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  • 1,3-Propanediol production in a two-step process fermentation from renewable feedstock
    Publication . Mendes, Filipa Soares; González-Pajuelo, Maria; Cordier, Hélène; François, Jean M.; Vasconcelos, Isabel
    In this work, the production of 1,3-propanediol from glucose and molasses was studied in a two-step process using two recombinant microorganisms. The first step of the process is the conversion of glucose or other sugar into glycerol by the metabolic engineered Saccharomyces cerevisiae strain HC42 adapted to high (>200 g l−1) glucose concentrations. The second step, carried out in the same bioreactor, was performed by the engineered strain Clostridium acetobutylicum DG1 (pSPD5) that converts glycerol to 1,3-propanediol. This two-step strategy led to a flexible process, resulting in a 1,3-propanediol production and yield that depended on the initial sugar concentration. Below 56.2 g l−1 of sugar concentration, cultivation on molasses or glucose showed no significant differences. However, at higher molasses concentrations, glycerol initially produced by yeast could not be totally converted into 1,3-propanediol by C. acetobutylicum and a lower 1,3-propanediol overall yield was observed. In our hand, the best results were obtained with an initial glucose concentration of 103 g l−1, leading to a final 1,3-propanediol concentration of 25.5 g l−1, a productivity of 0.16 g l−1 h−1 and 1,3-propanediol yields of 0.56 g g−1 glycerol and 0.24 g g−1 sugar, which is the highest value reported for a two-step process. For an initial sugar concentration (from molasses) of 56.2 g l−1, 27.4 g l−1 of glycerol were produced, leading to 14.6 g l−1 of 1.3-propanediol and similar values of productivity, 0.15 g l−1 h−1, and overall yield, 0.26 g g−1 sugar.
  • Relationship between nitrogen content in grape volatiles, namely heavy sulphur compounds, in wines
    Publication . Moreira, Nathalie; Pinho, Paula Guedes de; Santos, Cristina; Vasconcelos, Isabel
    Ammonium salts were added to white grape musts, before alcoholic fermentation, in order to evaluate their influence on the heavy sulphur compound and aliphatic higher alcohol composition of resulting wines. Six grape musts were used (Trajadura, Pedernã, Loureiro, Azal Branco, Avesso and Alvarinho). Ammonium supplementation of Trajadura and Pedernã grape musts, with the highest nitrogen level, did not influence the content of heavy sulphur compounds and aliphatic higher alcohols in wines; however, the addition of ammonium salts to grape musts with low initial nitrogen content, such as Loureiro, Azal Branco and Avesso, led to a higher production of 1-propanol and a lower production of isoamyl alcohols and sulphur compounds, e.g. S-methyl thioacetate, 2-mercaptoethanol, acetic acid-3-(methylthio)propyl ester, 3-mercapto-1-propanol, 4-(methylthio)-1-butanol, 3-(ethyithio)-1-propanol. 3-methylthiopropionic acid and N-3-(methylthiopropyl)acetamide. For Alvarinho grape must, a decrease in sulphur compound concentrations in wines was only observed for 3-methylthiopropionic acid, acetic acid-3-(methylthio)propyl ester and 2-mercaptoethanol.
  • Characterizing the potential of the non-conventional yeast Saccharomycodes ludwigii UTAD17 in winemaking
    Publication . Esteves, Marcos; Barbosa, Catarina; Vasconcelos, Isabel; Tavares, Maria João; Mendes-Faia, Arlete; Mira, Nuno Pereira; Mendes-Ferreira, Ana
    Non-Saccharomyces yeasts have received increased attention by researchers and winemakers, due to their particular contributions to the characteristics of wine. In this group, Saccharomycodes ludwigii is one of the less studied species. In the present study, a native S. ludwigii strain, UTAD17 isolated from the Douro wine region was characterized for relevant oenological traits. The genome of UTAD17 was recently sequenced. Its potential use in winemaking was further evaluated by conducting grape-juice fermentations, either in single or in mixed-cultures, with Saccharomyces cerevisiae, following two inoculation strategies (simultaneous and sequential). In a pure culture, S. ludwigii UTAD17 was able to ferment all sugars in a reasonable time without impairing the wine quality, producing low levels of acetic acid and ethyl acetate. The overall effects of S. ludwigii UTAD17 in a mixed-culture fermentation were highly dependent on the inoculation strategy which dictated the dominance of each yeast strain. Wines whose fermentation was governed by S. ludwigii UTAD17 presented low levels of secondary aroma compounds and were chemically distinct from those fermented by S. cerevisiae. Based on these results, a future use of this non-Saccharomyces yeast either in monoculture fermentations or as a co-starter culture with S. cerevisiae for the production of wines with greater expression of the grape varietal character and with flavor diversity could be foreseen. View Full-Text
  • Volatile compounds contribution of Hanseniaspora guilliermondii and Hanseniaspora uvarum during red wine vinifications
    Publication . Moreira, N.; Pina, C.; Mendes, F.; Couto, J.A.; Hogg, T.; Vasconcelos, I.
    This work aims to investigate the contribution of a selected non-Saccharomyces yeast species, Hanseniaspora guilliermondii, to higher alcohols, esters, fatty acids and heavy sulphur compounds composition of red wine. Red grape must vinifications of 100 l were performed and an inoculated fermentation with H. guilliermondii was compared to a spontaneous fermentation. The presence of apiculate yeasts was observed in both fermentations; however, Hanseniaspora uvarum was the only apiculate yeast isolated from the spontaneous fermentation. Apiculate yeasts dominated the fermentation until an ethanol concentration of 6% (v/v) was attained and remained in considerable high levels for an ethanol concentration of 12.5% (v/v). The grape must inoculated with H. guilliermondii led to the production of wine with higher concentrations of 1-propanol, 2-phenylethyl acetate and 3-(methylthio)propionic acid, and lower amounts of ethyl hexanoate, pentanoic acid, free fatty acids, 2-methyltetrahydrothiophen-3- one and acetic acid-3-(methylthio)propyl ester, than wine resulting from the spontaneous fermentation. The present study shows that the use of specific apiculate yeasts in grape must fermentations may lead to the production of wines with different chemical profiles, emphasising the importance of Hanseniaspora yeasts as mixed starter cultures with Saccharomyces cerevisiae, in winemaking.
  • Volatile sulphur compounds composition of monovarietal white wines
    Publication . Moreira, Nathalie; Pinho, Paula Guedes de; Santos, Cristina; Vasconcelos, Isabel
    The sulphur compounds composition of wines produced experimentally from six white cultivars (Alvarinho, Loureiro, Trajadura, Pedernã, Azal Branco and Avesso) was evaluated during two consecutive vintages. Results show that wines could be differentiated according to their sulphur compounds content. In general, Loureiro, Trajadura and Pedernã cultivars led to wines with low concentrations of sulphur compounds; however, Loureiro wines were characterised by significant amounts of dimethyl sulphone, whereas Trajadura wines possessed a high content of 3-(methylthio)propyl acetate and 4-(methylthio)-1-butanol. Alvarinho and Avesso wines showed high levels of S-methyl thioacetate, 3-mercapto-1-propanol, 3-(ethylthio)-1-propanol and 3-methylthiopropionic acid. Significant amounts of 2-methyltetrahydrothiophen-3-one, cis- and trans-2-methyltetrahydrothiophen-3-ol were also found in Avesso wines. Azal Branco wines were low in 3-(methylthio)propyl acetate and 2-(methylthio)ethanol, and high contents in S-methyl thioacetate, 3-mercapto-1-propanol and 2-mercaptoethanol. A linear discriminant analysis of sulphur compounds levels showed a differentiation of wines according to their varietal origin.
  • Effect of malolactic fermentation on the volatile profile of wines
    Publication . Moreira, N.; Ruão, P.; Couto, J. A.; Vasconcelos, I.; Pinho, P. Guedes de
  • H. guilliermondii impacts growth kinetics and metabolic activity of s. cerevisiae: the role of initial nitrogen concentration
    Publication . Lage, Patrícia; Barbosa, Catarina; Mateus, Beatriz; Vasconcelos, Isabel; Mendes-Faia, Arlete; Mendes-Ferreira, Ana
    Non-Saccharomyces yeasts include different species which comprise an ecologically and biochemically diverse group capable of altering fermentation dynamics and wine composition and flavour. In this study, single- and mixed-culture of Hanseniaspora guilliermondii and Saccharomyces cerevisiae were used to ferment natural grape-juice, under two nitrogen regimes. In single-culture the strain H. guilliermondii failed to complete total sugar breakdown even though the nitrogen available has not been a limiting factor of its growth or fermentative activity. In mixed-culture, that strain negatively interfered with the growth and fermentative performance of S. cerevisiae, resulting in lower fermentation rate and longer fermentation length, irrespective of the initial nitrogen concentration. The impact of co-inoculation on the volatile compounds profilewasmore evident in thewines obtained from DAP-supplementedmusts, characterised by increased levels of ethyl and acetate esters, associated with fruity and floral character of wines. Moreover, the levels of fatty acids and sulphur compounds which are responsible for unpleasant odours that depreciate wine sensory quality were significantly lower. Accordingly, data obtained suggests that the strain H. guilliermondii has potential to be used as adjunct of S. cerevisiae in wine industry, although possible interactions with S. cerevisiae still need to be elucidated.
  • Varietal thiols in white wines from portuguese grape varieties
    Publication . Ferreira, D.; Moreira, N.; Vasconcelos, I.; Silva, J. M. Ricardo da; Pinho, P. Guedes de
  • Varietal thiols in white wines from portuguese grape varieties
    Publication . Ferreira, D.; Moreira, N.; Vasconcelos, I.; Ricardo-da-Silva, J. R.; Pinho, P. Guedes de
    Wines from Antão Vaz, Arinto, Fernão Pires, Verdelho da Madeira and Viosinho grape cultivars, from 2009 and 2010 vintages, were studied in order to characterize the volatile thiols profile, namely 4-mercapto-4-methyl-pentan-2-one (4MMP), 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA). Volatile thiols profile of these wines was also compared to a Sauvignon blanc wine produced in Portugal. Besides sensory analysis, all wines were analyzed using GC coupled to different detectors, namely FID, FPD, MS and Olfactometry (GC-O). In all wines, GC-O analyses showed the existence of two olfactory zones related to the presence of 4MMP and 3MH. A third olfactory zone for 3MHA was only observed for the youngest wines. The wines produced from Portuguese cultivars presented aromatic intensities comparable or even higher than those obtained in the Sauvignon blanc wine. Indeed, 3MH was found in all wines and the highest amounts were observed in Antão Vaz, Arinto and Viosinho wines. In general, these findings are in agreement with those obtained by sensory analysis.