Method for extracting small-molecule polypeptides in chick embryos

A technology of small molecule polypeptide and extraction method, which is applied in the field of separation and extraction of Chinese medicinal materials

Active Publication Date: 2022-04-15
贵州中医药大学第一附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above technical problems, the present invention provides a method for extracting small molecular polypeptides from chicken embryos, which solves the problem of how to efficiently extract small molecular polypeptides from chicken embryos

Method used

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  • Method for extracting small-molecule polypeptides in chick embryos
  • Method for extracting small-molecule polypeptides in chick embryos
  • Method for extracting small-molecule polypeptides in chick embryos

Examples

Experimental program
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Effect test

Embodiment 1

[0031] a. Take Polygonum multiflorum resistant starch, add 2mol / L KOH aqueous solution to dissolve, add HCl at room temperature to adjust the pH value to 6.5, add water to make the resistant starch concentration 1mg / mL, and obtain the resistant starch solution, namely product A;

[0032] b. Dry chicken embryos under reduced pressure at 60-70°C, pulverize them into fine powder, use water as solvent, the ratio of chicken embryos to water is 1:9, stir evenly, add papain, the pH value is 5, The amount of enzyme added is 5%, the temperature is 53°C, and the enzymolysis time is 1.5 hours to obtain product B;

[0033] c. Take the above-mentioned product B and add neutral protease pH 6, the amount of enzyme added is 4%, the temperature is 52°C, and the enzymatic hydrolysis time is 1.2 hours to obtain product C;

[0034] d. Take the above product C, add alkaline protease pH8.2, enzyme addition 5%, and temperature 49°C. After enzymolysis, centrifuge at 1000rpm for 5 minutes, take the su...

Embodiment 2

[0041] a. Take Polygonum multiflorum resistant starch, add 2mol / L KOH aqueous solution to dissolve, add HCl at room temperature to adjust the pH value to 6.5, add water to make the resistant starch concentration 1mg / mL, and obtain the resistant starch solution, namely product A;

[0042] b. Dry chicken embryos under reduced pressure at 60-70°C, crush them to fine powder, use water as solvent, and the ratio of chicken embryos to water is 1:5, stir evenly, add papain, the pH value is 3, The amount of enzyme added is 3%, the temperature is 40°C, and the enzymolysis time is 1.0 hour, to obtain product B;

[0043] c. Take the above product B and add neutral protease pH 5, the enzyme addition amount is 3%, the temperature is 45°C, and the enzymatic hydrolysis time is 1.0 hour, to obtain product C;

[0044] d. Take the above product C, add alkaline protease pH 7.8, enzyme addition 5.5%, temperature 52°C, centrifuge at 1000rpm for 5 minutes after enzymolysis, take the supernatant to o...

Embodiment 3

[0051] a. Take Polygonum multiflorum resistant starch, add 2mol / L KOH aqueous solution to dissolve, add HCl at room temperature to adjust the pH value to 6.5, add water to make the resistant starch concentration 1mg / mL, and obtain the resistant starch solution, namely product A;

[0052] b. Dry the chicken embryos under reduced pressure at 70°C, crush them into fine powder, use water as the solvent, the ratio of chicken embryos to water is 1:10, stir well, add papain, the pH value is 6, add enzyme The amount is 6%, the temperature is 60°C, and the enzymolysis time is 2.0 hours to obtain product B;

[0053] c. Take the above-mentioned product B and add neutral protease pH 7, the enzyme addition amount is 4%, the temperature is 55°C, and the enzymolysis time is 1.2 hours to obtain product C;

[0054] d. Take the above product C, add alkaline protease pH 8.0, enzyme addition 6%, temperature 53°C, centrifuge at 1000rpm for 5 minutes after enzymolysis, take the supernatant to obtai...

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Abstract

The invention discloses a method for extracting small-molecule polypeptides in chick embryos. According to the method, the characteristics that resistant starch is large in molecular weight, high in polymerization degree and capable of spontaneously forming a V-shaped cavity structure with a large internal volume are utilized, chicken embryo protein obtained after enzymolysis and a resistant starch solution are subjected to inclusion, ultrasonic treatment and ultrafiltration tube centrifugation, and then the small-molecule polypeptide in the chicken embryo is obtained. Compared with a crushing impregnation extraction method and an enzymolysis impregnation extraction method, the micromolecule polypeptide extraction method disclosed by the invention is higher in extraction rate. The molecular weight of the micro-molecular polypeptide extracted by the method is 11-67 kDa, the structure is simple, the purity is 89.2 + / -3.1%, the extraction yield is 67 + / -2.2%, the micro-molecular polypeptide can quickly penetrate through a biological barrier, and animal experiments show that the micro-molecular polypeptide has a remarkable anti-inflammatory effect and a better treatment effect on osteoarthritis.

Description

technical field [0001] The invention relates to the field of separation and extraction of traditional Chinese medicinal materials, in particular to a method for extracting small-molecule polypeptides from chicken embryos. Background technique [0002] Chicken embryos are dried domestic chicken embryos (English name: chick embryo or embryonated egg) of pheasant family animals. Domestic eggs are hatched at 37.9±0.5°C for 17-18 days, the shells are removed, and dried. The taste is sweet, salty, fishy, ​​and warm in nature. Returning to the spleen and stomach meridians, it has the effects of warming the middle and replenishing qi, replenishing the essence and filling the marrow. It is used for physical weakness and thinness, fractures, tendon injuries, etc. It has been included in the 2019 edition of "Quality Standards for Traditional Chinese Medicinal Materials of Guizhou Province". In addition, the main component of chicken embryos is protein, which is similar to some componen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/14
CPCY02P20/10
Inventor 唐东昕辛加敏杨柱阮婧华范东生查鑫李佳璇谢丹
Owner 贵州中医药大学第一附属医院
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