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Browsing UB - Resumos em livros de actas by Subject "Alcoholic fermentation"
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- Antimicrobial peptides from Saccharomyces cerevisiae induce physiological changes in Hanseniaspora guilliermondiiPublication . Branco, Patricia; Albergaria, Helena; Arneborg, NilsSaccharomyces cerevisiae secretes antimicrobial peptides (AMPs) during alcoholic fermentation that are active against other wine-related yeasts (e.g. Hanseniaspora guilliermondii) (Albergaria et al., 2010) and bacteria (e.g. Oenococcus oeni) (Osborne and Edwards, 2007). In the present study we assessed the physiological changes induced by those AMPs on sensitive yeast cells of Hanseniaspora guilliermondii, namely membrane permeability and intracellular pH (pHi) alterations. Membrane permeability was evaluated by staining cells with propidium iodide (PI) and pHi by the fluorescence ratio imaging microscopy (FRIM) technique (Guldfeldt and Arneborg, 1998). Results showed that after 20 min of incubation with inhibitory concentrations of AMPs, the average pHi of cells dropped from 6.5 to 5.4. After 8 h of incubation, 32% of the cells had lost their ĢpH (=pHi-pHext) and after 24 h that percentage rose to 77%. The culturability (plating) and viability (PI staining) of the sensitive yeast cells also decreased in the presence of the AMPs. After 24 h of exposure to AMPs, 61% of the cells were dead (PI-stained) and the number of viable cells fell from 1 ~105 to 1.5 CFU/ml, which means that virtually all cells (99.999%) became unculturable but a sub-population of 39% of cells remained in a viable but non-culturable (VBNC) state. However, those VBNC cells were able to recover their culturability after incubation at optimal growth conditions. Our study revealed that the mode of action of these AMPs seems to be primarily targeted to the cell membranes, reducing their permeability and preventing cells to maintain pH homeostasis.
- Saccharomyces cerevisiae and Dekkera bruxellensis interactions in alcoholic fermentations: growth and 4-ethylphenol productionPublication . Coutinho, Rute; Branco, Patricia; Monteiro, Margarida; Malfeito-Ferreira, Manuel; Albergaria, HelenaThe yeast Dekkera/Brettanomyces bruxellensis can cause enormous economic losses both in wine industry and fuel-ethanol processes due to production of phenolic off-flavour compounds and low ethanol productivities. In winemaking this microbial hazard is usually tackled by the use of chemical preservatives such as sulphur dioxide. In spite of this, D. bruxellensis strains are frequently found in wines at low levels (ca 103 cells/ml) where they can metabolise residual sugars producing phenolic off-flavours compounds, such as 4-ethyl phenol. In the present work we investigated S. cerevisiae and D. bruxellensis interactions during alcoholic fermentations and evaluated the effectiveness of antimicrobial peptides secreted by S. cerevisiae to prevent growth of the main wine spoilage yeast and the production of 4-ethylphenol. Several fermentations were performed with single cultures of D. bruxellensis and mixed cultures of S. cerevisiae and D. bruxellensis, both in synthetic grape juice (SGJ) and grape must. Yeast growth (culturability and viability) and fermentation performance (i.e. sugars consumption, ethanol and 4-ethylphenol production) of those fermentations was accessed by different methods, namely by florescence in situ hybridization and flow cytometry. Results showed that S. cerevisiae significantly reduced the growth of D. bruxellensis and the production of 4-ethylphenol both in SGJ and grape must fermentations performed with mixed cultures. Moreover, our work also showed that antimicrobial peptides secreted by S. cerevisiae are effective to prevent growth of D. bruxellensis and production of phenolic off-flavor compounds in wine.