خالص سازي آنزيم پلي گالاكتوروناز از سويه 58 F قارچ فوزاريوم اگزيسيوروم

Purification of Polygalacturonase Enzyme from Strain F58 of Fusarium Oxysporum

Background and Objectives:Polygalacturonase enzyme is produced in pectin and polygalacturonic acid culture media by a wide variety of microorganisms, such as Fusarium oxysporum, which have important application in medicine, pharmacology and food industries. The purpose of this study was to extract and purify polygalacturonase enzyme by ion exchange chromatography techniques. Materials and Methods:Fusarium oxysporum strain F58 was obtained from "Jahad Daneshgahi" institute. The previous studies have shown that this strain can provide large quantities of enzyme. Purification of polygalacturonase enzymes was achieved by cation exchange chromatography on CM-Sepharose Fast Flow column equilibrated to pH 5.5 with 50mM sodium acetate buffer. Elution of the column was carried out with lmoi NaCl gradient. Results: The results of chromatography showed one enzyme activity peak between 0.2-0.4 of gradient salt in purification profile. Fractions with highest enzyme activity were pooled and the precipitated proteins were used for electrophoresis by SDS-PAGE methods. SDS-PAGE showed two distinct bands about 38 and 40 KDa. These two bands showed PG activity in gel containing pectin as substrate (Activity staining). Conclusion: Regarding the importance and value of this enzyme, scientific improvement and cost effectiveness of production processing, extraction, purification and production of this enzyme could be important due to its application in medicine and industry.