A Highly Active Keratinase from Bacillus sp. FJ-3-16 for Sustainable Feather Waste Valorization and Eco-Friendly Industrial Applications.

Keratinous biomass, such as feathers, wool, and hair, poses environmental challenges due to its insoluble and recalcitrant nature. In this study, we identified, purified and comprehensively characterized a previously uncharacterized extracellular alkaline keratinase, KerFJ, secreted by Bacillus sp. FJ-3-16, with broad industrial application potential. KerFJ was produced at high yield (1800 U/mL) in an optimized cost-effective medium and purified to homogeneity using ion-exchange chromatography. The enzyme exhibited optimal activity at pH 9.5 and 55 °C, with remarkable alkaline and thermal stability, and high tolerance to surfactants, oxidants, and metal ions. Sequence analysis revealed that KerFJ is a member of the serine peptidase S8 family, with a molecular weight of ~27.5 kDa. It efficiently degraded native keratin substrates, achieving 70.3 ± 2.1% feather, 39.7 ± 1.8% wool, and 15.4 ± 1.2% hair degradation, and the resulting feather hydrolysates exhibited strong antioxidant activities. KerFJ also demonstrated excellent compatibility with commercial detergents and enabled effective stain removal from fabrics without damage. Moreover, both laboratory- and pilot-scale trials showed that KerFJ facilitated non-destructive dehairing of sheep, donkey, and pig skins while preserving collagen integrity. These results highlight KerFJ as a robust and multifunctional biocatalyst suitable for keratin waste valorization, eco-friendly leather processing, and detergent formulations.