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Method for preparation of antioxidant polypeptide by enzymatic hydrolysis of eggshell membrane

A technology of antioxidant peptides and eggshell membranes, which is applied in the preparation method of peptides, chemical instruments and methods, peptides, etc., can solve the problems of low antioxidant capacity of antioxidant peptides, time-consuming enzymatic hydrolysis, toxic substances, etc., and achieve the realization of Waste utilization, high antioxidant capacity, high safety effect

Inactive Publication Date: 2018-10-26
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 1) The conventional enzymatic hydrolysis of eggshell membranes is very time-consuming, and usually requires at least 12 hours of enzymatic hydrolysis after high-temperature pretreatment with lye.
[0009] 2) The raw materials used in the prior art hydrolysis method are toxic substances;
[0010] 3) The total antioxidant capacity of the antioxidant polypeptide prepared by the existing method is low

Method used

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  • Method for preparation of antioxidant polypeptide by enzymatic hydrolysis of eggshell membrane
  • Method for preparation of antioxidant polypeptide by enzymatic hydrolysis of eggshell membrane
  • Method for preparation of antioxidant polypeptide by enzymatic hydrolysis of eggshell membrane

Examples

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Embodiment 1

[0093] The method for preparing antioxidant polypeptide 1 by enzymatic hydrolysis of eggshell membranes comprises the following steps:

[0094] 1) Weigh 50 g of eggshell membrane powder that has been cleaned and crushed to a size below 100 mesh, add poultry eggshell membrane powder to 1.0 kg of deionized water and mix evenly to obtain an aqueous membrane powder solution;

[0095] 2) Add 5.040 g of sodium sulfite to the membrane powder aqueous solution and mix uniformly to obtain a mixture; in the mixture, the reducing agent concentration is 40M.

[0096] 3) Add 14U / mg alkaline protease (enzyme / shell film powder quality) to the mixture, and enzymolyze it for 4.5h at a temperature of 55°C, a microwave power of 400W and a pH of 8.5 to obtain an enzymolyzed solution;

[0097] 4) Put the enzymolysis solution in hot water at 85°C for 15 minutes to inactivate the enzyme, then cool it down rapidly, centrifuge at a speed of 8000r / min for 10 minutes, collect the supernatant, and dialyze...

Embodiment 2

[0099] The method for preparing antioxidant polypeptide 2 by enzymatic hydrolysis of eggshell membranes comprises the following steps:

[0100] 1) Weigh 50 g of eggshell membrane powder that has been cleaned and crushed to a size below 100 mesh, add poultry eggshell membrane powder to 1.0 kg of deionized water and mix evenly to obtain an aqueous membrane powder solution;

[0101] 2) Add 5.040 g of sodium sulfite to the membrane powder aqueous solution and mix uniformly to obtain a mixture; in the mixture, the reducing agent concentration is 40 mM.

[0102] 3) Add 12U / mg alkaline protease (enzyme / shell film powder quality) to the mixture, and enzymolyze it for 4.0h at a temperature of 55°C, a microwave power of 200W, and a pH of 8.5 to obtain an enzymolyzed solution;

[0103] 4) Put the enzymolysis solution in hot water at 85°C for 15 minutes to inactivate the enzyme, then cool it down rapidly, centrifuge at a speed of 8000r / min for 10 minutes, collect the supernatant, and dial...

Embodiment 3

[0105] A method for preparing an antioxidant polypeptide 3 by enzymatic hydrolysis of eggshell membranes, comprising the following steps:

[0106] 1) Weigh 50 g of eggshell membrane powder that has been cleaned and crushed to a size below 100 mesh, add poultry eggshell membrane powder to 0.75 kg of deionized water and mix evenly to obtain an aqueous membrane powder solution;

[0107] 2) Add 5.040 g of sodium sulfite to the membrane powder aqueous solution and mix uniformly to obtain a mixture; in the mixture, the reducing agent concentration is 40 mM.

[0108] 3) Add 15U / mg papain (enzyme / shell film powder quality) to the mixture, and enzymolyze it for 3.0h at a temperature of 60°C, a microwave power of 600W, and a pH of 7.5 to obtain an enzymolyzed solution;

[0109] 4) Put the enzymolysis solution in hot water at 95°C to inactivate the enzyme for 10 minutes, then cool it down rapidly, centrifuge at a speed of 10,000 r / min for 5 minutes, collect the supernatant, and dialyze i...

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Abstract

The invention discloses a method for preparation of antioxidant polypeptide by enzymatic hydrolysis of eggshell membrane. The method includes the steps that first, the eggshell membrane powder is added into water to be evenly mixed and the aqueous solution of the membrane powder is obtained; the reductant is added to the aqueous solution of the membrane powder, evenly mixed and the mixture is obtained; then protease is added to the mixture and the hydrolysate is obtained; finally, the enzyme hydrolysate is treated in hot water at a temperature of 80-95 DEG C for 10-30min, then rapidly cooled and centrifuged for 5-20min at the speed of 8,000-10,000r / min, and the supernatant is collected, dialyzed against a bag of dialysis, and dried, an antioxidant polypeptide is obtain. The method is basedon searching for natural high-efficiency anti-oxidants, taking full advantage of the discarded eggshell membrane, using sodium sulfite to open the stubborn disulfide bond in the eggshell membrane, and the microwave assisted shortening the time of enzymatic hydrolysis, and the sodium sulfite used is low in toxicity and easy to remove. By controlling the cutting condition of alkaline protease, antioxidant activity can be efficiently achieved.

Description

technical field [0001] The invention relates to the technical field of poultry egg processing and utilization, in particular to a method for preparing antioxidant polypeptides by enzymatic hydrolysis of eggshell membranes. Background technique [0002] Oxidation is an important metabolic process for aerobic organisms, especially vertebrates and humans, but at the same time leads to the formation of free radicals. Free radicals are highly unstable atoms, molecules, and ions with unpaired electrons that are prone to chemical reactions with other molecules. Active oxygen free radicals can cause oxidative stress. Oxidative stress is involved in various human diseases such as cancer, atherosclerosis, diabetes, neurological diseases, etc. In food, oxygen free radicals can attack lipids or proteins resulting in unpleasant smell, dark color or poor texture. The application of antioxidants can protect the human body from the damage of oxidative stress and prevent the oxidative dete...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/14
CPCC07K1/14C12P21/06
Inventor 黄茜赵婕媛马美湖耿放李晓云
Owner HUAZHONG AGRI UNIV
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