Purification of two lipases with high phospholipase A1 activity from guinea-pig pancreas.
作者： Josette Fauvel , Marie-José Bonnefis , Louis Sarda , Hugues Chap , Jean-Paul Thouvenot
DOI: 10.1016/0005-2760(81)90173-9
关键词:
摘要: 1. 1. Two cationic lipases (Ia and Ib) were purified from homogenates of fresh guinea-pig pancreas by ion-exchange chromatography on DEAE-Sepharose CM-Sepharose (twice for the latter) followed gel filtration Sephadex G-100. 2. 2. Both enzymes homogeneous upon polyacrylamide electrophoresis. Their molecular weights are 37000 42000 Ia Ib, respectively, as determined G-100. Very close values isoelectric points found in pH range 9.3–9.4. 3. 3. The characterized a high phospholipase A activity (500 IU/mg protein using potentiometric assay with egg yolk lecithin substrate), resulting an unusual phospholipase/lipase ratio 1. 4. 4. Using doubly labelled phosphatidylcholine, specificity, A1, was described two enzymes, which unaffected N-ethylmaleimide, diisopropylfluorophosphate p-bromophenacylbromide. insensitive to EDTA slightly inhibited CaCl2and MgCl2, whereas sodium deoxycholate is required maximal activity.
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