Altered Patterns of Maltose and Glucose Fermentation by Brewing and Wine Yeasts Influenced by the Complexity of Nitrogen Source

Maltose and glucose fermentations by industrial brewing and wine yeasts strains were strongly affected by the structural complexity of the nitrogen source. In this study, four Saccharomyces cerevisiae strains, two brewing and two wine yeasts, were grown in a medium containing maltose or glucose supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low sugar concentration for brewing and wine strains, independent of nitrogen supplementation, and the type of sugar. At high sugar concentrations altered patterns of sugar fermentation were observed, and biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for brewing and wine strains. In maltose, high biomass production was observed under peptone and casamino acids for the brewing and wine strains, however efficient maltose utilization and high ethanol production was only observed in the presence of casamino acids for one brewing and one wine strain studied. Conversely, peptone and casamino acids induced higher biomass and ethanol production for the two other brewing and wine strains studied. With glucose, in general, peptone induced higher fermentation performance for all strains, and one brewing and wine strain produced the same amount of ethanol with peptone and casamino acids supplementation. Ammonium salts always induced poor yeast performance. The results described in this paper suggest that the complex nitrogen composition of the cultivation medium may create conditions resembling those responsible for inducing sluggish/stuck fermentation, and indicate that the kind and concentration of sugar, the complexity of nitrogen source and the yeast genetic background influence optimal industrial yeast fermentation performance.     

Effects of temperature, ammonium and glucose concentrations on yeast growth in a model wine system

In enology, alcoholic fermentation is a complex process involving several mechanisms. Slow and incomplete alcoholic fermentation is a chronic problem for the wine industry and factors leading to sluggish and stuck fermentations have been extensively studied and reviewed. The most studied cause of sluggish and stuck fermentation is the nitrogen content limitation. Nevertheless, other factors, such as temperature of fermentation and sugar concentration can affect the growth of yeasts. In this study we modelled the yeast growth‐cycle in wine model system as a function of temperature, sugar and ammonium concentrations; the individual effects and the interaction of these factors were analysed by means of a quadratic response surface methodology. Cell concentrations and weight loss were monitored in the whole wine fermentation process. The results of central composite design show that lower is the availability of nitrogen, higher is the cell growth rate; moreover, initial nitrogen concentration also influences survival time of Saccharomyces cerevisiae.     

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Glucose and fructose fermentations by industrial yeasts strains are strongly affected by both the structural complexity of the nitrogen source and the availability of oxygen. In this study two Saccharomyces cerevisiae industrial wine strains were grown, under shaken and static conditions, in a media containing either a) 20% (w/v) glucose, or b) 10% (w/v) fructose and 10% (w/v) glucose or c) 20% (w/v) fructose, all supplemented with nitrogen sources varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Data suggest that a complex structured nitrogen source is not submitted to the same control mechanisms as those involved in the utilization of simpler structured nitrogen sources, and mutual interaction between carbon and nitrogen sources, including the mechanisms involved in the regulation of aerobic/anaerobic metabolism, may play an important role in defining yeast fermentation performance and the differing response to the structural complexity of the nitrogen source, with a strong impact on fermentation performance.     

Sweet Wine Production by Two Osmotolerant Saccharomyces cerevisiae Strains

The use of Saccharomyces cerevisiae to produce sweet wine is difficult because yeast is affected by a hyperosmotic stress due to the high sugar concentrations in the fermenting must. One possible alternative could be the coimmobilization of the osmotolerant yeast strains S. cerevisiae X4 and X5 on Penicillium chrysogenum strain H3 (GRAS) for the partial fermentation of raisin musts. This immobilized has been, namely, as yeast biocapsules. Traditional sweet wine (that is, without fermentation of the must) and must partially fermented by free yeast cells were also used for comparison. Partially fermented sweet wines showed higher concentration of the volatile compounds than traditionally produced wines. The wines obtained by immobilized yeast cells reached minor concentrations of major alcohols than wines by free cells. The consumption of specific nitrogen compounds was dependent on yeast strain and the cellular immobilization. A principal component analysis shows that the compounds related to the response to osmotic stress (glycerol, acetaldehyde, acetoin, and butanediol) clearly differentiate the wines obtained with free yeasts but not the wines obtained with immobilized yeasts.     

Implications of nitrogen nutrition for grapes, fermentation and wine

This review discusses the impacts of nitrogen addition in the vineyard and winery, and establishes the effects that nitrogen has on grape berry and wine composition and the sensory attributes of wine. Nitrogen is the most abundant soil‐derived macronutrient in a grapevine, and plays a major role in many of the biological functions and processes of both grapevine and fermentative microorganisms. Manipulation of grapevine nitrogen nutrition has the potential to influence quality components in the grape and, ultimately, the wine. In addition, fermentation kinetics and formation of flavour‐active metabolites are also affected by the nitrogen status of the must, which can be further manipulated by addition of nitrogen in the winery. The only consistent effect of nitrogen application in the vineyard on grape berry quality components is an increase in the concentration of the major nitrogenous compounds, such as total nitrogen, total amino acids, arginine, proline and ammonium, and consequently yeast‐assimilable nitrogen (YAN). Both the form and amount of YAN have significant implications for wine quality. Low must YAN leads to low yeast populations and poor fermentation vigour, increased risk of sluggish/stuck/slow fermentations, increased production of undesirable thiols (e.g. hydrogen sulfide) and higher alcohols, and low production of esters and long chain volatile fatty acids. High must YAN leads to increased biomass and higher maximum heat output due to greater fermentation vigour, and increased formation of ethyl acetate, acetic acid and volatile acidity. Increased concentrations of haze‐causing proteins, urea and ethyl carbamate and biogenic amines are also associated with high YAN musts. The risk of microbial instability, potential taint from Botrytis‐infected fruit and possibly atypical ageing character is also increased. Intermediate must YAN favours the best balance between desirable and undesirable chemical and sensory wine attributes. ‘Macro tuning’, of berry nitrogen status can be achieved in the vineyard, given genetic constraints, but the final ‘micro tuning’ can be more readily achieved in the winery by the use of nitrogen supplements, such as diammonium phosphate (DAP) and the choice of fermentation conditions. This point highlights the need to monitor nitrogen not only in the vineyard but also in the must immediately before fermentation, so that appropriate additions can be made when required. Overall, optimisation of vineyard and fermentation nitrogen can contribute to quality factors in wine and hence affect its value. However, a better understanding of the effect of nitrogen on grape secondary metabolites and different types of nitrogen sources on yeast flavour metabolism and wine sensory properties is still required.     

Structural Complexity of the Nitrogen Source and Influence on Yeast Growth and Fermentation

The structural complexity of the nitrogen source strongly affects both biomass and ethanol production by industrial strains of Saccharomyces cerevisiae, during fermentation in media containing glucose or maltose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low glucose and maltose concentrations independent of nitrogen supplementation. At high sugar concentrations diauxie was not easily observed, and growth and ethanol production depended on the nature of the nitrogen source. This was different for baking and brewing ale and lager yeast strains. Sugar concentration had a strong effect on the shift from oxido‐fermentative to oxidative metabolism. At low sugar concentrations, biomass production was similar under both peptone and casamino acid supplementation. Under casamino acid supplementation, the time for metabolic shift increased with the glucose concentration, together with a decrease in the biomass production. This drastic effect on glucose fermentation resulted in the extinction of the second growth phase, probably due to the loss of cell viability. Ammonium salts always induced poor yeast performance. In general, supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, inducing higher biomass and ethanol production, and preserving yeast viability, in both glucose and maltose media, for baking and brewing ale and lager yeast strains. Determination of amino acid utilization showed that most free and peptide amino acids present, in peptone and casamino acids, were utilized by the yeast, suggesting that the results described in this work were not due to a nutritional status induced by nitrogen limitation.     

CONTROL OF AMMONIUM UPTAKE FROM MALT EXTRACT MEDIUM BY SACCHAROMYCES CEREVISIAE

When glucose was used as the energy source in the assay, the perceived ammonium permease activity of Saccharomyces cerevisiae NCYC 1108 remained relatively unchanged in yeast samples taken throughout the main period of fermentation of a malt extract medium. This was in contrast to the earlier work on the fermentation of a defined glucose/salts medium where the specific ammonium permease activity rose rapidly as fermentation proceeded. The present data suggest that the level of ammonium permease activity of cells fermenting a malt extract medium was low due to nitrogen catabolite repression and that inhibition of the carrier system was important in determining the actual rate of uptake from the medium. When ammonium permease in cells from a malt extract medium was assayed using maltose, essentially similar results were obtained. However, the actual values were lower and the results suggested that at some points during the fermentation the ability of the cells to take up maltose during the assay was an important factor in determination of the perceived ammonium permease activity.     

Influence of nitrogen on the biological aging of sherry wine

Nitrogen compounds are essential to the growth and metabolism of yeasts. The uptake and metabolism of nitrogen compounds by Saccharomyces cerevisiae depend not only on the strain and its physiological condition, but also on the chemical and physical properties of its environment. The effect of the addition of different amino acids (L ‐proline, L ‐threonine, L ‐arginine, L ‐glutamic acid, L ‐leucine and L ‐valine) to nitrogen‐depleted natural or nitrogen‐free synthetic wine on the cell growth, flor velum formation and sherry wine compound production was investigated under controlled biological aging by S. cerevisiae var. capensis strain G1 a typical flor yeast. The formation of flor velum was dependent on particular amino acid, oxygen availability and the composition of wine. Consumption of glycerol was related with the cell growth; in contrast, acetaldehyde tended to be released. Amino acid supplementation resulted in the release to wine of amino acids, esters and higher alcohols. The amino acid which was released in nearly all cases was L ‐leucine. Addition of L ‐glutamic acid resulted in the release mainly of ethyl acetate, in the case of L ‐leucine isoamyl alcohols were released, and for L ‐valine isobutanol. In the three cases, 1,1‐diethoxyethane was released in large quantities. The findings might indicate that the regulation of metabolism succeeds in the most efficient balancing of the redox potential. Copyright © 2006 Society of Chemical Industry     

Changes on free amino acids during the alcoholic fermentation of strawberry and persimmon

Changes in amino acids and ammonium were monitored during the alcoholic fermentation of strawberry and persimmon purees. Fermentations were carried out either by autochthonous or by commercial yeasts. The amino acid content in strawberry and persimmon was lower than that of grapes but enough to successfully perform the alcoholic fermentation, showing a different consumption pattern. Arginine, although is not present in the most strawberry substrates, appears in strawberry wines (2.75–3.36 mg L^?1). Additionally, as opposed to grape wine, an exceptional high consumption of proline was observed during the alcoholic fermentation of strawberry purees. The consumption of amino acids was highly influenced by the substrate and the S. cerevisiae strain used for the fermentation process. These results were confirmed by principal component analysis, which was able to group the samples based on substrate, harvest or yeast strain, considering the amino acids as variables.     

Effect of Sugar Catabolite Repression in Correlation with the Structural Complexity of the Nitrogen Source on Yeast Growth and Fermentation

Biomass and ethanol production by industrial Saccharomyces cerevisiae strains were strongly affected by the structural complexity of the nitrogen source during fermentation in media containing galactose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low galactose concentrations independent of nitrogen supplementation. At high sugar concentrations altered patterns of galactose utilisation were observed. Biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for baking and brewing ale and lager strains. Baking yeast showed improved galactose fermentation performance in the medium supplemented with casamino acids. High biomass production was observed with peptone and casamino acids for the ale brewing strain, however high ethanol production was observed only in the presence of casamino acids. Conversely, peptone was the nitrogen supplement that induced higher biomass and ethanol production for the lager brewing strain. Ammonium salts always induced poor yeast performance. The results with galactose differed from those obtained with glucose and maltose which indicated that supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, suggesting that sugar catabolite repression has a central role in yeast performance in a medium containing nitrogen sources with differing levels of structural complexity.     

Nitrogen requirements of commercial wine yeast strains during fermentation of a synthetic grape must

Nitrogen deficiencies in grape musts are one of the main causes of stuck or sluggish wine fermentations. Currently, the most common method for dealing with nitrogen-deficient fermentations is adding supplementary nitrogen (usually ammonium phosphate). However, it is important to know the specific nitrogen requirement of each strain, to avoid excessive addition that can lead to microbial instability and ethyl carbamate accumulation. In this study, we aimed to determine the effect of increasing nitrogen concentrations of three different nitrogen sources on growth and fermentation performance in four industrial wine yeast strains. This task was carried out using statistical modeling techniques. The strains PDM and RVA showed higher growth-rate and maximum population size and consumed nitrogen much more quickly than strains ARM and TTA. Likewise, the strains PDM and RVA were also the greatest nitrogen demanders. Thus, we can conclude that these differences in nitrogen demand positively correlated with higher growth rate and higher nitrogen uptake rate. The most direct effect of employing an adequate nitrogen concentration is the increase in biomass, which involves a higher fermentation rate. However, the impact of nitrogen on fermentation rate is not exclusively due to the increase in biomass because the strain TTA, which showed the worst growth behavior, had the best fermentation activity. Some strains may adapt a strategy whereby fewer cells with higher metabolic activity are produced. Regarding the nitrogen source used, all the strains showed the better and worse fermentation performance with arginine and ammonium, respectively.     

Effects of nitrogen catabolite repression‐related amino acids on the flavour of rice wine

The quality of rice wine is highly dependent on the content of the flavour compounds produced by the budding yeast Saccharomyces cerevisiae. In this study, the effects of three amino acids (arginine, glutamate and glutamine) related to nitrogen catabolite repression on the formation of flavour compounds were investigated. Each of these amino acids could promote the growth of S. cerevisiae, and a total of 83 flavour compounds were found in a model system of rice wine production. The effects of arginine, glutamate and glutamine on the content of the higher alcohols, amino acids and esters were significant, whereas the effects on the aldehydes and organic acids were slight. The results of this study could facilitate the development of new strategies to control the flavour pattern and improve the quality of rice wine. Copyright © 2015 The Institute of Brewing & Distilling     

Concentration of amino acids in wine after the end of fermentation by Saccharomyces cerevisiae strains

Both quantitative and qualitative differences in the utilization and release of assimilable nitrogen by two wine strains of Saccharomyces cerevisiae (cerevisiae and capensis) under different conditions of oxygen were observed. These differences were influenced by the presence of oxygen at the beginning of the fermentation, and by the strain of S cerevisiae. The release of some amino acids post‐fermentation may be the result of reoxidation of NAD(P)H in order to maintain a normal redox balance. Copyright © 2003 Society of Chemical Industry     

Alleviation of stuck wine fermentations using salt‐preconditioned yeast

The influence of salt (sodium chloride) on the cell physiology of wine yeast was investigated. Cellular viability and population growth of three wine‐making yeast strains of Saccharomyces cerevisiae, and two non‐Saccharomyces yeast strains associated with wine must microflora (Kluyveromyces thermotolerans and K. marxianus) were evaluated following salt pre‐treatments. Yeast cells growing in glucose defined media exposed to different sodium chloride concentrations (4, 6 and 10% w/v) exhibited enhanced viabilities compared with nontreated cultures in subsequent trial fermentations. Salt ‘preconditioning’ of wine yeast seed cultures was also shown to alleviate stuck and sluggish fermentations at the winery scale, indicating potential benefits for industrial fermentation processes. It is hypothesized that salt induces specific osmostress response genes to enable yeast cells to better tolerate the rigours of fermentation, particularly in high sugar and alcohol concentrations. Copyright © 2014 The Institute of Brewing & Distilling     

Screening of native S. cerevisiae strains in the production of Pajarete wine: a tradition of Atacama Region,Chile

Pajarete is a Chilean wine with an appellation of origin. Although it has organoleptic properties, intensive utilization of Saccharomyces cerevisiae commercial yeast through the years has presumably produced the loss of native strains that may be associated with Pajarete oenologic uniqueness. In order to evaluate the effect of re-incorporation of indigenous strains into Pajarete winemaking, native S. cerevisiae strains were isolated and selected based on their properties shown during small and large laboratory scale fermentation, and then evaluated in industrial bioreactors. From an initial set of 312 isolates, a single native strain was selected based on taxonomy, fermentation performance, aroma, residual sugars, and production of alcohol for incorporation into market scale.     

The Influence of Inoculum Level on Fermentation and Flavour Compounds of White Wines Made from cv. Emir

In this study, the influence of the addition of a commercial wine yeast (Saccharomyces cerevisiae) at inocula of 1 × 10⁴ to 1 × 10⁷ cells/ml in Emir must was investigated with a focus on yeast growth, fermentation rate, ethyl alcohol and flavour compound formation. Spontaneous fermentation without inoculation was also performed. Higher peak counts were observed with higher amounts of S. cerevisiae yeast. Addition of various amounts of yeast led to the earlier disappearance of non‐Saccharomyces yeasts. The fermentation rate was improved with higher amounts of yeast, but ethanol production was not affected. Concentrations of higher alcohols increased with increasing inoculum levels, especially inoculum sizes of 1 × 10⁶ cells/ml and 1 × 10⁷ cells/ml. The amount of ethyl acetate was reduced with increased inoculum levels.     

Production of Table Wine from Processed Tea Bags Using Different Strains of Saccharomyces cerevisiae

Wine samples were produced from locally available tea infusions (Lipton tea, Top tea and Highland tea) using baker’s yeast (Saccharomyces cerevisiae By1) and yeast cultures from pineapple (S. cerevisiae Py6) and cocoa (S. cerevisiae CY43). Physicochemical analyses and microbial evaluation were undertaken during fermentation. Lipton tea wine produced, using baker’s yeast, pineapple yeast and cocoa yeast had highest alcoholic contents of 7.88%, 6.25% and 7.20%, respectively. Top tea wine produced using the same set of yeasts had highest alcoholic contents of 9.78%, 5.43 and 8.15% respectively, while Highland tea wine produced highest alcoholic contents of 9.78%, 7.07% and 7.61% respectively. Physicochemical analyses for all the wines produced showed that the specific gravity, total solids and pH of the must decreased as fermentation progressed while the titratable acidity remained constant throughout the must fermentation. Colony counts showed a high biomass of yeast cells that decreased as it tended towards the end of fermentation. Sensory analysis of the wines showed that Lipton and Top tea are the most acceptable organoleptically when compared with the commercial wine used as control although the three tested teas were not significantly different statistically. Baker’s yeast was rated as the best yeast for wine fermentation irrespective of the type of tea used. All the wines produced were generally accepted as they were scored above average. This study highlights the potential of using different tea infusions as alternatives to grape and other fruit musts in wine making. It also confirms that commercial yeasts such as baker’s yeast can be used in homemade wine production.     

Evaluation of Brazilian ethanol production yeasts for maltose fermentation in media containing structurally complex nitrogen sources

Maltose and glucose fermentations are strongly affected by the structural complexity of the nitrogen source and by the presence of oxygen. In this study five industrial Saccharomyces cerevisiae strains were grown in synthetic medium, containing maltose or glucose, supplemented with different nitrogen sources, with or without agitation. All strains were able to grow and efficiently ferment glucose, but not all strains were able to grow and ferment maltose well. Peptone and ammonium sulfate induced improved fermentation for all strains and conditions. Under agitation, as expected, higher biomass accumulation was detected. Casamino acids supplementation induced efficient maltose fermentation for all of the strains under aerated conditions, but differing maltose utilization patterns were observed for the static cultures. Copyright © 2012 The Institute of Brewing & Distilling     

The Effect of Proanthocyanidins on Growth and Alcoholic Fermentation of Wine Yeast under Copper Stress

Proanthocyanidins (PAs) derived from the grape skin, as well as from grape seeds, grape stems, are an important group of polyphenols in wine. The aim of this study was to understand the effect of PAs (0.1, 1.0 g/L) on growth and alcoholic fermentation of 2 strains of Saccharomyces cerevisiae (commercial strain FREDDO and newly selected strain BH8) during copper‐stress fermentation, using a simple model fermentation system. Our results showed that both PAs and Cu²⁺ could pose significant inhibition effects on the growth of yeast cells, CO₂ release, sugar consumption, and ethanol production during the initial phase of the fermentation. Compared to PAs, Cu²⁺ performed more obvious inhibition on the yeast growth and fermentation. However, adding 1.0 g/L PAs increased in the vitality and metabolism activity of yeast cells at the mid‐exponential phase of fermentation in the mediums with no copper and 0.1 mM Cu²⁺ added, shortened the period of wine fermentation, and decreased the copper residues. It indicated that PAs could improve the ability of wine yeast to resist detrimental effects under copper‐stress fermentation condition, maintaining cells metabolic activity, and fermentation could be controlled by manipulating PAs supplementation.