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Lageiro, Maria Manuela

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  • Proteolytic Bacillus sp. Isolation and Identification from Tannery Alkaline Baths
    Publication . Lageiro, Maria Manuela; Simões, Fernanda; Alvarenga, Nuno; Delgado dos Reis, Alberto José
    ABSTRACT: The application of microbial alkaline proteases holds significant potential for eco-sustainable industrial processes by reducing chemical usage and lowering the costs of effluent treatment. In the search for novel proteases with industrial relevance, several microbial strains were isolated from alkaline baths of the Portuguese tannery agroindustry. The most promising protease-producing strains were selected for identification and further study. Two isolates demonstrated the highest proteolytic activity, reaching 0.51 +/- 0.01 U mL-1 and 0.70 +/- 0.01 U mL-1 after 7.5 h of submerged cultivation in nutrient broth. Based on API biochemical tests, molecular biology techniques, and GC-FAME analysis of membrane lipids, the isolates were identified as Bacillus subtilis and incorporated into INIAV's collection of industrial microbial cultures as B. subtilis CCMI 1253 (BMR2) and B. subtilis CCMI 1254 (BMR1). The most promising protease producer, B. subtilis CCMI 1253 (BMR2), exhibited a maximum specific growth rate of 0.88 +/- 0.10 h-1. The proteases produced exhibited good extracellular proteolytic activity, with adaptability to industrial conditions, indicating their suitability for agroindustry applications such as leather making, detergent formulations and the treatment of effluents and protein residues. The results support the potential of microbial proteases as valuable tools in the bioeconomy and green chemistry.
  • Applicability Assessment of a Microbial Proteolytic Fermentation Broth to Leather Processing and Protein Stain Removal
    Publication . Lageiro, Maria Manuela; Moura, Maria João; Simões, Fernanda; Alvarenga, Nuno; Reis, Alberto
    ABSTRACT: 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.