The Process Improvement On The Separation And Purification Of Superoxide Dismutase From Blood And Its Oxidation Resistance

The English abbreviation of Superoxide dismutase is SOD, which is a kind of important enzyme in organisms, it can catalyzed the Super oxide anion occur Qi Hua reaction and generate H2O2 and H2O , this can make the formation and elimination of O2– in a dynamic balance,resisting the toxicity of O2 - and eliminating the harmful substances which was produced by the biochemical metabolism of the organism, thus SOD has the Function of anti-oxidant, anti-inflammatory, resist radiation ,anti-tumor and anti-aging etc. For the first time since 1969 McCord isolated SOD from the ox erythrocyte, People have the SOD separated and purificated from the animal red erythrocyte, muscle such as cattle, pigs, sheep and plant .According to its different complementary metal SOD can be divided into three types: Fe-SOD, Mn-SOD CuZn-SOD, the Relative molecular weight ( RMW ) of Mn– SOD is 4000, purple, exists in the mitochondria of eukaryotic cells and prokaryote cells, When the mitochondrial DNA was attacked by oxygen free radicals,it produces 8 - hydroxyl deoxidation guanine nucleotides, The product is the cause of aging, Mn– SOD has the effect to Prevent the damage, The RMW of Fe-SOD is 38700, golden brown, exist in prokaryotes and plants, Cu Zn-SOD, Shallow blue, RMW 32000, exist in animal cells,the three-dimensional crystal structure of Cu Zn-SOD was analysised by 2AX rays, Each enzyme molecule consists of two came through the trap of covalent health association interactions dimers, synthetic peptide internal C55 cysteine Cl44 and by the sulphur (SH) disulfide bridge to associate plays an important role, Richardso determined its activity center and indicate that the active site is just Cu2 + ,Experiment proves that Cu2 + is the activity of Cu Zn– SOD, Zn2+ is connected with the stability of the enzymes.During the existing purification process of SOD, rough classification technoloy is inevitable, it must use a large number of organic solvent such as alcohol, chloroform, etc.) or inorganic phosphate (e.g., ammonium sulfate, etc.), Through the organic solvent and inorganic salt classification precipitation to remove most of the miscellaneous protein and gained coarse product SOD . Then you can use the next process, such as acetone precipitation, ultrafiltration enrichment, dialysis, column chromatography to do further purification. Firstly, using hierarchical precipitation to extracte the target protein, the waste liquid contains high levels of organic solvents or inorganic salts, the recovery of these poisonous and harmful material is very difficult.Once letting out will cause enormous pressure, secondly, the cycle of Organic solvent and inorganic salt precipitation operation is long, the extract must be stewing in low temperature for overnight , in addition, in order to Prompt protein deposition, The usage of inorganic and organic solvents are very huge , this will raise the cost of production, Therefore, economic and efficient, environmental purification technology has been the bottleneck of protein purification, in the Currently extraction process of SOD, The traditional classification precipitation technology is still used today, This makes the cost and output of SOD biochemical products be difficult to achieve the desired level.This paper studied the extraction process of SOD from animal blood, useing low molecular weight polyacrylic acid sodium and chloride copper as dispersant and efficient activators to purification SOD for the first time, and obtain coarse SOD via hemolysis, hot acetone precipitation, ultrafiltration concentrated, this make the activity of the coarse SOD reach the extraction separation effect of the traditional craft, this craft greatly simplifies the process of traditional craft and shorten the production cycle, in addition, chromatography is an important step to obtain the competitive products, The enzyme activity of SOD will be greatly enhanced after chromatography, After using the classification precipitation, Using Chromatography method to purify the coarse SOD, at present both in home and abroad, it mainly focus on gel filtering and ion exchange chromatography, or the combination of the two or more pace , With the improvement process after extraction of SOD , SephadexG-75 gel and DEAE -sepharose-fast flow ion exchange chromatography respectively is used to purify ultrafiltration liquid, the two kinds of purification methods are very easy to achieve electrophoresis pure , This greatly simplifies the traditional craft, it has some reference value to the mass production industry...