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Method for extracting and separating and purifying protein

A separation, purification and protein extraction technology, applied in the field of glycoproteins, can solve problems such as retention, slow research progress, and unrealized industrial production, and achieve the effects of fast filtration, reduced pollution, and easy membrane cleaning

Inactive Publication Date: 2016-09-21
JILIN HOUDE FOOD CO LTD
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  • Description
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Problems solved by technology

However, the research on the extraction of ovotransferrin from egg white is still at the laboratory scale, and industrial production has not been realized, and the research progress is relatively slow.

Method used

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  • Method for extracting and separating and purifying protein
  • Method for extracting and separating and purifying protein
  • Method for extracting and separating and purifying protein

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Embodiment Construction

[0034] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0035] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0036] 1. Ovomucin

[0037] 1.2 Separation and purification of ovomucin

[0038] 1.2.1 Process flow for separation and purification of ovomucin, such as figure 1 shown.

[0039] S101: After stirring the whole egg white evenly, add 5 times the volume of salt solution (0.05mol / LMgCl 2or 0.1mol / L NaCl), adjust the pH to 6.0 with 0.1mol / L HCl after homogenization, let stand overnight at 4°C, centrifuge at 15,000g for 10min (4°C), resuspend the precipitate with 0.5mol / L NaCl, and place ...

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Abstract

The invention discloses a method for extracting and separating and purifying protein. The method includes an ovomucin separating and purifying method, an ovotransferrin separating and purifying method and an egg immunoglobulin separating and purifying method. The method is characterized in that the method performs dilution before homogenization, egg white is diluted by normal saline and then homogenized, the viscosity of the egg white is lowered, the complex structure of the egg white is destroyed, and accordingly the egg white can be filtered fast, easy membrane cleaning is achieved, membrane pollution degree is lowered, and the membrane service life is prolonged; when operation pressure is 0.15MPa, a relatively-stable circulation state is achieved, and filtrate concentration polarization is inhibited to a certain degree; the method is based on a dilution method and uses polyethylene glycol and DEAE-Toyopearl 650M one-step elution ion-exchange column chromatography to perform purification, and high-yield and high-purity chicken egg yolk IgY.

Description

technical field [0001] The invention belongs to the technical field of glycoproteins, and in particular relates to a protein extraction, separation and purification method. Background technique [0002] Ovomucoid is a glycoprotein prepared from chicken egg white, which has a strong inhibitory effect on trypsin. It is often used in the study of the enzymatic properties of trypsin and the purification and preparation of trypsin by affinity chromatography. Molecular weight: 28000Da, the molecule is composed of four subunits with similar molecular weight, namely D-mannose, D-galactose, glucosamine, sialic acid; chemical stability, heat resistance: at 80 ℃, physical and chemical properties No change, resistance to organic solvents: in 50% acetone solution, it can maintain good solubility, resistance to precipitation agents: no precipitation occurs in 10% TCA solution. Isoelectric point properties, isoelectric point: pI between 3.9-4.5, state in solution: stable in a solution of ...

Claims

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Application Information

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IPC IPC(8): C07K14/465C07K16/02C07K1/34C07K1/30C07K1/18
CPCC07K14/465C07K16/02
Inventor 杨涛刘岩都韧宁于寒松
Owner JILIN HOUDE FOOD CO LTD
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