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Method for preparing corn superoxide dismutase from corn

A superoxide and dismutase technology, applied in biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of high cost, low purity of SOD products, and difficulty in reaching pharmaceutical grades, and achieve the effect of improving product purity.

Inactive Publication Date: 2012-07-04
李作红
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extraction from animal blood is not only expensive, but also the exclusivity of animal SOD, it is not easy to store at room temperature
Most of the superoxide dismutase extracted from corn is purified by ultrafiltration, which cannot effectively remove impurity proteins. The extracted SOD products have low purity and low active protein content, which is difficult to meet the requirements of pharmaceutical grade.

Method used

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  • Method for preparing corn superoxide dismutase from corn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation method of corn superoxide dismutase, concrete steps are:

[0027] (1) Selection, through winnowing, select high-quality corn with no mildew, no impurities, and full bracts.

[0028] (2) Disinfection soaking: wash 3-4 times with 1% sodium chloride solution, carry out sterilization and disinfection, soak for 25 hours with deionized water at 25°C-28°C, and drain;

[0029] (3) Germination: Control the temperature of the germination room at 25°C-28°C, water the corn every 2 hours, keep the water temperature at 25°C-28°C, check the germination situation at any time, wait for the germination to grow 1-2 mm, and germinate Finish;

[0030] (4) Cleaning: the germinated corn is cleaned 2-3 times with a 2% sodium chloride solution;

[0031] (5) Refining: Add 2 L / kg of germinated corn to 50 μmol / L of phosphate buffer solution for grinding, first rough grinding, then fine grinding, fully grinding;

[0032] (6) One-time enzymatic hydrolysis: add cellulase to the gro...

Embodiment 2

[0038] The preparation method of corn superoxide dismutase, concrete steps are:

[0039] (1) Selection, through winnowing, select high-quality corn with no mildew, no impurities, and full bracts.

[0040] (2) disinfection soaking: wash 4-5 times with the sodium chloride solution of concentration 2%, carry out sterilization, soak 26-28 hours with the deionized water of 28 ℃-30 ℃, drain;

[0041] (3) Germination: Control the temperature of the germination room at 28°C-30°C, water the corn every 2 hours, keep the water temperature at 28°C-30°C, and check the germination situation at any time. Finish;

[0042] (4) cleaning: the germinated corn is cleaned 2-3 times with a concentration of 3% sodium chloride solution;

[0043] (5) Refining: adding 3-4L / kg of germinated corn to 50umol / L of phosphate buffer solution for grinding, first coarse grinding, then fine grinding, and fully grinding;

[0044] (6) One-time enzymatic hydrolysis: add cellulase to the ground corn syrup at 0.2g / ...

Embodiment 3

[0050] The preparation method of corn superoxide dismutase, concrete steps are:

[0051](1) Selection, through winnowing, select high-quality corn with no mildew, no impurities, and full bracts.

[0052] (2) Disinfection soaking: wash 5-6 times with a sodium chloride solution with a concentration of 3%, carry out sterilization and disinfection, soak for 28-32 hours with deionized water at 30°C-33°C, and drain;

[0053] (3) Germination: Control the temperature of the germination room at 30°C-33°C, water the corn every 2 hours, keep the water temperature at 30°C-33°C, check the germination situation at any time, wait for the germination to grow 1-2 mm, and germinate Finish;

[0054] (4) Cleaning: the germinated corn was cleaned 3 times with a 2.5% sodium chloride solution;

[0055] (5) Refining: Grinding the germinated corn by adding 50umol / L of phosphate buffer solution at 5L / kg, first coarse grinding, then fine grinding, and fully grinding;

[0056] (6) One-time enzymatic h...

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Abstract

The invention relates to a method for preparing corn superoxide dismutasen, which mainly comprises the following steps of: (1) sterilization and impregnation; (2) sprouting; (3) cleaning; (4) jordaning; (5) primary enzymolysis, namely, uniformly mixing corn steep liquor obtained by the jordaning, cellulase and pectinase, and adding the solution of calcium chloride for precipitation; (6) primary separation, namely, performing prefiltration on the precipitated and demixed corn steep liquor, centrifugating the prefiltered corn steep liquor with a high-speed separator, and preserving supernate; (7) ultrafiltration and concentration, namely, performing ultrafiltration on the supernate with a hollow fiber superfiltration column to obtain filtrate, performing fine-filtration with a nanometer filter to obtain pure filtrate, and condensing the pure filtrate to a volume which is 10 percent of an original volume; (8) secondary enzymolysis; and (9) secondary separation, namely, performing the centrifugal separation again, performing the ultrafiltration to obtain ultra-pure filtrate, and performing quick-freezing to obtain the freeze-dried powder of the corn superoxide dismutase (SOD). The method has the advantages of maximally removing hybrid proteins and improving the purity of the corn SOD product by the secondary enzymolysis and the secondary separation so as to meet requirements on the medicinal grade.

Description

technical field [0001] The invention relates to a method for preparing superoxide dismutase from plants, in particular to a method for preparing superoxide dismutase from corn. Background technique [0002] SOD (superoxide dismutase) is an active substance derived from living organisms. It is an important antioxidant enzyme in organisms and is widely distributed in various organisms, such as animals, plants, and microorganisms. SOD has special physiological activity and is the primary substance for scavenging free radicals in organisms. The level of SOD in the living body means an intuitive indicator of aging and death; it has been confirmed that there are as many as 60 kinds of diseases caused by oxygen free radicals. It can resist and block the damage to cells caused by oxygen free radicals, repair damaged cells in time, and restore the damage to cells caused by free radicals. Due to the pressure of modern life, environmental pollution, various radiation and excessive ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08
Inventor 李作红徐殿英李洁
Owner 李作红