Isolation and Characterization of Proteases From the Viscera of Labeo Rohita
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Date
2011
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Abstract
A preliminary study on the isolation and characterization of proteases from the crude extract of
visceral organs o\' Labeo rohita were carried out. In order to estimate the number of proteases
and their mass, the crude protease extract were separated by SDS-PAGE and then the activity
was revealed by Zymogiain. The crude enzyme extract showed two clear bands on casein and
gelatin Zymogram, which indicated the presence of two major proteases. The optimal pH and
temperature of the protea,ses obtained in crude extract wore 8.0 and 40" C respectively, using
casein as a substrate. Furthermore, the crude enzymes uere characterized by protease inhibitors
namely phenylmethylsulphony l fluride (PMSF). EDTA and p mercaptoethanol. in which the
protease enzymes were strongly inhibited by PMSF (serine protease inhibitor). In addition, the
enzymes were found to be highly active in the presence of activators like Mg"^ Ca^\ Mn^"^ and
Na*. The protease kinetic constants, K„, and V,„„^ of the crude enzymes for casein, were
0.65 niM and 2.5 U/ml/min respectively. With respect to properties of the enzyme and its
capacity for degradation of different protein sources, these proteases finds potential application
for waste treatment used in detergent and leather industry