PURIFICATION OF GLUTAMINASE ENZYME PRODUCED FROM ERWINIA

The purpose of this study was to do Purification of the Glutaminase enzyme produced from free cells of Erwinia species at flask level. Glutaminase can be isolated from a number of sources such as plants, animals and microorganisms. Glutaminase is an important enzyme that serves many functions. It plays a key role in the energy and nitrogen metabolism of mammalian cells. Glutaminase is very important food enzyme used in food industries for flavor enhancement. Glutaminase, in combination with or as an alternative to asparaginase could be of great significance in enzyme therapy for cancer especially acute lymphocytic leukemia. Glutaminase enzyme was produced from free cells of Erwinia under optimized conditions such as Temperature, pH, Time, Inducer concentrations etc. After production of Glutaminase enzyme, Partial purification of enzyme was done with Ammonium Sulphate precipitation method. After isolation, the Glutaminase enzyme was purified with Gel filtration Chromatography & Ion Exchange chromatography. After purification by both methods, Purified samples were analyzed for enzyme activity & protein content. Enzyme activity was determined by Nessler's method & protein content was determined by Bradford method. It was found that after purification of crude sample by both methods, Gel Filtration chromatography shows maximum enzyme activity and specific activity than the samples purified with Ion Exchange Chromatography. Also %age recovery (97.59%) & purification fold (1.70) obtained was found maximum from the samples purified with Gel Filtration Chromatography. From above results it was concluded that Gel filtration method is Better method for the purification of Glutaminase enzyme than Ion exchange Chromatography.