1. Introduction
Yeasts have been used by humans to produce foods for thousands of years. Bread, wine, sake and beer are made with the essential contribution of yeasts, especially from the species
Nowadays, modern industries require very large amounts of selected yeasts to obtain high quality reproducible products and to ensure fast, complete fermentations. Around 0.4 million metric tonnes of yeast biomass, including 0.2 million tonnes baker's yeast alone, are produced each year worldwide. Efficient and profitable factory-scale processes have been developed to produce yeast biomass. The standard process was empirically optimised to obtain the highest yield by increasing biomass production and decreasing costs. However in recent years, several molecular and physiological studies have revealed that yeast undergoes diverse stressful situations along the biomass production process which can seriously affect its fermentative capacity and technological performance.
In this chapter, we review the yeast biomass production process, including substrates, growth configuration, yield optimisation and the particularities of brewing, baker- or wine-yeasts production. We summarise the new studies that describe the process from a molecular viewpoint to reveal yeast responses to different stressful situations. Finally, we highlight the key points to be optimised in order to obtain not only high yields, but also the best biomass fermentative efficiency, and we provide future directions in the field.
2. Molasses: A suitable substrate
Beet or cane molasses are the main substrate used in yeast production plants. These materials were selected for two main reasons: first, yeasts grow very well using the sugars present in the molasses and second, they are economically interesting since they are a waste product coming from sugar refineries without any other application. Usually, molasses contain between 65% and 75% of sugars, mainly sucrose (Hongisto and Laakso, 1978); but the composition is highly variable depending on the sucrose-refining procedure and on the weather conditions of that particular year. Sucrose is extracellularly hydrolysed by yeasts in two monosaccharides, glucose and fructose, which are transported to and incorporated into the yeast metabolism as carbon sources. However, molasses are deficient in other essential elements for yeast growth. One of them is nitrogen since its molasses content is very poor (less than 3%). Yeasts can use some of the amino acids present in molasses, but addition of nitrogen sources is needed, generally in the form of ammonium salts or urea. Magnesium and phosphate elements are also supplemented in salt forms. Finally, three vitamins (biotin, thiamine and pantothenic acid), required for fast growth, must be supplemented since their content in molasses is also very low (Oura, 1974; Woehrer and Roehr, 1981). Another negative aspect of molasses being used as a substrate to produce yeasts is the presence of different toxics that can affect yeast growth. Variable amounts of herbicides, insecticides, fungicides, fertilizers and heavy metals applied to beet or cane crops can be found in molasses and in different stocks. Moreover bactericides, which are added during sugar production in refinery plants, can be found (Reed and Nagodawithana, 1988). All these toxics can decrease yeast performance by inhibiting growth (Pérez-Torrado, 2004). In fact, a common practice in yeast plants is to mix different stocks to dilute potential toxics.
The effects of molasses composition on yeast growth have been recently analysed at molecular level by determining the transcriptional profile of yeast growing in beet molasses and by comparing it to complete synthetic media (Shima et al., 2005). The results revealed that yeast displays clear gene expression responses when grown in industrial media because of the induction of
In the last years, the price of molasses has increased because of their use in other industrial applications such as animal feeding or bioethanol production (Arshad et al., 2008; Kopsahelis et al. 2009; Xandé et al., 2010), thus rendering the evaluation of new substrates for yeast biomass propagation a trend topic for biomass producers’ research. New assayed substrates include molasses mixtures with corn steep liquor (20:80), different agricultural waste products (Vu and Kim, 2009) and other possibilities as date juice (Beiroti and Hosseini, 2007) or agricultural waste sources, also called wood molasses, that can be substrate only for yeast species capable of using xylose as a carbon source.
3. Scaling up: Bach and fed-bach
Nowadays, yeast biomass propagation of wine, distiller’s and brewer’s yeasts are usually produced in baker’s yeast plants. The procedure is designed as a multistage-based fermentation, previously defined for the production of baker´s yeast (Chen and Chiger, 1985; Reed and Nagodawithana, 1991) using supplemented molasses as growth media. The first stage (F1) is initiated with a flask culture containing molasses, which is inoculated with the selected yeast strain. Production cultures may be periodically renewed from the stock cultures maintained under more stringent control procedures in a central quality control laboratory. Then, the initial culture is used to inoculate the first fermentor, and cells grow in various transient stages during the batch (F2-F4) and fed-batch (F5-F6) phases of the process. In a sequence of consecutive fermentations, the yeast biomass grown in small fermentors is used to inoculate larger tanks (Reed, 1982; Chen and Chiger, 1985; Reed and Nagodawithana, 1991; Degre, 1993).
In the initial batch phase (F2), cells are exposed to the high sugars concentration present in molasses. All the other nutrients are also present in the fermentor, and pH must be adjusted to 4.5-5.0 after sterilisation to be then monitored during batch fermentation. Once the batch phase has started, the only controllable parameters are temperature and aeration. Yeast propagation typically involves continuous aeration or oxygenation, but a relatively short aeration period has been suggested to suffice (Maemura et al., 1998). However the presence of O2 from the beginning of the process allows yeast cells to synthesise lipids, thereby revitalising the sterol-deficient cell population and ensuring that fermentation can proceed efficiently. Besides, those propagation experiments carried out in non-oxygenated media considerably reduce yeast growth and increase internal oxidative stress (Boulton, 2000; Pérez-Torrado et al., 2009).
During batch fermentation (F2-F4), a growth lag phase takes place in which cells synthesise the enzymes involved in gluconeogenesis and the glyoxylate cycle (Haarasilta and Oura, 1975). During the subsequent exponential phase, a very small amount of glucose is oxidised in the mitochondria, but when the sugar concentration drops below a strain-specific level or the specific growth rate in aerobic cultures exceeds a critical value (μcrit), a mixed respiro-fermentative metabolism occurs. This phenomenon has been described as the ”Crabtree effect” (De Deken, 1966; Pronk et al., 1996) and was originally considered a consequence of the catabolite repression and limited respiratory capacity of
Alcoholic fermentation leads to a suboptimal biomass concentration because the ATP yield is much lower than the yield obtained during respiratory carbohydrate degradation (Verduyn, 1991; Rizzi et al., 1997). However, pre-adaptation to large amounts of glucose during the batch phase is necessary to ensure the produced biomass’ optimal fermentative capacity by accumulating several necessary reserve metabolites to be used in the fed-batch phase (Dombek and Ingram, 1987; Rizzi et al., 1997; Pérez-Torrado et al., 2009). In addition, prolonged growth in aerobic, glucose-limited chemostat cultures of
Optimisation of biomass productivity requires an increase in both the specific growth rate and the biomass yield during the fed-batch phase to the highest values possible under sugar-limited cultivation. Generally, the growth rate profile during fed-batch cultivation is controlled primarily by the carbohydrate feedstock feed rate (Beudeker et al., 1990). The control of optimum dissolved oxygen during the fed-batch phase is also essential to obtain a high biomass yield, and important studies have been done to optimise aeration control (Blanco et al., 2008). Therefore sugar-limited cultivation in the presence of O2 allows the full respiratory growth of
Many research efforts have focused on optimising fed-batch processes for baker´s yeast production with different aims (productivity, yeast quality, or energy saving) and most have been commonly done under laboratory conditions (Van Hoek et al., 1998; Van Hoek et al., 2000; Jansen et al., 2005; Henes and Sonnleitner, 2007; Cheng et al., 2008), but rarely under pilot plant conditions (Di Serio et al., 2001; Lei et al., 2001; Gibson et al., 2007; Gibson et al., 2008). They have all been designed to mainly analyse the fed-batch phase without considering the whole process. The first published study on the complete industrial process was the simulation of wine yeast biomass propagation by performing batch and fed-batch phases in only one bioreactor (Pérez-Torrado et al., 2005). This simplification of the process enabled the study of yeast physiology from a molecular point of view with a bench-top design (Fig. 1), whose results display a good correlation with those obtained from pilot plants and this set of parameters for further investigation.
The parameters employed throughout the process (sucrose and ethanol production / consumption, dissolved O2, cell density and feed rate) have been adapted from Gómez-Pastor et al., 2010b. The lower panel shows representative cellular states, along with the most relevant metabolites, proteins and gene expressions throughout biomass propagation.
4. Desiccation of wine yeasts
In contrast to baker’s and brewer’s yeast, seasonal wine production requires the development of highly stable dry yeast products. At the end of biomass propagation, wine yeast cells are recovered and dehydrated to obtain ADY (Chen and Chiger, 1985; Degre, 1993; Gonzalez et al., 2005). After the maturation step, yeast cells are separated from fermented media by centrifugation, and are subjected to washing separations to reduce non-yeast solids, a necessary step because they affect the proper rehydration process of ADY for must fermentation. The separation process yields a slightly coloured yeast cream containing up to 22% yeast solids. After this step, the yeast cream can be stored at 4C after adjusting the pH to 3.5 to avoid microbial contaminations. The cream yeast is further dehydrated to 30-35% solids by means of rotary vacuum filters or filter presses. The filtered yeast is usually mixed with emulsifiers prior to its extrusion into yeast strands. The yeast cake is extruded through a perforated plate, while particles are loaded into the dryer and dehydrated to obtain a product with very low residual moisture. Although several types of dryers exist (roto-louvre, belt dryers, spray dryers), the one most commonly used in industry is the fluidized-bed dryer. In this dryer, heated air is blown from the bottom through yeast particles at velocities which keep them in suspension. Air is treated to reduce its water content and to ensure that the yeast temperature does not exceed 35C or 41C during drying. Drying times may vary from 15 to 60 min depending on the mass volume and the used conditions. Finally, ADY with less than 8% residual moisture is vacuum-packaged or placed in an inert atmosphere, such as nitrogen and CO2, to reduce oxidation. Depending on the strain, loss of viability is estimated at between 10% and 25% per year at 20C. For this reason, manufacturers recommend storing ADY at 4C in a dry atmosphere for a maximum 3-year period.
In order to produce an ADY product with acceptable fermentative activity and storage stability, several factors must be taken into account. The drying temperature and rate can be critical for yeast resistance to dehydration and rehydration (Beney et al., 2000; Beney et al., 2001; Laroche and Gervais, 2003). Some studies have shown that cell death during desiccation is strongly related to membrane integrity loss, leading to cell lysis during rehydration (Beney and Gervais, 2001; Laroche et al., 2001; Simonin et al. 2007; Dupont et al., 2010). A gradual dehydration kinetics, which allows a slow water efflux through the plasmatic membrane and homogenous desiccation, followed by a progressive rehydration during the starter preparation, have been related with high cell viability (Gervais et al., 1992; Gervais and Marechal, 1994¸ Dupont et al., 2010). The amount of cell constituents leaked during rehydration can also be reduced by adding emulsifiers, such as sorbitan monostearate (Chen and Chiger, 1985). Moreover, biomass propagation conditions have a major influence on yeast resistance to dehydration-rehydration. Several cultivation factors can affect cell resistance to desiccation, such as the substrate, growth phase and ion availability (Trofimova et al., 2010).
5. Yeast stress along biomass production
Several classic studies have evaluated the energy, kinetic and yield parameters of the yeast biomass production process (Reed, 1982; Chen and Chiger, 1985; Reed and Nagodawithana, 1991; Degre, 1993). However, the biochemical and molecular aspects of yeast adaptation to adverse industrial growth conditions have been poorly characterised. In recent years, a substantial effort has been made to gain insight into yeast responses during the process. It was believed that industrial conditions were optimised to obtain the best performing yeast cells, but now we know that yeast cells endure several stressful situations that induce multiple intracellular changes and challenge their technological fitness (Attfield, 1997; Pretorius, 1997; Pérez-Torrado et al., 2005). With wine yeast, moreover, the biomass is concentrated and dehydrated at the end of the process to obtain ADY yeasts that can be stored for long periods of time (Degre, 1993). Subsequently in a period of several hours during maturation and final drying processing, cells undergo nutrient limitation and a complex mixture of different stresses (thermic, osmotic, oxidative, etc.) (Garre et al., 2010). As a result, these dynamic environmental injuries seriously affect biomass yield, fermentative capacity, vitality, and cell viability (Attfield, 1997; Pretorius, 1997; Pérez-Torrado et al., 2005; Pérez-Torrado et al., 2009).
Eukaryotic cells have developed molecular mechanisms to sense stressful situations, transfer information to the nucleus and adapt to new conditions (Hohmann and Mager, 1997; Estruch, 2000; Hohmann, 2002). Protective molecules are rapidly synthesised in stressful situations and transcriptional factors are activated, thus changing the transcriptional profile of cells. Many stress response genes are induced under several adverse conditions through sequence element STRE (stress-responsive element), which targets the main transcriptional factors Msn2p and Msn4p (Kobayashi and McEntee, 1993; Martinez-Pastor et al., 1996). This pathway, also known as the “general stress response pathway”, increases the expression of many different genes, including the well-studied
Given the antiquity of yeast fermentation processes, these microorganisms have evolved in natural stressing environments, which have favoured the selection of “domesticated” yeast that displays high stress resistance (Jamieson, 1998). Studies of brewing yeast under industrial fermentations have demonstrated the suitability of the marker gene expression as a tool to study yeast stress responses in industrial processes (Higgins et al., 2003a). Monitoring stress-related marker genes, such as
As mentioned earlier, an oxygen supply is necessary to generate yeast biomass and to ensure optimal physiological conditions for effective fermentation (Chen and Chiger, 1985; Reed and Nagodawithana, 1991; Hulse, 2008). Oxygen is required for lipid synthesis, which is necessary to maintain plasma membrane integrity and function, and consequently for both cell replication and the biosynthesis of sterols and unsaturated fatty acids. Despite its potential toxicity, eliminating oxygen in the first part of the batch phase diminishes biomass yield (Boulton et al., 2000; Pérez-Torrado et al., 2009) and avoids the expression of those genes related to oxidative stress response, such as
During an industrial-scale propagation of wine and brewing yeasts, catalase and Mn superoxide dismutase activities increase as propagation proceeds (Martin et al., 2003; Gómez-Pastor et al., 2010a), indicating the importance of oxidative stress response throughout the process, whereas Sod1p (Cu/Zn superoxide dismutase) transiently accumulates at the end of the batch phase when ethanol is consumed (Gómez-Pastor et al., 2010a). A study of different types of oxidative damage during wine yeast biomass propagation has revealed that lipid peroxidation considerably increases during the metabolic transition from fermentation to respiration, which decreases to basal levels during the fed-batch phase (Gómez-Pastor et al., 2010a). Besides, the protein carbonylation analysis, one of the most important oxidative damages (Stadtman and Levine, 2000), has revealed different protein oxidation patterns during biomass propagation, which reach maximum global carbonylation levels at the end of the batch phase (Gómez-Pastor et al., 2010a). As protein oxidation causes the loss of catalytic or structural integrity, further research into the specific oxidised proteins during biomass production should be done to correlate the detriment in fermentative capacity with specific damaged proteins. In addition, reduced glutathione, an important antioxidant molecule, varies during the process as is lowers during the metabolic transition, while oxidised glutathione increases. Then, reduced glutathione increases constantly in different stages of the process (Gibson et al., 2006; Gómez-Pastor et al., 2010a). Whether glutathione is directly affected by O2 during biomass propagation remains unknown and requires further investigation.
The fed-bath phase is characterised by the accumulation of other important antioxidant molecules, such as trehalose and thioredoxin (Trx2p) (Pérez-Torrado, 2004; Gómez-Pastor, 2010), although the mRNA levels for the
Proteomic studies have also been carried out to gain a better understanding of the fluctuations in the stress-related gene mRNA levels during biomass propagation and to correlate glycolytic enzyme activities with their corresponding protein levels. However, the proteomic data available from industrial processes are very limited and usually centre on bioethanol production (Cot et al., 2007; Cheng et al., 2008) or wine and beer fermentations (Trabalzini et al., 2003; Zuzuarregui et al., 2006; Salvadó et al., 2008; Rossignol et al., 2009). Recent proteomic studies performed by 2D-gel electrophoresis during wine yeast biomass propagation have revealed that several glycolytic enzyme isoforms increase during biomass production. This is probably due to the post-translational modifications after oxidative stress exposure (Gómez-Pastor et al., 2010b; Costa et al., 2002). Trabalzini et al. (2003) suggested that some specific isoforms of glycolytic/gluconeogenic pathway enzymes in wine strains of
The industrial yeast biomass dehydration process also involves damaging environmental changes. As the biomass is being concentrated, water molecules are removed and temperature increases, all of which affect the viability and vitality of cells (Matthews and Webb, 1991). Dehydration is known to cause both cell growth arrest and severe damage to membranes and proteins (Potts, 2001; Singh et al., 2005). Removal of water molecules causes protein denaturalisation, aggregation, and loss of activity in an irreversible manner (Prestrelski et al., 1993). Additionally at the membrane level, desiccation is associated with an increased package of polar groups of phospholipids, and with the formation of endovesicles leading to cell lysis during rehydration (Crowe et al., 1992; Simonin et al., 2007). Yeasts have several strategies to maintain membrane fluidity (Beney and Gervais, 2001). One of them is to accumulate ergosterol, this being the predominant sterol in
Recent phenomic and transcriptomic analyses during the desiccation of a laboratory strain have indicated that this process represents a complex stress involving changes in about 12% of the yeast genome (Ratnakumar et al., 2011). Under these conditions, the induction of 71 genes grouped into the “environmental stress response” category was observed, suggesting a role of the general stress transcription factors Msn2p and Msn4p in the desiccation stress response. Furthermore, the phenomic screen looking for genes that are beneficial to desiccation tolerance has identified several of the transcriptional regulators or protein kinases involved in oxidative (
The accumulation of some metabolites has been related to yeasts’ resistance to drying and subsequent rehydration. One of them is the amino acid proline. This amino acid exhibits multiple functions
Interestingly, wine yeasts accumulate large amounts of disaccharide trehalose, usually in the 12-20% range of cell dry weight (Degre, 1993) although higher percentages have been detected in industrial stocks (Garre et al., 2010). Trehalose content has been proposed as one of the most important factors to affect dehydration survival. Baker’s yeasts with 5% of trehalose are 3 times more sensitive to desiccation than those cells accumulating 20% of trehalose (Cerrutti et al., 2000). The main function of this metabolite is to act as a protective molecule in stress response. This effect can be achieved in two ways: by protecting membrane integrity through the union with phospholipids (reviewed in Crowe et al., 1992); by preserving the native conformation of proteins and preventing the aggregation of partially denatured proteins (Singer and Lindquist, 1998a). The indispensability of this metabolite to survive dehydration is a controversial subject. Some studies have suggested that its presence is essential and needed in both sides of the membrane to confer suitable protection (Eleuterio et al., 1993; Sales et al, 2000). However, these results are argued alongside the
6. Conclusions
In the last few decades, the yeast biomass production industry has contributed with many advanced approaches to traditional technological tools with a view to studying the physiology, biochemistry and gene expression of yeast cells during biomass growth and processing. This has provided a picture of the determinant factors for the commercial product’s high yield and fermentative fitness. Cell adaptation to adverse industrial conditions is a key element for good progress to be made in biomass propagation and desiccation, and towards the characterisation of specific stress responses during industrial processes to clearly indicate the main injuries affecting cell survival and growth. One major aspect of relevance in the complex pattern of molecular responses displayed by yeast cells is oxidative stress response, a network of mechanisms ensuring cellular redox balance by minimising structural damages under oxidant insults. Different components of this machinery have been identified as being involved in cellular adaptation to industrial growth and dehydration, including redox protein thioredoxin, redox buffer glutathione and several detoxifying enzymes such as catalase and superoxide dismutase, plus protective molecules like trehalose which play a relevant role in dehydration.
7. Future prospects
In spite of the sound knowledge available on molecular responses to exogenous oxidants, the endogenous origin of oxidative stress in yeast biomass production, given the metabolic transitions required for growth under the described multistage-based fermentation conditions and desiccation, makes it challenging to search for the specific targets undergoing oxidative damage during both biomass propagation and desiccation, and to correlate this damage with physiologically detrimental effects. Based on the currently global data available and the use of potent analytical and genetic manipulation tools, further research has to be conducted to (i) define specific oxidised proteins and to know how this oxidation affects fermentative efficiency, (ii) identify new key elements in stress response, which can be manipulated to improve it and can be also used as markers to select suitable strains for biomass production, (iii) analyse the effects of potential beneficial additives, such as antioxidants, on yeast cells’ ability to adapt to stress, and then yeast biomass’ yield and fermentative fitness in industrial production processes.
Acknowledgments
This work has been supported by grants AGL 2008-00060 from the Spanish Ministry of Education and Science (MEC). E.G. was a fellow of the FPI program of the Spanish Ministry of Education and Science, R.G-P was a predoctoral fellow of the I3P program from the CSIC (Spanish National Research Council).
References
- 1.
Alexander M. A. Jeffries T. W. 1990 Respiratory efficiency and metabolize partitioning as regulatory phenomena in yeasts.12 2 29 - 2.
Aranda A. Querol A. del Olmo M. 2002 Correlation between acetaldehyde and ethanol resistance and expression of HSP genes in yeast strains isolated during the biological aging of sherry wines. ,177 4 304 312 - 3.
Arshad M. Khan Z. M. Khalil-ur-Rehman Shah. F. A. Rajoka M. I. 2008 Optimization of process variables for minimization of byproduct formation during fermentation of blackstrap molasses to ethanol at industrial scale. .47 5 410 414 - 4.
Attfield P. V. 1997 Stress tolerance: the key to effective strains of industrial baker’s yeast. ,15 13 1351 1357 - 5.
Bauer F. F. Pretorius I. S. 2000 Yeast stress response and fermentation efficiency: how to survive the making of wine- A review.21 27 51 - 6.
Beiroti A. Hosseini S. N. 2007 Production of baker’s yeast using date juice. .23 4 746 750 - 7.
Benaroudj N. Lee D. H. Goldberg A. L. 2001 Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. ,276 26 24261 24267 - 8.
Beney L. Martinez d. M. Marechal I. P. A. Gervais P. 2000 Influence of thermal and osmotic stresses on the viability of the yeast . Int.J.Food Microbiol.,55 1-3 275 279 - 9.
Beney L. Gervais P. 2001 Influence of the fluidity of the membrane on the response of microorganisms to environmental stresses.57 1-2 34 42 - 10.
Beney L. Marechal P. A. Gervais P. 2001 Coupling effects of osmotic pressure and temperature on the viability of . Appl.Microbiol.Biotechnol.,56 3-4 513 516 - 11.
Beudeker R. F. van Dam H. W. van der Plaat J. B. Vellenga K. 1990 Developments in bakers’ yeast production. In: (Verachtert, H. and De Mot, R., Eds.),103 146 Marcel Dekker, New York, NY. - 12.
Bisson L. F. Karpel Ramakrishnan J. E. V. Joseph L. 2007 Functional genomics of wine yeast . Adv Food Nutr Res.,53 65 121 - 13.
Blanco C. A. Rayo J. Giralda J. M. 2008 Improving industrial full-scale production of baker’s yeast by optimizing aeration control. ,91 3 607 613 - 14.
Boulton C. 2000 Trehalose, glycogen and sterol. In: . 1st ed (SmartKA, Ed),10 19 Blackwell Science, Oxford, UK. - 15.
Boy-Marcote E. Perrot M. Bussereau F. Boucherie H. Jacquet M. 1998 Msn2p and Msn4p control a large number of genes induced at the diauxic transition which are repressed by cyclic AMP in . J. Bacteriol180 5 1044 1052 - 16.
Brewster J. L. de V. T. Dwyer N. D. Winter E. Gustin M. C. 1993 An osmosensing signal transduction pathway in yeast. ,259 5102 1760 1763 - 17.
Bruckmann A. Hensbergen P. J. Balog C. I. Deelder A. M. de Steensma H. Y. van Heusden G. P. 2007 Post-transcriptional control of the proteome by 14-3-3 proteins. J.Proteome.Res.,6 5 1689 1699 - 18.
Caesar R. Palmfeldt J. Gustafsson J. S. Pettersson E. Hashemi S. H. Blomberg A. 2007 Comparative proteomics of industrial lager yeast reveals differential expression of the cerevisiae and non-cerevisiae parts of their genomes. ,7 22 4135 4147 - 19.
Cerrutti P. Segovia de H. M. Galvagno M. Schebor C. del Pilar B. M. 2000 Commercial baker’s yeast stability as affected by intracellular content of trehalose, dehydration procedure and the physical properties of external matrices. ,54 4 575 580 - 20.
Production of baker’s yeast. (Chen S. L. Chiger M. 1985 In ed. Moo-Young, M., Blarch, H.W., Drew, S. and Wang, D.I.C.429 462 New York: Pergamon Press. - 21.
Cheng J. S. Qiao B. Yuan Y. J. 2008 Comparative proteome analysis of robust insights into industrial continuous and batch fermentation. Appl.Microbiol.Biotechnol.,81 2 327 338 - 22.
Clarkson S. P. Large P. J. Boulton C. A. Bamforth C. W. 1991 Synthesis of superoxide dismutase, catalase and other enzymes and oxygen and superoxide toxicity during changes in oxygen concentration in cultures of brewing yeast.7 2 91 103 - 23.
Coote P. J. Cole M. B. Jones M. V. 1991 Induction of increased thermotolerance in may be triggered by a mechanism involving intracellular pH. J.Gen.Microbiol.,137 7 1701 1708 - 24.
Costa V. M. Amorim M. A. Quintanilha A. Moradas-Ferreira P. 2002 Hydrogen peroxide-induced carbonylation of key metabolic enzymes in : the involvement of the oxidative stress response regulators Yap1 and Skn7. Free Radic.Biol.Med.,33 11 1507 1515 - 25.
Cot M. Loret M. O. Francois J. Benbadis L. 2007 Physiological behaviour of in aerated fed-batch fermentation for high level production of bioethanol. FEMS Yeast Res.,7 1 22 32 - 26.
Crowe J. H. Hoekstra F. A. Crowe L. M. 1992 Anhydrobiosis. ,54 579 599 - 27.
da Costa. Morato N. D. da Silva C. G. Mariani D. Fernandes P. N. Pereira M. D. Panek A. D. Eleutherio E. C. 2008 The role of trehalose and its transporter in protection against reactive oxygen species. ,1780 12 1408 1411 - 28.
De Deken R. H. 1966 The Crabtree effect: a regulatory system in yeast. ,44 2 149 156 - 29.
Degre R. 1993 Selection and commercial cultivation of wine yeast and bacteria. In: (Fleet GH, Ed.)421 448 Harwood Academic Publishers, Chur, Switzerland. - 30.
Di Serio M. Tesser R. Santacesaria E. 2001 A kinetic and mass transfer model to simulate the growth of baker’s yeast in industrial bioreactors. J82 347 354 - 31.
Dombek K. M. Ingram L. O. 1987 Ethanol production during batch fermentation with : changes in glycolytic enzymes and internal pH. Appl.Environ.Microbiol.,53 6 1286 1291 - 32.
Dupont S. Beney L. Ritt J. F. Lherminier J. Gervais P. 2010 Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration. B,1798 5 975 985 - 33.
Dupont S. Beney L. Ferreira T. Gervais P. 2010 Nature of sterols affects plasma membrane behavior and yeast survival during dehydration. . - 34.
Eleutherio E. C. Araujo P. S. Panek A. D. 1993 Role of the trehalose carrier in dehydration resistance of Saccharomyces cerevisiae. ,1156 3 263 266 - 35.
Eleutherio E. C. Maia F. M. Pereira M. D. Degre R. Cameron D. Panek A. D. 1997 Induction of desiccation tolerance by osmotic treatment in . Can.J.Microbiol.,43 5 495 498 - 36.
Ertugay N. Hamamci H. 1997 Continuous cultivation of bakers’ yeast: change in cell composition at different dilution rates and effect of heat stress on trehalose level. ,42 5 463 467 - 37.
Espindola A. S. Gomes D. S. Panek A. D. Eleutherio E. C. 2003 The role of glutathione in yeast dehydration tolerance. ,47 3 236 241 - 38.
Estruch F. 2000 Stress-controlled transcription factors, stress-induced genes and stress tolerance in budding yeast. ,24 4 469 486 - 39.
França M. B. Panek A. D. Eleutherio E. C. 2005 The role of cytoplasmic catalase in dehydration tolerance of . Cell Stress.Chaperones.,10 3 167 170 - 40.
França M. B. Panek A. D. Eleutherio E. C. 2007 Oxidative stress and its effects during dehydration. ,146 4 621 631 - 41.
François J. Parrou J. L. 2001 Reserve carbohydrates metabolism in the yeast . FEMS Microbiol.Rev.,25 1 125 145 - 42.
Garre E. Pérez-Torrado R. Gimeno-Alcañíz J. V. Matallana E. 2009 Acid trehalase is involved in intracellular trehalose mobilization during postdiauxic growth and severe saline stress in . FEMS Yeast Res.,9 1 52 62 - 43.
Garre E. Raginel F. Palacios A. Julien A. Matallana E. 2010 Oxidative stress responses and lipid peroxidation damage are induced during dehydration in the production of dry active wine yeasts. ,136 3 295 303 - 44.
Gasch A. P. Werner-Washburne M. 2002 The genomics of yeast responses to environmental stress and starvation. ,2 4-5 181 192 - 45.
Gervais P. Marechal P. A. Molin P. 1992 Effects of the kinetics of osmotic pressure variation on yeast viability. ,40 11 1435 1439 - 46.
Gervais P. Marechal P. A. 1994 Yeast resistance to high-levels of osmotic-pressure-influence of kinetics. ,22 399 407 - 47.
Gibson B. R. Lawrence S. J. Boulton C. A. Box W. G. Graham N. S. Linforth R. S. Smart K. A. 2008 The oxidative stress response of a lager brewing yeast strain during industrial propagation and fermentation. ,8 4 574 585 - 48.
Gibson B. R. Smith J. M. Lawrence S. J. Shelton N. Smith J. M. Smart K. A. 2006 Oxygen as toxin: oxidative stress and brewing yeast physiology.31 1 25 36 - 49.
Gibson B. R. Lawrence S. J. Leclaire P. R. Powell C. D. Smart K. A. 2007 Yeast responses to stresses associated with industrial brewering handling.31 5 535 569 - 50.
Godon C. Lagniel G. Lee J. Buhler J. M. Kieffer S. Perrot M. Boucherie H. Toledano M. B. Labarre J. 1998 The H2O2 stimulon in . J.Biol.Chem.,273 35 22480 22489 - 51.
Gómez-Pastor R. 2010 Estrés oxidativo en la producción de levaduras vínicas. Implicación del gel TRX2. Universitat de València. Valencia; PhD Thesis. - 52.
Gómez-Pastor R. Pérez-Torrado R. Matallana E. 2010ª Improving yield of industrial biomass propagation by increasing the Trx2p dosage. ,1 5 352 353 - 53.
Gómez-Pastor R. Pérez-Torrado R. Cabiscol E. Matallana E. 2010b Transcriptomic and proteomic insights of the wine yeast biomass propagation process. ,10 7 870 884 - 54.
Gómez-Pastor R. Pérez-Torrado R. Cabiscol E. Ros J. Matallana E. 2010c Reduction of oxidative cellular damage by overexpression of the thioredoxin TRX2 gene improves yield and quality of wine yeast dry active biomass. ,9 9 - 55.
González R. Muñoz R. Carrascosa A. V. 2005 Producción de cultivos iniciadores para elaborar el vino. (Carrascosa, A.V, Muñoz, R. &. González, R, Ed.)318 341 AMV Ediciones, España. - 56.
Haarasilta S. Oura E. 1975 Effect of aeration on the activity of gluconeogenetic enzymes in growing under glucose limitation. Arch.Microbiol.,106 3 271 273 - 57.
Henes B. Sonnleitner B. 2007 Controlled fed-batch by tracking the maximal culture capacity. ,132 2 118 126 - 58.
Herdeiro R. S. Pereira M. D. Panek A. D. Eleutherio E. C. 2006 Trehalose protects from lipid peroxidation during oxidative stress. Biochim.Biophys.Acta,1760 3 340 346 - 59.
Higgins V. J. Beckhouse A. G. Oliver A. D. Rogers P. J. Dawes I. W. 2003b Yeast genome-wide expression analysis identifies a strong ergosterol and oxidative stress response during the initial stages of an industrial lager fermentation. ,69 8 4777 4787 - 60.
Higgins V. J. Rogers P. J. Dawes I. W. 2003a Application of genome-wide expression analysis to identify molecular markers useful in monitoring industrial fermentations. ,69 12 7535 7540 - 61.
Hohmann S. Mager W. H. 1997 Yeast stress responses. MBIU R.G. Landes Company, USA. - 62.
Hohmann S. Mager W. H. 2003 Yeast stress responses. Springer, New York, USA. - 63.
Hohmann S. 2002 Osmotic stress signaling and osmoadaptation in yeasts. ,66 2 300 372 - 64.
Hongisto H. J. Laakso P. 1978 th General Meeting, American Society of Sugar Beet Technologists, San Diego, 26 February-2 March. - 65.
Hulse G. 2008 Yeast Propagation, in Brewing Yeast Fermentation Performance, Second Edition (ed K. Smart), Blackwell Science, Oxford, UK,doi: ch23. - 66.
Ivorra C. Pérez-Ortin J. E. del Olmo M. 1999 An inverse correlation between stress resistance and stuck fermentations in wine yeasts. A molecular study. ,64 6 698 708 - 67.
Jamieson D. J. 1998 Oxidative stress responses of the yeast Saccharomyces cerevisiae. Yeast,14 16 1511 1527 - 68.
Jansen M. L. Diderich J. A. Mashego M. Hassane A. de Winde J. H. ran-Lapujade P. Pronk J. T. 2005 Prolonged selection in aerobic, glucose-limited chemostat cultures of causes a partial loss of glycolytic capacity. Microbiology,151 5 1657 1669 - 69.
Kaino T. Takagi H. 2009 Proline as a stress protectant in the yeast : effects of trehalose and PRO1 gene expression on stress tolerance. H.Biosci Biotechnol Biochem,73 9 2131 2135 - 70.
Kaino T. Tateiwa T. Mizukami-Murata S. Shima J. Takagi H. 2008 Self-cloning baker’s yeasts that accumulate proline enhance freeze tolerance in doughs. ,74 18 5845 5849 - 71.
Kim J. Alizadeh P. Harding T. Hefner-Gravink A. Klionsky D. J. 1996 Disruption of the yeast gene confers better survival after dehydration, freezing, and ethanol shock: potential commercial applications. Appl.Environ.Microbiol.,62 5 1563 1569 - 72.
Kobayashi N. Mc Entee K. 1993 Identification of cis and trans components of a novel heat shock stress regulatory pathway in . Mol.Cell Biol.,13 1 248 256 - 73.
Kopsahelis N. Nisiotou A. Kourkoutas Y. Panas P. Nychas G. J. Kanellaki M. 2009 Molecular characterization and molasses fermentation performance of a wild yeast strain operating in an extremely wide temperature range. .100 20 4854 4862 - 74.
Landolfo S. Zara G. Zara S. Budroni M. Ciani M. Mannazzu I. 2010 Oleic acid and ergosterol supplementation mitigates oxidative stress in wine strains of . Int J Food Microbiol,141 3 229 235 - 75.
Laroche C. Beney L. Marechal P. A. Gervais P. 2001 The effect of osmotic pressure on the membrane fluidity of at different physiological temperatures. Appl.Microbiol.Biotechnol.,56 1-2 249 254 - 76.
Laroche C. Gervais P. 2003 Achievement of rapid osmotic dehydration at specific temperatures could maintain high viability. Appl.Microbiol.Biotechnol.,60 6 743 747 - 77.
Le Moan N. Clement G. Le Moan S. Tacnet F. Toledano M. B. 2006 The Saccharomyces cerevisiae proteome of oxidized protein thiols: contrasted functions for the thioredoxin and glutathione pathways. J.Biol.Chem.,281 15 10420 10430 - 78.
Lei F. Rotbøll F. Jørgensen S. B. 2001 A biochemically structured model for . J. Biotechnol88 3 205 221 - 79.
Lindquist S. Craig E. A. 1988 The heat-shock proteins. ,22 631 677 - 80.
Maemura H. Morimura S. Kida K. 1998 Effects of aeration during the cultivation of pitching yeast on its characteristics during the subsequent fermentation of wort.104 207 211 - 81.
Martin V. Quain D. E. Smart K. A. 2003 Brewing yeast oxidative stress responses: impact of brewery handling. . 2nd edn (SmartKA, ed),61 73 Blackwell Science, Oxford, UK. - 82.
Martinez-Pastor M. T. Marchler G. Schuller C. Marchler-Bauer A. Ruis H. Estruch F. 1996 The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). EMBO J.,15 9 2227 2235 - 83.
Matthews T. M. Webb C. 1991 Culture systems. In: (Tuite, M.F., Oliver, S.G., Eds.)249 282 Plenum Press. - 84.
Miskiewicz T. Borowiak D. 2005 A logistic feeding profile for a Fed-Batch baker’s yeast process. Electronic Journal of Polish Agricultural University8 4 1 13 - 85.
Oku K. Watanabe H. Kubota M. Fukuda S. Kurimoto M. Tsujisaka Y. Komori M. Inoue Y. Sakurai M. 2003 NMR and quantum chemical study on the OH...pi and CH...O interactions between trehalose and unsaturated fatty acids: implication for the mechanism of antioxidant function of trehalose.125 42 12739 12748 - 86.
Oura E. 1974 Effect of aeration intensity on the biochemical composition of baker’s yeast. I. Factors affecting the type of metabolism. ,16 9 1197 1212 - 87.
Pereira E. J. Panek A. D. Eleutherio E. C. 2003 Protection against oxidation during dehydration of yeast. ,8 2 120 124 - 88.
Pérez-Torrado R. 2004 Estudio y mejora del proceso de producción industrial de levaduras vínicas. Universitat de València. Valencisa ; PhDThesis. - 89.
Pérez-Torrado R. Bruno-Barcena J. M. Matallana E. 2005 Monitoring stress-related genes during the process of biomass propagation of strains used for wine making. Appl.Environ.Microbiol.,71 11 6831 6837 - 90.
Pérez-Torrado R. Gimeno-Alcaniz J. V. Matallana E. 2002 Wine yeast strains engineered for glycogen overproduction display enhanced viability under glucose deprivation conditions. ,68 7 3339 3344 - 91.
Pérez-Torrado R. Gómez-Pastor R. Larsson C. Matallana E. 2009 Fermentative capacity of dry active wine yeast requires a specific oxidative stress response during industrial biomass growth. ,81 5 951 960 - 92.
Piper P. W. 1995 The heat shock and ethanol stress responses of yeast exhibit extensive similarity and functional overlap. ,134 2-3 121 127 - 93.
Potts M. 2001 Desiccation tolerance: a simple process? .9 11 553 559 - 94.
Postma E. Verduyn C. Scheffers W. A. Van Dijken J. P. 1989 Enzymic analysis of the crabtree effect in glucose-limited chemostat cultures of . Appl.Environ.Microbiol.,55 2 468 477 - 95.
Prestrelski S. J. Tedeschi N. Arakawa T. Carpenter J. F. 1993 Dehydration-induced conformational transitions in proteins and their inhibition by stabilizers.65 2 661 671 - 96.
Utilization of polysaccharides by . (Pretorius I. S. 1997 In: Yeast Sugar Metabolism. F.K. Zimmermann & K.-D. Entian (Eds.),459 501 Technomic Publishing Co., Inc., Lancaster, Pennsylvania, USA. - 97.
Pronk J. T. Yde S. H. Van Dijken J. P. 1996 Pyruvate metabolism in Yeast,12 16 1607 1633 - 98.
Querol A. Fernández-Espinar M. T. del Olmo M. Barrio E. 2003 Adaptive evolution of wine yeast.86 3 10 - 99.
Ratnakumar S. Tunnacliffe A. 2006 Intracellular trehalose is neither necessary nor sufficient for desiccation tolerance in yeast. ,6 6 902 913 - 100.
Ratnakumar S. Hesketh A. Gkargkas K. Wilson M. Rash B. M. Hayes A. Tunnacliffe A. Oliver S. G. 2011 Phenomic and transcriptomic analyses reveal that autophagy plays a major role in desiccation tolerance in . Mol.Biosyst.,7 1 139 149 - 101.
Reed G. Nagodawithana T. W. 1988 Technology of Yeast Usage in Winemaking. Am. J. Enol. Vitic.,39 1 83 90 - 102.
Reed G. Nagodawithana T. W. 1991 Baker’s Yeast Production. In: (Reed, G. & Nagodawithana,T. W., Eds.),261 314 Van Nostrand Reinhold, New York. - 103.
Reed S. I. 1982 Yeast genetics. ,215 4537 1233 1234 - 104.
Rizzi M. Baltes M. Theobald U. Reuss M. 1997 In vivo analysis of metabolic dynamics in : II. Mathematical model. Biotechnol.Bioeng.,55 4 592 608 - 105.
Rossignol T. Kobi D. Jacquet-Gutfreund L. Blondin B. 2009 The proteome of a wine yeast strain during fermentation, correlation with the transcriptome. ,107 1 47 55 - 106.
Rossignol T. Postaire O. Storaï J. Blondin B. 2006 Analysis of the genomic response of a wine yeast to rehydration and inoculation.71 5 699 712 - 107.
Sales K. Brandt W. Rumbak E. Lindsey G. 2000 The LEA-like protein HSP 12 in has a plasma membrane location and protects membranes against desiccation and ethanol-induced stress. Biochim.Biophys.Acta,1463 2 267 278 - 108.
Salvadó Z. Chiva R. Rodríguez-Vargas S. Rández-Gil F. Mas A. Guillamón J. M. 2008 Proteomic evolution of a wine yeast during the first hours of fermentation. ,8 7 1137 1146 - 109.
Shamrock V. J. Lindsey G. G. 2008 A compensatory increase in trehalose synthesis in response to desiccation stress in Saccharomyces cerevisiae cells lacking the heat shock protein Hsp12p. ,54 7 559 568 - 110.
Shima J. Hino A. Yamada-Iyo C. Suzuki Y. Nakajima R. Watanabe H. Mori K. Takano H. 1999 Stress tolerance in doughs of trehalase mutants derived from commercial Baker’s yeast. Appl.Environ.Microbiol.,65 7 2841 2846 - 111.
Shima J. Kuwazaki S. Tanaka F. Watanabe H. Yamamoto H. Nakajima R. Tokashiki T. Tamura H. 2005 Identification of genes whose expressions are enhanced or reduced in baker’s yeast during fed-batch culture process using molasses medium by DNA microarray analysis. ,102 1 63 71 - 112.
Simonin H. Beney L. Gervais P. 2007 Sequence of occurring damages in yeast plasma membrane during dehydration and rehydration: mechanisms of cell death. ,1768 6 1600 1610 - 113.
Simons K. Ikonen E. 1997 Functional rafts in cell membranes. ,5 6633 569 572 - 114.
Singer M. A. Lindquist S. 1998 Thermotolerance in : the Yin and Yang of trehalose. Trends Biotechnol.16 11 460 468 - 115.
Singh J. Kumar D. Ramakrishnan N. Singhal V. Jervis J. Garst J. F. Slaughter S. M. De Santis A. M. Potts M. Helm R. F. 2005 Transcriptional response of to desiccation and rehydration. Appl.Environ.Microbiol.,71 12 8752 8763 - 116.
Sorger P. K. 1991 Heat shock factor and the heat shock response. ,65 3 363 366 - 117.
Stadtman E. R. Levine R. L. 2000 Protein oxidation. ,899 191 208 - 118.
Swan T. M. Watson K. 1998 Stress tolerance in a yeast sterol auxotroph: role of ergosterol, heat shock proteins and trehalose. .1 1 191 197 - 119.
Takagi H. 2008 Proline as a stress protectant in yeast: physiological functions, metabolic regulations, and biotechnological applications. ,81 2 211 223 - 120.
Trabalzini L. Paffetti A. Ferro E. Scaloni A. Talamo F. Millucci L. Martelli P. Santucci A. 2003 Proteomic characterization of a wild-type wine strain of . Ital.J.Biochem.,52 4 145 153 - 121.
Trevisol E. T. Panek A. D. Mannarino S. C. Eleutherio E. C. 2011 The effect of trehalose on the fermentation performance of aged cells of . Appl.Microbiol.Biotechnol. - 122.
Trofimova Y. Walker G. Rapoport A. 2010 Anhydrobiosis in yeast: influence of calcium and magnesium ions on yeast resistance to dehydration-rehydration. ,308 1 55 61 - 123.
Trollmo C. Andre L. Blomberg A. Adler L. 1988 Physiological overlap between osmotolerance and thermotolerance in. FEMS Microbiol Lett56 3 321 326 - 124.
Valentinotti S. Holmberg U. Srinivasan B. Cannizzaro C. Rhiel M. von Stockar U. Bonvin D. 2002 Optimal operation of fed-batch fermentations via adaptive control of overflow metabolite.11 6 665 674 - 125.
Van der Aar P. C. Van Verseveld H. W. Stouthamer A. H. 1990 Stimulated glycolytic flux increases the oxygen uptake rate and aerobic ethanol production, during oxido-reductive growth of . J. Biotechnol13 347 359 - 126.
Van H. P. de H. E. Van Dijken J. P. Pronk J. T. 2000 Fermentative capacity in high-cell-density fed-batch cultures of baker’s yeast. ,68 5 517 523 - 127.
Van H. P. Van Dijken J. P. Pronk J. T. 1998 Effect of specific growth rate on fermentative capacity of baker’s yeast. ,64 11 4226 4233 - 128.
Varela J. C. van B. C. Planta R. J. Mager W. H. 1992 Osmostress-induced changes in yeast gene expression. Mol.Microbiol.,6 15 2183 2190 - 129.
Vemuri G. N. Eiteman M. A. Mc Ewen J. E. Olsson L. Nielsen J. 2007 Increasing NADH oxidation reduces overflow metabolism in Proc. Natl. Acad. Sci.,104 7 2402 2407 - 130.
Verduyn C. 1991 Physiology of yeasts in relation to biomass yields. ,60 3-4 325 353 - 131.
Vu V. H. Kim K. 2009 High-cell-density fed-batch culture of Saccharomyces cerevisiae KV-25 using molasses and corn steep liquor. .19 12 1603 1611 - 132.
Woehrer W. Roehr M. 1981 Regulatory aspects of bakers’ yeast metabolism in aerobic fed-batch cultures.23 3 567 581 - 133.
Xandé X. Archimède H. Gourdine J. L. Anais C. Renaudeau D. 2010 Effects of the level of sugarcane molasses on growth and carcass performance of Caribbean growing pigs reared under a ground sugarcane stalks feeding system. .42 1 13 20 - 134.
Xu Z. Tsurugi K. 2006 A potential mechanism of energy-metabolism oscillation in an aerobic chemostat culture of the yeast . FEBS J.,273 8 1696 1709 - 135.
Zuzuarregui A. Carrasco P. Palacios A. Julien A. del Olmo M. 2005 Analysis of the expression of some stress induced genes in several commercial wine yeast strains at the beginning of vinification. ,98 2 299 307 - 136.
Zuzuarregui A. Monteoliva L. Gil C. del Olmo M. 2006 Transcriptomic and proteomic approach for understanding the molecular basis of adaptation of to wine fermentation. Appl.Environ.Microbiol.,72 1 836 847