In the present study, we have purified the group V phospholipase from the heart of cartilaginous fish stingray Dasyatis pastinaca and compared its biochemical properties with group IIA (sPLA 2 -IIA) and IB (sPLA 2 -IB) phospholipases previously purified from pancreas and intestine, respectively. Group V phospholipase (sPLA 2 -V) was purified to homogeneity by heat treatment, ammonium sulphate precipitation and RP-HPLC. The N-terminal sequence of the purified sPLA 2 -V exhibits a high degree of homology with those of mammal. The enzyme was found to be monomeric with a molecular mass estimation of 14kDa. The specific activity of the purified enzyme, measured at pH 8 and 37°C was 52U/mg. Like sPLA 2 -IB and sPLA 2 -IIA, the sPLA 2 -V is found to be stable between pH 3 and 11 after 30min of incubation. The purified sPLA 2 -V retained 65% of its activity after 10min of incubation at 70°C and it absolutely requires Ca 2+ for enzymatic activity. In addition it displayed high tolerance to organic solvents. Kinetic parameters K mapp , k cat and the deduced catalytic efficiency (k cat /K mapp ) of the purified group-V, -IB and -IIA PLA 2 s were determined using phosphatidylethanolamine (PE), phosphatidylcholine (PC) or phosphatidylserine (PS) as substrate. The three enzymes hydrolyze the zwiterionic PE and PC substrates more efficiently than anionic PS substrate.