Purification and Characterization of Lactoperoxidase from Goat Milk, Investigation of Inhibition Effects of Cefotaxime Sodium

İt has been shown in the literature that LPO is important for natural immunity. The purpose of this study is to purify the lactoperoxidase (LPO) enzyme from goat milk and to investigate the inhibition property of cefotaxime sodium on the enzyme. In this study, the LPO enzyme was purified from goat milk and the inhibition effect of cefotaxime sodium was examined. Enzyme purification processes were carried out by various chromatographic methods and the enzyme inhibitory effect was analyzed with Lineweaver-Burk graphs. İnhibition of cefotaxime sodium on LPO was determined as competitive inhibition. The enzyme was purified by first partially purifying milk using Amberlite CG-50 H+ resins, CM-Sephadex C-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography. The 2.2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) substrate was used to determine the enzyme’s Km and Vmax values. Enzyme purity was determined using SDS-PAGE electrophoresis. A specific activity of 7.21 EU mg-1 of protein per liter was detected. The LPO was purified 13.35 times from 1 liter of defatted milk, and 1.9 mg of enzyme was obtained. ABTS substrate (ε412nm=32400M-1 cm-1) was used for the enzyme-activity assays.