Purification and biochemical characterization of pancreatic phospholipase A2 from the common stingray Dasyatis pastinaca

Table 1 Flow sheet of stingray PLA2 purification.

Purification step	Total ^(a) activity (units)	Protein ^(b) (mg)	Specific activity (U/mg)	Activity recovery (%)	Purification factor
Extraction	9000	3500	2.6	100	1
Heat and acidic treatment	7650	1050	7.3	72	2.8
(NH ₄ ) ₂ SO ₄ Precipitation (30-70%)	6120	500	12.3	68	4.7
Ethanol fractionation (50-90%)	5250	150	35	58	13.46
Mono-S Sepharose	4050	8.1	250	45	96
Mono-Q Sepharose	2900	5.3	550	32	211.5
RP-HPLC	2100	2.8	750	23	288.5

(a) 1 Unit: μmole of fatty acid released per min using PC emulsion as substrate in the presence of 4 mM NaTDC and in the presence of 8 mM CaCl₂.
(b) Proteins were estimated by Bradford method (32). The experiments were conducted three times.

ISSN: 1476-511X