Method for extracting superoxide dismutase (SOD) from pig blood

A technology of superoxide and dismutase, applied in the field of bioengineering, can solve the problems of long cycle, high cost, and environmental pollution, and achieve the effects of short cycle, low pollution, and simple production method

Inactive Publication Date: 2012-02-01
天津宝迪农业科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of our country uses biochemical methods to extract SOD, such as gene synthesis, microbial fermentation and chemical extraction. At the same time, most of the SOD purification methods use agarose gel method and anion-cation resin exchange method. These methods generally have complex product

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Add 10% volume of 2-5% potassium oxalate solution to pig blood, centrifuge at 3000rpm for 20min to remove plasma, and collect red blood cells. Then add 5 times of 0.9% NaCl solution to wash, and then centrifuge at 4000rpm for 20min after stirring constantly. After discarding the supernatant, collect the erythrocytes and add 2 times the volume ratio of deionized water (pH = 9), and obtain hemolysis after standing in a water bath at 4°C for 10 hours; After 0.25 volume of 95% ethanol, slowly add 0.2 volume of pre-cooled chloroform, and stir for 15 min. After standing still for 0.5h, centrifuge at 4000rmp for 20min, collect the supernatant to obtain crude enzyme solution; add 3 times the volume of acetone to the crude enzyme solution, stir for 10min, centrifuge at 4000rmp for 20min, and take the precipitate; add the precipitate to 2 times volume of deionized water to dissolve, and after heat treatment at 60°C for 15min, quickly ice-bathed to below 4°C. Then, add 20 volume...

Embodiment 2

[0023] Add 10% volume of 2-5% potassium oxalate solution to pig blood, centrifuge at 3000rpm for 30min to remove plasma, and collect red blood cells. Then add 3 times of 0.9% NaCl solution to wash, and then centrifuge at 4000rpm for 20min after stirring constantly. After discarding the supernatant, collect the erythrocytes and add 3 times the volume ratio of deionized water (pH=9), let stand in a water bath at 4°C for 10 hours to obtain hemolysis; slowly add cold pre-cooled at 4°C to the hemolysis After 0.5 volume of 95% ethanol, slowly add 0.5 volume of pre-cooled chloroform, stir for 15min, let stand for 0.5h, centrifuge at 4000rmp for 30min, collect the supernatant to obtain crude enzyme solution; add 2 times to the crude enzyme solution volume of acetone, stirred for 10 minutes, centrifuged at 4000rmp for 20 minutes, and the precipitate was taken; the precipitate was dissolved by adding 2 times the volume of deionized water, and after heat treatment at 50°C for 20 minutes,...

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PUM

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Abstract

The invention discloses a method for extracting superoxide dismutase (SOD) from pig blood. The method comprises the following steps of separating blood cells from blood plasma by a separation-cleaning-disruption integrated one-step separation technology and preparing a blood solution, treating the blood solution by an ethanol-chloroform dissolution and precipitation technology to obtain a crude enzyme solution, treating the crude enzyme solution by an acetone precipitation method to obtain a SOD crude protein, heating supernate obtained by the previous step for degeneration, collecting supernate, removing precipitates, carrying out further separation and purification of the supernate by a spiral gradient elution technology to improve enzyme unit activity and obtain a purified SOD enzyme solution, filtering the purified SOD enzyme solution by a membrane for desalination, and carrying out freeze drying to obtain a purified SOD product. The method simplifies production technology and production equipment, and satisfies requirements of large-scale industrialized production.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering, in particular to a method for extracting superoxide dismutase SOD from pig blood. Background technique: [0002] Superoxide dismutase (SOD) is a metalloenzyme that can catalyze the dismutation reaction of superoxide anion radicals. Its main function is to specifically remove superoxide anion radicals generated in the process of biological oxidation. The key line of defense against oxygen toxicity can prevent or treat various diseases caused by superoxide anion free radicals. It has therapeutic effect, anti-aging, anti-radiation and preventing cell mutation, etc. It has the characteristics of being able to remove excess oxygen free radicals in the body, so it has been widely used in medicine, food, cosmetics and other fields. [0003] Since SOD was first isolated from bovine red blood cells in 1969, people have successively isolated and purified SOD from red blood cells, muscle, liver tis...

Claims

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Application Information

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IPC IPC(8): C12N9/08
Inventor 宋古全卢进峰吴明文王雅静
Owner 天津宝迪农业科技股份有限公司
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