Abstract

Aslam Farheen*, Siraj Asma, Tabassum Savera and Abdul Manan Laiba

ABSTRACT

Collagenase play a crucial role in various industrial applications, including food processing and biomedical research. This research focuses upon the isolation, purification and in-silico analysis of collagenase producing bacteria. Collagen is an abundant protien in many higher organisms and due to its unique amino acid compostion it can only be cleaved by speicialized proteases like collagenase enzyme secreted by some bacteria such as Baccilus Cereus and Clostrodium Histolyticum. In this research collagenase was purified from beef by repeated streaking method. This research work concludes that the bacterial collagenase displayed best activty at pH 8, with optimum substrate concentration of 2% gelatin and the optimum time of incubation is 48h. The results show that maximum enzyme activity was recorded with Peptone, Maltose, and MgSO4 which were the best sources for nitrogen, carbon, and metal ions respectively. PredictProtein tool was utilized to determine the secondary structure of enzyme and it categorized the protein into 33.26% alpha helices, 16.48% beta strands, and 50.26% loops/turns. Phyre2 and Swiss model were used for determining 3D structure of the enzyme.