Enhanced Synthesis, Purification, and Characterization of a Marine Bacterial Consortium-Derived Protease Enzyme With Destaining and Keratinolytic Activity.

Two marine-derived bacteria, Bacillus paralicheniformis (HR-1) and Bacillus haynesii (HR-5), were isolated from sediments and identified using 16S ribosomal RNA gene amplification and sequencing as well as biochemical analysis. The development of a bacterial consortium (HR-1 & HR-5) from these two bacteria was used to increase the production of the protease enzyme under various conditions, including fermentation media, carbon and nitrogen sources (1% w/v), different pH levels, incubation time, and the obtained enzyme, were detected using SDS-PAGE followed by purification. Bacterial consortium HR-1 & HR-5 exhibited maximum protease production (330.42 ± 4.47 U/mL) than the individual isolates HR-1 (156.32 ± 2.14 U/mL) and HR-5 (185.73 ± 5.14 U/mL) on supplementing peptone (1% w/v), 2.8% skim milk + N-broth, pH 9, and dextrose (1% w/v) after 48 h of incubation time. The purified enzyme showed increased activity at alkaline pH 9.0 and also in the presence of ions such as Ca⁺², Fe⁺³, Mg⁺², and Mn⁺². The purified protease obtained from the consortium HR-1 and HR-5 shows improved efficiency for stain removal from cloth as well as high keratinolytic efficiency for poultry feather degradation, making this enzyme suitable for industrial use, particularly in the textile and tannery sectors.