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Alternative Glycerol Balance Strategies among Saccharomyces Species in Response to Winemaking Stress.

Pérez-Torrado R, Oliveira BM, Zemančíková J, Sychrová H, Querol A - Front Microbiol (2016)

Bottom Line: The results show different pattern and level of expression among the different species, especially for STL1.These experiments also revealed a different functionality of the glycerol transporters among the different species studied.All these data point to different strategies to handle glycerol accumulation in response to winemaking stresses as hyperosmotic or cold-hyperosmotic stress in the different species, with variable emphasis in the production, influx, or efflux of glycerol.

View Article: PubMed Central - PubMed

Affiliation: Food Biotechnology Department, Systems Biology in Yeast of Biotechnological Interest, Instituto de Agroquímica y Tecnología de los Alimentos, IATA-CSIC Valencia, Spain.

ABSTRACT
Production and balance of glycerol is essential for the survival of yeast cells in certain stressful conditions as hyperosmotic or cold shock that occur during industrial processes as winemaking. These stress responses are well-known in S. cerevisiae, however, little is known in other phylogenetically close related Saccharomyces species associated with natural or fermentation environments such as S. uvarum, S. paradoxus or S. kudriavzevii. In this work we have investigated the expression of four genes (GPD1, GPD2, STL1, and FPS1) crucial in the glycerol pool balance in the four species with a biotechnological potential (S. cerevisiae; S. paradoxus; S. uvarum; and S. kudriavzevii), and the ability of strains to grow under osmotic and cold stresses. The results show different pattern and level of expression among the different species, especially for STL1. We also studied the function of Stl1 glycerol symporter in the survival to osmotic changes and cell growth capacity in winemaking environments. These experiments also revealed a different functionality of the glycerol transporters among the different species studied. All these data point to different strategies to handle glycerol accumulation in response to winemaking stresses as hyperosmotic or cold-hyperosmotic stress in the different species, with variable emphasis in the production, influx, or efflux of glycerol.

No MeSH data available.


Related in: MedlinePlus

Microvinification experiments in synthetic must at low temperature with S. cerevisiae T73 (dark red) and FCry (light red), S. paradoxus Chr16.2 (light green) and 108 (dark green), S. uvarum 12600 (dark purple) and BMV58 (light purple), and S. kudriavzevii CR85 (light blue) and IFO1802 (dark blue) strains. Precultured cells were inoculated in synthetic must at 12°C and samples were taken after 0, 1, 4, 24 and 48 h to determine extra (A) and intracellular (B) glycerol content for each strain. Three independent microvinification bottles were used for each strain and average ± standard deviation is shown.
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Figure 2: Microvinification experiments in synthetic must at low temperature with S. cerevisiae T73 (dark red) and FCry (light red), S. paradoxus Chr16.2 (light green) and 108 (dark green), S. uvarum 12600 (dark purple) and BMV58 (light purple), and S. kudriavzevii CR85 (light blue) and IFO1802 (dark blue) strains. Precultured cells were inoculated in synthetic must at 12°C and samples were taken after 0, 1, 4, 24 and 48 h to determine extra (A) and intracellular (B) glycerol content for each strain. Three independent microvinification bottles were used for each strain and average ± standard deviation is shown.

Mentions: Since hyperosmotic and also cold stress responses are unequivocally related to glycerol accumulation we wanted to determine glycerol levels during hyperosmotic-cold stress in wine fermentations. Thus we performed wine fermentations in synthetic must with the studied Saccharomyces species and strains, and we measured intra- and extracellular amount of glycerol during the first hours and days of the fermentation. In the results presented in Figure 2 we observed two steps regarding glycerol accumulation in S. cerevisiae strains. In the first step, glycerol starts to accumulate inside the cell (Figure 2B) immediately after inoculating into the cold-hyperosmotic condition, reaching a maximal value after 24 h. Also, minimal glycerol levels are accumulated in extracellular media in the beginning of our experiment (Figure 2A). In the next 2 days, intracellular glycerol is reduced and tends to recover its original levels whereas extracellular glycerol increases with the time. In the case of S. paradoxus and S. kudriavzevii, maximal intracellular glycerol accumulation, which are approximately half of those in S. cerevisiae strains, occurs in the first hours and levels are maintained during 48 h. Analyzing the intracellular glycerol level (Figure 2B), it is interesting to note that, comparing with the other species, S. cerevisiae strains accumulated the higher levels of glycerol between 4 and 48 h of incubation. The S. uvarum strains showed the lowest values of intracellular glycerol with a maximal level after 1 h in the case of BMV58 and after 48 h in the case of 12600. Regarding extracellular glycerol (Figure 2A), S. paradoxus presented similar levels and accumulation pattern as S. cerevisiae and S. uvarum and, in addition, S. kudriavzevii showed a similar pattern but higher accumulation levels (around five times more). Is interesting to emphasize that S. uvarum and S. kudriavzevii showed a higher extracellular glycerol accumulation rate compared to the other two species. Interestingly, no extracellular glycerol was observed at time 0 in any species. It should be noted that strains do not show significant growth after 1 or 4 h and maximal yeast biomass was observed at the 24 or 48 h time point except for the IFO1802 that show very low growth level in grape must (Supplementary Figure 1), in concordance with data observed in Figure 1 in osmotic and cold stress conditions.


Alternative Glycerol Balance Strategies among Saccharomyces Species in Response to Winemaking Stress.

Pérez-Torrado R, Oliveira BM, Zemančíková J, Sychrová H, Querol A - Front Microbiol (2016)

Microvinification experiments in synthetic must at low temperature with S. cerevisiae T73 (dark red) and FCry (light red), S. paradoxus Chr16.2 (light green) and 108 (dark green), S. uvarum 12600 (dark purple) and BMV58 (light purple), and S. kudriavzevii CR85 (light blue) and IFO1802 (dark blue) strains. Precultured cells were inoculated in synthetic must at 12°C and samples were taken after 0, 1, 4, 24 and 48 h to determine extra (A) and intracellular (B) glycerol content for each strain. Three independent microvinification bottles were used for each strain and average ± standard deviation is shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4814467&req=5

Figure 2: Microvinification experiments in synthetic must at low temperature with S. cerevisiae T73 (dark red) and FCry (light red), S. paradoxus Chr16.2 (light green) and 108 (dark green), S. uvarum 12600 (dark purple) and BMV58 (light purple), and S. kudriavzevii CR85 (light blue) and IFO1802 (dark blue) strains. Precultured cells were inoculated in synthetic must at 12°C and samples were taken after 0, 1, 4, 24 and 48 h to determine extra (A) and intracellular (B) glycerol content for each strain. Three independent microvinification bottles were used for each strain and average ± standard deviation is shown.
Mentions: Since hyperosmotic and also cold stress responses are unequivocally related to glycerol accumulation we wanted to determine glycerol levels during hyperosmotic-cold stress in wine fermentations. Thus we performed wine fermentations in synthetic must with the studied Saccharomyces species and strains, and we measured intra- and extracellular amount of glycerol during the first hours and days of the fermentation. In the results presented in Figure 2 we observed two steps regarding glycerol accumulation in S. cerevisiae strains. In the first step, glycerol starts to accumulate inside the cell (Figure 2B) immediately after inoculating into the cold-hyperosmotic condition, reaching a maximal value after 24 h. Also, minimal glycerol levels are accumulated in extracellular media in the beginning of our experiment (Figure 2A). In the next 2 days, intracellular glycerol is reduced and tends to recover its original levels whereas extracellular glycerol increases with the time. In the case of S. paradoxus and S. kudriavzevii, maximal intracellular glycerol accumulation, which are approximately half of those in S. cerevisiae strains, occurs in the first hours and levels are maintained during 48 h. Analyzing the intracellular glycerol level (Figure 2B), it is interesting to note that, comparing with the other species, S. cerevisiae strains accumulated the higher levels of glycerol between 4 and 48 h of incubation. The S. uvarum strains showed the lowest values of intracellular glycerol with a maximal level after 1 h in the case of BMV58 and after 48 h in the case of 12600. Regarding extracellular glycerol (Figure 2A), S. paradoxus presented similar levels and accumulation pattern as S. cerevisiae and S. uvarum and, in addition, S. kudriavzevii showed a similar pattern but higher accumulation levels (around five times more). Is interesting to emphasize that S. uvarum and S. kudriavzevii showed a higher extracellular glycerol accumulation rate compared to the other two species. Interestingly, no extracellular glycerol was observed at time 0 in any species. It should be noted that strains do not show significant growth after 1 or 4 h and maximal yeast biomass was observed at the 24 or 48 h time point except for the IFO1802 that show very low growth level in grape must (Supplementary Figure 1), in concordance with data observed in Figure 1 in osmotic and cold stress conditions.

Bottom Line: The results show different pattern and level of expression among the different species, especially for STL1.These experiments also revealed a different functionality of the glycerol transporters among the different species studied.All these data point to different strategies to handle glycerol accumulation in response to winemaking stresses as hyperosmotic or cold-hyperosmotic stress in the different species, with variable emphasis in the production, influx, or efflux of glycerol.

View Article: PubMed Central - PubMed

Affiliation: Food Biotechnology Department, Systems Biology in Yeast of Biotechnological Interest, Instituto de Agroquímica y Tecnología de los Alimentos, IATA-CSIC Valencia, Spain.

ABSTRACT
Production and balance of glycerol is essential for the survival of yeast cells in certain stressful conditions as hyperosmotic or cold shock that occur during industrial processes as winemaking. These stress responses are well-known in S. cerevisiae, however, little is known in other phylogenetically close related Saccharomyces species associated with natural or fermentation environments such as S. uvarum, S. paradoxus or S. kudriavzevii. In this work we have investigated the expression of four genes (GPD1, GPD2, STL1, and FPS1) crucial in the glycerol pool balance in the four species with a biotechnological potential (S. cerevisiae; S. paradoxus; S. uvarum; and S. kudriavzevii), and the ability of strains to grow under osmotic and cold stresses. The results show different pattern and level of expression among the different species, especially for STL1. We also studied the function of Stl1 glycerol symporter in the survival to osmotic changes and cell growth capacity in winemaking environments. These experiments also revealed a different functionality of the glycerol transporters among the different species studied. All these data point to different strategies to handle glycerol accumulation in response to winemaking stresses as hyperosmotic or cold-hyperosmotic stress in the different species, with variable emphasis in the production, influx, or efflux of glycerol.

No MeSH data available.


Related in: MedlinePlus