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Method for extracting water-soluble protein component of yolk

An extraction method and protein technology, which can be used in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., and can solve problems such as few functional proteins, laboratory scale, and complex protein types.

Inactive Publication Date: 2009-04-08
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the field of bioprocessing, ultrafiltration technology is often used for protein purification and concentration, and has been widely used in industry. There are very few examples of isolated functional proteins of high purity, and research in this area is still at the laboratory scale

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. A method for extracting water-soluble protein components of yolk from yellow-skinned eggs, the steps are:

[0021] 1. Take eight fresh yellow eggs, wash the eggshells with tap water, and dry them with paper. After removing the eggshells and egg whites, gently puncture the yolk membrane with a syringe needle to collect the yolk liquid, about 100mL;

[0022] 2. Use deionized water to dilute the egg yolk solution obtained in the above steps to 700 mL, put in a stir bar, stir well for 20 minutes, freeze at -20°C for 8 hours, and then thaw;

[0023] 3. Take 400 mL of the above thawing solution, centrifuge at 3000 rpm at 4°C, discard the precipitate, and take the supernatant, about 350 mL;

[0024] 4. Use a commercially available polysulfone membrane with a molecular weight cut-off of 30kDa to perform ultrafiltration separation on the supernatant obtained in step 3, the separation conditions are: 4°C, pH 6.0, and the filtrate is a yolk phosphoprotein solution with a pu...

Embodiment 2

[0029] Example 2. A method for extracting water-soluble protein components of egg yolk from white-skinned eggs, the steps are as follows:

[0030] 1. Take six fresh white-skinned eggs, wash the egg shells with tap water, and dry them with paper. After removing the egg shells and egg whites, gently puncture the yolk membrane with a syringe needle to collect the yolk liquid, about 76 mL;

[0031] 2. Use deionized water to dilute the egg yolk solution obtained in the above steps to 760 mL, put it in a stir bar, fully stir for 30 minutes, freeze at -20°C for 12 hours, and then thaw;

[0032] 3. Take 420 mL of the above thawing solution, centrifuge at 6000 rpm at 4°C, discard the precipitate, and take the supernatant, about 360 mL;

[0033] 4. Use a commercially available polysulfone membrane with a molecular weight cut-off of 100kDa to perform ultrafiltration separation on the supernatant obtained in step 3, and the separation conditions are: 4°C, pH 8.0, and the filtrate is a yolk ph...

Embodiment 3

[0038] Embodiment 3, the method for extracting water-soluble protein components of egg yolk from eggs, the steps are:

[0039]1. Take ten fresh eggs, wash the eggshells with tap water, and dry them with paper. After removing the eggshells and egg whites, gently puncture the yolk membrane with a syringe needle to collect the yolk liquid, about 122mL;

[0040] 2. Use a pH5.0, 200mM phosphate buffer solution to dilute the egg yolk solution obtained in the above steps to 1000mL, put it in a stir bar, fully stir for 25 minutes, freeze at -20°C for 10 hours, and then thaw;

[0041] 3. Take 600 mL of the above thawing solution, centrifuge at 5000 rpm at 4°C, discard the precipitate, and take the supernatant, about 515 mL;

[0042] 4. Use a commercially available polysulfone membrane with a molecular weight cut-off of 100kDa to perform ultrafiltration separation on the supernatant obtained in step 3. The separation conditions are: 4°C, pH 7.0, and the filtrate is a yolk phosphoprotein sol...

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Abstract

The invention provides a method for extracting water soluble protein of yolk, pertaining to the technical field of bioseparation engineering. The method is realized mainly by the application of ultrafiltration separation technology, and four types of water soluble protein of yolk with high purity, which include immunoglobulin of yolk, vitellin, phosvitin and unknown protein with the molecular weight of 50kDa, and can be extracted from the yolk through five simple steps of dilution, freeze thawing, centrifugation, filtration and ultrafiltration. During the extracting process, any organic solvent is not used, thus effectively avoiding the protein denaturation caused by the organic solvent and precluding hidden danger caused by solvent residues on product security.

Description

Technical field [0001] The invention belongs to the field of biological separation engineering and technology, and specifically is a method for separating and extracting four water-soluble protein components from chicken egg yolk by applying ultrafiltration technology. The purity of the products is all over 85%, and the purity of some products is higher than 95%. Background technique [0002] Chicken egg yolk contains 119 kinds of protein, accounting for 17.8% of the total mass. Among them, most of them are combined with fat to form lipoproteins, while the water-soluble proteins are relatively few. Common types include yolk immunoglobulin, yolk protein, yolk phosphoprotein, and yolk phosphoprotein. [0003] Yolk immunoglobulin refers to the main immunoglobulin present in the yolk of chickens. It is 7 to 10 days after the hen is immunized, that is, the yolk maturity period, which is selectively transferred from the immunoglobulin IgG in the hen's blood to the yolk. Formed in. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/34
Inventor 刘建国杨娟王晶冰
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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