Purification and invesigation of kinetic properties of lipase enzyme from pinon nut seed ( Pinus pinea L. )

Lipase enzmies (triacylglycerol açilhidrolaz are EC 3.1.1.3) are catalyzing the synthesis of esters and hydrolysis that consisting of glycerol and long chain fatty acids in living things. The use of lipases in biological processes in the industry is increasing day by day. In his study, purification of lipase enzyme and examination of the kinetic properties have been aimed from kınds of pine nut seeds (Pinus pinea) related to the family of Pinaceae. To purify Lipase from pine nut seeds non phytic acid and protein determination was performed. In the folowing stages, some methods have been performed such as ammonium sulfate precipitation, dialysis, gel filtration chromatography. To determine the purity of the obtained enzymes, activity and electro-phoresis determination (SDS-PAGE) was performed. As a result, the enzyme's (purified from the seeds of pine nuts) optimal PH 4,8 and optimum temperature 50 ° C have been determined. Storage stability of lipase enzyme derived from pine nuts seed at 4 °C was observed that specific activity of the enzyme failure to protect and also enzyme is weak. Km and Vmax values of the pinon nuts seed lipase were found as Km=0,0773mM and Vmax 1,5013 U/dk.mg. Key Words: Pinon nuts seeds, Enzyme kinetics, Characterization, Lipase, Purification.