Method for extracting ovotransferrin from egg white

An extraction method and ferritin technology, which are applied in the field of ultrafiltration technology to extract and purify ovotransferrin from egg white, can solve the problems of complex protein types, laboratory scale, and few functional proteins, and achieve good application prospects and products. The effect of high recovery rate and simple operation

Inactive Publication Date: 2012-07-18
CHINA UNIV OF PETROLEUM (EAST CHINA)
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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 ar

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1, the method for extracting ovotransferrin from egg white, steps:

[0014] (1) Take 200 mL of fresh egg white, add an equal volume of 100 mM Tris-HCl buffer solution, use 1 M hydrochloric acid to adjust the pH of the solution to 4.0, stir at 4°C for 1 hour, centrifuge at 2000 g, and take about 368 mL of the supernatant;

[0015] (2) Using 100 mM Tris-HCl buffer solution (pH 4.0), dilute the supernatant (368 mL) obtained in step (1) to 6.0 L, and use it as a crude enzyme solution for ovotransferrin;

[0016] (3) Utilize the polyethersulfone ultrafiltration membrane pack (0.09m 2 ), the 6.0L ovotransferrin crude enzyme solution obtained in step (2) was concentrated and separated until the retentate volume was 200mL. The operating pressure of the ultrafiltration membrane inlet and outlet are: 0.03MPa and 0.07MPa respectively, and the flow rate is: 3.8L / h;

[0017] (4) After lyophilizing the 200 mL retentate obtained in step (3), about 0.76 g of ovotransferrin...

Embodiment 2

[0018] Embodiment 2, the method for extracting ovotransferrin from egg white, steps:

[0019] (1) Take 300 mL of fresh egg white, add an equal volume of 10 mM Tris-HCl buffer solution, use 1 M hydrochloric acid to adjust the pH value of the solution to 4.8, stir at 4 ° C for 5 hours, centrifuge at 3000 g, and take about 526 mL of the supernatant;

[0020] (2) Using 100 mM Tris-HCl buffer solution (pH 4.8), dilute the supernatant (526 mL) obtained in step (1) to 10.5 L, and use it as a crude enzyme solution for ovotransferrin;

[0021] (3) Utilize the polyethersulfone ultrafiltration membrane pack (0.09m 2 ×2), the 10.5L ovotransferrin crude enzyme solution obtained in step (2) was concentrated and separated until the volume of the retentate was 260mL. The operating pressures of the ultrafiltration membrane inlet and outlet are: 0.03MPa and 0.07MPa respectively, and the flow rate is: 5.6L / h;

[0022] (4) After lyophilizing 260 mL of the retentate obtained in step (3), about 1...

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Abstract

The invention provides a method for extracting ovotransferrin from egg white. The method is characterized by comprising the steps of: directly extracting the ovotransferrin from ovotransferrin crude enzyme liquid by using an ultra-filtration membrane of which the molecular weight cut off is 70kDa, wherein the ovotransferrin crude enzyme liquid is obtained by diluting the egg white, regulating the pH value, stirring, centrifuging, retaining the supernatant and further diluting. The method has the characteristics of simple process, short extraction cycle, easiness in procedure amplification and the like, and has good application prospect.

Description

technical field [0001] The invention belongs to the field of biological separation engineering and technology, in particular to a method for extracting and purifying ovotransferrin from egg white by using ultrafiltration technology. Background technique [0002] Ovotransferrin is an acidic glycoprotein, which accounts for about 12-13% of the total protein content of egg whites. It is also an iron-binding protein and plays an important role in enhancing the body's immunity (Biochemical Cell Biology 2007, 85: 150-155). It plays an important role in antibacterial, antiviral and antioxidative aspects (Biochimica et Biophysica Acta-General subjects 2006, 1760: 347-355; International Journal of Biological Macromolecules 2007, 41: 631-640). Based on its multiple functions, people continue to explore various extraction methods. Since Fraenkel-Conrat and Feeney first isolated ovotransferrin from egg whites by ammonium sulfate precipitation in 1950 (Archives of Biochemistry 1950, 29:...

Claims

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Application Information

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IPC IPC(8): C07K14/79C07K1/34
Inventor 刘建国曲剑波肖梓军吕建仁崔占峰
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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