Abstract
Nitrogen is an important nutrient in alcoholic fermentation because its starvation affects both fermentation kinetics and the formation of yeast metabolites. In most alcoholic fermentations, yeasts have to ferment in nitrogen-starved conditions, which requires modifications of cell functions to maintain a high sugar flux and enable cell survival for long periods in stressful conditions. In this review, we present an overview of our current understanding of the responses of the wine yeast Saccharomyces cerevisiae to variations of nitrogen availability. Adaptation to nitrogen starvation involves changes in the activity of signaling pathways such as target of rapamycin (TOR) and nitrogen catabolite repression (NCR), which are important for the remodeling of gene expression and the establishment of stress responses. Upon starvation, protein degradation pathways involving autophagy and the proteasome play a major role in nitrogen recycling and the adjustment of cellular activity. Recent progress in the understanding of the role of these mechanisms should enable advances in fermentation management and the design of novel targets for the selection or improvement of yeast strains.
Similar content being viewed by others
References
Agenbach WA (1977) A study of must nitrogen content in relation to incomplete fermentations, yeast production and fermentation activity. In: Proceedings South African Society of Enology and Viticulture, Cape Town, South Africa. Stellenbosh, S. A, pp. 66-87
Albers A, Larsson C, Andlid T, Walsh MC, Gustafsson L (2007) Effect of nutrient starvation on the cellular composition and metabolic capacity of Saccharomyces cerevisiae. Appl Environ Microbiol 73:4839–4848
Backhus LE, DeRisi J, Brown PO, Bisson LF (2001) Functional genomic analysis of a commercial wine strain of Saccharomyces cerevisiae under differing nitrogen conditions. FEMS Yeast Res 1:111–125
Barbosa C, Falco V, Mendes-Faia A, Mendes-Ferreira A (2009) Nitrogen addition influences formation of aroma compounds, volatile acidity and ethanol in nitrogen deficient media fermented by Saccharomyces cerevisiae wine strains. J Biosci Bioeng 108:99–104
Bell SJ, Henschke PA (2005) Implications of nitrogen nutrition for grapes, fermentation and wine. Aust J Grape Wine Res 11:242–295
Beltran G, Novo M, Rozès N, Mas A, Guillamon JM (2004) Nitrogen catabolite repression in Saccharomyces cerevisiae during wine fermentation. FEMS Yeast Res 4:625–632
Beltran G, Esteve-Zarzoso B, Rozès N, Mas A, Guillamon JM (2005) Influence in the timing of nitrogen additions during synthetic grape must fermentations on fermentation kinetics and nitrogen consumption. J Agric Food Chem 53:996–1002
Bely M, Sablayrolles JM, Barre P (1990a) Automatic control of assimilable nitrogen addition during alcoholic fermentation in enological conditions. J Ferm Bioeng 70:1–6
Bely M, Sablayrolles JM, Barre P (1990b) Description of alcoholic fermentation kinetics: its variability and significance. Am J Enol Vitic 40:319–324
Bely M, Sablayrolles JM, Barre P (1990c) Automatic detection of assimilable nitrogen deficiencies during alcoholic fermentation in enological conditions. J Ferm Bioeng 70:246–252
Bertram PG, Choi JH, Carvalho J, Ai W, Zeng C, Chan TF, Zheng XF (2000) Tripartite regulation of Gln3p by TOR, Ure2p, and phosphatases. J Biol Chem 275:35727–35733
Beyer A, Hollunder J, Nasheuer HP, Wilhelm T (2004) Post-transcriptional expression regulation in the yeast Saccharomyces cerevisiae on a genomic scale. Mol Cell Proteomics 3:1083–1092
Bisson LF (1991) Influence of nitrogen on yeast and fermentation of grapes. Proceedings of the International Symposium on Nitrogen in Grapes and Wine. Am J Enol Vitic 42:78–89
Blateyron L, Sablayrolles JM (2001) Stuck and slow fermentations in enology: statistical study of causes and effectiveness of combined additions of oxygen and diammonium phosphate. J Biosci Bioeng 91:184–189
Boer VM, de Winde JH, Pronk JT, Piper MDW (2003) The genome-wide transcriptional responses of Saccharomyces cerevisiae grown on glucose in aerobic chemostat cultures limited for carbon, nitrogen, phosphorus, or sulfur. J Biol Chem 278:3265–3274
Breker M, Gymrek M, Schuldiner M (2013) A novel single-cell screening platform reveals proteome plasticity during yeast stress responses. J Cell Biol 200:839–850
Brice C, Sanchez I, Tesnière C, Blondin B (2014a) Assessing the mechanisms responsible for differences in nitrogen requirements between Saccharomyces cerevisiae wine yeasts in alcoholic fermentation. Appl Environ Microbiol 80:1330–1339
Brice C, Sanchez I, Bigey F, Legras JL, Blondin B (2014b) A QTL approach for deciphering the genetics bases of variability in nitrogen requirement of yeast. BMC Genomics 15:495
Broach JR (2012) Nutritional control of growth and development in yeast. Genetics 192:73–105
Butzke CE (1998) Study of yeast assimilable N status in musts from California, Oregon and Washington. Am J Enol Vitic 49:220–224
Carrasco P, Perez-Ortin JE, delOlmo M (2003) Arginase activity is a useful marker of nitrogen limitation during alcoholic fermentations. Syst Appl Microbiol 26:471–479
Carrau FM, Neirotti E, Gioia O (1993) Stuck wine fermentations: effect of killer/sensitive yeast interactions. J Ferm Bioeng 76:67–69
Carrau FM, Medina K, Farina L, Boido E, Henschke PA, Dellacassa E (2008) Production of fermentation aroma compounds by Saccharomyces cerevisiae wine yeasts: effects of yeast assimilable nitrogen on two model strains. FEMS Yeast Res 8:1196–1207
Cebollero E, Gonzales R (2006) Induction of autophagy by second-fermentation yeasts during elaboration of sparkling wines. Appl Environ Microbiol 72:4121–4127
Chiva R, Baiges I, Mas A, Guillamon JM (2009) The role of GAP1 gene in the nitrogen metabolism of Saccharomyces cerevisiae during wine fermentation. J Appl Microbiol 107:235–244
Contreras A, García V, Salinas F, Urzúa U, Ganga MA, Martínez C (2012) Identification of genes related to nitrogen uptake in wine strains of Saccharomyces cerevisiae. World J Microbiol Biotechnol 28:1107–1113
Cramer AC, Vlassides S, Block DE (2002) Kinetic model for nitrogen-limited wine fermentations. Biotechnol Bioeng 77:49–60
Crépin L, Nidelet T, Sanchez I, Dequin C, Camarasa C (2012) Sequential use of nitrogen compounds by Saccharomyces cerevisiae during wine fermentation: a model based on kinetic and regulation characteristics of nitrogen permeases. Appl Environ Microbiol 78:8102–8111
Cutler NS, Pan X, Heitman J, Cardenas ME (2001) The TOR signal transduction cascade controls cellular differentiation in response to nutrient. Mol Biol Cell 12:4103–4113
Dubois C, Manginot C, Roustan JL, Sablayrolles JM, Barre P (1996) Effect of variety, year, and grape maturity on the kinetics of alcoholic fermentation. Am J Enol Vitic 47:363–368
Dupré S, Volland C, Haguenauer-Tsapis R (2001) Membrane transport: ubiquitylation in endosomal sorting. Curr Biol 11:R932–R934
Finley D, Ulrich HD, Sommer T, Kaiser P (2012) The ubiquitin-proteasome system of Saccharomyces cerevisiae. Genetics 192:319–360
Gasch AP, Spellman PT, Kao CM, Carmel-Harel O, Eisen MB, Storz G, Bolstein D, Brown PO (2000) Genomic expression programs in the response of yeast cells to environmental changes. Mol Biol Cell 11:4241–4257
Godard P, Urrestarazu A, Vissers S, Kontos K, Bontempi G, van Helden J, André B (2007) Effect of 21 different nitrogen sources on global gene expression in the yeast Saccharomyces cerevisiae. Mol Cell Biol 27:3065–3086
Grundmann O, Mösch HU, Braus GH (2001) Repression of GCN4 mRNA translation by nitrogen starvation in Saccharomyces cerevisiae. J Biol Chem 276:25661–25671
Gutiérrez A, Chiva R, Sancho M, Beltran G, Arroyo-López FN, Guillamon JM (2012) Nitrogen requirements of commercial wine yeast strains during fermentation of a synthetic grape must. Food Microbiol 31:25–32
Gutiérrez A, Beltran G, Warringer J, Guillamon JM (2013) Genetic basis of variations in nitrogen source utilization in four wine commercial yeast strains. PLoS ONE 8:e67166
Henschke P, Jiranek V (1993) Yeasts metabolism of nitrogen compounds. In: Fleet GH (ed) Wine. Microbiology and biotechnology. Harwood Academic, Chur pp 77-164
Herraiz T, Ough CS (1993) Formation of ethyl esters of amino acids by yeasts during the alcoholic fermentation of grape juice. Am J Enol Vitic 44:41–48
Hinnebusch AG (2005) Translational regulation of GCN4 and the general amino acid control of yeast. Annu Rev Microbiol 59:407–450
Hofman-Bang J (1999) Nitrogen catabolite repression in Saccharomyces cerevisiae. Mol Biotechnol 12:35–73
Huang H, Kawamata T, Horie T, Tsugawa H, Nakayama Y, Ohsumi Y, Fukusaki E (2015) Bulk RNA degradation by nitrogen starvation‐induced autophagy in yeast. EMBO J34:154–168
Jiranek V, Langridge P, Henschke PA (1991) Yeast nitrogen demand: selection criterion for wine yeasts for fermenting low nitrogen musts. In: Ranz JM (ed) Proceedings of the international symposium on nitrogen in grapes and wine, pp 266-269
Jiranek V, Langridge P, Henschke PA (1995) Amino acid and ammonium utilization by Saccharomyces cerevisiae wine yeasts from a chemically defined medium. Am J Enol Vitic 46:75–83
Julien A, Roustan JL, Dulau L, Sablayrolles JM (2000) Comparison of nitrogen and oxygen demands of enological yeasts: technological consequences. Am J Enol Vitic 51:215–222
Kelly SP, Bedwell (2015) Both the autophagy and proteasomal pathways facilitate the Ubp3p-dependent depletion of a subset of translation and RNA turnover factors during nitrogen starvation in Saccharomyces cerevisiae. RNA 21:1–13
Kraft C, Deplazes A, Sohrmann M, Peter M (2008) Mature ribosomes are selectively degraded upon starvation by an autophagy pathway requiring the Ubp3/Bre5p ubiquitin protease. Nat Cell Biol 10:602–610
Kraft C, Peter M, Hofmann K (2010) Selective autophagy: ubiquitin-mediated recognition and beyond. Nat Cell Biol 12:836–841
Krampe S, Boles E (2002) Starvation-induced degradation of yeast hexose transporter Hxt7p is dependent on endocytosis, autophagy and the terminal sequences of the permease. FEBS Lett 513:193–196
Magasanik B, Kaiser CA (2002) Nitrogen regulation in Saccharomyces cerevisiae. Gene 290:1–18
Maisonnave P, Sanchez I, Moine V, Dequin S, Galeote V (2013) Stuck fermentation: development of a synthetic stuck wine and study of a restart procedure. Int J Food Microbiol 163:239–247
Malherbe S (2003) Modélisation de la fermentation alcoolique en conditions oenologiques. Sciences et procédés biologiques et industriels. Thesis. Montpellier II University
Manginot C, Sablayrolles JM, Roustan JL, Barre P (1997) Use of constant rate alcoholic fermentations to compare the effectiveness of different nitrogen sources added during the stationary phase. Enzym Microb Technol 20:373–380
Manginot C, Roustan JL, Sablayrolles JM (1998) Nitrogen demand of different yeast strains during alcoholic fermentation. Importance of the stationary phase. Enzym Microb Technol 23:511–517
Marks VD, van der Merwe GK, van Vuuren HJJ (2003) Transcriptional profiling of wine yeast in fermenting grape juice: regulatory effect of diammonium phosphate. FEMS Yeast Res 3:269–287
Marks VD, Ho Sui SJ, Erasmus D, van der Merwe GK, Brumm J, Wasserma WW, Bryan J, van Vuuren JJ (2008) Dynamics of the yeast transcriptome during wine fermentation reveals a novel fermentation stress response. FEMS Yeast Res 8:35–52
Martínez-Moreno R, Morales P, Gonzalez R, Mas A, Beltran G (2012) Biomass production and alcoholic fermentation performance of Saccharomyces cerevisiae as a function of nitrogen source. FEMS Yeast Res 12:477–485
Martínez-Moreno R, Quirós M, Morales P, Gonzalez R (2014) New insights into the advantages of ammonium as a winemaking nutrient. Int J Food Microbiol 177:128–135
Medina K, Carrau FM, Gioia O, Bracesco N (1997) Nitrogen availability of grape juice limits killer yeast growth and fermentation activity during mixed-culture fermentation with sensitive commercial yeast strains. Appl Environ Microbiol 63:2821–2825
Mendes-Ferreira A, Mendes-Faia A, Leão C (2004) Growth and fermentation patterns of Saccharomyces cerevisiae under different ammonium concentrations and its implications in winemaking industry. J Appl Microbiol 97:540–545
Mendes-Ferreira A, del Olmo M, Garcia-Matinez J, Jimenez-Marti E, Mendes-Faia A, Perez-Ortin JE, Leão C (2007) Transcriptional response of Saccharomyces cerevisiae to different nitrogen concentrations during alcoholic fermentation. Appl Environ Microbiol 73:3049–3060
Mendes-Ferreira A, Barbosa C, Falco V, Leão C, Mendes-Faia A (2009) The production of hydrogen sulphide and other aroma compounds by wine strains of Saccharomyces cerevisiae in synthetic media with different nitrogen concentrations. J Ind Microbiol Biotechnol 36:571–583
Monteiro F, Bisson LF (1992) Nitrogen supplementation of grape juice. I. Effect on amino acid utilization during fermentation. Am J Enol Vitic 43:1–10
Narayanaswamy R, Levy M, Tsechansky M, Stovall GW, O’Connell JD, Mirrielees J, Ellington AD, Marcotte EM (2009) Widespread reorganization of metabolic enzymes into reversible assemblies upon nutrient starvation. Proc Natl Acad Sci USA 106:10147–10152
Natarajan K, Meyer MR, Jackson BM, Slade D, Roberts C, Hinnebusch AG, Marton MJ (2001) Transcriptional profiling shows that Gcn4p is a master regulator of gene expression during amino acid starvation in yeast. Mol Cell Biol 21:4347–4368
Nicolini G, Larcher R, Versini G (2004) Status of yeast assimilable nitrogen in Italian grape musts and effects of variety, ripening and vintage. Vitis 43:89–96
Nomura W, Maeta K, Kita K, Izawa S, Inoue Y (2010) Methylglyoxal activates Gcn2 to phosphorylate eIF2 α independently of the TOR pathway in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 86:1887–1894
Novo MT, Beltran G, Rozès N, Guillamon JM, Mas A (2005) Effect of nitrogen limitation and surplus upon trehalose metabolism in wine yeast. Appl Microbiol Biotechnol 66:560–566
Nykanen L (1986) Formation and occurrence of flavor compounds in wine and distilled alcoholic beverage. Am J Enol Vitic 37:84–96
Onodera J, Oshimi Y (2005) Autophagy is required for maintenance of amino acid levels and protein synthesis under nitrogen starvation. J Biol Chem 280:31582–31586
Orlova M, Kanter E, Krakovich D, Kuchin S (2006) Nitrogen availability and TOR regulate the Snf1 protein kinase in Saccharomyces cerevisiae. Eukaryot Cell 5:1831–1837
Perez M, Luyten K, Michel R, Riou C, Blondin B (2005) Analysis of Saccharomyces cerevisiae hexose carrier expression during wine fermentation: both low- and high-affinity Hxt transporters are expressed. FEMS Yeast Res 5:351–361
Piggott N, Cook MA, Tyers M, Measday V (2011) Genome-wide fitness profiles reveal a requirement for autophagy during yeast fermentation. G3 5:353-367
Rohde J, Heitman J, Cardenas ME (2001) The TOR kinases link nutrient sensing to cell growth. J Biol Chem 276:9583–9586
Rossignol T, Dulau L, Julien A, Blondin B (2003) Genome-wide monitoring of wine yeast gene expression during alcoholic fermentation. Yeast 20:1369–1385
Rossignol T (2004) Analyse de l’expression du génome des levures oenologiques en fermentation alcoolique par des approches post-génomiques. Thesis, Montpellier II University, Montpellier
Rossignol T, Kobi D, Jacquet-Gutfreund L, Blondin B (2009) The proteome of a wine yeast strain during fermentation, correlation with the transcriptome. J Appl Microbiol 107:47–55
Saint-Prix F, Bönquist L, Dequin S (2004) Functional analysis of the ALD gene family of Saccharomyces cerevisiae during anaerobic growth on glucose: the NADP+-dependent Ald6p and Ald5p isoforms play a major role in acetate formation. Microbiol 150:2209–2220
Salmon JM (1989) Effect of sugar transport inactivation in Saccharomyces cerevisiae on sluggish and stuck enological fermentations. Appl Environ Microbiol 55:953–958
Salmon JM, Barre P (1998) Improvement of nitrogen assimilation and fermentation kinetics under enological conditions by derepression of alternative nitrogen-assimilatory pathways in an industrial Saccharomyces cerevisiae strain. Appl Environ Microbiol 64:3831–3837
Shirra MK, McCartney RR, Zhang C, Shokat KM, Schmidt MC, Arndt KM (2008) A chemical genomics study identifies Snf1 as a repressor of GCN4 translation. J Biol Chem 283:35889–35898
Snowdon C, van der Merwe G (2012) Regulation of Hxt3 and Hxt7 turnover converges on the Vid30 complex and requires inactivation of the Ras/cAMP/PKA pathway in Saccharomyces cerevisiae. PLoS ONE 7:e50458
Stephan JS, Yeh YY, Ramachandran V, Deminoff SJ, Herman PK (2009) The Tor and PKA signaling pathways independently target the Atg1/Atg13 protein kinase complex to control autophagy. Proc Natl Acad Sci U S A 106:17049–17054
Swinnen E, Wanke V, Roosen J, Smets B, Dubouloz F, Pedruzzi I, Cameroni E, De Virgilio C, Winderickx J (2006) Rim15 and the crossroads of nutrient signalling pathways in Saccharomyces cerevisiae. Cell Div 1:3
TerSchure EG, Van Riel NAV, Verrips CT (2000) The role of ammonia metabolism in nitrogen catabolite repression in Saccharomyces cerevisiae. FEMS Microbiol Rev 24:67–83
Tesnière C, Delobel P, Pradal M, Blondin B (2013) Impact of nutrient imbalance on wine alcoholic fermentations: nitrogen excess enhances yeast cell death in lipid-limited must. PLoS ONE 8:e61645
Trabalzini L, Paffetti A, Scaloni A, Talamo F, Ferro E, Coratza G, Bovalini L, Lusini P, Martelli P, Santucci A (2003) Proteomic response to physiological fermentation stresses in a wild-type wine strain of Saccharomyces cerevisiae. Biochem J 370:35–46
Urban J, Soulard A, Huber A, Lippman S, Mukhopadhyay D, Deloche O, Wanke V, Anrather D, Ammerer G, Riezman H, Broach JR, De Virgilio C, Hall MN, Loewith R (2007) Sch9 is a major target of TORC1 in Saccharomyces cerevisiae. Mol Cell 26:663–674
Uttenweiler A, Mayer A (2008) Microautophagy in the yeast Saccharomyces cerevisiae. Methods Mol Biol 445:245–259
Varela C, Pizarro F, Agosin E (2004) Biomass content governs fermentation rate in nitrogen-deficient wine musts. Appl Environ Microbiol 6:3392–3400
Vilanova M, Ugliano M, Varela C, Siebert T, Pretorius IS, Henschke PA (2007) Assimilable nitrogen utilisation and production of volatile and non-volatile compounds in chemically defined medium by Saccharomyces cerevisiae wine yeasts. Appl Microbiol Cell Physiol 77:145–157
Vinod PK, Sengupta N, Bhat PJ, Venkatesh KV (2008) Integration of global signaling pathways, cAMP-PKA, MAPK and TOR in the regulation of FLO11. PLoS ONE 3:e1663
Walker ME, Nguyen TD, Liccioli T, Schmid F, Kalatzis N, Sundstrom JF, Gardner JM, Jiranek V (2014) Genome-wide identification of the fermentome; genes required for successful and timely completion of wine-like fermentation by Saccharomyces cerevisiae. BMC Genomics 15:552
Warner JR (1999) The economics of ribosome biosynthesis in yeast. Trends Biochem Sci 24:437–440
Zaman S, Lippman SI, Zhao X, Broach JR (2008) How Saccharomyces responds to nutrients. Annu Rev Genet 42:27–81
Zhang J, Vaga S, Chumnanpuen P, Kumar R, Vemuri GN, Aebersold R, Nielsen J (2011) Mapping the interaction of Snf1 with TORC1 in Saccharomyces cerevisiae. Mol Syst Biol 7:545
Zhao X, Zou H, Fu J, Chen J, Zhou J, Du G (2013) Nitrogen regulation involved in the accumulation of urea in Saccharomyces cerevisiae. Yeast 30:437–447
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tesnière, C., Brice, C. & Blondin, B. Responses of Saccharomyces cerevisiae to nitrogen starvation in wine alcoholic fermentation. Appl Microbiol Biotechnol 99, 7025–7034 (2015). https://doi.org/10.1007/s00253-015-6810-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-015-6810-z