Efficient co-fermentation of glucose and xylose remains a critical hurdle in second-generation bioethanol production. In this study, we evaluated two non-Saccharomyces yeasts—Wickerhamomyces anomalus UEMG-LF-Y2 and Diutina rugosa UEMG-LF-Y4—under mixed-sugar conditions. D. rugosa exhibited superior xylose metabolism and ethanol productivity, achieving a maximum volumetric productivity (Q_P) of 0.55 g/L·h in a medium containing 20 g/L glucose and 40 g/L xylose. Its highest ethanol yield (Y_P/S) reached 0.45 g EtOH/g sugar, comparable to results from engineered Saccharomyces cerevisiae strains. By contrast, W. anomalus displayed lower ethanol yields (0.24–0.34 g/g) and greater sensitivity to catabolite repression induced by 2-deoxyglucose (2-DG). Xylose consumption by D. rugosa exceeded 80% in high-xylose media, while W. anomalus left residual xylose under all tested conditions. A strong inverse correlation (r < −0.98) between ethanol accumulation and xylose uptake was observed, especially for W. anomalus, indicating ethanol-induced inhibition as a key challenge. These findings highlight the potential of D. rugosa as a robust non-Saccharomyces platform for lignocellulosic bioethanol processes, whereas W. anomalus may benefit from further metabolic or process optimizations. Future research should address ethanol tolerance, inhibitory byproducts, and large-scale feasibility to fully exploit these strains for second-generation bioethanol production.

Pre- and post-milking teat antisepsis is one of the most effective methods for reducing the incidence of mastitis. Plant extracts have proven effective in reducing microorganisms, providing a natural alternative for antisepsis, along with additional benefits for teat health. This study aimed to evaluate the antimicrobial and antiseptic effects of two newly formulated products, used as pre-dipping and post-dipping agents, respectively, based on plant extracts from papain (Carica papaya), aloe vera (Aloe barbadensis), andiroba (Carapa guianensis), copaiba (Copaifera officinalis), tea tree (Melaleuca alternifolia), and barbatimão (Stryphnodendron barbatiman). Minimum inhibitory concentration tests were performed in vitro, along with microbiological analyses of different bacterial groups and in vivo compliance assessments to evaluate the antimicrobial activity of the formulations, as well as the molecular identification of Staphylococcus aureus. The results demonstrated that the products were effective in vitro against the main pathogens that cause mastitis, including Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. In the in vivo assays, similar performance was observed between the tested products and the control products, indicating that the plant-extract-based formulations, applied as pre-dipping and post-dipping agents, effectively reduced the microbial load on the teats. The effects were equivalent to those of hydrogen-peroxide- and iodine-based products, being able to reduce the total microbial load by more than 80%.

Lectins are glycan-binding proteins involved in diverse biological processes and have gained attention for their potential applications in biotechnology and immunomodulation. BOL (Brassica oleracea lectin) is a unique ~34 kDa lectin isolated from Brassica oleracea var. botrytis, composed exclusively of TRAF-like domains, where TRAF stands for tumor necrosis factor receptor–associated factor. To overcome the limitations of plant-based extraction, we aimed to produce recombinant BOL in Escherichia coli. Various strains and expression vectors were tested under distinct induction conditions to optimize solubility and yield. While expression using pET28a was unsuccessful, GST-tagged BOL was efficiently expressed in E. coli BL21-R3-pRARE2(DE3) and purified using affinity chromatography. Functional assays demonstrated that the recombinant protein retained lectin activity, as evidenced by hemagglutination of goat erythrocytes. Protein identity was confirmed by MALDI-TOF/TOF mass spectrometry, with tryptic peptides matching the BOL lectin sequence in the National Center for Biotechnology Information (NCBI) database. Our findings highlight the importance of codon optimization, temperature modulation, and fusion tag selection for the successful expression of eukaryotic lectins in E. coli. This work provides a platform for future functional studies of BOL and supports its potential application in plant immunity and biomedical research.