Purification and characterization of lipase enzyme from black mustard seed (Brassica nigra)

When lipases are investigated, they are known as enzymes which act as catalysts in the formation of ester bonds, which are separated into fatty acids and glycerol-derived esters as a result of interaction with water or lipid structures. In this study, it is aimed that the mustard seed is an oily seed and that the obtained lipase is obtained pure and characterized. An alternative to lipase-purified plant sources was established by the study and the first time the enzyme of the lipase enzyme from mustard seed was realized. The lipase enzyme purified from mustard seeds is initiated by de-oiling the proteins found in mustard seeds. The mustard seed lipase was purified using gel filtration chromatography. In all purification steps protein quantity was measured according to Bradford method and lipase activity was measured by titrimetric method. Specific activity was calculated and it was determined that lipase was purified 37.89 fold. Maximum acitivity of cotton seed lipase was observed at pH 5.6 and 60 ˚C. It was found that stabil pH and stabil temperature 6.8 and 4 ˚C, respectively. To determine storage stability of purified lipase, activities were measured for two weeks. After two weeks lipases in 4 ˚C were protect their catalytic activity. To calculate Km and Vmax of mustard seed lipase, triolein was used substrate and Km value was calculated as 0,1222 mM, Vmax value was calculated as 1.0195 U/dk.mg.