PURIFICATION AND CHARACTERIZATION OF BILE SALT-ACTIVATED LIPASE FROM THE HEPATOPANCREAS OF RED-SEA BREAM, PAGRUS-MAJOR

A lipase was purified from the extract of the delipidated powder of re d sea bream hepatopancreas to near homogeneity by fractional precipita tion with ammonium sulfate and sequential chromatography on first anio n-exchange-, hydrophobic- and second anion-exchange columns followed b y gel filtration and anion-exchange HPLC. The final enzyme preparation showed a single band with an apparent molecular mass of approx. 64 kD a by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The pu rified enzyme had a pH optimum in the range of pH 7.0-9.0. Using rho-n itrophenyl myristate or triolein as a substrate? the enzyme required t he presence of sodium taurocholate or sodium cholate for its activity. No activity was observed in the presence of sodium deoxycholate. The enzyme preferentially hydrolyzed ethyl esters of polyunsaturated fatty acid, such as arachidonic acid and eicosapentaenoic acid which were r esistant to porcine pancreatic lipase. These results strongly suggest that the enzyme purified from the hepatopancreas of red sea bream is h omologous to mammalian bile salt-activated lipase.