Purification and Kinetics of Lipase of Pseudomonas fluorescens from Vegetable Oil Polluted Soil

B. Popoola and C. Olateru

Background and Objective: Bacterial lipases occupy a place of prominence among biocatalysts and are used for various biotechnological processes. This study was designed to observe the pure lipases and to enable the establishment of the structure-function relationships. Materials and Methods: The lipase of Pseudomonas fluorescens was partially purified using ammonium sulphate, dialysis, column and ion-exchange chromatography. The purity and molecular weight of lipase was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and their kinetics was studied. Results: The enzyme was purified 8.005-fold from Pseudomonas fluorescens. The molecular mass of the partially purified Pseudomonas fluorescens lipase was 45 KDa. Enzyme kinetics displayed Michaelis-Menten constant K_m 1.25 mg mL^–1 and V_max value of 0.7 μg sec^–1. Conclusion: Hence, knowledge of the kinetics, molecular weights of purified lipases, as well as the three-dimensional structure of lipases plays an important role in designing and engineering lipases for specific purposes.

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