CN114350732A - Egg white protein peptide with oxidation resistance and inflammation resistance - Google Patents
Egg white protein peptide with oxidation resistance and inflammation resistance Download PDFInfo
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses an egg white protein peptide with oxidation resistance and anti-inflammation, which is prepared by the following method: pre-treating; carrying out enzymolysis; membrane separation; modification treatment; and (5) freeze drying. The invention combines enzymolysis and graft modification to process egg white protein, has higher oxidation resistance compared with a single enzyme method, and further improves the processing property and the digestion property of the egg white protein peptide after modification. Meanwhile, the preparation method has simple process and low cost, and is suitable for industrial production.
Description
Technical Field
The invention relates to the field of food processing, in particular to an egg white protein peptide with oxidation resistance and inflammation resistance.
Background
The specific weights of the mass of the eggshell (containing the eggshell membrane), the egg white and the egg yolk in the eggs and the total mass of the eggs are respectively about 9.5%, 63% and 27.5%. Eggs contain mainly water (75%), protein (12%) and lipids (12%), while also containing small amounts of carbohydrates and mineral elements. Wherein the protein is mainly distributed in egg white and egg yolk, the lipid is mainly distributed in the egg yolk in the form of lipoprotein, and the trace elements are mainly distributed in egg shell. Eggs are rich in protein, balanced in nutrition, easy to digest and absorb and long considered as one of the foods with the highest nutritional value, and in recent years, researches show that egg white protein and egg white peptide also have diversified physiological activities.
The egg white contains 9.7-10.6% of protein, the protein mode of the egg white is most similar to that of a human body, the nutritive value is high, and the egg white is one of animal foods acceptable for people with different dietary habits and different religious beliefs in all countries in the world. In addition, the egg white protein structure contains a plurality of functional segments, and is a high-quality raw material for producing active peptide.
Bioactive peptides are special protein fragments, can have positive effects on the functions and the states of human bodies, and finally affect the health of the human bodies. The peptide fragments generally contain 2-20 amino acid residues, and the physiological activity of the peptide fragments is closely related to the types and sequences of the amino acid residues. Bioactive peptides are generally present in the parent protein in an inactive state, and when the parent protein is hydrolyzed or fermented, different peptide fragments with various physiological activities can be released, and the bioactive peptides can influence the physiological activities of the cardiovascular system, the endocrine system, the immune system and the nervous system of a human body and can influence the utilization rate of nutrients by the body. The peptide fragment produced by the albumen protein after being hydrolyzed by specific enzyme has a series of biological activities such as antioxidation, antihypertensive, antidiabetic, antimicrobial activity and immunological activity.
Chinese patent CN 104313099A discloses a preparation method of egg white protein peptide with high antioxidant activity, which comprises the steps of denaturing egg white protein suspension at 85-95 ℃, cooling to room temperature, hydrolyzing with alkaline protease and pepsin in sequence, inactivating enzyme, cooling, centrifuging, taking supernatant, ultrafiltering, performing gradient elution with ethanol, collecting, vacuum concentrating and freeze drying; chinese patent CN 103146791A discloses a method for hydrolyzing egg white protein by a plurality of proteases, which comprises the steps of firstly obtaining egg white, heating and denaturing, and then simultaneously or in stages hydrolyzing the egg white protein by two or more of compound protease, neutral protease, alkaline protease and flavourzyme. In the prior art, the research on egg white protein mainly focuses on in-vitro enzymolysis preparation and in-vitro oxidation resistance evaluation, and the problems of low preparation efficiency and single functional activity of the prepared egg white protein peptide still exist.
Disclosure of Invention
In order to solve the technical problems, the invention provides an egg white protein peptide with oxidation resistance and inflammation resistance, and the technical scheme is as follows:
an egg white protein peptide with oxidation resistance and anti-inflammation is prepared by the following method:
s1 preprocessing;
s2 enzymolysis;
s3 membrane separation;
s4 modification treatment;
s5 freeze drying.
Further, the egg white protein peptide with oxidation resistance and anti-inflammatory is prepared by adopting the following method:
s1 pretreatment: taking 400-500 g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 6-8 wt%, heating and denaturing at 90-95 ℃ for 30-40 min, and homogenizing the egg white suspension at 10000-11000 rpm for 2-3 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.05-0.1 mol/L sodium hydroxide aqueous solution, heating to 48-52 ℃, adding alkaline protease for enzymolysis, inactivating the enzyme at 90-95 ℃ for 10-15 min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting the pH of the solution after the enzyme deactivation in the step S2 to 7.0 by using 0.1-0.2 mol/L hydrochloric acid aqueous solution, then carrying out centrifugation at 6000-8000 r/min at 2-4 ℃ for 10-15 min, taking supernate and using 1 x 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: adding fructose into the filtrate obtained in the step S3, adjusting the pH value to 8-8.5 by using 0.05-0.1 mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55-60 ℃, stirring at the rotating speed of 150-300 rpm for 3-4 h, and performing rotary evaporation and concentration on the solution at the temperature of 40-45 ℃ to obtain 1/8-1/5 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 to-18 ℃ for 10 to 12 hours, and freeze-drying at-40 to-30 ℃ for 14 to 18 hours to obtain the egg white protein peptide with the functions of resisting oxidation and inflammation.
Further, the amount of alkaline protease added in step S2 is 2.6-3 wt%.
Further, the enzymolysis time in the step S2 is 6-8 h.
Further, the mass ratio of the filtrate to the fructose in the step S4 is (10-15): 1.
the invention carries out enzymolysis treatment on an egg white suspension by using alkaline protease to obtain egg white protein peptide, and then carries out modification treatment by utilizing the combination of fructose and peptide to glycosylate the egg white protein peptide, compared with the egg white protein peptide obtained by single enzyme method treatment, the egg white protein peptide has higher inoxidizability, and the processing property and the digestion property of the egg white protein peptide are further improved after modification, and the invention is probably related to introducing more hydroxyl groups after the egg white protein peptide is treated by combining grafting modification with the enzyme method to enhance the acting force between molecules.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The operations referred to in the examples are, unless otherwise specified, all those of ordinary skill in the art.
Some raw material parameters in the comparative examples and examples of the invention are as follows:
egg white protein powder purchased from western Ann Shown Biotech limited;
alkaline protease with enzyme activity of 100000U/g is purchased from Oboxing biotechnology limited of Beijing.
Example 1
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 2.6 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: and (4) taking the filtrate obtained in the step S3, and adding fructose into the filtrate, wherein the mass ratio of the filtrate to the fructose is 15: 1, adjusting the pH value to 8 by using a 0.1mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55 ℃, stirring for 3 hours at the rotating speed of 200rpm, and performing rotary evaporation and concentration on the solution at 45 ℃ until the volume of the solution is 1/8 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Example 2
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 2.7 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: and (4) taking the filtrate obtained in the step S3, and adding fructose into the filtrate, wherein the mass ratio of the filtrate to the fructose is 14: 1, adjusting the pH value to 8 by using a 0.1mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55 ℃, stirring for 3 hours at the rotating speed of 200rpm, and performing rotary evaporation and concentration on the solution at 45 ℃ until the volume of the solution is 1/8 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Example 3
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 2.8 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: and (4) taking the filtrate obtained in the step S3, and adding fructose into the filtrate, wherein the mass ratio of the filtrate to the fructose is 13: 1, adjusting the pH value to 8 by using a 0.1mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55 ℃, stirring for 3 hours at the rotating speed of 200rpm, and performing rotary evaporation and concentration on the solution at 45 ℃ until the volume of the solution is 1/8 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Example 4
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 2.9 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: and (4) taking the filtrate obtained in the step S3, and adding fructose into the filtrate, wherein the mass ratio of the filtrate to the fructose is 12: 1, adjusting the pH value to 8 by using a 0.1mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55 ℃, stirring for 3 hours at the rotating speed of 200rpm, and performing rotary evaporation and concentration on the solution at 45 ℃ until the volume of the solution is 1/8 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Example 5
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 3.0 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 modification treatment: and (4) taking the filtrate obtained in the step S3, and adding fructose into the filtrate, wherein the mass ratio of the filtrate to the fructose is 11: 1, adjusting the pH value to 8 by using a 0.1mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55 ℃, stirring for 3 hours at the rotating speed of 200rpm, and performing rotary evaporation and concentration on the solution at 45 ℃ until the volume of the solution is 1/8 of the volume of the original solution;
s5 freeze-drying: and S4, cooling the concentrated solution, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Comparative example 1
The preparation method of the egg white protein peptide with the functions of resisting oxidation and resisting inflammation comprises the following steps:
s1 pretreatment: taking 500g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 8 wt%, heating and denaturing at 95 ℃ for 30min, and homogenizing the egg white suspension at 11000rpm for 2 min;
s2 enzymolysis: adjusting the pH value of the pretreated homogeneous liquid obtained in the step S1 to 10.5 by using 0.1mol/L sodium hydroxide aqueous solution, heating to 50 ℃, adding alkaline protease for enzymolysis for 6 hours, wherein the addition of the alkaline protease is 2.6 wt%, inactivating the enzyme at 90 ℃ for 15min after the enzymolysis is finished, and cooling to room temperature;
s3 membrane separation: adjusting pH of the solution obtained in step S2 to 7.0 with 0.1mol/L hydrochloric acid aqueous solution, centrifuging at 4 deg.C for 15min at 6000r/min, collecting supernatant, and purifying with 1 × 104Ultrafiltration is carried out on an ultrafiltration membrane of Da;
s4 freeze-drying: and S3, cooling the filtrate, pre-freezing at-20 deg.C for 12h, and freeze-drying at-40 deg.C for 18h to obtain the antioxidant and anti-inflammatory egg white protein peptide.
Test example
Anti-inflammatory testing: the test method comprises the following steps: RAW264.7 cells were placed at 37 ℃ and CO2The cells were cultured in DMEM cell culture medium under the environment, and then divided into LPS model group (i.e. model injury group) (1. mu.g/mL LPS action for 24h), and egg albumin peptide addition group, each of which was added with LPS. And (3) detecting the concentration of the proinflammatory factor TNF-alpha according to a kit method. The method mainly comprises the following steps: diluting the cell supernatant to the detection range of an Elisa kit, adding the diluted cell supernatant into a 96-well plate coated with an antibody for incubation, respectively adding the antibody of a target detection object and a color development solution TMB, adding a reaction stopping solution, detecting the reading of the reaction stopping solution by using a microplate reader, drawing a standard curve according to a standard provided by the kit, and calculating the concentration of proinflammatory factors in the supernatant.
DPPH free radical clearance test: dissolving 1g egg white protein peptide in 50mL water, taking 3mL of the same volume of the solution to be detected and 2X 10﹣4mixing the solution of DPPH in mol/L (A1 tube); taking equal volume of absolute ethyl alcohol and 2 multiplied by 10﹣4mixing the solution of DPPH in mol/L (A2 tube); taking absolute ethyl alcohol with the same volume and uniformly mixing the absolute ethyl alcohol with a solution to be detected (A3 tube); after 30min of dark reaction at 30 ℃, the absorbance values of tubes A1, A2 and A3 were measured at 517nm using distilled water as a blank and recorded as ODA1, ODA2 and ODA3, respectively. DPPH radical clearance was calculated according to the following formula.
TABLE 1 anti-inflammatory test results table for egg white protein peptide
In the process of occurrence and development of diseases of the body, inflammation at different parts has an important promoting effect. When tissues or an immune system are inflamed, immune related cells can generate a large number of signal molecules to enable the immune system to make corresponding response, the signal molecules comprise various proinflammatory factors such as TNF-alpha and the like, the anti-inflammatory activity of active substances can be evaluated by measuring the expression level of the TNF-alpha, and as can be seen from the table, the egg white protein peptide prepared by the method has certain anti-inflammatory activity, and the anti-inflammatory activity of the egg white protein peptide is enhanced along with the increase of the enzyme dosage.
The stronger the DPPH free radical clearance rate is, the better the oxidation resistance of the albumen peptide is, and the DPPH free radical clearance rate of the albumen peptide in the example 5 is more than 90%, which shows that the enzyme method combined with the modification treatment can effectively improve the oxidation resistance of the albumen peptide, and the oxidation resistance of the albumen peptide is changed along with the change of the ratio of the albumen peptide to the fructose.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. The egg white protein peptide with the functions of resisting oxidation and resisting inflammation is characterized by being prepared by adopting the following method:
s1 preprocessing;
s2 enzymolysis;
s3 membrane separation;
s4 modification treatment;
s5 freeze drying.
2. The egg white protein peptide with antioxidant and anti-inflammatory functions as claimed in claim 1, wherein the pretreatment step comprises: taking 400-500 g of egg white protein powder raw material, adding water to prepare an egg white suspension with the concentration of 6-8 wt%, heating and denaturing at 90-95 ℃ for 30-40 min, and homogenizing the egg white suspension at 10000-11000 rpm for 2-3 min.
3. The egg white protein peptide with antioxidant and anti-inflammatory effects as claimed in claim 1, wherein the enzymolysis step is as follows: and (4) adjusting the pH value of the pretreated homogeneous liquid obtained in the step (S1) to 10-10.5 by using 0.05-0.1 mol/L sodium hydroxide aqueous solution, heating to 48-52 ℃, adding alkaline protease for enzymolysis, inactivating the enzyme at 90-95 ℃ for 10-15 min after the enzymolysis is finished, and cooling to room temperature.
4. The egg white protein peptide with antioxidant and anti-inflammatory effects as claimed in claim 1, wherein the membrane separation step comprises: adjusting the pH of the solution after enzyme deactivation to 7.0 by using 0.1-0.2 mol/L hydrochloric acid aqueous solution, then centrifuging at 6000-8000 r/min at 2-4 ℃ for 10-15 min, and taking supernatant for ultrafiltration by using an ultrafiltration membrane.
5. The egg white protein peptide with oxidation resistance and anti-inflammatory property as claimed in claim 1, wherein the modification treatment step comprises: and (4) adding fructose into the filtrate obtained in the step (S3), adjusting the pH value to 8-8.5 by using 0.05-0.1 mol/L sodium hydroxide aqueous solution, heating the mixed solution to 55-60 ℃, stirring at the rotating speed of 150-300 rpm for 3-4 h, and performing rotary evaporation and concentration on the solution at the temperature of 40-45 ℃ to obtain 1/8-1/5 of the volume of the original solution.
6. The egg white protein peptide with antioxidant and anti-inflammatory effects as claimed in claim 1, wherein the freeze drying step comprises: and S4, cooling the concentrated solution, pre-freezing at-20 to-18 ℃ for 10 to 12 hours, and freeze-drying at-40 to-30 ℃ for 14 to 18 hours to obtain the egg white protein peptide with the functions of resisting oxidation and inflammation.
7. The antioxidant and anti-inflammatory egg white protein peptide as claimed in claim 3, wherein the peptide comprises the following components: the addition amount of the alkaline protease in the enzymolysis process is 2.6-3 wt%.
8. The antioxidant and anti-inflammatory egg white protein peptide as claimed in claim 3, wherein the peptide comprises the following components: and the enzymolysis time in the enzymolysis process is 6-8 h.
9. The antioxidant and anti-inflammatory egg white protein peptide as claimed in claim 4, wherein the peptide comprises the following components: the pore diameter of the ultrafiltration membrane is 1 multiplied by 104Da。
10. The antioxidant and anti-inflammatory egg white protein peptide as claimed in claim 5, wherein: the mass ratio of the filtrate to the fructose is (10-15): 1.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586376A (en) * | 2012-02-24 | 2012-07-18 | 大连工业大学 | Albumen antioxidant peptide and preparation method thereof |
| CN102628073A (en) * | 2012-03-28 | 2012-08-08 | 东北农业大学 | Glycosylated lactalbumin polypeptide with antioxidant activity as well as preparation method and application of glycosylated lactalbumin polypeptide |
| CN105594786A (en) * | 2015-12-19 | 2016-05-25 | 仇颖超 | Preparation method of egg white protein powder bread additive |
| CN108651690A (en) * | 2018-04-25 | 2018-10-16 | 河南科技大学 | A kind of process of enzymolysis and the functional egg-white powder of glycosylation modified synergic preparation |
| CN110115347A (en) * | 2019-05-13 | 2019-08-13 | 扬州大学 | A method of the modified preparation egg white powder with high foamability of enzymatic hydrolysis cooperative fermentation |
| US20210115488A1 (en) * | 2019-02-28 | 2021-04-22 | Jiangnan University | Preparation Method and Application Thereof of Peptides with Anti-lipid-oxidation Capability |
-
2021
- 2021-12-02 CN CN202111457044.3A patent/CN114350732B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586376A (en) * | 2012-02-24 | 2012-07-18 | 大连工业大学 | Albumen antioxidant peptide and preparation method thereof |
| CN102628073A (en) * | 2012-03-28 | 2012-08-08 | 东北农业大学 | Glycosylated lactalbumin polypeptide with antioxidant activity as well as preparation method and application of glycosylated lactalbumin polypeptide |
| CN105594786A (en) * | 2015-12-19 | 2016-05-25 | 仇颖超 | Preparation method of egg white protein powder bread additive |
| CN108651690A (en) * | 2018-04-25 | 2018-10-16 | 河南科技大学 | A kind of process of enzymolysis and the functional egg-white powder of glycosylation modified synergic preparation |
| US20210115488A1 (en) * | 2019-02-28 | 2021-04-22 | Jiangnan University | Preparation Method and Application Thereof of Peptides with Anti-lipid-oxidation Capability |
| CN110115347A (en) * | 2019-05-13 | 2019-08-13 | 扬州大学 | A method of the modified preparation egg white powder with high foamability of enzymatic hydrolysis cooperative fermentation |
Non-Patent Citations (3)
| Title |
|---|
| LIXIAN MOU 等: "Comparison of bovine serum albumin glycation by ribose and fructose in vitro and in vivo", BIOCHIM BIOPHYS ACTA MOL BASIS DIS ., vol. 1868, no. 1, pages 166283, XP086875074, DOI: 10.1016/j.bbadis.2021.166283 * |
| 于滨 等: "糖基化改善蛋清蛋白功能性的研究", 中国家禽, vol. 31, no. 7, pages 15 - 18 * |
| 吴烨婷 等: "蛋清酶解多肽的制备及其清除自由基活性", 食品工业科技, vol. 39, no. 11, pages 90 - 98 * |
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Application publication date: 20220415 Assignee: Hangzhou Haihuan Nutritional Food Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000936 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Chendaokou E-commerce Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000935 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Baibeiyou Nutritional Food Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000934 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Guangzhou Hangyi Trading Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000933 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Baibeiyou E-commerce Technology Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000932 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Ailiwei Trading Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000931 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Shutikang Biotechnology Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000943 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Guangyoujiu Enterprise Management Partnership (L.P.) Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000942 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Guangzhou Xuyang Trading Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000941 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Ximazhai Nutritional Food Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000940 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Shimengze Biotechnology Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000939 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Shanlibai Trading Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000938 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 Application publication date: 20220415 Assignee: Hangzhou Liangmeiyan Trading Co.,Ltd. Assignor: Hangzhou baibeiyou Biotechnology Co.,Ltd. Contract record no.: X2023330000937 Denomination of invention: An egg white protein peptide with antioxidant and anti-inflammatory properties Granted publication date: 20230905 License type: Common License Record date: 20231221 |