Isolation and characterization of lipase enzyme from microorganisms isolated from soil

The area of usage, the place, and importance of Lipase enzymes in biological processes is increasing day by day. In this study it was aimed to obtain an alternative (lipase enzyme) to lipase enzymes used in industrial processes. The microorganism isolated from the soil and determined to produce lipase was determined by 16S rRNA sequence analysis and the lipase enzyme it produced was partially purified by using methods such as ammonium sulfate precipitation, dialysis, etc. As a result of the procedures, it was found that lipase was purified 1.74 times by dialysis and 2.59 times by ammonium sulfate. Characterization and molecular weight determination of the obtained lipase were performed by SDS-PAGE. The molecular weight of the enzyme was calculated as 72 kDa. The purified lipase showed maximum hydrolytic activity at 50 °C and pH 10.0. The effect of detergent additives such as EDTA, SDS and PMSF on enzyme activity was determined. The Km and Vmax values for the lipase using p - Nitrophenyl palmitate ( P-NPP) as a substrate were calculated as; Km = 1,458 mM and Vmax = 22,47 U/ml. These results indicate that the lipaze obtained has very suitable properties for industrial applications in terms of its thermal and pH stability and other properties,