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- Applicability Assessment of a Microbial Proteolytic Fermentation Broth to Leather Processing and Protein Stain RemovalPublication . Lageiro, Maria Manuela; Moura, Maria João; Simões, Fernanda; Alvarenga, Nuno; Reis, AlbertoABSTRACT: Microbial proteases are fundamental towards the eco-sustainability of proteolysis at the industrial scale. A proteolytic broth was obtained from a bioreactor fermentation of a proteolytic Bacillus strain isolated from an industrial alkaline bath. Broth proteolytic activity was applied to leather tanning and to the removal of protein stains. The hide tanned with the microbial proteolytic fermentation broth showed better physical properties than the one tanned with commercial pancreatic proteases of the same activity (780 LVU). Proteinaceous stains on cotton fabric were removed more efficiently using the Bacillus proteolytic broth than water or a commercial detergent. Blood and egg yolk disappeared in less than 30 min. The removal of soya and English sauce stains was even faster. Broth proteolytic activity was characterised by caseinolytic (5200 LVU), collagenolytic (10.0 U mg-1), elastolytic (3.7 U mg-1), and keratinolytic (0.7 U mg-1) activities, which were compared with those of a commonly used commercial protease. Alkaline protease activity in the broth was demonstrated by a 20% increase in caseinolytic activity from pH 5 to 8. Besides the demonstrated applications in the leather and detergent industries, the produced alkaline microbial proteases can also be used in the treatment of proteinaceous wastes and effluents, offering potential environmental benefits reinforcing and impacting the bioeconomy.