Influence of sequential inoculum of Starmerella bacillaris and Saccharomyces cerevisiae on flavonoid composition of monovarietal Sangiovese wines

The selection of Starmerella bacillaris strains to be used with Saccharomyces cerevisiae as mixed cultures has been recently suggested in order to produce wines containing lower ethanol and higher glycerol concentrations and to promote fructose degradation due to their fructophilic character. However, studies about effects of such mixed starter cultures on phenolic compounds, which are responsible for the colour and health-enhancing properties in red wines, are currently lacking. Therefore, in this work, the influence of sequential inoculated fermentation (SIF) with Starm. bacillaris and S. cerevisiae on phenolic content of monovarietal Sangiovese wine was evaluated by fermentations at laboratory scale. Axenic fermentations (AXFs) with S. cerevisiae were performed as control. S. cerevisiae attained higher cell densities in AXF compared with SIF. The experimental wines obtained by SIF showed significant lower ethanol and higher glycerol concentrations, whereas no significant difference was detected in colour intensity. The total phenol index reached significantly lower values in SIF. Furthermore, the wines produced by SIF contained higher concentrations of vitisin A that has a greater colour stability than the anthocyanin monomer. Finally, a lower content of both free anthocyanins and flavan-3-ols, key compounds for wine quality possessing also health-enhancing properties, was found in wines obtained by SIF. On the contrary, no significant difference was detected on flavonol concentration between SIF and AXF. This study highlighted that the use of sequential inoculum of Starm. bacillaris and S. cerevisiae can contribute to increasing the colour stability of red wines, even if at the expense of compounds with health properties.     

Physiological features of Schizosaccharomyces pombe of interest in making of white wines

This work studies the physiology of Schizosaccharomyces pombe strain 938 in the production of white wine with high malic acid levels as the sole fermentative yeast, as well as in mixed and sequential fermentations with Saccharomyces cerevisiae Cru Blanc. The induction of controlled maloalcoholic fermentation through the use of Schizosaccharomyces spp. is now being viewed with much interest. The acetic, malic and pyruvic acid concentrations, relative density and pH of the musts were measured over the entire fermentation period. In all fermentations in which Schizo. pombe 938 was involved, nearly all the malic acid was consumed and moderate acetic concentrations produced. The urea content and alcohol level of these wines were notably lower than in those made with Sacch. cerevisiae Cru Blanc alone. The pyruvic acid concentration was significantly higher in Schizo. pombe fermentations. The sensorial properties of the different final wines varied widely.     

Influence of alcoholic and malolactic starter cultures on bioactive amines in Merlot wines

The influence of alcoholic and malolactic fermentations on the levels of amines in Merlot wines was investigated. Saccharomyces bayanus, S. cerevisiae, Lactobacillus plantarum, Oenococcus oeni (DSM 7008 and 12923) and spontaneous fermentations were used. Four of the 10 amines investigated were detected: spermidine, serotonin, putrescine and cadaverine. When considering the factors independently, the malolactic bacteria significantly affected the levels of serotonin and total amines, whereas the fermentation yeasts significantly affected the levels of spermidine (two way Kruskal–Wallis, p ? 0.05). Spermidine levels were significantly higher in wines produced with S. cerevisiae. Significantly higher serotonin levels were found in wines made with L. plantarum. Putrescine and cadaverine were not detected in wines produced by spontaneous alcoholic fermentation or by L. plantarum. There were significant differences in alcohol content, total and volatile acidity, sulphite levels and taste quality among wines (Tukey test, p ? 0.05).     

Monitoring a mixed starter of Hanseniaspora vineae-Saccharomyces cerevisiae in natural must: impact on 2-phenylethyl acetate production

The effect of simultaneous or sequential inoculation of Hanseniaspora vineae CECT 1471 and Saccharomyces cerevisiae T73 in non-sterile must on 2-phenylethyl acetate production has been examined. In both treatments tested, no significant differences in Saccharomyces yeast growth were found, whereas non-Saccharomyces yeast growth was significantly different during all days of fermentation. Independently of the type of inoculation, S. cerevisiae was the predominant species from day 3 till the end of the fermentation. The dynamics of indigenous and inoculated yeast populations showed H. vineae to be the predominant non-Saccharomyces species at the beginning of fermentation in sequentially inoculated wines, whereas the simultaneous inoculation of S. cerevisiae did not permit any non-Saccharomyces species to become predominant. Differences found in non-Saccharomyces yeast growth in both fermentations influenced the analytical profiles of final wines and specifically 2-phenylethyl acetate concentration which was two-fold increased in sequentially inoculated wines in comparison to those co-inoculated. In conclusion we have shown that H. vineae inoculated as part of a sequential mixed starter is able to compete with native yeasts present in non-sterile must and modify the wine aroma profile.     

Lachancea thermotolerans and Saccharomyces cerevisiae in simultaneous and sequential co-fermentation: A strategy to enhance acidity and improve the overall quality of wine

In the last few years there is an increasing interest on the use of mixed fermentation of Saccharomyces and non-Saccharomyces wine yeasts for inoculation of wine fermentations to enhance the quality and improve complexity of wines. In the present work Lachancea (Kluyveromyces) thermotolerans and Saccharomyces cerevisiae were evaluated in simultaneous and sequential fermentation with the aim to enhance acidity and improve the quality of wine.     

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.     

Increasing the levels of 2-phenylethyl acetate in wine through the use of a mixed culture of Hanseniaspora osmophila and Saccharomyces cerevisiae

The impact of mixed cultures of Hanseniaspora osmophila and Saccharomyces cerevisiae with different initial yeast ratios on wine composition has been examined. The mixed culture significantly affected sugar consumption, the main enological parameters and ester concentrations, with the exception of glycerol, isoamyl acetate and diethyl succinate levels. Remarkably, in wines obtained with mixed cultures the concentration of 2-phenylethyl acetate was approximately 3- to 9-fold greater than that produced by S. cerevisiae pure culture. Moreover sensory evaluation revealed a stronger fruity character in wines fermented with mixed cultures than in control wines. Independently of the mixed culture used, all wines showed concentrations of acetic acid and ethyl acetate within the ranges described for wines. Our data suggest that a mixed culture of H. osmophila and S. cerevisiae can be used as a tool to increase 2-phenylethyl acetate in wine and that its concentration can be controlled by modulating the initial yeast ratio in the culture.     

胶红酵母与酿酒酵母共发酵对干红葡萄酒香气与色泽的影响

研究优选胶红酵母与酿酒酵母不同混合发酵方案对季风性气候区干红葡萄酒的香气质量、多酚及颜色的影响,优化增香酿造技术方案.实验以陕西合阳梅鹿辄葡萄为试材,设计2种菌株不同比例同时接种和顺序接种策略酿造干红葡萄酒.酿造完成后,对葡萄酒样品的香气成分进行顶空固相微萃取结合气相色谱-质谱联用法定性定量分析,香气特征通过感官量化品...     

Indigenous Saccharomyces cerevisiae strains and their influence on the quality of Cataratto, Inzolia and Grillo white wines

The present paper deals with three new strains of Saccharomyces cerevisiae, isolated in old wineries of Sicily, which were microbiologically and molecularly characterized and tested for their ability to produce white wines. Examined in terms of their growth pattern, fermentation vigour, sulphite tolerance, fermentative power, spore formation, and production of acetic acid, hydrogen sulphide and phenolic off-flavours, the strains were utilized as starters in experimental fermentations of musts obtained from the cultivars Inzolia, Grillo and Catarratto. Further, the three musts were also fermented using two commercial S. cerevisiae strains. The quality of the wines produced was confirmed by their principal oenochemical parameters, by sensory analysis and qualitative and quantitative determination of the volatile aroma constituents. All the data were statistically elaborated. Interestingly, the new selected yeasts were able to increase the pear notes (Z)-ethyl-4-decenoate, (E)-ethyl-3-decenoate, and (Z)-ethyl-3-decenoate which are fundamental for the aroma of these Sicilian wines. From our results, the new yeast strains were found to produce white wines of a quality which was not inferior to those obtainable with the best commercial strains selected in other geographical areas, but also with a distinctive aromatic profile.     

Selected non-Saccharomyces wine yeasts in controlled multistarter fermentations with Saccharomyces cerevisiae

Non-Saccharomyces yeasts are metabolically active during spontaneous and inoculated must fermentations, and by producing a plethora of by-products, they can contribute to the definition of the wine aroma. Thus, use of Saccharomyces and non-Saccharomyces yeasts as mixed starter cultures for inoculation of wine fermentations is of increasing interest for quality enhancement and improved complexity of wines. We initially characterized 34 non-Saccharomyces yeasts of the genera Candida, Lachancea (Kluyveromyces), Metschnikowia and Torulaspora, and evaluated their enological potential. This confirmed that non-Saccharomyces yeasts from wine-related environments represent a rich sink of unexplored biodiversity for the winemaking industry. From these, we selected four non-Saccharomyces yeasts to combine with starter cultures of Saccharomyces cerevisiae in mixed fermentation trials. The kinetics of growth and fermentation, and the analytical profiles of the wines produced indicate that these non-Saccharomyces strains can be used with S. cerevisiae starter cultures to increase polysaccharide, glycerol and volatile compound production, to reduce volatile acidity, and to increase or reduce the total acidity of the final wines, depending on yeast species and inoculum ratio used. The overall effects of the non-Saccharomyces yeasts on fermentation and wine quality were strictly dependent on the Saccharomyces/non-Saccharomyces inoculum ratio that mimicked the differences of fermentation conditions (natural or simultaneous inoculated fermentation).     

Increased flavour diversity of Chardonnay wines by spontaneous fermentation and co-fermentation with Hanseniaspora vineae

Discovery, characterisation and use of novel yeast strains for winemaking is increasingly regarded as a way for improving quality and to provide variation, including subtle characteristic differences in fine wines. The objective of this work was to evaluate the use of a native apiculate strain, selected from grapes, Hanseniaspora vineae (H. vineae) 02/5A. Fermentations were done in triplicate, working with 225 L oak barrels, using a Chardonnay grape must. Three yeast fermentation strategies were compared: conventional inoculation with a commercial Saccharomyces cerevisiae strain, ALG 804, sequential inoculation with H. vineae and then strain ALG 804 and spontaneous fermentation. Yeast strain identification was performed during fermentation, in which the apiculate strain was found to be active, until 9% of alcohol in volume, for the co-fermentation and the spontaneous fermentation was completed by three native S. cerevisiae strains. Basic winemaking parameters and some key chemical analysis, such as concentration of glycerol, biogenic amines, organic acids, and aroma compounds were analysed. Sensory analysis was done using a trained panel and further evaluated with professional winemakers. Sequential inoculation with H. vineae followed by S. cerevisiae resulted in relatively dry wines, with increased aroma and flavour diversity compared with wines resulting from inoculation with S. cerevisiae alone. Wines produced from sequential inoculations were considered, by a winemaker’s panel, to have an increased palate length and body. Characteristics of wines derived from sequential inoculation could be explained due to significant increases in glycerol and acetyl and ethyl ester flavour compounds and relative decreases in alcohols and fatty acids. Aroma sensory analysis of wine character and flavour, attributed to winemaking using H. vineae, indicated a significant increase in fruit intensity described as banana, pear, apple, citric fruits and guava. GC analysis of the relative accumulation of 23 compounds to significantly different concentrations for the three fermentation strategies is discussed in relation to aroma compound composition.     

Effect of pure and mixed cultures of the main wine yeast species on grape must fermentations

Mixed inoculation of non-Saccharomyces yeasts and S. cerevisiae is of interest for the wine industry for technological and sensory reasons. We have analysed how mixed inocula of the main non-Saccharomyces yeasts and S. cerevisiae affect fermentation performance, nitrogen consumption and volatile compound production in a natural Macabeo grape must. Sterile must was fermented in triplicates and under the following six conditions: three pure cultures of S. cerevisiae, Hanseniaspora uvarum and Candida zemplinina and the mixtures of H. uvarum:S. cerevisiae (90:10), C. zemplinina:S. cerevisiae (90:10) and H. uvarum:C. zemplinina:S. cerevisiae (45:45:10). The presence of non-Saccharomyces yeasts slowed down the fermentations and produced higher levels of glycerol and acetic acid. Only the pure H. uvarum fermentations were unable to finish. Mixed fermentations consumed more of the available amino acids and were more complex and thus better able to synthesise volatile compounds. However, the amount of acetic acid was well above the admissible levels and compromises the immediate application of mixed cultures.     

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.     

Fermentation kinetics and chemical characterisation of vino tostado,a traditional sweet wine from Galicia (NW Spain)

BACKGROUND: Grapes after harvesting are air dried and pressed in order to concentrate sugars, acids and flavour compounds to produce vino tostado (toasted wine), a wine with intense aroma and flavour notes and high residual sugar concentration. In order to get a better knowledge of the difficulties involved, several fermentations were conducted at 12 and 28 °C using 0, 15 and 30 g hL^?1 ammonium sulfate and 0, 25 and 50 g hL^?1 exogenous commercial yeast (Saccharomyces cerevisiae var. bayanus) to study the kinetics of sugar consumption and ethanol, acetic acid and glycerol production. RESULTS: Fermentation kinetic parameters were calculated and metal concentrations and antioxidant activity were analysed. CONCLUSION: The spontaneous fermentation at 12 °C and all fermentations conducted with the commercial yeast gave vino tostado of adequate quality, while the spontaneous fermentation at 28 °C was sluggish. High‐temperature fermentations led to sweeter wines with higher volumetric productivities, although low‐temperature fermentations produced better wines in terms of higher glycerol and lower acetic acid levels. Fructose was the only sugar to be consumed during spontaneous fermentations, while both glucose and fructose were consumed during fermentations of the inoculated musts, with preference for each monosaccharide depending on temperature. Copyright © 2009 Society of Chemical Industry     

Selection of indigenous Saccharomyces cerevisiae strains according to their oenological characteristics and vinification results

Xynomavro and Muscat Hamburg grapes were fermented spontaneously, producing red and white wine, respectively. The indigenous yeast flora, isolated at three phases of these fermentations, was studied. One hundred and ten strains were isolated, eighty-four of which were identified as Saccharomyces cerevisiae. Six of these strains were selected for their satisfactory oenological properties to be tested as starters in wine fermentations. The quality of the produced wines was evaluated after determination of their aromatic compounds and sensory analysis. S. cerevisiae strain BT4, as well as the AM4+BM3 combination, were considered the best as they produced wines characterized by fruity aromas and were highly accepted among panelists.     

耐热克鲁维酵母和粟酒裂殖酵母顺序接种发酵对美乐干红葡萄酒品质的影响

研究耐热克鲁维酵母（Lachancea thermotolerans）与粟酒裂殖酵母（Schizosaccharomyces pombe）顺序接种发酵替代苹果酸-乳酸发酵对美乐干红葡萄酒品质的影响,为利用非酿酒酵母提升干红葡萄酒品质提供技术支持。以甘肃武威产区美乐葡萄为原料,将3 株不同的L. thermotolerans菌株分别与1 株S. pombe菌株间隔4 d顺序接种发酵及S. pombe菌株单独接种发酵为处理组,以酿酒酵母（Saccharomyces cerevisiae）单独接种发酵为对照,对发酵过程中菌种的生长动态、葡萄酒主要理化指标、风味成分及感官品质进行分析。结果表明：处理组完成乙醇发酵均需16 d,在顺序接种发酵过程中,S. pombe菌体生长稳定,L. thermotolerans菌株在第4天菌体数量达到最大,但随着S. pombe菌体的加入而明显减少,并在10～12 d完全消亡。3 株L. thermotolerans和S. pombe菌株以顺序接种方式发酵产生的乳酸含量无显著差异,其质量浓度为2.46～2.86 g/L,但均显著高于对照组苹果酸-乳酸发酵后的乳酸含量（P＜0.05）。顺序接种发酵能够使挥发酸质量浓度降低0.11～0.16 g/L,总酸质量浓度增加0.96～1.97 g/L,也能够增加酒体色度,降低酒体色调。顺序接种发酵酒样中酯类、高级醇类和酸类化合物的含量分别下降了38.46%～49.34%、13.36%～19.42%、46.84%～49.71%,但增加了苯乙醇、萜烯类和大马酮等挥发性化合物的产出,提高了香气的复杂性。感官分析表明顺序发酵改善了葡萄酒的色泽,酸度偏高,花果香气明显。表明L. thermotolerans菌株与S. pombe菌株的顺序接种方式发酵具有替代苹果酸-乳酸发酵的潜力,对葡萄酒增酸和感官品质的提升也具有积极影响。     

Monoterpene alcohols release and bioconversion by Saccharomyces species and hybrids

Terpene profile of Muscat wines fermented by Saccharomyces species and hybrid yeasts was investigated. The amount of geraniol decreased in most wines with respect to the initial must except for Saccharomyces bayanus wines. On the other hand, alpha-terpineol amount was higher in wines fermented by Saccharomyces cerevisiae and hybrid yeasts. The amount of linalool was similar in all wines and comparable to the amount in the initial must. Lower levels of beta-d-glucosidase activity were found in the hybrid yeasts with respect to S. cerevisiae. Moreover, no relationship between beta-d-glucosidase activity and terpenes profile in Muscat wines fermented with Saccharomyces species and hybrids was found. Growth of yeasts on minimum medium supplemented with geraniol showed bioconversion of geraniol into linalool and alpha-terpineol. Percentages of geraniol uptake and bioconversion were different between Saccharomyces species and hybrids. Strains within S. bayanus, Saccharomyces kudriavzevii and hybrids showed higher geraniol uptake than S. cerevisiae, whereas the percentage of produced linalool and alpha-terpineol was higher in S. cerevisiae and hybrid yeasts than in S. bayanus and S. kudriavzevii. The relationship between geraniol uptake and adaptation of Saccharomyces species to grow at low temperature is discussed.     

Deacidification of red and white wines by a mutant ofSchizosaccharomyces malidevoransunder commercial winemaking conditions

A mutant ofSchizosaccharomyces malidevorans, which utilizes malic acid more rapidly than the wild-type strain and does not utilize significant amounts of glucose or fructose, was used in commercial scale deacidification of grape juice during two vintages in a New Zealand winery. Four trials were conducted in Chardonnay, Semillon and Cabernet Sauvignon juices, 1000–4500 l. In the first two trials, the juices were inoculated with the alcoholic fermentation yeast,Saccharomyces cerevisiae, 33–48 h after the mutant inoculation. In the remaining two trials, the juices were inoculated simultaneously with the mutant andS. cerevisiae. Malic acid, ranging from 3.5–10 g l⁻¹in the juices, was completely degraded within 21–73 h. TheSchiz. malidevoransmutant utilized malic acid at sulfur dioxide levels and temperatures used in red and white winemaking. The wines produced in the trials lacked obvious organoleptic defects. They were blended with untreated wines for acid balance and retailed as varietal wines.