ABSTRACT
An aminopeptidase from the skeletal muscle of fish, Tilapia mossambica, was partially purified to 96-fold using salt precipitation, ion-exchange chromatography and molecular sieve chromatography. The enzyme showed optimum activity between pH 6.5-7.5 at 43 degrees C and Vmax and Km of 14.36 units/mg and 0.059 mM respectively with alanine beta-naphthylamide as the substrate. The aminopeptidase having a molecular weight of 305 kDa was activated by sulphydryl compounds and Co2+ and inhibited by bestatin, puromycin and metal chelators. Inhibition caused by metal chelators could be reversed by the addition of Co2+. Inclusion of L-amino acids, particularly isoleucine and leucine, in the assay medium caused inhibition of the enzyme activity. Substrate specificity together with inhibition and activation pattern indicated that the enzyme is alanine aminopeptidase.