جداسازي و خالص سازي ايزوآنزيم هاي پراكسيداز از تربچه

Separation and Purification of Peroxidase Isoenzymes from Cultivated Radish

Introduction: Peroxides is a glycoprotein that contains hens group and variable cationic and anionic is enzymes. ~.: . This enzyme is used in laboratory diagnosis for determination of hormones and bacterial toxins with ELISA method , detection of tissue antigens and macrophage activation against tumoral cells. In addition , this enzyme is used for measurement of glucose, urea, uric acid and cholesterol. Although peroxides is widely seen in available sources of plants, this enzyme for meeting the consuming needs of our country is imported from abroad with high foreign currency costs. Objective: This experimental study was conducted for separation and purification of peroxides is enzymes from cultivated radish which is cheap and abundant in all seasons in Iran. Materials and Methods: Extraction was carried out by homogenization of plant roots and concentration of Ammonium sulfate. Then, purification method was followed by gel filtration, cationic exchange chromatography. Finally, separation of basic and acidic is enzymes was conducted by different and repeated column chromatography and SDS-PAGE. Results: As a result of these procedures, Purification degree obtained was 168.5 folds more than the raw extract and specific activity was comparable with commercial peroxides . Conclusion: As this enzyme is stable at wide range of PH and temperature, we can economize its purification methods to a great extent and achieve the maximum required foreign currency savings. It is hoped that with the use of wide and cheap sources , we can take an effective step toward large scale production of the enzyme in our country and avoid spending our financial resources in purchasing peroxides from foreign countries.