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Method for extracting superoxide dismutase (SOD) from porcine blood red cells

A technology for red blood cells and superoxide, which is applied in the field of superoxide dismutase extraction, can solve the problems of unguaranteed residual safety of reagents, unfavorable large-scale production, reduction of SOD yield, etc., so as to avoid chromatographic column production. Lower rate, shorten production cycle, reduce the effect of co-precipitation

Inactive Publication Date: 2011-09-14
HEILONGJIANG BAODI MEAT FOOD
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Problems solved by technology

Most of the miscellaneous protein denaturation in the later stage adopts chloroform and ethanol method to remove hemoglobin, then the supernatant is precipitated with acetone, and the chromatographic column (Sephadex G-100 or DE-32 or metal chelation chromatography) on the precipitate, its shortcoming is: (1 ) The cost of removing hemoglobin with chloroform and ethanol is higher, while chloroform is still a toxic reagent, and the safety of reagent residues is not guaranteed; (2) in the process of removing hemoglobin with chloroform-ethanol method, due to the coprecipitation of SOD and hemoglobin, SOD The yield is reduced; (3) due to the limitation of the chromatographic column volume, the purification capacity is small, and the equipment cost is high, which is not conducive to large-scale production

Method used

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Embodiment Construction

[0029] A method for extracting copper-zinc superoxide dismutase SOD from pig red blood cells is completed according to the following steps:

[0030] 1. Collect fresh and healthy pig blood; fresh and healthy pig blood means that the processed animal blood all comes from designated large-scale slaughterhouses, and all pigs must pass the inspection of the national animal quarantine department for disease-free.

[0031] 2. Anticoagulant treatment of pig blood: the anticoagulant used is food-grade sodium citrate, and the addition amount is 4.0-6.0g / kg pig blood.

[0032] 3. The anticoagulated pig blood was centrifuged at a high speed in a tube centrifuge to obtain blood cells with a volume of 50 L. The rotating speed of the tube centrifuge is 14000r / min-16000r / min.

[0033] 4. Hemolyze and crush the blood cells; the blood cells are crushed twice by colloid mill, then add 2 times the volume of distilled water or high-purity water to swell overnight at 0°C-4°C;

[0034] 5. First ad...

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Abstract

The invention relates to a method for extracting superoxide dismutase (SOD) from porcine blood red cells, in the method, anticoagulation treatment is adopted for preventing blood coagulation and breaking of blood cells. Colloid mill breaking and low-temperature swelling are adopted for releasing matters dissolved in the blood cells to the maximum extent so as to get hemolysate. Thermal change is performed on the hemolysate twice for removing impurities, removing hemoglobin and a large number of impurity proteins so as to get SOD crude enzyme liquid, and ultrafiltration concentration, gradient elution, dialysis, salt removal and freeze-drying are further performed on the SOD crude enzyme liquid so as to get an SOD product. The method is simple to operate and easy to master; furthermore, by adopting the method, the product quality can be effectively improved, the production cost can be reduced, the production cycle can be shortened, the production efficiency can be improved, and the method is applicable to industrialized production.

Description

Technical field: [0001] The invention relates to a method for extracting superoxide dismutase, in particular to a method for extracting superoxide dismutase SOD from pig red blood cells. Background technique: [0002] Superoxide dismutase (Superoxide Dismutase, hereinafter referred to as SOD) is a metalloenzyme capable of removing excess free radicals in the body, and has been found to exist widely in various tissues in humans and animals. SOD has the functions of anti-aging, preventing pigmentation, anti-inflammation and protecting the immune function of the body. At present, it has been widely used in food, health products, cosmetics and agriculture. [0003] The existing methods for extracting SOD from blood cells are similar in steps such as blood collection, anticoagulation, washing with normal saline, and low-temperature hemolysis. Most of the miscellaneous protein denaturation in the later stage adopts chloroform and ethanol method to remove hemoglobin, then the sup...

Claims

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Application Information

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IPC IPC(8): C12N9/08
Inventor 夏继华张立娟宋古全沈峰王长维
Owner HEILONGJIANG BAODI MEAT FOOD
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