Method for separating ovalbumin from egg white

A technology for ovalbumin and egg white is applied in the field of separating ovalbumin, which can solve the problems of complex chromatography separation equipment, high separation cost, and complicated chromatography separation process, and achieves improved separation efficiency, high separation efficiency and simple process. Effect

Inactive Publication Date: 2017-12-05
HUZHOU TEACHERS COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The salting-out method combined with ion-exchange chromatography requires not only desalination treatment, but also complex chrom

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  • Method for separating ovalbumin from egg white
  • Method for separating ovalbumin from egg white

Examples

Experimental program
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Example Embodiment

[0028] Example 1

[0029] Take fresh eggs, wash, separate the egg white and egg yolk, collect 50ml of egg white stock solution; add 250ml of pH 5 acetic acid-sodium acetate dilution buffer containing 0M NaCl, mix well to obtain 300ml egg white dilution A; add 7.5ml caprylic acid, 4 Let stand for 2 hours at ℃; add 15.375g of hydrophobic charge-inducing magnetic microspheres and shake for 10 minutes to obtain a mixture of hydrophobic charge-inducing magnetic microspheres and egg white that adsorb ovalbumin; hold the microspheres with a magnet and discard the supernatant Wash the microspheres with the dilution buffer for several times to obtain the hydrophobic charge-inducing magnetic microspheres that adsorb ovalbumin; add 30.75ml of pH 3 acetic acid-sodium acetate elution buffer, shake for 5 minutes, and hold it with a magnet Microspheres, collect the supernatant to obtain an ovalbumin solution; freeze-dry to obtain a solid product of ovalbumin with a purity of >98%.

Example Embodiment

[0030] Example 2

[0031] Take fresh eggs, wash them, separate the egg whites from the yolks, collect 50ml egg white stock solution; add 250ml acetic acid-sodium acetate dilution buffer with pH 5 containing 0.5M NaCl, mix well to obtain 300ml egg white dilution A; add 7.5ml caprylic acid, Let stand at 30°C for 12 hours; add 15.375g of hydrophobic charge-inducing magnetic microspheres and shake for 60 minutes to obtain a mixture of hydrophobic charge-inducing magnetic microspheres and egg white that adsorb ovalbumin; hold the microspheres with a magnet and discard Clear and precipitate, wash the microspheres with dilution buffer for several times to obtain hydrophobic charge-inducing magnetic microspheres that adsorb ovalbumin; add 30.75ml pH4 acetic acid-sodium acetate elution buffer, shake for 10 minutes, and hold it with a magnet Microspheres, collect the supernatant to obtain an ovalbumin solution; freeze-dry to obtain a solid product of ovalbumin with a purity of >98%.

Example Embodiment

[0032] Example 3

[0033] Take fresh eggs, wash them, separate the egg whites from the yolks, collect 50ml egg white stock solution; add 250ml of 0.25M NaCl pH 5 acetic acid-sodium acetate dilution buffer, mix well to obtain 300ml egg white dilution A; add 7.5ml caprylic acid, Let stand at 8°C for 6 hours; add 15.375g of hydrophobic charge-inducing magnetic microspheres and shake for 30 minutes to obtain a mixture of hydrophobic charge-inducing magnetic microspheres and egg white that adsorb ovalbumin; hold the microspheres with a magnet and discard Clear and precipitate, wash the microspheres with dilution buffer several times to obtain hydrophobic charge-inducing magnetic microspheres that adsorb ovalbumin; add 30.75ml pH3.5 acetic acid-sodium acetate elution buffer, shake for 8 minutes, use a magnet The microspheres are sucked and the supernatant is collected to obtain an ovalbumin solution; freeze-dried to obtain a solid product of ovalbumin with a purity of >98%.

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Abstract

The invention discloses a method for separating ovalbumin from egg white. The method comprises 1) taking fresh eggs and collecting an egg white stock solution, 2) adding a dilution buffer into the solution to obtain an egg white diluent A, 3) adding octanoic acid into the egg white diluent A and carrying out standing to obtain an egg white diluent B, 4) adding hydrophobic charge-induced agnetic beads into the egg white diluent B and carrying out shaking to adsorb ovalbumin, 5) washing the beads with the dilution buffer multiple times to obtain the hydrophobic charge-induced magnetic beads adsorbing the ovalbumin, 6) adding an elution buffer into the magnetic beads, carrying out shaking and collecting an ovalbumin solution, and 7) carrying out freeze-drying to obtain an ovalbumin solid product having purity greater than 98%. The method designs a simple and fast separation process, realizes separation of ovalbumin from egg white, is characterized by directly separating ovalbumin from egg white and has the advantages of simple operation processes and high separation efficiency.

Description

technical field [0001] The invention relates to a method for separating ovalbumin from egg white, and belongs to the technical field of separation and purification of biologically active substances. Background technique [0002] my country is rich in egg resources, and eggs contain a large amount of active protein with high nutritional value. With the continuous deepening of the research on the active ingredients of eggs, the utilization of eggs has developed from the simple initial processing stage to the development of physiologically active ingredients with higher added value. The separation, extraction and functional research of active proteins such as ovalbumin, ovotransferrin and lysozyme in egg white have attracted more and more attention. [0003] Ovalbumin is one of the main active ingredients in egg white, containing 386 amino acids, with a relative molecular mass of 45KDa and an isoelectric point of 4.5. Ovalbumin is very similar to human albumin in terms of mol...

Claims

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Application Information

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IPC IPC(8): C07K14/77C07K1/22
CPCC07K14/77
Inventor 顾佳黎童红飞林志坚
Owner HUZHOU TEACHERS COLLEGE
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