Study On Purification And Characteristic Of SOD From Cordyceps Militaris By Solid Cultur

Superoxide Dismutase also called Liver protein or Orgotein,Referred to as SOD. There are three main types, namely Cu、 Zn-SOD、Fe-SOD、Mn-SOD. They are widely present in tissue cells of animals, plants and microorganisms. SOD as an active substance derived from Organism, it not only can remove the superoxide anion radical generated from Organism metabolism,but also can protect the body cells from the damage of aerobic metabolite as a substrate induced enzyme. Constitute the first line of defense against oxidative damage. SOD supplement to the human body constantly produces the special effects of anti-aging.Therefore, SOD has been widely used in clinical medicine, food and chemical industry.Cordyceps also known as Cordyceps Militaris,it has a variety of active ingredients,such as cordyceps polysaccharide, cordyceps acid, cordycepin, superoxide dismutase and trace elements. It is good for human heath、tonic and disease treatment。There are reports illustrate, the pharmacological properties of Cordyceps associated with the it’s superoxide dismutase activity.The SOD of Cordyceps mainly distributed in it’s mycelium, there are still plenty of mycelium residues in the medium after the fruiting bodies were harvested. This experiment focused on the methods of extraction, separation and purification of superoxide dismutase from Cordyceps fruiting and solid culture residues medium and the stability of Cordyceps SOD..The experiment also studies effect of different components of the solid medium for SOD content,and contrast the content and enzyme activity of Cordyceps fruiting and solid culture residues medium.The experiment has first crude extracted the SOD of Cordyceps fruiting and solid culture residues medium.Then use sulfate salting classification> DEAE-32anion exchange chromatography and CM-52cation exchange chromatography to further separation and purification.Specific activity of the SOD has increased respectively from39.0U/mg and29.5U/mg to939.5U/mg and897.4U/mg.The purified SOD has appeared two bands through the non-denaturing polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis, and then measured the two Cordyceps SOD subunit relative molecular are16.71KDa and15.97KDa. The thermal stability of the Cordyceps SOD from the experiment is very well. The experiment shows that the activity remained unchanged after incubated120min under40-60℃.It still maintains20%activity after incubated20min under90℃.Experiments on Cordyceps SOD with different concentrations of H202and chloroform-ethanol,it shows that Cordyceps SOD is sensitive to H2O2,and not sensitive to chloroform-ethanol.Continuously sweep the spectrum of200-700nm, it shows that the maximum absorption occurs at260nm,it also has Characteristic absorption peaks at690nm.The experiment proofs the Cordyceps SOD is Cu、 Zn-SOD.By the comparison,the activity of SOD from Cordyceps fruiting is two times of SOD from solid culture residues medium. But it can be seen that there are still contains amount of SOD, and has high activity, it has a certain value for development. This provides the basis for the reuse of Cordyceps solid culture residues medium.