title: Purification and biochemical characterization of phospholipase A2 from dromedary pancreas.
abstract: Dromedary pancreatic PLA2 (DrPLA2) was purified from delipidated pancreases. Pure protein was obtained after heat and acidic treatment (70 degrees C; pH 3.0), precipitation by ammonium sulphate and ethanol respectively, followed by sequential column chromatographies on Sephadex G-50, MonoS Sepharose, MonoQ Sepharose and C-8 reverse phase high pressure liquid chromatography. Purified DrPLA2, which is not glycosylated protein, was found to be monomeric protein with a molecular mass of 13748.55 Da. A specific activity of 600 U/mg for purified DrPLA2 was measured at optimal conditions (pH 8.0 and 37 degrees C) in the presence of 3 mM NaTDC and 7 mM CaCl(2) using PC as substrate. The sequence of the first fourteen amino-acid residues at the N-terminal extremity of DrPLA2 was determined by automatic Edman degradation. One single sequence was obtained and shows a close similarity with all other known pancreatic secreted phospholipases A2.
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS route de Soukra, 3038 Sfax, Tunisia.
authors.forename: Abir Ben
authors.initials: AB
authors.lastname: Bacha
authors.forename: Youssef
authors.initials: Y
authors.lastname: Gargouri
authors.forename: Sofiane
authors.initials: S
authors.lastname: Bezzine
authors.forename: Hafedh
authors.initials: H
authors.lastname: Mejdoub
mesh_terms: D000818:Animals; D002162:Camelus; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D010179:Pancreas / Q000201:enzymology*; D010741:Phospholipases A / Q000302:isolation & purification* / Q000378:metabolism*; D054467:Phospholipases A2; D013696:Temperature
chemical_list: D010741:Phospholipases A; D054467:Phospholipases A2