title: Purification and characterization of a phospholipase A2-IIA from common stingray (Dasyatis pastinaca) intestine.
abstract: A phospholipase A2 belonging to IIA group secretory PLA2 was isolated and purified to homogeneity from the intestine of common stingray (Dasyatis pastinaca) using acidic treatment (pH 1.5) and ammonium sulphate precipitation methods combined with single-column ion-exchange chromatography. The purified enzyme was found to be a glycosylated monomeric protein with a molecular mass of about 14 kDa. The stingray sPLA2-IIA had optimum activity at 45 degrees C, unlike known mammalian PLA2-IIAs, which show optimum activity at 37 degrees C. The purified enzyme exhibited a specific activity of 290 U/mg at optimal conditions (pH 9.5 and 45 degrees C) in the presence of 6 mM NaDC and 8 mM CaCl2 with egg yolk as substrate. The NH2-terminal sequence of the enzyme and some protein fragments obtained from its tryptic digestion were also determined. All sequences obtained were similar to those of sPLA2-IIA. The enzyme also showed good stability in the presence of organic solvents, acidic and alkaline pH media and high temperature conditions. Thus, the purified enzyme exhibited a number of unique and promising properties, making it a potential possible candidate for future applications in the treatment of phospholipid-rich industrial effluents and synthesis of useful preparations for the food production and processing industry.
authors.affiliation: Biochemistry Department, Science College, King Saud University, PO Box 22452, Riyadh 11495, Saudi Arabia. abirghanouchibenbacha@yahoo.fr
authors.forename: Abir
authors.initials: A
authors.lastname: Ben Bacha
authors.forename: Sooad K
authors.initials: SK
authors.lastname: Daihan
authors.forename: Nadine M S
authors.initials: NM
authors.lastname: Moubayed
authors.forename: Hafedh
authors.initials: H
authors.lastname: Mejdoub
mesh_terms: D000818:Animals; D018940:Elasmobranchii / Q000378:metabolism*; D004789:Enzyme Activation; D004795:Enzyme Stability; D054501:Group II Phospholipases A2 / Q000737:chemistry* / Q000302:isolation & purification*; D007422:Intestines / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D054501:Group II Phospholipases A2