Method for extracting superoxide dismutase from blood of mammal

A mammalian, superoxide technology, applied in the field of food processing, can solve the problems of damage to superoxide dismutase, trouble, low enzyme yield, etc., and achieve the effect of simplifying the process and improving the efficiency

Inactive Publication Date: 2013-02-13
景志刚 +1
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Problems solved by technology

[0005] Ethanol and chloroform are used in the McCord-Fridovich method. Some scholars suspect that precipitating hemoglobin with ethanol-chloroform can change the protein properties of superoxide dismutase. For example, Harts and Deutch think that the Tsuchihashi method will damage superoxide dismutase, so they use other methods. method to remove hemoglobin, but it is troublesome, and the enzyme yield is low. Bannister et al. pointed out that there is no qualitative difference between the superoxide dismutase obtained by ethanol chloroform treatment and non-treatment

Method used

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Embodiment Construction

[0022] Below in conjunction with specific embodiment the present invention is described in further detail:

[0023] The method for extracting superoxide dismutase from mammalian blood comprises the following main preparation steps: preparing food-grade superoxide dismutase from mammalian blood. The main content includes establishing a relevant quality and safety control system, realizing vacuum blood collection and simultaneous quarantine, adding a certain amount of ideal anticoagulant, adopting low-temperature continuous separation technology, improving and perfecting the separation conditions, improving the separation effect of blood, and realizing mammalian blood plasma Continuous separation of blood cells and blood cells; the main preparation process steps are: separate mammalian blood into plasma and blood cells with a centrifuge, and after the blood cells are treated with microwave-enzyme double-effect wall breaking, add water to the plasma or mix them with water to disso...

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Abstract

The invention discloses a method for extracting superoxide dismutase from the blood of a mammal, belonging to the technical field of food processing. The method is used to solve the problems of high cost, low benefit and low purity in extracting superoxide dismutase from the blood of the mammal. The method for extracting superoxide dismutase from the blood of the mammal comprises the following steps of: (a) collection of the blood of the mammal; (b) hemolysis; (c) thermal denaturation in the presence of copper salt; (d)ultrafiltration; (e) precipitation of acetone and recovery of acetone with a rotary evaporator; (f)column chromatography; and (g)freeze-drying. The method for extracting superoxide dismutase from the blood of the mammal can be widely used in extracting superoxide dismutase from the blood of any mammal.

Description

technical field [0001] The invention relates to the technical field of food processing, in particular to a method for extracting superoxide dismutase from mammalian blood. Background technique [0002] The existing classic McCord-Fridovich method: the first step is to remove hemoglobin. Adopt the Tsuchihashi method, that is, add 0.25 times the volume of ethanol and 0.15 times the volume of chloroform to the hemolysis, stir continuously at 4°C for 15 minutes to denature and precipitate the hemoglobin, then add 0.1 times the volume of water, and stir evenly. The ethanol-chloroform phase was separated by centrifugation at 4°C. [0003] The second step is organic solvent precipitation. The ethanol-chloroform extract is placed at room temperature to balance, and 300 grams of anhydrous dipotassium hydrogen phosphate solution and chloroform are added in every 1000 milliliters, while the upper phase contains less dipotassium hydrogen phosphate, mainly ethanol. Superoxide dismutas...

Claims

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Application Information

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IPC IPC(8): C12N9/08
Inventor 景志刚杨洋
Owner 景志刚
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