PURIFICATION AND CHARACTERIZATION OF LIPASE FROM PSEUDOMONAS-FLUORESCENS NO 33

Extracellular lipase from Pseudomonas fluorescens No. 33 was purified to electrophoretic homogeneity by a procedure including acid precipita tion, hydrophobic-interaction chromatography, ion-exchange chromatogra phy and gel filtration. The enzyme was purified about 4200- fold givin g a yield of 35 % of the original activity. The molecular weight of th e enzyme was 52,000 by SDS-PAGE: The optimum pH and temperature for th e hydrolysis of butter oil emulsion were 7.5-8.5 and 45-degrees-C, res pectively. The enzyme was stable over the pH range 5.5-7.5. It was rem arkably more heat-labile at 40-degrees-C than at 50-60-degrees-C. The activity was inhibited by Zn2+ and Hg2+.