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The potential of cold-active proteases are greater than thermostable proteases in view of their high catalytic efficiency at low and moderate temperatures. However, these enzymes along with their producing organisms are remains unexplored. The aim of present work was to isolate, purify and characterize novel cold-active extracellular proteases and explore its biotechnological potential. As a result, two potential cold-adapted bacteria viz. C. luteum and S. maltophilia are isolated that producing cold-active metallo-proteases and alkaline protease, respectively. The enzyme from S. maltophilia showed excellent compatibility with commercial detergents and exhibited high efficiency for the removal of blood & grass stains at low temperature. The present dissertation is specially meant for undergraduate/postgraduate students and research scholars of biotechnology/biochemistry and applied sciences; and will be very helpful in developing interest to pursue studies and research in enzyme biotechnology. Key features of the book include theoretical & practical approach to understand production, purification and characterization of cold-active proteases and their biotechnological applications.