Effect of added sulphur dioxide levels on the fermentation characteristics of strawberry wine

The aim of this study to evaluate the effects of sulphur dioxide (SO₂) on strawberry wine fermentation and on the quality of the final wine product. Major aroma compounds, reducing sugars, ethanol, titratable acid and microflora were analyzed during the fermentation of strawberry wine supplemented with 0–100 mg/L SO₂. As the amount of added SO₂ increased, the consumption of reducing sugars and soluble solids and ethanol production decreased during early fermentation, but increased during late fermentation. During the fermentation process, the concentrations of 2‐phenethanol, isobutanol and isopentanol significantly increased and those of n‐propanol, isoamyl acetate and ethyl lactate decreased with increasing amount of added SO₂. The production of n‐butanol, ethyl acetate and ethyl butyrate was slightly dependent on the amount of added SO₂. Yeast cells were the dominant microbe in the fermenting strawberry pulp, and indigenous bacteria and fungi populations decreased rapidly or disappeared because of their sensitivity to SO₂. It was concluded that 60–80 mg/L SO₂ should be added during strawberry wine fermentations to improve wine quality. Copyright © 2016 The Institute of Brewing & Distilling     

Dissolved oxygen content in apple must: technological implications in cider processing

Oxygen can influence the composition and quality of apple must and cider. In this study, the effect of unit operations on oxygen dissolution during apple must processing was investigated and the technological implications discussed. Oxygen dissolution after different apple must processing unit operations, and also the effect of enzymatic browning reaction on dissolved oxygen uptake were analysed. During pressing, the highest and first oxygen dissolution occurred in the must at 6.8 mg/L, on average. The enzymatic browning reaction after 10 min was responsible for an oxygen uptake above 80%. The addition of sulphur dioxide or maceration of crushed apple results in a must with more oxygen content, however the beverage colour is negatively affected. The incorporation of oxygen into the must by bubbling oxygen or the movement of liquid may not be effective owing to consumption by the reaction of residual polyphenol oxidase. Moreover, this can affect the alcoholic fermentation because of the oxidation of nutrients such as vitamins. If the apple must is depectinized, racked and filtered, there will be enough oxygen to obtain a quality cider. These results and observations can be useful for selecting unit operations in cider companies in order to improve the beverage quality. Copyright © 2014 The Institute of Brewing & Distilling     

Apple Aminoacid Profile and Yeast Strains in the Formation of Fusel Alcohols and Esters in Cider Production

The amino acid profile in dessert apple must and its effect on the synthesis of fusel alcohols and esters in cider were established by instrumental analysis. The amino acid profile was performed in nine apple musts. Two apple musts with high (>150 mg/L) and low (<75 mg/L) nitrogen content, and four enological yeast strains, were used in cider fermentation. The aspartic acid, asparagine and glutamic acid amino acids were the majority in all the apple juices, representing 57.10% to 81.95%. These three amino acids provided a high consumption (>90%) during fermentation in all the ciders. Principal component analysis (PCA) explained 81.42% of data variability and the separation of three groups for the analyzed samples was verified. The ciders manufactured with low nitrogen content showed sluggish fermentation and around 50% less content of volatile compounds (independent of the yeast strain used), which were mainly 3‐methyl‐1‐butanol (isoamyl alcohol) and esters. However, in the presence of amino acids (asparagine, aspartic acid, glutamic acid and alanine) there was a greater differentiation between the yeasts in the production of fusel alcohols and ethyl esters. High contents of these aminoacids in dessert apple musts are essential for the production of fusel alcohols and most of esters by aromatic yeasts during cider fermentation.     

Influence of SO<Subscript>2</Subscript> on the evolution of volatile compounds through alcoholic fermentation of must stabilized by pulsed electric fields

The aim of this work was to study the influence of sulphur dioxide (SO₂) on the formation of volatile compounds by yeast through wine alcoholic fermentation. Thus Parellada must was microbiologically stabilized using a pulsed electric field (PEF) treatment and inoculated with Saccharomyces cerevisiae Na33 strain. Fermentation was carried out with or without SO₂ and the results showed that the evolution of the volatile compounds profile throughout the process was similar. The content of volatile acids in wine obtained by using sulphur dioxide was not significantly different from that fermented without adding the compound. However, the final content of total alcohols and esters was significantly different even thought the differences were small. Consequently, when grape must is treated by PEF the sulphur dioxide concentration usually used in winemaking could be reduced to safer levels or even eliminated without an important effect on the volatile compounds content of the final product. Therefore, the absence of sulphur dioxide should not have a negative impact on the sensory characteristics of wine.     

Influence of a malolactic starter on the quality of the cider produced on an industrial scale

The application of starter culture technologies to induce the malolactic fermentation in cidermaking are not yet established, as is already the case in winemaking. In this work, a selected indigenous cider malolactic strain was tested as a starter culture to assess its implementation and its influence on cider production on an industrial scale (10,000 l). The results showed that the culture, under the fermentation conditions tested, allowed successful achievement of the malolactic fermentation. Analysis of L-malic acid in the fermenting apple must medium revealed a highly significant difference in the rate of consumption of this acid from the moment of bacterial inoculation when compared with that of the control. The concentration of the malolactic starter culture maintained the inoculation level (10⁶ cfu/ml) during the process, the level necessary to ensure the malolactic fermentation. Thus, the cider malolactic starter culture showed good adaptation to the harsh environmental conditions found in the fermenting apple must, such as low temperature and the presence of SO₂ and ethanol. Several parameters were monitored during the process (specific gravity, temperature, pH, free SO₂ content, microbiological count, malic acid degradation rate, lactic acid formation, other major organic acids and volatile compounds) under inoculated and not inoculated conditions.     

Replacement of sulfur dioxide by lysozyme and oenological tannins during fermentation: influence on volatile composition of white wines

BACKGROUND: In recent years the use of sulfur dioxide, a commonly used additive in winemaking, has been questioned because of its toxic effects on human health. Studies have been conducted to find alternatives that can effectively substitute for this additive in all its various technological functions. In previous work, lysozyme and oenological tannins were found as possible substitutes in controlling bacterial undesirable fermentations and phenolic oxidation. However, data on the volatile composition of wines obtained by that protocol are lacking. In this work, the effects on volatile composition of white wines by the substitution of SO₂ during fermentation with lysozyme and tannin were studied. At the same time, the technological performance of two strains of yeast that produce low amounts of SO₂ were evaluated. RESULTS: The results showed that both SO₂ and lysozyme prevented the development of undesirable bacterial fermentations. The study of volatile compounds shows differences in the alcohol, acid and ester contents among the final products: wines fermented with strain 1042 and lysozyme had higher total alcohol concentration, while the addition of SO₂ promoted higher production of 3‐methyl‐1‐butanol, 3‐methylthio‐1‐propanol, phenylethyl alcohol and 4‐hydroxy‐benzenethanol. Esters, as a total, were influenced by the different strain and tannins addition, while amounts of medium‐chain fatty acid ethyl esters and their corresponding fatty acids were found in higher amounts in wines coming from fermentations with lysozyme. The sensory analysis revealed a preference for wines to which lysozyme and tannins had been added. CONCLUSION: The data suggest that the addition of lysozyme and oenological tannins during alcoholic fermentation could represent a promising alternative to the use of SO₂ and for the production of wines with reduced content of SO₂. The composition of the volatiles in the final wines was affected by the different vinification protocols (mainly with regards to alcohols and ethyl esters). Copyright © 2009 Society of Chemical Industry     

Hydrogen sulphide production during cider fermentation is moderated by pre‐fermentation methionine addition

Yeast assimilable nitrogen (YAN) concentration and composition impact hydrogen sulphide (H₂S) production and fermentation kinetics during wine fermentation, but this phenomenon has not been extensively studied in cider fermentation. Our hypothesis was that H₂S production during cider fermentation could be decreased through pre‐fermentation modification of concentrations of individual amino acids. Apple juice (53 mg L⁻¹ YAN) was supplemented with asparagine, arginine, methionine or ammonium and fermented with EC1118 and UCD522 yeast strains. No difference in H₂S production among fermentations was observed with addition of asparagine, arginine or ammonium. Methionine addition of 5 mg L⁻¹ decreased H₂S production by yeast strain EC1118 at 53 mg L⁻¹ YAN. With 153 mg L⁻¹ initial YAN, only methionine addition of 50 mg L⁻¹ decreased H₂S production, and no tested methionine rates decreased H₂S production with 253 mg L⁻¹ initial YAN. Supplementation to 153 mg L⁻¹ YAN resulted in increased H₂S production at all methionine concentrations tested. Sensory differences in aroma were detected in samples supplemented with ammonium and methionine, and these differences were correlated with observed differences in H₂S production. Our results indicate that supplementing cider fermentations with methionine leads to lower H₂S formation, especially in apple juice containing low YAN. © 2017 The Authors Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

Influence of Enzymatic Clarification with a Pectin Methylesterase on Cider Fermentation

In this study, the effect of two apple juice treatments (clarification with a pectin methylesterase in the presence and absence of sulphiting) on the cidermaking process was investigated. Pre‐fermentative clarification with the pectin methylesterase (Rapidase CPE) slowed the alcoholic fermentation in respect to traditional fermentation, the greatest effect being found for the combination of the enzymatic treatment with sulphur dioxide addition. With this treatment the start of the fermentation was delayed seven days. Enzymatic clarification delayed malolactic fermentation (MLF) and the interaction of both treatments produced a cider in which malolactic fermentation had not occurred. Both treatments ensured that the volatile acid levels during the maturation phase were significantly lower than in the traditional fermentation. Enzymatic clarification led to lower levels of yeasts during the active phase of alcoholic fermentation with respect to the control. When this treatment was combined with sulphiting, a decrease in apiculate yeast numbers during alcoholic fermentation was observed and a better survival of these species was noted after 59 days of fermentation.     

发酵过程中添加磷酸氢二铵对苹果酒香气物质的影响

以富士苹果为原料,分别在发酵前、酵母生长对数期和稳定期前期添加280 mg/L的磷酸氢二铵（DAP）,研究DAP的添加对酵母发酵性能和苹果酒香气物质的影响。结果表明,发酵过程中添加DAP可加快酵母发酵速率,在酵母发酵前和酵母生长对数期添加DAP显著降低了琥珀酸含量（P＜0.05）;在稳定期前期添加DAP显著降低了苹果酸含量（P＜0.05）,总香气含量最高（11.09 mg/L）,更有利于苹果酒中挥发性香气化合物的合成,但对酵母数影响不大。     

Improvement of wine aromatic quality using mixtures of lysozyme and dimethyl dicarbonate,with low SO2 concentration

The use of sulphur dioxide (SO₂) in the treatment of foodstuffs presents some problems as it could lead to pseudo-allergies in some people. The aim of this research work was to study the addition of different preservative mixtures and their influence on the concentration of volatile compounds and sensorial quality in wine. To do so, vinifications were carried out using Garnacha must to which lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO₂ were added at different doses (25 and 50 mg l^?1). The results were compared with a control sample to which only SO₂ had been added (50 mg l^?1). In general, mixtures of SO₂ with lysozyme and DMDC favoured the formation of volatile compounds in the wines. Wines obtained from the mixtures of lysozyme and DMDC with 25 mg l^?1 of SO₂ had better sensorial quality than the wines obtained with 50 mg l^?1 as the only preservative used.     

Inactivation of different strains of Escherichia coli O157:H7 in various apple ciders treated with dimethyl dicarbonate (DMDC) and sulfur dioxide (SO2) as an alternative method

Escherichia coli has been identified as the causative agent in numerous foodborne illness outbreaks associated with the consumption of fresh apple cider. Apple cider has a pH which is normally below 4.0 and would not be considered a medium capable of supporting the growth of foodborne pathogens. The association of unpasteurized apple cider with foodborne illness due to E. coli O157:H7 has however, led to increased interest in potential alternative methods to produce pathogen free cider. Apple cider was prepared from eight different apple cultivars, inoculated with approximately 10⁶–10⁷ CFU of three strains of E. coli O157:H7 per ml (933, ATCC 43889, and ATCC 43895) and tested to determine the effectiveness of sulfur dioxide (SO₂) and dimethyl dicarbonate (DMDC). Bacterial populations for treated and untreated samples were then enumerated by using non-selective media. Eight different ciders were treated with DMDC (125 and 250 ppm) and SO₂ (25, 50, 75, 100 ppm). Greater than a 5-log reduction was achieved at room temperature with 250 ppm of DMDC and 50 ppm of SO₂ after the incubation time of 6 h and 24 h, respectively. Addition of DMDC and/or SO₂ may offer an inexpensive alternative to thermal pasteurization for the production of safe apple cider for small apple cider producers.     

Effectiveness of dimethyldicarbonate to stop alcoholic fermentation in wine

The alcoholic fermentation of Botrytis-affected wines is stopped by the addition of high concentrations of sulfur dioxide (SO₂). The natural microbial unstability of these wines and the binding phenomena forces winemakers to periodically add sulfur dioxide during maturation, leading to a high concentration of a maximum of 400 mg/l in the bottled wine. Dimethyldicarbonate (DMDC) is now considered as a reliable fungicide which could be partially used instead of SO₂, especially just before bottling. This study investigated the use of DMDC to stop alcoholic fermentation. The experiment was carried out on pure cultures of three yeast species present in this type of wine (Saccharomyces cerevisiae, Candida stellata and Zygosaccharomyces bailii). The results were very promising and suggested that DMDC was more effective than SO₂. The yeast cells died after the addition of DMDC whereas they partially entered into a viable but non-culturable (VBNC) state with SO₂. However, the same experiment carried out on botrytized must, whose fermentation was carried out using indigenous microflora, was less conclusive. It pointed out that DMDC, used in a concentration of 200 mg/l, was more effective than SO₂ but leading to the same results: the entering of a part of the cells into a VBNC state. DMDC could be used to stop alcoholic fermentation, but could not replace SO₂. Nevertheless, the concentrations of SO₂ added in this type of wine could be reduced in this way.     

Effects of SO2 on lactic acid bacteria physiology when used as a preservative compound in malolactic fermentation

SO₂ is a preservative compound widely used at an industrial scale to control the cider‐making processes. However, little is known about the effect that sulfur dioxide exerts on the cell physiology of lactic acid bacteria (LAB) during malolactic fermentation, taking into account the cellular heterogeneity of the microbial populations. Analysis of flow cytometry results has shown how increasing SO₂ concentrations were not related to a significant rise in the dead cell subpopulation. SO₂ affected malolactic activity and cell viability, giving rise to different levels of LAB tolerance to the fermentation conditions. Malate consumption in the bioreactor and the enzymatic potential of whole cells were monitored during the bioconversion process under the different assayed conditions. After SO₂ exposure, specific malolactic activity of whole cells measured by enzymatic assays in which SO₂ or other inhibitory compounds were absent showed higher values than the malolactic activity determined in the bioreactor, suggesting that inhibition was not irreversible, at least not at the concentrations tested. A correlation between total malolactic activity and viable and viable but nonculturable cell concentrations was established. Increasing amounts of SO₂ in the fermentation media also affected the malic acid/lactic acid ratio. Changes observed on LAB physiology at different sulfiting levels could be useful for practical cider‐making applications. Copyright © 2012 The Institute of Brewing & Distilling     

Supplementation of amino acids in apple must for the standardization of volatile compounds in ciders

The aim of this study was to identify the main amino acids responsible for the production of esters in ciders. The experiments were carried out in apple must and synthetic must. The amino acids were analysed by high‐performance liquid chromatography and the volatile compounds by gas chromatography. Aspartate, asparagine and glutamate positively influenced the production of esters in the cider models and were used in the experiments with mixtures. The response surface generated by the proposed model yielded two regions that maximized the production of esters. In addition, the combination of aspartate and glutamate predicted a higher production. The optimal suggested concentrations were 43.4% of aspartate and 56.6% of glutamate for 120 mg/L of total nitrogen. The apple must supplemented with these two amino acids resulted in four times more esters than in the same cider without supplementation, demonstrating the effectiveness of the proposed model.     

啤酒糟苹果醋不同发酵阶段挥发性香气成分及抗氧化性变化分析

采用顶空固相微萃取（HS-SPME）结合气质联用（GC-MS）法测定啤酒糟苹果汁、苹果酒、苹果醋的挥发性成分,探究啤酒糟苹果醋不同发酵阶段挥发性香气成分及抗氧化性变化。结果表明,苹果醋不同发酵阶段样品共检测出50种挥发性化合物,其中酯类16种、醇类8种、醛类8种、酸类7种、酚类4种、其他类4种、酮类2种、醚类1种。啤酒糟苹果汁、苹果酒、苹果醋中分别共检测出12种、15种、24种挥发性物质。抗氧化活性试验结果表明,DPPH自由基的清除率顺序为啤酒糟苹果酒＞啤酒糟苹果汁＞啤酒糟苹果醋,Fe3+还原能力顺序为啤酒糟苹果汁＞啤酒糟苹果酒＞啤酒糟苹果醋。     

Influence of yeast strain on binding of sulphur dioxide in wines,and on its formation during fermentation

The amounts of sulphur dioxide bound by acetaldehyde, pyruvic acid and α-ketoglutaric acid during fermentation of three grape juices by eight wine yeasts (Saccharomyces sp.) are reported. These constituents accounted for 49–83 % (mean 69) of the measured bound SO₂, depending on the yeast strain and juice. the maximum range of concentrations of the binding components for individual wines were 10–48 ppm for acetaldehyde, 9–77 ppm for pyruvic acid and 5–63 ppm for α-ketoglutaric acid, depending on yeast strain and grape juice. the validity of the calculations was verified by an experiment with SO₂ and the three binding compounds in a multicomponent model system. The acetaldehyde content was related to the total SO₂ present, which itself was determined by the strain of yeast. SOz bound in the wines after a further SO₂ addition was correlated significantly with pyruvic and α-ketoglutaric acids, but not with acetaldehyde. Certain yeasts produced SO₂ during fermentation in grape juice and in synthetic media with defined sulphur sources. More SO₂ was produced at pH 3.6 than 3.0 in the absence of added sulphate in grape juice. Sulphate was the best sulphur source for SO₂ production in synthetic media, although some yeasts were able to produce smaller amounts of SO₂ from l-cysteine and reduced glutathione.     

Characterisation of Asturian cider apples on the basis of their aromatic profile by high-speed gas chromatography and solid-phase microextraction

Headspace solid-phase microextraction (SPME) coupled to high-speed gas chromatography with FID detector (HSGC-FID) and with quadrupole mass spectrometric detector (GC–MS) was employed to study the volatile compounds in Asturian cider apples. The optimised conditions used for characterisation of cider apple were: sample equilibration at 25 °C for 12 h, followed by PDMS–DVB fibre exposure to the headspace above the sample for 5 min and finally thermal desorption of the adsorbed substances in the injector port for GC analysis. The use of high-speed gas chromatography allowed the separation of 27 compounds in less than 8 min, reducing the analysis time in 80% compared to conventional gas chromatography. A simple, rapid and reliable method to analyze volatile compounds in cider apples has been developed. The aromatic profile of 59 apple samples included in eight apple varieties was analysed. The different apples were obtained from two consecutive harvests (2005 and 2006). The apples volatile compounds together with chemometric techniques such as principal components analysis (PCA), Bayesian analysis and linear discriminant analysis (LDA), allowed us to differentiate apples on the basis of the sweet or sharp category to which the cider apple variety belongs. Volatile compounds such as ethyl heptanoate, E-farnesene, ethyl butyrate and hexyl caprylate are closely related to cider apple cultivars of the sweet category, while propyl butyrate and butyl acetate are related to the sharp class.     

The development of free amino acids and volatile compounds in cheese ‘Oloumoucké tvarůžky’ (PGI) during ripening

Twenty‐two free amino acid (FAA) concentrations were observed during manufacture (7 days) and ripening period (42 days follow‐up) of Olomoucké tvar??ky (PGI, smear acid cheese). During the ripening period, the amounts of volatile compounds and selected sensory attributes were also analysed. The free amino acids were determined by means of ion‐exchange chromatography, and the volatile compounds were detected and identified using HS‐SPME coupled with GC/MS method. The development of the individual FAA content positively correlated with the ripening period (r = 0.7734–0.9229; P < 0.01). Forty‐six volatile compounds (14 alcohols; 10 esters; 7 ketones; 5 acids; 4 aldehydes; 3 sulphur compounds; 2 phenol compounds; and 1 terpene) were identified. Concentrations of several compounds increased (e.g. 3‐methylbutanal, 2‐butanol, 2‐butanone; P < 0.05) or decreased (e.g. dimethyldisulphide, methanethiol, 2‐phenylethylacetate, methylbutyric acid; P < 0.05) over cheese ageing. The results gave information about the development of sensory active substances and its precursors in Olomoucké tvar??ky during ripening. In conclusion, we found that free amino acid concentrations and sulphur compounds are positive with improved flavour.     

Influence of cider‐making process parameters on the odourant volatile composition of hard ciders

This study was performed to explore the relationships between some parameters of the French cider‐making process and the odourant compounds of cider. Sixteen ciders were prepared on a pilot plant scale using experimental design and varying according to apple blends, pressing conditions, pre‐fermentation clarification implementation and conditions, and biomass reduction during fermentation. Odourant compounds were extracted from final ciders by headspace solid‐phase microextraction with a CAR/PDMS fibre, a method previously shown to provide extracts representative of the studied cider. Extracts were analysed by gas chromatography coupled with mass spectrometry and flame ionization detection. All of the parameters tested had at least a slight effect on the odourant composition and particularly on the esters, which bring fruity notes that are appreciated by consumers. Clarification and biomass reduction had a greater impact than apple blend and pressing conditions. This could be explained by the influence of the nitrogen content on fermentation rate and efficiency, which affects the production of secondary metabolites. Under the conditions tested, a juice obtained from a bitter blend of apples by a slow pressing of the pulp at low temperature, after 1 h of cuvage, clarified by keeving and fermented without biomass reduction, produced a cider with the highest quantity of esters. These results could help cider‐makers enhance product quality according to consumer expectations. Copyright © 2015 The Institute of Brewing & Distilling     

A New Analytical Method to Volatile Compounds in Cider Apples: Application to Evaluate the Starch Index

Headspace solid-phase microextraction coupled to Fast gas chromatography (GC) with flame ionization detector and with quadrupole mass spectrometric detector was employed to study the volatile compounds in Asturian cider apples. The best conditions used for volatile analysis of apple were: sample equilibration at 25 °C for 60 min, followed by polydimethylsiloxane-divinylbenzene fiber exposure to the headspace above the sample for 5 min, and finally thermal desorption of the adsorbed substances in the injector port for GC analysis. The use of Fast gas chromatography allowed the separation of 24 compounds in less than 8 min, reducing the analysis time in 80% compared with conventional gas chromatography. The aromatic profile of 26 crushed apple samples included in Designation of Origin Cider from Asturias was analyzed. The chemometric technique named multiple linear regression allowed us to establish a relationship between apple volatile compounds and the starch index. The results obtained are consistent with the known apple ripening model.