Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for extracting superoxide dismutase from animal blood

A technology of superoxide and dismutase, applied in the direction of oxidoreductase, etc., can solve the problems of physical damage to operators, serious environmental pollution, complicated process, etc., and achieve the effects of low cost, increased ionic strength, and improved yield

Inactive Publication Date: 2013-12-11
浙江迈康生物工程有限公司
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this process is that the process is complicated, the cost is high, and organic solvents such as chloroform and acetone have strong toxicity, which will seriously pollute the environment and cause immeasurable damage to the operator's body.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Add 50L of 8% sodium citrate solution to 1000L of fresh pig blood for anticoagulation, and use a continuous flow centrifuge to separate 450L of red blood cells and 600L of plasma. Plasma is spray-dried to become plasma protein powder. The red blood cells are pumped into the reaction tank, and at the same time, 1200 L of distilled water is added and stirred with a mixer for 30 minutes, so that the red blood cells are fully broken and hemolyzed. After heating to 60°C and keeping it warm for 15 minutes, add 37Kg of sodium chloride under stirring, raise the temperature to 65°C and keep it warm for 30 minutes, then raise the temperature to 72°C, continue heating for 20 minutes, and quickly water-cool to below 30°C. Press filter with 800 mesh filter cloth to obtain a total of 1300L of SOD crude extract, with a unit activity of 220U / ml. The SOD crude extract is pumped into the ultrafiltration storage tank, first pretreated by the ultrafiltration membrane to intercept macromol...

Embodiment 2

[0025] 2000L of fresh duck blood was anticoagulated with 100L of 10% sodium citrate solution, and 830L of red blood cells and 1270L of plasma were separated by a continuous flow centrifuge. Put the red blood cells in the reaction tank, add 1660L distilled water, and homogenize under high pressure for 20 minutes to completely break the red blood cells. Continue to stir and raise the temperature to 60°C for 15 minutes, add 58Kg of sodium chloride, raise the temperature to 65°C and continue to heat for 30 minutes, then increase the temperature to 70°C, keep the heat for 20 minutes, and quickly discharge the material to cool. 800-mesh filter cloth was mechanically pressed to obtain 1900L of SOD crude extract, with a unit activity of 270U / ml. The SOD crude extract is pumped into the ultrafiltration storage tank, first pretreated by the ultrafiltration membrane to intercept macromolecular proteins and cell membrane fragments with a molecular weight greater than 1 million, and then r...

Embodiment 3

[0029] A method for extracting superoxide dismutase from animal blood, comprising the following steps:

[0030] A. Blood separation: take fresh chicken blood, add a sodium citrate solution with a mass concentration of 10%, the volume of the sodium citrate solution is 6.5% of the blood volume, collect red blood cells by centrifugation in a continuous flow centrifuge after anticoagulant treatment, and pump into the hydrolysis reaction tank;

[0031] B. Hemolysis and broken cells: add 1.5 times the volume of pure water to the red blood cells collected in step A, and homogenize under high pressure to completely break the red blood cells to obtain red blood cells for hemolysis;

[0032] C. Thermal denaturation: heat the erythrocyte hemolysis to 60°C and keep it warm for 10 minutes, then add NaCl to the erythrocyte hemolysis to increase the ionic strength of the solution until the NaCl concentration is 0.55mol / L, and continue heating to 65°C. After keeping warm for 50 minutes, heat...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for extracting superoxide dismutase from animal blood. The method comprises the following steps: A. blood separation; B. hemolytic cell breaking; C. heat denaturation; D. filter pressing; E. ultrafiltration and concentration; F. precipitation for removing hybrid protein. Such organic solvents as chloroform, ethanol and acetone, and the like, are completely eliminated, and a rough product SOD with high activity is simply and conveniently separated out through a heat denaturation and metal salt precipitation method according to the structure characteristics and physicochemical properties of SOD molecules, which is a revolutionary improvement of SOD production. Since no inorganic solvent is used, the method is low in cost and is more suitable for industrial production. Hemoglobin is removed by thermal denaturation in the invention, so that hemolysis can be realized at normal temperature, the operation is simpler and more convenient and the energy consumption is reduced.

Description

(1) Technical field [0001] The invention relates to a method for extracting superoxide dismutase from animal blood. (2) Background technology [0002] Superoxide dismutase (SOD) is a metalloenzyme ubiquitous in organisms that can scavenge superoxide anion free radicals. It has many functions such as anti-inflammation, anti-radiation, and anti-tumor. It has been widely used in medicine, food, health food and cosmetics. [0003] At present, the technological process of extracting superoxide dismutase from animal blood is mainly blood anticoagulation - washing with normal saline - low temperature hemolysis - chloroform ethanol extraction of hemoglobin - potassium phosphate extraction - heat denaturation - acetone precipitation - - ion exchange chromatography - molecular sieve column chromatography. The disadvantage of this process is that the process is complicated, the cost is high, and organic solvents such as chloroform and acetone have strong toxicity, which seriously pol...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/02
Inventor 龚兴国秦勇聂颢韩斌应育权
Owner 浙江迈康生物工程有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com