Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1216078 | Journal of Chromatography B | 2007 | 7 Pages |
Abstract
Ostrich pancreatic phospholipase A2 (OPLA2) was purified from delipidated pancreases. Pure protein was obtained after heat treatment (70 °C), precipitation by ammonium sulphate and ethanol, respectively followed by sequential column chromatography on MonoQ Sepharose and size exclusion HPLC column. Purified OPLA2, which is not a glycosylated protein, was found to be monomeric protein with a molecular mass of 13773.93 Da. A specific activity of 840 U/mg for purified OPLA2 was measured at optimal conditions (pH 8.2 and 37 °C) in the presence of 4 mM NaTDC and 10 mM CaCl2 using PC as substrate. This enzyme was also found to be able to hydrolyze, at low surface pressure, 1,2-dilauroyl-sn-glycero-3 phosphocholine (di C12-PC) monolayers. Maximal activity was measured at 5-8 mN mâ1. The sequence of the first 22 amino-acid residues at the N-terminal extremity of purified bird PLA2 was determined by automatic Edman degradation and showed a high sequence homology with known mammal pancreatic secreted phospholipases A2.
Keywords
NaDCNaTDCsPLA2DTTEGTABSAbovine serum albuminephospholipase A2EDTAethylene diamine tetraacetic acidsodium dodecyl sulfate-polyacrylamide gel electrophoresisSDS-PAGELipid monolayerdithiothreitolsodium taurodeoxycholateSodium deoxycholatephosphatidylcholineBile saltsPAFarbitrary unitsHigh pressure liquid chromatographyHPLC
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Abir Ben Bacha, Youssef Gargouri, Sofiane Bezzine, Habib Mosbah, Hafedh Mejdoub,