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Method for extracting superoxide dismutase with high yield from corn

A technology of superoxide and dismutase, applied in oxidoreductase and other directions, can solve the problems of complex, difficult SOD, and high difficulty in industrialization, and achieve the effects of improving yield, good enzyme stability, and reducing the harm of copper ions

Inactive Publication Date: 2013-06-26
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when obtaining SOD from plants, because the tissue is more complex than animal raw materials, it is difficult to obtain high-purity SOD by conventional processes on the one hand, and on the other hand, some processes that can obtain high-purity SOD are more difficult to industrialize.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for extracting superoxide dismutase from corn, the steps are as follows:

[0021] Step 1: Take 120Kg of dry corn kernels, wash them, soak them in warm water (30°C) for 24 hours, take them out and drain them to obtain 140Kg of wet corn. The corn was refined with 180L of water to obtain 350L of corn slurry containing superoxide dismutase.

[0022] Step 2: Add 24.5 g of anhydrous copper sulfate to the above corn slurry, adjust the pH to 7-9 with ammonia water, and perform solid-liquid separation by pressure filtration to obtain 210 L of crude enzyme solution with a superoxide dismutase activity of 468 U / mL.

[0023] The third step: the superoxide dismutase crude enzyme solution obtained in the previous step is sequentially filtered through a ceramic membrane and concentrated by an ultrafiltration membrane. The ceramic membrane removes impurities with a molecular weight > 120,000, and the ultrafiltration membrane removes impurities with a molecular weight < 10,000....

Embodiment 2

[0025] A method for extracting superoxide dismutase with high yield from corn, on the basis of Example 1, the following steps are carried out.

[0026] The fourth step: add acetone to the concentrated solution obtained in the third step, separate the precipitate, and then dry the precipitate; then dissolve the dried precipitate in water, add acetone to the solution, separate the precipitate, and dry The precipitate was obtained as a crude product of superoxide dismutase, wherein the activity retention rate of superoxide dismutase was 90.3%.

Embodiment 3

[0028] A method for extracting superoxide dismutase with high yield from corn, on the basis of Example 2, for further purification, the following steps are carried out.

[0029] Step 5: Dissolve the crude product of superoxide dismutase obtained in step 4 in water, centrifuge, take the supernatant, and pass it through: macroporous resin——DEAE-cellulose——Sephadex G-75 chromatographic column , high-purity superoxide dismutase enzyme liquid can be obtained after three-stage chromatography, wherein the activity retention rate of superoxide dismutase is

[0030] 78.3%. After the above enzyme liquid is freeze-dried, the superoxide dismutase dry powder with an activity of 6058 U / mg can be obtained.

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PUM

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Abstract

The invention provides a method for extracting superoxide dismutase with a high yield from corn. The method comprises the following steps of: corn slurry-adding copper salt and separating-membrane filtering-precipitating organic liquid-drying-dissolving-conducting chromatography-freezing and drying and the like. According to the method, the copper salt is adopted to improve about 20% of activity of superoxide dismutase, and copper ion damage caused by the copper salt can be removed by means of subsequent membrane filtering as well as precipitating organic liquid for many times and drying. According to the invention, almost impurities in SOD (Superoxide Dismutase) are removed by combined steps of precipitating-drying-dissolving for multiple times by using time difference when superoxide dismutase and almost impurities (heteroprotein, nucleic acid and soluble starch) are completely dissolved in water, so that the purity is improved.

Description

technical field [0001] The invention relates to a method for extracting biologically active substances from plants, in particular to a method for extracting superoxide dismutase from corn. Background technique [0002] Superoxide dismutase (hereinafter referred to as: SOD) is a very important metalloenzyme that defends against oxidative damage in organisms. As a specific scavenger for superoxide anion free radicals, it is used in the treatment of diseases, food, cosmetics, Agriculture and other aspects have a wide range of applications. But at present the production of superoxide dismutase mainly comes from animal blood and liver. Although pure SOD can be obtained by a simple process when extracting SOD from animal blood, some viruses in the blood may inevitably be brought into the SOD product during the extraction process, resulting in cross-infection between humans and animals. Since 1996, the European Union has banned the use of blood products in the treatment of human ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02
Inventor 詹拥共梁姣高建军刘西洋
Owner HUNAN UNIV
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