PRODUCTION, PURIFICATION AND KINETIC CHARACTERIZATION OF L-ASPARAGINASE FROM PSEUDOMONAS FLUORESCENS

Objective: L-asparaginase has been isolated from a wide variety of microbial as well as plant and animal sources. In the study the production, partial purification and kinetic characterization of L-asparaginase from Pseudomonas fluorescens were carried.Methods: L-asparaginase was produced by submerged fermentation method. The purification of L-asparaginase was done through ammonium sulphate precipitation as well as three phase partitions (TPP) methods. Partially purified enzyme was further subjected to kinetic characterization and optimum pH and temperature determination.Results: The purification by ammonium sulphate precipitation followed TPP produced 2.34 and 4.137 fold purified enzyme respectively. The optimum pH and temperature for the enzyme activity were found to be 7 and 37°C respectively. The Km and Vmax as determined by Lineweaver-Burk plot analysis, which was found to be 0.00451 mM and 28.57 µM/ml/min at 370C.Conclusion: The higher activity and better kinetic properties of an enzyme in Pseudomonas fluorescens fermentation broth reflects its potential application in the field of medical and food industries.Â