Kestane (Castanea sativa) tohumundan li̇paz enzi̇mi̇ni̇n saflaştirilmasi ve ki̇neti̇k özelli̇kleri̇ni̇n i̇ncelenmesi̇

In this study, lipase was firstly purified from chestnut (Castanea sativa) seeds, being abundant in Giresun (Turkey) and surroundings and the kinetic properties of the enzyme were investigated. Chestnut seeds, being fat-free with petroleum ether extraction, were homogenized with 0.06 M sodium phosphate buffer (pH=7.0) 90% of (NH4)2SO4 fraction of lipase esterase activity homogenate was obtained. After dialysis, this fraction was applied respectively to hydroxyapatite and DEAE-cellulose columns, and lipase was purified 186.87 times. In the purification process, the protein content was determined by using Lowry and E280/E260 Warburg methods, and lipase esterase activtiy was assayed with Erlanson's method. The purified enzyme, which showed SDS-PAGE, had a molecular weight of 30 kDa. The optimum pH and temperature values, pH and temperature stabilities, optimum reaction time, appropriate concentrations of the enzyme and substrate were investigated. It was also found that the lipase purified from chestnut seeds had an optimum pH value of 9.0 and an optimum temperature of 30C. When high affinity for various substrates of the enzyme is analyzed, it was found that most of the lipase of interest in myristate and 1.017, respectively p-nitrophenyl is the Km and Vmax values against the substrate unit is mm and 163.874. Key Words: Chestnut (Castanea Sativa) Seed, Lipase, Purification, Characterization.