PURIFICATION OF CHYMOTRYPSIN FROM FISH WASTE USING REVERSE MICELLES
Reverse micelles systems AOT/isooctane was used for the concentration chymotrypsin from crude aqueous extract of red perch (intestine). The effects of pH and AOT concentration in the forward extraction step and pH and KCl concentration in the backward extraction step on the enzyme activity, purification fold and recovery yield were studied. The optimum conditions for the forward extraction were AOT concentration 20 mM and pH of 7.0 and optimum conditions for backward extraction were KCl concentration 1.0 M and pH of 7.5 which gave a good recovery yield (102.24%) and a purification (32.24-fold). The addition of 15% v/v alcohol in backward extraction dramatically improved recovery yield by 4.5 times and purification by 2.5 times. The enzyme activity and recovery yield obtained using reverse micelles method under its optimal conditions were 2 fold higher than those obtained using the ammonium sulphate precipitation method, while purification fold were 3 fold higher.