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 LineweaverBurk plot analysis, which was found to be 0.00451 mM and 28.57 μM/ml/min at 370 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. C.