The results showed that
the collagenase enzyme was produced as a latent enzyme
and was activated with bovine trypsin and potassium
thiocyanate (KSCN). The enzyme activity was
affected by pH and temperature. Optimal enzyme
activities were found at 35˚C and a pH of 7.5 when
insoluble collagene type I was used as substrate and
the liberated amino acids were measured in relation
to L-leucine in the presence of ninhydrin. The enzyme
activity was completely inhibited by the action of
ethylenediaminetetraacetic acid (EDTA) suggesting
that the collagenase enzyme isolated from the fish
waste is a metalloproteinase enzyme requiring metal
ions for enzyme activity. Dialysis against KSCN decreased
the enzyme total activity and increased its
specific activity. Sodium dodecyl sulphate polyacrylamide
gel electrophoresis (SDS-PAGE) indicated that
the purified procollagenase enzyme have only one
band at molecular weight of 50 kilodaltons (kDa).
When the enzyme was cleaved with trypsin, it was
found to consist of two subunits: a large unit with a
molecular weight of 50 kDa and a small unit with a
molecular weight of 10 kDa.
Keywords: Fish Waste; Collagenase Enzyme;
Extraction; Purification; Inhibitors; Activators