Bioenergia - UB
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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.
- Design optimisation of five pilot-scale two-stage vertical flow-constructed wetlands for piggery wastewater treatmentPublication . Karan, N.; Gogoi, Jayanta; Ganguly, Anasuya; Brito, António; Marques dos Santos, C.; de Oliveira Corrêa, Diego; Gouveia, Luisa; Mutnuri, SrikanthABSTRACT: With growing pig farming, sustainable piggery wastewater treatment methods are essential for environmental protection. This study evaluated five pilot-scale two-stage vertical flow-constructed wetlands (VFCWs) with varying configurations of aeration, plantation, and saturation zones. Three VFCW configurations (1VFCW, 2VFCW, and 3VFCW) were unsaturated, while 4VFCW and 5VFCW were saturated in the second stage (up to 60 and 90 cm, respectively). The 5VFCW featured a stacked configuration with no space between its two stages. Passive aeration was selectively applied in 2VFCW, 3VFCW, 4VFCW, and 5VFCW, while plants were present in most configurations except the control. Saturated 4VFCW achieved the highest removal efficiency for TN (77.03 ± 16.24%) and NO3− (46.06 ± 45.96%), while the stacked 5VFCW showed the highest removal for chemical oxygen demand (COD) (94.17 ± 4.85%) and Total ammoniacal nitrogen (TOC) (86.35 ± 6.78%). Unsaturated 1VFCW excelled in TAN removal (98.89 ± 0.33%), and the control system (C) showed the highest removal efficiency for PO43− (90.38 ± 6.52%) and TOC (87.52 ± 9.83%). Overall, 4VFCW emerged as the most balanced and effective system, supported by an optimal combination of aerobic and anaerobic conditions that facilitated sequential nitrification and denitrification, along with an extended hydraulic retention time due to saturation.
- Influence of Inhibitors Generated in Lignocellulosic Hydrolysates from Group of Acids on the Growth of Strains TG1 and Tuner of Escherichia coliPublication . Gaspar, Suelen S.; Alves Ferreira Caturra, Júnia Aparecida; Moniz, Patricia; Silva-Fernandes, Talita; Silvestre, Adriana I. R; Torrado, Ivone; Pesce, Gaetano R.; Carvalheiro, Florbela; Duarte, Luís; Fernandes, Maria da ConceiçãoABSTRACT: Concerns over fossil fuels are of increasing interest in biorefineries that utilize lignocellulosic residues. Besides sugars, inhibitors are formed during biomass pretreatment, including acetic acid (AI) and formic acid (FI), which can hinder microbial fermentation. The TG1 and Tuner strains of Escherichia coli were subjected to various acid concentrations. Samples were taken during fermentation to monitor growth, sugar consumption, biomass yield, and product yield. With increasing AI, the TG1 strain maintained stable growth (0.102 1/h), while xylose consumption decreased, and product formation improved, making it better suited for high-acetic-acid industrial applications. In contrast, the Tuner strain performed better under low-inhibitor conditions but suffered metabolic inhibition at high AI levels, compensating by increasing lactic acid production-an adaptation absent in TG1. However, Tuner showed greater resistance to formic acid stress, sustaining higher growth and ethanol production, whereas TG1 experienced a greater metabolic decline but maintained stable acetic acid output. Both strains experienced inhibition in formic acid metabolism, but TG1 had a higher yield despite its lower overall robustness in formic acid conditions. The use of TG1 for value-added compounds such as ethanol or formic acid may help to avoid the use of chemicals that eliminate acetic acid. Tuner could be used for lactic acid production, especially in hydrolysates with under moderate concentration.
- Monitoring C. vulgaris Cultivations Grown on Winery Wastewater Using Flow CytometryPublication . Lopes da Silva, Teresa; Silva, Thiago; França, Bruna Thomazinho; Ribeiro, Belina; Reis, AlbertoABSTRACT: Winery wastewater (WWW), if released untreated, poses a serious environmental threat due to its high organic load. In this study, Chlorella vulgaris was cultivated in diluted WWW to assess its suitability as a culture medium. Two outdoor cultivation systems-a 270 L raceway and a 40 L bubble column-were operated over 33 days using synthetic medium (control) and WWW. A flow cytometry (FC) protocol was implemented to monitor key physiological parameters in near-real time, including cell concentration, membrane integrity, chlorophyll content, cell size, and internal complexity. At the end of cultivation, the bubble column yielded the highest cell concentrations: 2.85 x 106 cells/mL (control) and 2.30 x 106 cells/mL (WWW), though with lower proportions of intact cells (25% and 31%, respectively). Raceway cultures showed lower cell concentrations: 1.64 x 106 (control) and 1.54 x 106 cells/mL (WWW), but higher membrane integrity (76% and 36% for control and WWW cultures, respectively). On average, cells grown in the bubble column had a 22% larger radius than those in the raceway, favouring sedimentation. Heterotrophic cells were more abundant in WWW cultures, due to the presence of organic carbon, indicating its potential for use as animal feed. This study demonstrates that FC is a powerful, real-time tool for monitoring microalgae physiology and optimising cultivation in complex effluents like WWW.
- Recovery of Nd3+ and Dy3+ from E-Waste Using Adsorbents from Spent Tyre Rubbers: Batch and Column Dynamic AssaysPublication . Nogueira, Miguel; Matos, Inês; Bernardo, Maria; Pinto, Filomena; Fonseca, Isabel Maria; Lapa, NunoABSTRACT: This paper investigates the use of spent tyre rubber as a precursor for synthesising adsorbents to recover rare earth elements. Through pyrolysis and CO2 activation, tyre rubber is converted into porous carbonaceous materials with surface properties suited for rare earth element adsorption. The study also examines the efficiency of leaching rare earth elements from NdFeB magnets using optimised acid leaching methods, providing insights into recovery processes. The adsorption capacity of the materials was assessed through batch adsorption assays targeting neodymium (Nd3+) and dysprosium (Dy3+) ions. Results highlight the superior performance of activated carbon derived from tyre rubber following CO2 activation, with the best-performing adsorbent achieving maximum uptake capacities of 24.7 mg.g(-1) for Nd3+ and 34.4 mg.g(-1) for Dy3+. Column studies revealed efficient adsorption of Nd3+ and Dy3+ from synthetic and real magnet leachates with a maximum uptake capacity of 1.36 mg.g(-1) for Nd3+ in real leachates and breakthrough times of 25 min. Bi-component assays showed no adverse effects when both ions were present, supporting their potential for simultaneous recovery. Furthermore, the adsorbents effectively recovered rare earth elements from e-waste magnet leachates, demonstrating practical applicability. This research underscores the potential of tyre rubber-derived adsorbents to enhance sustainability in critical raw material supply chains. By repurposing waste tyre rubber, these materials offer a sustainable solution for rare earth recovery, addressing resource scarcity while aligning with circular economy principles by diverting waste from landfills and creating value-added products.
- Sustainable Production of Poly(3-hydroxybutyrate) Using Eucalyptus Bark: Integration with Green Downstream ProcessingPublication . Matias, João; Rodrigues, Thomas; Torres, Cristiana A. V.; Marques, Susana; Ribeiro, Belina; Gírio, Francisco; Reis, Maria A.; Freitas, FilomenaABSTRACT: This study integrates the valorization of a lignocellulose material into poly(3-hydroxybutyrate), P(3HB), with biopolymer extraction from bacterial cells with the enzyme alcalase. The work focused on Burkholderia thailandensis DSM 13276 as the P(3HB) producer and on eucalyptus bark, a byproduct from the pulp industry, as the sole feedstock for bacterial cultivation. The eucalyptus bark was hydrolyzed by a cellulolytic enzymatic cocktail following steam explosion and further subjected to ultrafiltration for enzyme recovery. The resulting hydrolysate supported good cell growth, achieving a cell dry weight of 7.67 +/- 0.16 g/L within 72 h of cultivation, and high P(3HB) content (60.0 +/- 2.19 wt %) in the bacterial cells, clearly favoring biopolymer synthesis over cell growth, as demonstrated by the polymer and growth yields (0.190 gP(3HB)/gsugar and 0.026 gX/gsugar, respectively). High extraction efficiency (96%) and biopolymer purity (100 +/- 3.38%) were reached by enzymatic treatment, resulting in a sample with properties aligned with those of commercial P(3HB) in terms of molecular mass distribution, crystallinity, and thermal properties. These findings demonstrate the successful use of a sustainable feedstock together with the application of environmentally friendly technologies based on the use of enzymes for both lignocellulosic saccharification and biopolymer recovery to develop high-quality bioplastics, advancing the goals of a circular bioeconomy.
