light absorption value of the solution at 325nm, measuring the light absorption value once every 30 seconds, continuously measuring for 4.5min, drawing by taking time As an x axis and the light absorption value As a y axis, wherein the slope of a straight line is the pyrogallol autooxidation rate (A0), then taking 4.5mL Tris-HCl buffer solution, respectively adding 1mL sample solutions prepared in advance with different concentrations, measuring the light absorption value of the sample solution at 325nm by using an ultraviolet spectrophotometer by the same method, drawing, wherein the slope of the straight line is the pyrogallol autooxidation rate (As), and the clearance rate is according to a formula O 2 - The clearance was calculated As (a0-As)/a0 × 100%, and fig. 4 was obtained. Wherein A0 is the absorbance of the control group, and As is the absorbance of the sample group.
As can be seen from FIG. 4, the donkey whey protein and donkey whey proteolytic peptide pair O 2 - The clearance rate of the whey protein increases along with the increase of concentration, and the clearance rate of the whey protein enzymolysis peptide is higher than that of the whey protein to O under the same concentration 2 - The clearance of (2) is 10mg/mL, the clearance of enzymolysis peptide is 53.79%, and the clearance of whey protein is 36.82%. The results show that the antioxidant activity of the donkey whey protein after enzymolysis by alpha-chymotrypsin is higher than that of the donkey whey protein without enzymolysis, which is probably because peptide bonds of the protein are broken after enzymolysis, peptide segments with antioxidant activity and free amino acids are released from a protein sequence.
Example 8
Reducing power of enzymolysis peptide
Respectively putting 1mL of sample solution with the concentration of 0, 2, 4, 6, 8, 10 and 12mg/mL into a test tube, respectively adding 2.5mL of 0.2M phosphate buffer solution and 2.5mL of 1% potassium ferricyanide solution, uniformly mixing, standing in a water bath at 50 ℃ for 20min, taking out, rapidly cooling to room temperature, adding 2.5mL of 10% trichloroacetic acid, centrifuging at 4 ℃ at 3000r/min for 10min, taking out 2.5mL of supernate, adding 0.5mL of 0.1% ferric trichloride solution and 2.5mL of distilled water, uniformly mixing, taking distilled water as a blank control, and measuring the absorbance of the sample at OD700 to obtain the graph 5.
The above results reflect the influence of donkey whey protein, donkey whey proteolytic peptide and ascorbic acid of different concentrations on the iron ion reducing ability. It can be seen that, with the increase of the concentration, the reducing power of both the whey protein and the whey protein enzymolysis peptide is gradually enhanced, and the reducing power of the whey protein enzymolysis peptide is higher than that of the whey protein, and the reducing power of the whey protein enzymolysis peptide is 0.46 and 0.36 at 10 mg/mL.
Example 9
Influence of donkey whey protein enzymolysis peptide on cell life of saccharomyces cerevisiae
After the saccharomyces cerevisiae is cultured to the balance period, 5 parts of the saccharomyces cerevisiae are averagely divided into centrifugal tubes and labeled, and the prepared 1, 2, 4mg/mL donkey whey protein enzymolysis peptide is expressed by the ratio of 1: adding 10 proportions into each bacterial solution, mixing, continuously culturing at 30 deg.C and 190r/min for 12 days, taking out small amount of bacterial solution on days 0, 3, 6, 9 and 12, centrifuging at high speed, removing supernatant, washing the precipitated bacteria with 1 × PBS buffer solution, resuspending, and adjusting bacterial solution concentration to 9.3 × 10 by counting with hemocytometer 6 cfu/mL, yeast plate count was performed to obtain FIG. 6. Simultaneous luwen's basic methylene blue staining was performed to give figure 7. Wherein the positive control is 4mg/mL L-carnosine, and the blank control is sterile water.
From the above results, it can be seen that the survival rate of the Saccharomyces cerevisiae cells gradually decreased with the increase of the culture time. In the first 3 days, the survival rates of the cells of the groups are not greatly different, and the descending trend starts to obviously change at the 6 th day, wherein the survival rate of the cells of the positive control group is 65.43%, the survival rate of the high-dose group is 64.36%, the survival rate of the medium-dose group is 61.32%, and the survival rate of the low-dose group is 52.31%, which are all higher than the survival rate of the blank group by 50.61%, which indicates that the donkey whey protein zymolytic peptide can delay the decay of the saccharomyces cerevisiae cells to a certain extent, and probably because the zymolytic peptide regulates the activity of the intracellular antioxidase, the antioxidase is promoted to play a role and the time sequence life of the saccharomyces cerevisiae cells is delayed. Meanwhile, the enzymolysis peptide concentration and the anti-saccharomyces cerevisiae cell apoptosis effect are shown to be in a positive correlation relationship.
The activity of the yeast cells can be distinguished by staining with a Lu's alkaline Melanin staining solution, and as can be seen from FIG. 7, the cells of each group are not stained blue at day 0, which indicates that the yeast cells are active, the blue-stained cells begin to appear in each group at day 6, and the blank group is relatively more, the blue cells gradually decrease with the increase of the concentration in the sample group, and the positive control group is the least. The proportion of blue-stained cells in each group on day 12 is far more than that on day 6, but the proportion of blue-stained cells in the sample group and the positive control group is still less than that in the blank group, so that the donkey whey protein enzymolysis peptide has a certain anti-aging effect on the saccharomyces cerevisiae cells.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims (9)

1. The preparation method of the donkey milk antioxidant peptide is characterized by comprising the following steps:
carrying out enzymolysis on donkey milk whey protein by using alpha-chymotrypsin to obtain the donkey milk antioxidant peptide; the temperature of the enzymolysis is 30-60 ℃, the time of the enzymolysis is 2-8h, and the pH value of the enzymolysis is 6-10.
2. The preparation method according to claim 1, wherein the donkey milk whey protein is donkey milk whey protein solution, and the concentration of the whey protein in the donkey milk whey protein solution is 2-6%.
3. The method according to claim 1, wherein the mass ratio of the alpha-chymotrypsin to donkey milk whey protein is 2-6: 100.
4. The preparation method of claim 1, wherein the donkey milk antioxidant peptide is obtained by adjusting pH to neutral after enzymolysis, centrifuging and drying.
5. The method according to claim 4, wherein the solvent for adjusting pH is hydrochloric acid or sodium hydroxide.
6. The method of claim 4, wherein the centrifugation is freeze centrifugation; the rotating speed of the centrifugation is 4000-4500rmp, and the time of the centrifugation is 5-15 min.
7. The method of claim 4, wherein the drying is freeze-drying.
8. The donkey milk antioxidant peptide prepared by the preparation method according to any one of claims 1 to 7.
9. The use of donkey milk antioxidant peptide according to claim 8 in an antioxidant functional product.
CN202210711742.XA 2022-06-22 2022-06-22 Donkey milk antioxidant peptide and preparation method thereof Pending CN114958950A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101928742A (en) * 2009-06-23 2010-12-29 光明乳业股份有限公司 Whey protein active peptide with antioxidant activity and preparation method thereof
CN110643660A (en) * 2019-08-29 2020-01-03 北京化工大学 Method for preparing antioxidant peptide by donkey hide protease hydrolysis under assistance of ultrasound
CN112075529A (en) * 2020-09-15 2020-12-15 中国食品发酵工业研究院有限公司 Method for preparing antioxidant peptide powder by enzymolysis of whey protein powder and application of antioxidant peptide powder in milk tea powder

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101928742A (en) * 2009-06-23 2010-12-29 光明乳业股份有限公司 Whey protein active peptide with antioxidant activity and preparation method thereof
CN110643660A (en) * 2019-08-29 2020-01-03 北京化工大学 Method for preparing antioxidant peptide by donkey hide protease hydrolysis under assistance of ultrasound
CN112075529A (en) * 2020-09-15 2020-12-15 中国食品发酵工业研究院有限公司 Method for preparing antioxidant peptide powder by enzymolysis of whey protein powder and application of antioxidant peptide powder in milk tea powder

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
梁星: "驴乳清蛋白酶解肽的抗氧化活性及应用研究", 中国优秀硕士学位论文全文数据库工程科技Ⅰ辑, no. 12, pages 024 - 233 *
梁星等: "计算机模拟酶解制备驴乳清蛋白抗氧化肽的研究", 《天然产物研究与开发》, vol. 34, pages 93 - 101 *

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