Purification of bovine pancreatic phospholipase A2 by an affinity ultrafiltration technique

Phospholipase A2 (PLA2; EC 3.1.1.4) is a lipolytic enzyme that hydrolyze phosphoglycerides at the acyl ester bond at the sn-2-position into the corresponding lysophospholipid and fatty acid. Mammalian PLA2 enzymes are subdivided into three groups: low molecular weight Ca2+-dependent enzymes (sPLA2), high molecular weight Ca2+-dependent enzymes (cPLA2) and the Ca2+-independent isoforms (iPLA2). The object of this study is to discover the synthesis of an affinity complex which could be used in an ultrafiltration system for the biospecific affinity isolation of phospholipase A2 from mixtures containing other proteins. For this purpose, first, the development of a macromolecular water soluble resin was attempted, based on chitosan bearing phosphatidylethanolamine, a phospholipase A2 substrate. Then, together with ultrafiltration techniques, the macroligand was employed to purify phospholipase A2 from bovine pancreas. The enzyme was purified 79-fold and had a high activity yield 76.3%. The enzymatic properties obtained and showed nearly similarity in terms of optimum temperature and pH, molecular weight, Ca2+-dependence with bovine and other mammalian pancreatic PLA2's. © 2008 Elsevier B.V. All rights reserved.Firat University Scientific Research Projects Management Unit: 2002-FEN-007This work was supported by a grant from Ege University Scientific Research Project (2002-FEN-007). -