Production of Alkaline Protease from Bacillus Subtilis Isolated from Traditional Fermented Foods.

ABSTRACT

The demand for environmentally friendly and efficient biocatalysts has spurred significant interest in alkaline proteases, especially for their applications in the detergent, pharmaceutical, food, and leather industries. This study focuses on the isolation, screening, characterization, and optimization of alkaline protease production from Bacillus subtilis isolated from traditionally fermented foods. The study aims to harness indigenous strains from local fermented foods to enhance enzyme production and stability in an environmentally sustainable way. Samples of traditionally fermented foods were collected and screened for bacterial isolates, from which Bacillus subtilis was identified using colonial characteristics, gram staining, and biochemical tests. Initial screenings for alkaline protease production were conducted using skim milk agar plates, where the zone of clearance around bacterial colonies indicated protease activity. Enzyme production was optimized under varying conditions, including different temperatures, pH levels, carbon and nitrogen sources, and agitation speeds.

The optimal conditions for peak alkaline protease activity were found to include a pH range of 9-11 and a temperature of approximately 37°C. Enzyme activity was further characterized through assays that examined its stability across a range of pH and temperature conditions, revealing robust activity conducive to industrial applications. The findings suggest that Bacillus subtilis isolated from fermented foods has significant potential as a source of alkaline protease suitable for eco-friendly industrial processes. Optimized production conditions enhanced enzyme yield and stability, making the process cost-effective and scalable. This research highlights the value of utilizing locally sourced microorganisms, contributing to sustainable biotechnology practices while supporting industrial needs for efficient, resilient enzymes.