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Impact of nutrient imbalance on wine alcoholic fermentations: nitrogen excess enhances yeast cell death in lipid-limited must.

ABSTRACT: We evaluated the consequences of nutritional imbalances, particularly lipid/nitrogen imbalances, on wine yeast survival during alcoholic fermentation. We report that lipid limitation (ergosterol limitation in our model) led to a rapid loss of viability during the stationary phase of fermentation and that the cell death rate is strongly modulated by nitrogen availability and nature. Yeast survival was reduced in the presence of excess nitrogen in lipid-limited fermentations. The rapidly dying yeast cells in fermentations in high nitrogen and lipid-limited conditions displayed a lower storage of the carbohydrates trehalose and glycogen than observed in nitrogen-limited cells. We studied the cell stress response using HSP12 promoter-driven GFP expression as a marker, and found that lipid limitation triggered a weaker stress response than nitrogen limitation. We used a SCH9-deleted strain to assess the involvement of nitrogen signalling pathways in the triggering of cell death. Deletion of SCH9 increased yeast viability in the presence of excess nitrogen, indicating that a signalling pathway acting through Sch9p is involved in this nitrogen-triggered cell death. We also show that various nitrogen sources, but not histidine or proline, provoked cell death. Our various findings indicate that lipid limitation does not elicit a transcriptional programme that leads to a stress response protecting yeast cells and that nitrogen excess triggers cell death by modulating this stress response, but not through HSP12. These results reveal a possibly negative role of nitrogen in fermentation, with reported effects referring to ergosterol limitation conditions. These effects should be taken into account in the management of alcoholic fermentations.

SUBMITTER: Tesniere C

PROVIDER: S-EPMC3637302 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Impact of nutrient imbalance on wine alcoholic fermentations: nitrogen excess enhances yeast cell death in lipid-limited must.

Tesnière Catherine C Delobel Pierre P Pradal Martine M Blondin Bruno B

PloS one 20130426 4

We evaluated the consequences of nutritional imbalances, particularly lipid/nitrogen imbalances, on wine yeast survival during alcoholic fermentation. We report that lipid limitation (ergosterol limitation in our model) led to a rapid loss of viability during the stationary phase of fermentation and that the cell death rate is strongly modulated by nitrogen availability and nature. Yeast survival was reduced in the presence of excess nitrogen in lipid-limited fermentations. The rapidly dying yea ...[more]

PMID: 23658613

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Project description:Alcoholic fermentation is fundamentally an adaptation process, in which the yeast Saccharomyces cerevisiae outperforms its competitors and takes over the fermentation process itself. Although wine yeast strains appear to be adapted to the stressful conditions of alcoholic fermentation, nitrogen limitations in grape must cause stuck or slow fermentations, generating significant economic losses for the wine industry. One way to discover the genetic bases that promote yeast adaptation to nitrogen-deficient environments are selection experiments, where a yeast population undergoes selection under conditions of nitrogen restriction for a number of generations, to then identify by sequencing the molecular characteristics that promote this adaptation. In this work, we carried out selection experiments in bioreactors imitating wine fermentation under nitrogen-limited fermentation conditions (SM60), using the heterogeneous SGRP-4X yeast population, to then sequence the transcriptome and the genome of the population at different time points of the selection process. The transcriptomic results showed an overexpression of genes from the NA strain (North American/YPS128), a wild, non-domesticated isolate. In addition, genome sequencing and allele frequency results allowed several QTLs to be mapped for adaptation to nitrogen-limited fermentation. Finally, we validated the ECM38 allele of NA strain as responsible for higher growth efficiency under nitrogen-limited conditions. Taken together, our results revealed a complex pattern of molecular signatures favouring adaptation of the yeast population to nitrogen-limited fermentations, including differential gene expression, allele frequency changes and loss of the mitochondrial genome. Finally, the results suggest that wild alleles from a non-domesticated isolate (NA) may have a relevant role in the adaptation to the assayed fermentation conditions, with the consequent potential of these alleles for the genetic improvement of wine yeast strains.

Project description:Multi-starter wine fermentations employing non-Saccharomyces (NS) yeasts are becoming an emerging trend in winemaking. It is therefore important to determine the impacts of different NS strains in the wine phenotype and in particular the aroma outputs in different inoculation schemes and fermentation conditions. Here, two native NS yeasts, Lachancea thermotolerans LtMM7 and Hanseniaspora uvarum HuMM19, were assessed for their ability to improve the quality of Moschofilero, a Greek aromatic white wine. The NS strains were initially examined in laboratory scale fermentations in mixed inoculations with ScMM23, a native Saccharomyces cerevisiae strain. LtMM7 was selected to be further evaluated in pilot scale fermentations. Five different inoculation schemes were considered: single inoculation of ScMM23 (IS), simultaneous inoculation of ScMM23 with HuMM19 (SMH) or LtMM7 (SML), and sequential inoculation of HuMM19 (SQH) or LtMM7 (SQL) followed by ScMM23. At laboratory scale fermentations, the chemical profiles were largely affected by both the NS species and the inoculation scheme applied. The sequential inoculation using HuMM19 produced the most divergent wine phenotype. However, HuMM19 caused significant increases in acetic acid and ethyl acetate levels that impeded its use in pilot scale trials. LtMM7 significantly affected the chemical profiles of wines produced at the winery, especially in the sequential inoculation scheme. Importantly, LtMM7 significantly increased the levels of acetate esters or ethyl esters, depending on the inoculation method applied. In particular, acetate esters like isobutyl acetate, hexyl acetate, and 2-phenylethyl acetate, which all impart fruity or floral aromas, were significantly increased in SQL. On the other hand, higher levels of total ethyl esters were associated with SML. The most striking differences were observed in the levels of fruit-impair esters like ethyl decanoate, 3-methylbutyl octanoate, and isoamyl hexanoate. This is the first study to report a significant increase in the ethyl ester fraction by L. thermotolerans. Interestingly, L. thermotolerans in SQL also increased the concentrations of damascenone and geraniol, the major teprenic compound of Moschofilero, which are associated with several typical floral and fruity aromas of the variety. Present results show that L. thermotolerans may enhance the varietal character and increase the chemical complexity of Moschofilero wines.

Dataset Information

Impact of nutrient imbalance on wine alcoholic fermentations: nitrogen excess enhances yeast cell death in lipid-limited must.

Publications

Impact of nutrient imbalance on wine alcoholic fermentations: nitrogen excess enhances yeast cell death in lipid-limited must.

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