Biochemical characterization of a semi-purified aspartic protease from sea catfish Bagre panamensis with milk-clotting activity.

Abstract

Pepsin from stomach of Bagre panamensis was semi-purified and biochemically characterized. The acid proteolytic activity and purification fold were 3875U/mg protein and 91.85, respectively, after purification process. The optimum pH and temperature for semi-purified protease were 2-3 and 65°C, respectively. The enzyme activity was stable after heating proteases at 50°C for 120min, but only 30% residual activity was detected after heating at 65°C for 30min. SDS-PAGE analysis showed two proteins bands after dialysis (26.1 and 38.6kDa). Only the band of 38.6kDa had proteolytic activity, which was inhibited using pepstatin A. Organic solvents, surfactants and reducing agents affect the proteolytic activity at different extent; however, metal ions or EDTA have no impact on protease activity. The semi-purified protease exhibited milk coagulant activity, with a maximum activity at 45°C. The obtained results highlight the potential biotechnological use of B. panamensis pepsin.