Yeast cells can be affected by several causes of osmotic stress, such as high salt, sorbitol or glucose concentrations. The last condition is particularly interesting during natural processes where this microorganism participates. Response to osmostress requires the HOG (High Osmolarity Glycerol) pathway and several transcription factors, including Hot1, which plays a key role in high glucose concentrations. In this work, we describe how the yeast response to osmotic stress shows differences in accordance with the stress agent responsible for it. Compared with other conditions, under high glucose stress, delocalization of MAPK (Mitogen-Activated Protein Kinase) Hog1 is slower, induction of HOT1 expression is higher and Msn2/4 transcription factors are involved to a lesser extent. The transcriptomic analyses carried out with samples incubated for 30 min in the presence of high glucose or sorbitol reveal the presence of two functional categories with a differential expression between these conditions: glycogen biosynthesis and mobilization, and membrane-anchored proteins. We present data to demonstrate that the cells treated with 20% (w/v) (1.11 M) glucose contain higher chitin levels and are more sensitive to calcofluor white and ethanol.
Certain bioactive compounds that derive from tryptophan have been determined in 33 fermented foods, which suggests that the role of yeast is putative in origin. Melatonin is a 34 neurohormone in humans that plays an important role in health. The activity of other 35 compounds, such as 3-indoleacetic acid, has been recently highlighted, and interest in 36 elucidating the conditions of their production has grown. 37 However, the biosynthesis of melatonin by yeasts remains unclear to a large extent. 38 Therefore, this work was undertaken to demonstrate the unequivocally synthesis of 39 melatonin and other compounds that derive from tryptophan metabolism by yeast by 40 determining them in the intracellular compartment. By high resolution mass spectrometry 41 and a validated method, tryptophan itself, melatonin, serotonin, N-acetyl-5-42 hydroxytryptamine and 3-indoleacetic acid, were identified in the intracellular compartment 43 of Saccharomyces and non-Saccharomyces wine yeasts.
Melatonin is a bioactive compound that is present in fermented beverages and synthesized by yeast during alcoholic fermentation. Many studies have shown that melatonin interacts with some mammalian proteins, such as sirtuins or orphan receptor family proteins. The aim of this study was to determine the intracellular synthesis profile of melatonin in Saccharomyces cerevisiae and to identify the proteins that may interact with this molecule in yeast cells. Melatonin from fermentation samples was analyzed by liquid chromatography mass spectrometry, and proteins bound to melatonin were immunopurified by melatonin-IgG-Dynabeads. Melatonin was produced intracellularly in the lag phase of yeast growth and was exported to the extracellular media during the stationary phase. During this period, melatonin was bound to six proteins with molecular weights from 55 to 35 kDa. Sequence analysis showed that most proteins shared high levels of homology with glycolytic enzymes. An RNA-binding protein was also identified, the elongation alpha factor, which is related to mitochondria. This study reports for the first time the interaction of melatonin and proteins inside yeast cells. These results highlight the possible role of melatonin as a signal molecule and provide a new perspective for understanding its role in yeast.
Melatonin is a bioactive compound that is present in fermented beverages and has been described to be synthesized by yeast during alcoholic fermentation. The aim of this study was to assess the capacity of intracellular and extracellular melatonin production by different Saccharomyces strains from diverse food origin and to study the effects of different fermentation parameters, such as sugar and nitrogen concentration, temperature or initial population, on melatonin production using a synthetic grape must medium. Melatonin from fermentation samples was analyzed by liquid chromatography mass spectrometry. Intracellular melatonin synthesis profile did not present differences between yeast strains. However, extracellular melatonin production depended on the yeast origin. Thus, we suggest that melatonin production and secretion during the different yeast growth phases follows a species-specific pattern. Other parameters that affected the fermentation process such as sugar content and low temperature had an impact on intracellular melatonin production profile, as well as the melatonin content within the cell. This study reports the effect of several conditions on the melatonin synthesis profile, highlighting its possible role as a signal molecule.
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