Purification, characterization, and investigation of in vitro inhibition by metals of paraoxonase from different sheep breeds

Abstract

Paraoxonase (PON) was purified and characterized from the Merino and Kivircik sheep's blood serums by a two-step procedure using ammonium sulphate precipitation and Sepharose-4B-L-tyrosine-1-napthylamine hydrophobic interaction chromatography for the first time. On SDS-polyacyrilamide gel electrophoresis, purified human serum paraoxonase yielded a single band of 66 kDa on SDS-PAGE. The K-M and V-max were 0.482 mM and 41.348 U/mL.dak for Merino PON enzyme, 0.153 mM and 70.289 U/mL.dak for Kivircik PON, respectively. The effect of Mn2+, Hg2+, Co2+, Cd2+, Ni2+ and Cu2+ heavy metals on purified Merino and Kivircik serum PON in vitro was determined.