Extracellular Protease Production, Optimization, and Partial Purification from Bacillus nakamurai PL4 and its Applications

Abstract

The major goal of the study was to isolate bacteria synthesizing protease enzyme from a soil sample taken in Dandeli, Karnataka, India. Furthermore, screening, production, and optimization of medium components for maximum protease activity, partial purification of crude enzymes, and application of protease produced by Bacillus nakamurai were carried out. At 72 hrs and pH 6, the optimum incubation time, pH, and temperature were evaluated (acidic and thermophilic). As a substrate, casein was employed. Plackett-Burman screening was performed, and KH2PO4, xylose, MnCl2, and peptone were discovered to be essential components in protease production media. Following the production and optimization processes, partial purification was performed using ammonium sulfate precipitation, with the maximum protease activity at 60% ammonium sulfate, and further dialysis was performed using precipitated enzyme, yielding enzyme activity of 0.747 U/mL. The protease enzyme proved effective at removing egg yolk stains as well as degrading (dehairing) chicken feathers and hairs from goat skin. Bacillus nakamurai PL4 can be utilized for the industrial-scale production of proteases to fulfill current demands. Thus, such optimized parameters can optimize protease production and their application across various industries.