Enzymatic Characterization of Alkaline Protease from a Novel Microorganism Isolated from a Halophilic Environment

Introduction: Microbial enzymes, especially bacterial alkaline proteases, are essential to many industrial processes, including the manufacturing of detergents, food processing, bioremediation, medicines, and tanneries. Because of its possible industrial benefits, this study focuses on the purification and characterisation of a halophilic alkaline protease generated by Bacillus sp. strain SPII-4. Methods: The bacteria SPII-4's 16S rRNA gene was sequenced and subjected to phylogenetic analysis. Casein was used as a substrate to measure the extracellular crude enzyme's proteolytic activity. Temperature, pH, salinity, metal ions, and chemical solvents were all used to assess enzymatic activity. Every experiment was run in triplicate, and Student's t-tests with unequal variances in Microsoft Excel were used to assess statistical significance. Results: The 16S rRNA sequencing matched Bacillus sp. strain 2S4 with 100% identity and 99% coverage. The protease was most active at 40°C, in the alkaline pH range of 9-11, and at concentrations of up to 5% NaCl. The enzyme had the maximum activity (14.64 U/mg) among the metal ions examined when BaCl2 was present. Additionally, it maintained its activity in the presence of the surfactant Triton-X and in a variety of chemical solvents. The observed differences were statistically significant (p < 0.001) Discussion: The Bacillus SPII-4 protease showed exceptional stability and activity in the presence of surfactants and solvents, as well as in extremely high and low salinity and alkalinity conditions. These characteristics point to the protease's potential for widespread industrial use and are in line with research on related halophilic bacterial enzymes. To maximize its commercial usage, more purification and scale-up research are necessary. Conclusion: Bacillus sp. SPII-4's halo-alkaline protease exhibits considerable industrial promise because of its stability in conditions that are high in salt, alkalinity, and solvents. These qualities make it a viable option for use in the food, detergent, and pharmaceutical sectors as well as in bioremediation.