Lipase purification from the chamomile (Matricaria chamomilla L.) and its investigation of some kinetic properties

In this study, lipase was firstly purified from chamomile (Matricaria chamomilla L.) being abundant in Giresun (Turkey) and surroundings and the kinetic properties of the enzyme were investigated. Chamomile, being fat-free with petroleum ether extraction, were homogenized with 0.06 M sodium phosphate buffer (pH=7.0) 70% 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 26 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 chamomile had an optimum pH value of 12 and an optimum temperature of 50°C. When the high affinity of the enzyme agains different substrates was examined, the highest value was obtained with p-nitrophenyl palmitate, with respectively Km and Vmax values being 0.2899 mM and 144.93 Units.