ABSTRACT.Components with phospholipase A 2 activity were isolated by gel filtration and cationic exchange chromatography from the venom of Bothrops asper snakes from Chiriguaná, Colombia (9°22´N; 73°37´W). Five fractions were obtained by the gel filtration, and PLA 2 activity was found in fraction 3 (F3). In the cationic exchange chromatography, F3showed eight components with PLA 2 activity. Six of these components appeared as one band in polyacrylamide gel electrophoresis (SDS-PAGE). Fractions II and VII exhibited an optimal activity at pH 9 and 52ºC. The optimum calcium concentration for fraction II was 48 mM and for fraction VII, 384 mM. Both fractions showed thermal stability. Fraction II was stable at pH values between 2.5 and 9, and fraction VII, between 2.5 and 8. The Michaelis Menten constant (K M ) was 3.5x10 -3 M for fraction II and 1.6x10 -3 M for fraction VII. The molecular weight was 16,000 Dalton for fraction II and 17,000 Dalton for fraction VII. Both isoenzymes did not show any toxic activity (DL 50 ) at 5.3 and 4 µg/g. The two fractions showed different kinetic constant (K M ), calcium requirement, and substrate specificity for haemolytic activity.
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