Characterization and purification of lipase enzyme from aspir (Carthamus tinctorius L.) seed

Lipase is an enzyme that acts as a catalyst in the hydrolysis of triacylglycerols to glycerol and fatty acids. Today, the acceleration of biotechnological developments increases the usage areas of lipase enzyme. In this study, it was aimed to contribute to the literature by purifying and characterizing lipase from safflower (Carthamus tinctorius L.) seed for the first time. In the steps of defatting of safflower seed proteins, lipase purification was carried out using oil removal, ammonium sulfate precipitation, dialysis, gel filtration chromatography methods, respectively. In the purification steps, protein determination was performed in accordance with the Lowry method and titrematic procedures were performed for activity determinations. As a result of the procedures, it was determined that safflower seed was purified 8,47 times. Specific activity values were also calculated as a result of the procedures. After purification of the lipase enzyme, characterization was performed by SDS-PAGE. The results obtained as a result of the procedures showed that the best activity enzyme pH: 10.4 and temperature 60℃ were noted as the values. However, the stable pH value was found to be 13.2 and the stable temperature level was 50℃. The storage stability of the lipase enzyme was also determined and a 70% decrease in enzyme activity was observed in measurements made for 5 consecutive days (60℃, pH: 10.4). The Km= 25.49 mM and Vmax value was found to be approximately 5.6 U/min.mg of enzyme in olive oil and triolein substrate. Key words: Safflower seed, Enzyme, Characterization, Lipase, Purification