Purified fish proteases with high specific activities and its process of production

Fish proteases isolated from cod fish viscera comprising an average specific Trypsin activity of 240±40 U/mg, Chymotrypsin activity of 4±2 U/mg, Collagenase activity of 0.04±0.02 U/mg and a Protease activity of 65±10 U/mg obtained by a process comprising:

• a) extracting crude enzyme from the cod fish viscera with a calcium chloride buffer at pH 7, filtrating and ultrafiltrating to obtain an ultrafiltrate;
• b) extracting the ultrafiltrate with an aqueous solution of CaCl2 having conductivity from 52 to 62 mS in a pH range of 7.2 to 8.2 followed by depth filtration to obtain a filtrate;
• c) purifying by hydrophobic interaction chromatography the filtrate obtained in step b) using as stationary phases an agarose base matrix with straight chain alkyl ligands or aryl ligands, and eluting with buffers of low salt content and then with an aqueous mixture of water miscible organic solvents and a polyol to obtain a purified fraction with trypsin activity;
• d) dialyzing the purified fraction obtained from step c);
• and
• e) freeze-drying the dialysate from step d) to obtained the fish proteases.