Ethanol yield and volatile compound content in fermentation of agave must by Kluyveromyces marxianus UMPe-1 comparing with Saccharomyces cerevisiae baker's yeast used in tequila production

In tequila production, fermentation is an important step. Fermentation determines the ethanol productivity and organoleptic properties of the beverage. In this study, a yeast isolated from native residual agave must was identified as Kluyveromyces marxianus UMPe-1 by 26S rRNA sequencing. This yeast was compared with the baker's yeast Saccharomyces cerevisiae Pan1. Our findings demonstrate that the UMPe-1 yeast was able to support the sugar content of agave must and glucose up to 22% (w/v) and tolerated 10% (v/v) ethanol concentration in the medium with 50% cells survival. Pilot and industrial fermentation of agave must tests showed that the K. marxianus UMPe-1 yeast produced ethanol with yields of 94% and 96% with respect to fermentable sugar content (glucose and fructose, constituting 98%). The S. cerevisiae Pan1 baker's yeast, however, which is commonly used in some tequila factories, showed 76% and 70% yield. At the industrial level, UMPe-1 yeast shows a maximum velocity of fermentable sugar consumption of 2.27g·L(-1)·h(-1) and ethanol production of 1.38g·L(-1)·h(-1), providing 58.78g ethanol·L(-1) at 72h fermentation, which corresponds to 96% yield. In addition, the major and minor volatile compounds in the tequila beverage obtained from UMPe-1 yeast were increased. Importantly, 29 volatile compounds were identified, while the beverage obtained from Pan1-yeast contained fewer compounds and in lower concentrations. The results suggest that the K. marxianus UMPe-1 is a suitable yeast for agave must fermentation, showing high ethanol productivity and increased volatile compound content comparing with a S. cerevisiae baker's yeast used in tequila production.     

Production of Dairy-Based,Natural Sulphur Flavor Concentrate by Yeast Fermentation

Production of sulphur flavor concentrate by yeast fermentation was investigated using dairy media (milk and cream) spiked with L-methionine. The yeasts studied included so-called dairy yeasts Candida kefyr, Kluyveromyces marxianus, Debaryomyces hansenii, Geotrichum candidum, and Yarrowia lipolytica as well as wine yeasts Saccharomyces cerevisiae and Saccharomyces bayanus. Methionol was produced as the predominant volatile sulphur flavor compound. Other volatile sulphur flavor compounds produced were dimethyl disulphide, S-methyl thioacetate, 3-methylthio-1-propanoic acid, methional, 3-methylthio-1-propene, and ethyl 3-methylthio-1-propanoate. In addition, numerous other volatile flavor compounds were produced, including ethanol, branched-chain alcohols, aromatic alcohol, ethyl esters, branched-chain esters, branched-chain fatty acids, and branched-chain aldehydes. The sensory attribute of a methionol sulphur flavor concentrate in cream is concentration-dependent, ranging from a savory-slight potato note to a strong potato-savory note to a cooked cheese-potato note.     

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.     

Influence of carbon and nitrogen source on production of volatile fragrance and flavour metabolites by the yeast Kluyveromyces marxianus

The yeast Kluyveromyces marxianus produces a range of volatile molecules with applications as fragrances or flavours. The purpose of this study was to establish how nutritional conditions influence the production of these metabolites. Four strains were grown on synthetic media, using a variety of carbon and nitrogen sources and volatile metabolites analysed using gas chromatography–mass spectrometry (GC–MS). The nitrogen source had pronounced effects on metabolite production: levels of the fusel alcohols 2‐phenylethanol and isoamyl alcohol were highest when yeast extract was the nitrogen source, and ammonium had a strong repressing effect on production of 2‐phenylethyl acetate. In contrast, the nitrogen source did not affect production of isoamyl acetate or ethyl acetate, indicating that more than one alcohol acetyl transferase activity is present in K. marxianus. Production of all acetate esters was low when cells were growing on lactose (as opposed to glucose or fructose), with a lower intracellular pool of acetyl CoA being one explanation for this observation. Bioinformatic and phylogenetic analysis of the known yeast alcohol acetyl transferases ATF1 and ATF2 suggests that the ancestral protein Atf2p may not be involved in synthesis of volatile acetate esters in K. marxianus, and raises interesting questions as to what other genes encode this activity in non‐Saccharomyces yeasts. Identification of all the genes involved in ester synthesis will be important for development of the K. marxianus platform for flavour and fragrance production. Copyright © 2014 John Wiley & Sons, Ltd.     

Headspace flavour compounds produced by yeasts and lactobacilli during fermentation of preferments and bread doughs.

Production of volatile flavour compounds during fermentation with pure cultures of Saccharomyces cerevisiae and Candida guilliermondii, Lactobacillus brevis and Lactobacillus plantarum have been investigated, using wheat doughs and several preferements as substrates. For yeast, preferments consisted of 10% (w/v) glucose, maltose and sucrose solutions, whereas for lactobacilli they consisted of supplemented and unsupplemented (3% and 10% (w/v)) glucose solutions, and a 10% (w/v) wheat flour slurry. Seven volatile compounds (acetaldehyde, acetone, ethyl acetate, ethanol, hexanal+isobutyl alcohol, and propanol) were detected when using yeasts. All these compounds, except propanol, appeared for all the substrates assayed, with ethanol as the predominant component. Generally, S. cerevisiae produced higher amounts of the different components than C. guilliermondii. Both yeasts produced larger amounts of volatile flavour compounds during fermentation in glucose and sucrose solutions than in maltose or wheat dough. In general the yeasts examined produced more flavour components than the lactobacilli. For the lactobacilli the highest number of volatile flavour compounds were observed for substrates containing flour.     

Effects of yeast fermentation on transforming the volatile compounds of unsalted pork hydrolysate

This study investigated the possibility of transforming unsalted pork hydrolysate into a liquid seasoning by studying volatile compound production through yeast fermentation. The yeasts used included a typical soya sauce yeast and three wine yeasts. The yeasts inoculated in the glucose supplementary pork hydrolysate increased by approximately 2.0 log of CFU mL⁻¹ within 24 hours at 30°C without adding salt. Yeast fermentation largely depleted the abundant aldehydes (hexenal) present in unfermented pork hydrolysate to trace levels with the formation of ethanol, corresponding alcohols and carboxylic acids. The wine yeasts showed higher production of fruity esters such as ethyl acetate and isoamyl acetate, whereas the soya sauce yeast produced more ketones such as acetone and 2-heptanone. This study revealed a potential method of producing a value-added meat hydrolysate from pork by-product.     

Performance of indigenous yeasts in the processing of Chinese strong‐flavoured liquor during spontaneous mixed solid‐state or submerged fermentation

To explore the in situ metabolic characteristics of yeasts involved in the spontaneous fermentation process of Chinese strong‐flavoured liquor, a comparison was conducted between solid‐state fermentation (SSF) and submerged fermentation (SmF) when supplemented with 24 indigenous yeast strains, with a focus on the production of ethanol and a broad range of volatile compounds responsible for the characteristics of Chinese strong‐flavoured liquor. Under the various experimental conditions, the 24 indigenous yeast strains showed different influences on the mixed fermentation system. The fluctuations caused by different yeast strains in the mixed system were less than those caused by the different fermentation modes relative to the formation of flavour compounds. SSF was found to be more suitable for the production of ethanol, methanol and ethyl lactate, whereas SmF was more suitable for the production of 10 higher alcohols, four esters and four acids. This study revealed the relationships amongst the indigenous yeasts, SSF, and the distinctive flavour profiles of Chinese strong‐flavoured liquor. This work provides evidence of the existence of internal stability in spontaneous SSF, thereby facilitating a better understanding of the fermentative mechanism in the SSF process for Chinese strong‐flavoured liquor production Copyright © 2015 The Institute of Brewing & Distilling     

Utilization of post‐fermentation yeasts for yeast extract production by autolysis: the effect of yeast strain and saponin from Quillaja saponaria

Spent yeasts, a co‐product from fermentation processes, are a source for unconvential autolysis processes. In this study, five post‐fermentation yeast strains that are often used in fermentation processes were used: Saccharomyces cerevisiae Ethanol Red (Lessafre), Kluyveromyces marxianus LOCK 0026, K. marxianus NCYC 179, Scheffersomyces stipitis NCYC 1541 and Pichia angusta NCYC 495. Autolysis was conducted at 50°C for 48 hours in the presence of saponins from Quillaja saponaria . The concentrations of proteins and free amino acids in the yeast autolysates were evaluated using IR spectroscopy and chromatography. The lysates were found to be good sources of essential amino acids, which constituted between 29.3% (S. cerevisiae ) and 40.7% (K. marxianus LOCK 0026) of the amino acid pools. The largest pools of free amino acids were found in autolysates of S. cerevisiae Ethanol Red (44.9 g/L) and P. angusta NCYC 495 (40.53 g/L). Saponin can be used as an auxiliary or alternative to conventional methods of cell lysis, especially since Q. saponaria extracts are approved for use in foods and could have significant health benefits. The usability of five post‐fermentation yeast strains as a source of valuable nitrogen compounds in unconventional salt‐free lysates was demonstrated for the first time in the present study. Copyright © 2017 The Institute of Brewing & Distilling     

THE EFFECT OF YEAST TREHALOSE CONTENT AT PITCHING ON FERMENTATION PERFORMANCE DURING BREWING FERMENTATIONS

The effect of yeast trehalose content at pitching on the fermentation performance during brewing fermentations was studied using a commercial strain of lager yeast, Saccharomyces cerevisiae (AJL 2155). Pitching yeasts with different trehalose contents were obtained by collecting cells in suspension after 96 h and 144 h of fermentation in EBC tubes in 10.8°P brewers wort at 14°C. The trehalose content of the pitching yeast had no effect on growth, specific gravity and ethanol production during the subsequent fermentation. A high trehalose content of the pitching yeast, however, sustained cell viability during the initial stage of fermentation, increased the carbohydrate utilisation rate and increased the production of isoamyl alcohol and isobutanol. For these aspects of fermentation performance, the trehalose content of the pitching yeast may prove useful in evaluating the vitality of pitching yeasts within the brewery .     

Fermentation Characteristics and Aromatic Profile of Plum Wines Produced with Indigenous Microbiota and Pure Cultures of Selected Yeast

The aim of this study was to assess and compare fermentation characteristics and aromatic profile of plum wines produced with indigenous microbiota and pure cultures of different selected yeast. Experiments were carried out with plum (Prunus domestica L.) varieties of different fruit ripening times (?a?anska rana, ?a?anska lepotica, and Po?ega?a). Wine fermentations were conducted by the activity of indigenous microbiota, commercially available Saccharomyces cerevisiae and Saccharomyces bayanus yeast strains and joint activity of Schizosaccharomyces pombe and S. cerevisiae (sequential inoculation). Statistically significant differences in fermentative characteristics and the content of certain volatile compounds were observed as a result of metabolic activity of various indigenous and/or selected yeasts during fermentation of plum pomace. Minimal duration of fermentation (4 to 5 d) and fastest ethanol production rate (from 12.3 to 15.5 g/L/d) were the characteristics of the studied S. cerevisiae strains. Isobutanol, 3‐methyl‐1‐butanol, 1‐heptanol, and 1‐octanol were the most prevalent higher alcohols in the tested plum wine samples. The predominant ester in plum wines was ethyl acetate, ethyl lactate, amyl acetate, isoamyl acetate, and ethyl palmitate, esters responsible for the floral and fruity olfactory tones, were also present in large amounts. Also, the use of S. cerevisiae strains resulted in the production of plum wines with better sensory characteristics than ones produced with other investigated yeasts. Obtained results are significant since there is limited data on the compounds responsible for the unique flavor of plum wine, as well as on the impact of different yeast starter cultures application on the overall quality of fruit wines.     

Impact of mixed Torulaspora delbrueckii-Saccharomyces cerevisiae culture on high-sugar fermentation

Conventional wine yeasts produce high concentrations of volatile acidity, mainly acetic acid, during high-sugar fermentation. This alcoholic fermentation by-product is highly detrimental to wine quality and, in some cases, levels may even exceed legal limits. In this study, a non-conventional species, Torulaspora delbrueckii, was used, in pure cultures and mixed with Saccharomyces cerevisiae yeast, to ferment botrytized musts. Fermentation rate, biomass growth, and the formation of volatile acidity, acetaldehyde, and glycerol were considered. This study demonstrated that T. delbrueckii, often described as a low acetic producer under standard conditions, retained this quality even in a high-sugar medium. Unlike S. cerevisiae, this species did not respond to the hyper-osmotic medium by increasing acetic production as soon as it is inoculated into the must. Nevertheless, this yeast produced low ethanol and biomass yields, and the fermentation was sluggish. As a result, T. delbrueckii fermentations do not reach the required ethanol content (14%vol.), although this species can survive at this concentration. A mixed culture of T. delbrueckii and S. cerevisiae was the best combination for improving the analytical profile of sweet wine, particularly volatile acidity and acetaldehyde production. A mixed T. delbrueckii/S. cerevisiae culture at a 20:1 ratio produced 53% less in volatile acidity and 60% less acetaldehyde than a pure culture of S. cerevisiae. Inoculating S. cerevisiae after 5 days' fermentation by T. delbrueckii had less effect on volatile acidity and acetaldehyde production and resulted in stuck fermentation. These results contribute to a better understanding of the behaviour of non-Saccharomyces and their potential application in wine industry.     

Outlining a future for non-Saccharomyces yeasts: selection of putative spoilage wine strains to be used in association with Saccharomyces cerevisiae for grape juice fermentation

The use of non-Saccharomyces yeasts that are generally considered as spoilage yeasts, in association with Saccharomyces cerevisiae for grape must fermentation was here evaluated. Analysis of the main oenological characteristics of pure cultures of 55 yeasts belonging to the genera Hanseniaspora, Pichia, Saccharomycodes and Zygosaccharomyces revealed wide biodiversity within each genus. Moreover, many of these non-Saccharomyces strains had interesting oenological properties in terms of fermentation purity, and ethanol and secondary metabolite production. The use of four non-Saccharomyces yeasts (one per genus) in mixed cultures with a commercial S. cerevisiae strain at different S. cerevisiae/non-Saccharomyces inoculum ratios was investigated. This revealed that most of the compounds normally produced at high concentrations by pure cultures of non-Saccharomyces, and which are considered detrimental to wine quality, do not reach threshold taste levels in these mixed fermentations. On the other hand, the analytical profiles of the wines produced by these mixed cultures indicated that depending on the yeast species and the S. cerevisiae/non-Saccharomyces inoculum ratio, these non-Saccharomyces yeasts can be used to increase production of polysaccharides and to modulate the final concentrations of acetic acid and volatile compounds, such as ethyl acetate, phenyl-ethyl acetate, 2-phenyl ethanol, and 2-methyl 1-butanol.     

Fermentative stress adaptation of hybrids within the Saccharomyces sensu stricto complex

Along the fermentation process yeasts are affected by a succession of stress conditions that affect their viability and fermentation efficiency. Among the stress conditions the most relevant are high sugar concentration and low pH in musts, temperature and, as fermentation progresses, ethanol accumulation. Nowadays, due to the demanding nature of modern winemaking practices and sophisticated wine markets, there is an ever-growing search for particular wine yeast strains possessing a wide range of optimized, improved or novel enological characteristics. Traditionally, the species S. cerevisiae and S. bayanus within the Saccharomyces sensu stricto species are considered some of the most important yeast species involved in fermentation processes. However, in the last years, hybrid strains between the species S. cerevisiae, S. bayanus and S. kudriavzevii have been described as yeasts conducting the alcoholic fermentations and some of them are commercially available. Our results indicate that yeasts in the Saccharomyces sensu stricto complex were not affected by low pH or high glucose content in the media; however temperature and ethanol concentration variables appreciably affected their growth. The strains pertaining to S. cerevisiae were able to tolerate high temperature stress, whereas strains within S. bayanus and S. kudriavzevii were better adapted to growth at lower temperatures. Regarding to alcohol tolerance, S. cerevisiae is tolerating alcohol better than S. bayanus or S. kudriavzevii. Surprisingly, the natural hybrids between these species have adapted to growth under ethanol and temperature stress by inheriting competitive traits from one or another parental species. These results open new perspectives in the construction of new hybrid strains with biotechnological interest, as the characteristics of the parents may result in interesting combinations in the hybrids.     

Probiotic potential of γ-aminobutyric acid (GABA)–producing yeast and its influence on the quality of cheese

Kazakh cheese is a traditional dairy product in Xinjiang, China. To study the function and potential probiotic characteristics of yeast in Kazakh cheese and its contribution to cheese fermentation, we screened the γ-aminobutyric acid (GABA)–producing yeasts Pichia kudriavzevii 1–21, Kluyveromyces marxianus B13–5, Saccharomyces cerevisiae DL6–20, and Kluyveromyces lactis DY1–10. We investigated the potential probiotic properties of these strains and their use in cheese fermentation (cheeses designated CSP, CSM, CSS, and CSI, respectively); a control with no added yeast was designated CS. The results showed that the 4 yeast strains all showed high self-polymerization (2- and 24-h autoaggregation capacity of >80 and 90%, respectively), hydrophobicity (40–92% variation, low hydrophobicity in xylene, but within the range of probiotics), and the ability to survive the gastrointestinal tract (survival rate >75% after simulation), indicating the probiotic ability of the strains in vitro. The GABA production capacity of the CSM cheese increased (to 95.6 mg/100 g), but its protein content did not change significantly, and amino acid degradation was obvious. The GABA production capacity of the CSS cheese decreased (to 450 mg/kg); its protein content declined, and its amino acid content increased. Except for water and protein, we found no obvious differences in most physical and chemical indicators. Kluyveromyces marxianus B13–5 helped to form the desired texture. Multivariate statistical analysis showed that fermentation of the cheese with the 4 yeasts improved the production of esters and alcohols. The CSS cheese had good aroma production performance, because S. cerevisiae DL6–20 produced high concentrations of isoamyl alcohol, hexanoic acid ethyl ester, benzyl alcohol, octanoic acid ethyl ester, 3-hydroxy-2-butanone, and hexanoic acid; the content of 2-methyl-propanoic acid was low. Compared with the CSP cheese, the CSI and CSM cheeses had a fruitier aroma and a milder odor, but the CSI and CSM cheeses had high concentrations of ethyl acetate, butanoic acid, ethyl ester, 3-methyl-1-butanol-acetate, ethyl hexanoate, ethyl octanoate, acetic acid 2-phenylethyl ester, and ethyl lactate; concentrations of 3-methyl-butanoic acid, propanoic acid, acetic acid, and butanoic acid were low. The CSP cheese had stronger acid-producing ability. The order of fragrance production performance was CSS > CSI, CSM > CSP > CS. Research into the fermentation mechanisms of GABA-producing yeast in cheese will provide a theoretical basis for the quality control and industrial production of Kazakh cheese.     

Evaluation of the potential of commercial non-Saccharomyces yeast strains of Torulaspora delbrueckii and Lachancea thermotolerans in beer fermentation

This work evaluated the potential of three commercial non-Saccharomyces yeast strains Torulaspora delbrueckii (Biodiva and Prelude) and Lachancea thermotolerans (Concerto) for beer fermentation. The fermentation performance, volatile and non-volatile profiles were compared. Growth behaviours of all three yeast strains exhibited similar trends during the initial fermentation phase although a marked population decline was detected in strains Prelude and Concerto, which also showed a rapid utilisation of maltose, while strain Biodiva was unable to consume maltose and consumed lesser amounts of amino acids. Additionally, terpenoids inherently absent in the wort such as β-caryophyllene and geranyl acetone were produced in all beers, significantly higher in beers fermented with strain Prelude. For volatile profiles, Prelude and Concerto produced more ethanol and significantly higher amounts of acetate esters and long-chained ethyl esters. Strain Biodiva, on the other hand, produced higher amounts of isoamyl alcohol and ethyl butanoate.     

Influence of yeast strains on the physicochemical characteristics,methanol and acetaldehyde profiles and volatile compounds for Korean rice distilled spirit

The objective of this study was to investigate the influence of yeast strains on the physicochemical characteristics, methanol and acetaldehyde profiles, and volatile compounds of Korean rice distilled spirits. Ten yeast strains were employed for the brewing of distilled spirits and the resulting products were filtered and distilled twice. The amounts of methanol and acetaldehyde for the ten yeasts showed different profiles. Higher amounts of methanol were detected for strains CL, CY, DV, BD, ED and LP, while EC, D2, D4 and RH had <2 mg L^?1 methanol content. Strains D2, BD and ED produced the lowest amounts of acetaldehyde. The head portions of the spirits, which started in the fraction that contained <5 mg L^?1 of acetaldehyde, were between 7.7% (BD) and 18.2% (LP) of the total fractions. Strains D2, CL and CY produced more alcohol in the body fraction than the other yeasts. The major volatile compounds were esters in the form of fatty acid ethyl esters, such as ethyl palmitate, ethyl myristate and ethyl oleate. Isoamyl alcohol, which is an important volatile compound for rice wine, occupied 0.91–2.24% of the relative peak areas. Strain D2, of the strains tested, appeared to be the most appropriate yeast for Korean distilled spirit based on alcohol production and the high relative peak area of volatile compounds, except for ethanol. Strains CL and CY could also be considered for producing high‐quality Korean rice distilled spirits with efficiency and flavour. Copyright © 2015 The Institute of Brewing & Distilling     

优良本土酿酒酵母的酿酒特性及产香能力初析

酿酒酵母产生的多种挥发性物质影响葡萄酒的香气风格,筛选本土酿酒酵母对改善葡萄酒同质化有积极作用.以13株不同来源的酿酒酵母和1株商业对照为试验菌株,对供试酵母的酒精耐受性、嗜杀性、产硫化氢能力及生长曲线进行了测定.随后在模拟汁中探究酵母的发酵特性,用固相微萃取-气相色谱/质谱(solid-phase microextr...     

Isolation and Characterization of Shochu Yeasts with Superior Brewing Ability from Shochu Mashes

Seven yeasts with superior ability in terms of alcohol fermentation and aromatic ingredient generation were isolated from 272 wild yeasts obtained from shochu mashes of shochu breweries. These seven yeasts were examined for their fermentation ability with rice and sweet potato using small scale of fermentation tests. Moreover, their thermotolerance was evaluated by growth tests and fermentation tests with barley koji. Among the isolated seven yeasts, the shochu yeast “MF062” was superior in the characteristics of fast fermentation, high alcohol production, aromatic ingredient generation and thermotolerance. It was named the “Heisei Miyazaki Yeast”. From the results of analyses of physiological and genetic characteristics of MF062, it was shown to be a Saccharomyces cerevisiae, but it showed different characteristics from the industrial yeasts used in shochu production. Using a sweet potato shochu brewing test, with 60 kg of raw materials and yeast MF062, it was confirmed that a high alcohol production yield and a high quality shochu could be obtained.     

三株野生型耐高温酵母的耐性对比研究

对三株野生型产酒精耐高温马克斯克鲁维酵母(Kluyveromyces marxianus)在不同温度、pH、酒精浓度、糖浓度下的生理耐性进行了研究,并与安琪耐高温酵母进行了比较,得出的结论为:三株野生型耐高温酵母K. marxianus GX-17,K. marxianus HN-79,K.marxianus HN-138高温条件下的生长和产酒精能力明显高于安碘酵母;K marxianus GX-17对pH变化的适应力较安琪酵母强;野生型耐高温酵母的耐糖能力略低于安琪酵母;安琪酵母的耐酒精能力明显高于野生型耐高温酵母.     

Impact of ammonium additions on volatile acidity, ethanol, and aromatic compound production by different Saccharomyces cerevisiae strains during fermentation in controlled synthetic media

Variations in ethanol, volatile acidity, and aromatic compounds produced by different Saccharomyces cerevisiae strains were studied in controlled synthetic medium (CSM). Different amounts of assimilable nitrogen (in the form of ammonium sulfate) were added at two fermentation stages (i.e. the beginning of fermentation and the halfway point). There were significant differences in the amount of ethanol produced when ammonia was added to the CSM, although this depended on the yeast strain used. When assimilable nitrogen was added, ethanol production either increased (with Fermicru AR2 and Stellevin NT116 yeast strains) or decreased (with LW LVCB CT1+ yeast strain). The degree of variation also depended on the time that the ammonia was added, with differences of up to 0.7% (v/v). Adding ammonium to the CSM always resulted in lower volatile acidity in the fermentation product. Different yeast strains ( P < 0.0001) and varying amounts of ammonium produced significant differences. Maximum impact – up to 70% less volatile acidity – was obtained using the Stellevin NT116 yeast strain and adding 280 mgN/L ammonium at the halfway point in fermentation. Acetoin production increased at higher concentrations of added ammonium, most markedly when it was added halfway through fermentation. The total amount of esters increased when 140 mgN/L was added to all three yeasts strains tested, irrespective of the fermentation stage. Smaller amounts of higher alcohols were produced following larger ammonium additions, especially at the beginning of fermentation.