The molecular characterization of new types of Saccharomyces cerevisiae × S. kudriavzevii hybrid yeasts unveils a high genetic diversity

New double‐ and triple‐hybrid Saccharomyces yeasts were characterized using PCR‐restriction fragment length polymorphism of 35 nuclear genes, located on different chromosome arms, and the sequencing of one nuclear and one mitochondrial gene. Most of these new hybrids were originally isolated from fermentations; however, two of them correspond to clinical and dietary supplement isolates. This is the first time that the presence of double‐hybrid S. cerevisiae × S. kudriavzevii in non‐fermentative substrates has been reported and investigated. Phylogenetic analysis of the MET6 nuclear gene confirmed the double or triple parental origin of the new hybrids. Restriction analysis of gene regions in these hybrids revealed a high diversity of genome types. From these molecular characterizations, a reduction of the S. kudriavzevii fraction of the hybrid genomes is observed in most hybrids. Mitochondrial inheritance in hybrids was deduced from the analysis of mitochondrial COX2 gene sequences, which showed that most hybrids received the mitochondrial genome from the S. kudriavzevii parent. However, two strains inherited a S. cerevisiae COX2, being the first report of S. cerevisiae × S. kudriavzevii hybrids with S. cerevisiae mitochondrial genomes. These two strains are those showing a higher S. kudriavzevii nuclear genome reduction, especially in the wine hybrid AMH. This may be due to the release of selective pressures acting on the other hybrids to maintain kudriavzevii mitochondria‐interacting genes. Copyright © 2011 John Wiley & Sons, Ltd.     

Performance of wild Saccharomyces and Non-Saccharomyces yeasts as starter cultures in dough fermentation and bread making

The use of wild Saccharomyces and non-Saccharomyces yeasts might result in bread with different and attractive sensory characteristics. This study aimed to evaluate the performance of Saccharomyces and non-Saccharomyces yeasts as starter culture in dough fermentation to bread making and the physicochemical parameters and aromatic profile of bread. All 26 wild yeasts strains isolated from Brazilian Cerrado fruit and tree bark were osmotolerant, and 19.4% were able to ferment maltose. Candida tropicalis ART101.3 and Saccharomyces cerevisiae SC5952 had the best growth capacity under high concentrations of glucose and maltose. Also, they were resistant to lyophilisation. Kinetic parameters of bioreactor cultivations showed high cell growth and lower generation time with 10 g L⁻¹ maltose. Bread produced with C. tropicalis ART101.3 and the control bread had similar physicochemical properties and acceptance of consumers. Bread with S. cerevisiae SC5951 had a lower specific volume and a different colour than control bread; however, the consumers found no significant difference. More than 70% of the consumers demonstrated purchase intention of bread produced with both wild yeasts. The present study shows the potential of native Cerrado yeasts to be used and exploited in industrial processes and contributes to the diversification of bread starter cultures.     

Screening of non-Saccharomyces wine yeasts for the presence of extracellular hydrolytic enzymes

Twenty-six strains of yeasts, belonging to the genera Candida, Debaryomyces, Hanseniaspora, Hansenula, Kloeckera, Metschnikowia, Pichia, Saccharomyces and Torulaspora previously isolated from wines, were screened for the production of extracellular pectinases, amylases, lipases, proteases and β-glucosidases. Some strains of Candida species and Hanseniaspora uvarum/Kloeckera apiculata produced extracellular proteolytic or lipolytic activities. Most yeasts exhibited β-glucosidase activity, but particularly high activity was observed in strains of Pichia anomala/Candida pelliculosa (formerly Hansenula anomala ) and Hanseniaspora uvarum/Kloeckera apiculata . The potential impact of these enzymes on wine quality is discussed.     

Amino acid uptake by wild and commercial yeasts in single fermentations and co-fermentations

Musts require nitrogen-containing compounds in order to ensure yeast development. This study examined the nitrogen-nutrient requirements of two commercial yeasts and three wild strains isolated from inoculated fermentations. The results showed that wild strains generally consumed lower amounts of amino acids than commercial yeasts. Most amino acids were assimilated during the exponential growth phase; only a few – including asparagine and histidine – were metabolized until the end of fermentation. The study also sought to determine whether industrial drying affected yeast nitrogen requirements.     

Cork-borne bacteria and yeasts as potential producers of off-flavours in wine

Bacteria and yeasts were found to be present within cork lenticels, covered by mucous or fibrous substances. They survived heating, peroxide treatment and contact with the alcohol and sulfur dioxide of wine. 187 bacteria and 36 yeast strains were isolated from cork stoppers of wine bottles and, during various stages of production, from corkwood and new cork stoppers. After culturing, a number of isolates showed the ability to modify the aroma of model systems consisting of dilute or full strength wine and pulverised cork. The aromas produced by isolates of varying cork origin are tabled. A small number of isolates methylated 2,4,6-trichlorophenol, yielding 2,4,6-trichloroanisole, responsible for the typical cork taint. During the boiling of cork slabs, the internal temperature on the inside of a box made from cork slices did not exceed 87°C.     

Antimicrobial activity of weak acids in liquid feed fermentations, and its effects on yeasts and lactic acid bacteria

BACKGROUND: The efficacy of weak organic acids in the control of yeasts in pig liquid feed was studied taking into account the effects on lactic acid bacteria (LAB) responsible for beneficial fermentation. RESULTS: The yeast population in pig liquid feed was taxonomically identified. Kazachstania exigua, Debaryomyces hansenii and Pichia deserticola dominated the fermentation in liquid feed and whey. Pichia deserticola was found in whey and foaming liquid feed and dominated the fermentation after incubation. The sensitivity of the isolates against weak acids was measured in culture medium as well as in fermented and non‐fermented liquid feed. Formic acid and potassium sorbate successfully reduced the growth of yeasts in all media without interfering with LAB development. Both of these organic acids showed an increased antifungal effect when used in liquid feed fermented by a Lactobacillus plantarum strain. CONCLUSION: The loss of energy, reduced palatability and other practical problems due to the high growth of yeasts in fermented liquid diets can be reduced by organic acids without affecting lactic acid fermentation. Copyright © 2011 Society of Chemical Industry     

Experimental approaches to study evolutionary cell biology using yeasts

The past century has witnessed tremendous advances in understanding how cells function. Nevertheless, how cellular processes have evolved is still poorly understood. Many studies have highlighted surprising molecular diversity in how cells from diverse species execute the same processes, and advances in comparative genomics are likely to reveal much more molecular diversity than was believed possible until recently. Extant cells remain therefore the product of an evolutionary history that we vastly ignore. Evolutionary cell biology has emerged as a discipline aiming to address this knowledge gap by combining evolutionary, molecular, and cellular biology thinking. Recent studies have shown how even essential molecular processes, such as DNA replication, can undergo fast adaptive evolution under certain laboratory conditions. These developments open new lines of research where the evolution of cellular processes can be investigated experimentally. Yeasts naturally find themselves at the forefront of this research line. Not only do they allow the observation of fast evolutionary adaptation, but they also provide numerous genomic, synthetic, and cellular biology tools already developed by a large community. Here we propose that yeasts can serve as an “evolutionary cell lab” to test hypotheses, principles, and ideas in evolutionary cell biology. We discuss various experimental approaches available for this purpose, and how biology at large can benefit from them.     

不同发酵条件对酿酒酵母甘油产量的影响

为提高酿酒酵母的甘油产量,分别考察不同初始葡萄糖和果糖质量浓度、pH值、发酵温度及SO2添加量对酿酒酵母D254甘油产量的影响。对酿酒酵母D254不同发酵初始条件进行单因素试验,其他因素固定条件下,葡萄糖质量浓度180g/L时酵母菌体生长平稳、生长量最高;果糖质量浓度108g/L时酵母甘油产量最高;pH值为3.5更适宜酵母菌体生长和合成甘油;在发酵温度和SO2添加量的单因素试验中也分别得出适宜发酵温度为28℃和适宜SO2添加量为 20mg/L。通过单因素试验,筛选出最利于酿酒酵母D254生长和产甘油的各因素的最佳质量浓度,进行Plackett-Burman发酵条件组合试验,得到发酵条件最佳组合为：初始葡萄糖质量浓度216g/L、果糖质量浓度144g/L、发酵温度32℃、pH 3.0、SO2添加量40mg/L,此条件下,酿酒酵母D254获得最高甘油产量达655.64μmol/L。     

Volatile composition and aromatic attributes of wine made with Vitisvinifera L.cv Cabernet Sauvignon grapes in the Xinjiang region of China: effect of different commercial yeasts

The Xinjiang region is a major grape- and wine-production area in China, but the region’s notably high temperatures in the summer and year-round intense sun exposure play negative roles in the aroma, complexity, and elegance of Cabernet Sauvignon wine. In this study, Cabernet Sauvignon grapes harvested in this region were fermented on an industrial scale using four commercial yeast strains (L2323, D254, RVA, and CECA) and spontaneous yeast (NF). The results showed that a total of 123 volatile compounds were detected and 15 volatile compounds significantly contributed their flavor notes to the wine’s overall aroma. The use of RVA and CECA strains resulted in wine with higher concentrations of higher alcohols, terpenes and norisoprenoids. However, the D254-fermented wine showed high level of esters and carbonyl compounds. Wine fermented with the L2323 and D254 strain possessed a stronger fruity aroma, whereas the RVA strain enhanced the herbaceous, chemical, and fatty aromas in wine. Principal component analysis revealed that a significant aromatic feature difference was observed in these wines after alcoholic and malolactic fermentation. The use of different commercial yeast strains altered the aromatic profile of Cabernet Sauvignon wine.     

昌黎产区产酶酵母多样性及其应用潜力分析

利用优选酵母混菌发酵,是改善葡萄酒品质及增香的有效途径。以赖氨酸培养基、WL培养基和产酶筛选培养基为基础,分析了昌黎4个葡萄园土样中产酶酵母的多样性,结果表明,该地区产酶酵母丰富,50%的产酶酵母能同时产2种或2种以上的酶。对产酶酵母酿造因子耐性分析表明,菌株GY1、M9和J24具有高糖耐性,适合于高糖低醇果酒酿造;菌株GY13和M9具有高酒精耐性,酒精含量为15.000%时活性较强;菌株J24和GY1具有高酸耐性,pH 2.5时具有较高活性;7株菌15℃时具有较高活性。综合分析发现,菌株M9(Torulaspora delbrueckii)同时产β-葡萄糖苷酶、果胶酶和纤维素酶,除酸耐性较差外,具有高糖、高酒精度和低温耐性,在高糖低醇果酒酿造中具有很大应用潜力。     

产香酵母的筛选及其苹果酒发酵特性

从分离自赤霞珠葡萄酿酒不同时期发酵液中的酵母菌中筛选产香菌株,并对其苹果酒发酵特性进行研究。结果表明,经感官评定初筛,获得7株产香较优的酵母菌,并对其进行耐酒精、耐SO2和耐盐性能分析,其中菌株B2-13具有较好的发酵性能,经分子生物学鉴定菌株B2-13为葡萄酒有孢汉逊酵母（Hanseniaspora uvarum）。菌株B2-13发酵苹果酒酒精度为4.2%vol～5.5%vol,糖度为2.6～44 g/L,共鉴定出32种香气成分,主要有酯类（6种）、醛酮类（4种）、醇类（6种）、酸类（8种）等,其中异戊醇、苯乙醇含量最高,分别为0.36 mg/mL和0.16 mg/mL,另外还有赤霞珠干红葡萄酒的特有香气物质3-羟基-2-丁酮（0.16 mg/mL）。菌株B2-13是一株具有较好的苹果酒发酵和产香性能的酵母菌株,有良好的工业化生产应用前景。     

Nectar yeasts: a natural microcosm for ecology

The species of yeasts that colonize floral nectar can modify the mutualistic relationships between plants and pollinators by changing the chemical properties of nectar. Recent evidence supporting this possibility has led to increased interest among ecologists in studying these fungi as well as the bacteria that interact with them in nectar. Although not fully explored, nectar yeasts also constitute a promising natural microcosm that can be used to facilitate development of general ecological theory. We discuss the methodological and conceptual advantages of using nectar yeasts from this perspective, including simplicity of communities, tractability of dispersal, replicability of community assembly, and the ease with which the mechanisms of species interactions can be studied in complementary experiments conducted in the field and the laboratory. To illustrate the power of nectar yeasts as a study system, we discuss several topics in community ecology, including environmental filtering, priority effects, and metacommunity dynamics. An exciting new direction is to integrate metagenomics and comparative genomics into nectar yeast research to address these fundamental ecological topics.     

外源褪黑素对酿酒酵母发酵及抗氧化体系的影响

本文研究了外源褪黑素对酿酒酵母发酵能力、主发酵产物、抗氧化体系和抗氧化能力的影响,并测定了褪黑素含量的变化。发现对照组在发酵的第8 d发酵基本停止,处理组直到发酵的第14 d发酵终止,但最终的CO2总失重量、残糖和酒精含量无显著影响。褪黑素在发酵的第8 d及以后能显著地降低海藻糖、甘油的生成量,发酵第8 d时,对照组甘油含量达到最大值6.25 g/L,而低、中、高浓度褪黑素处理组甘油含量比对照组分别低7.57%、12.29%、11.99%。低浓度的褪黑素能刺激酵母超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性提高,减少丙二醛(MDA)的积累,发酵结束时低、中浓度褪黑素处理组SOD酶活分别比对照组高68.15%和59.89%,低、中、高褪黑素处理组CAT酶活分别是对照组的2.85、2.87、1.84倍,同时高浓度褪黑素处理组的MDA含量只有对照组的71.44%。发酵液抗氧化能力随褪黑素浓度增加而增强强,褪黑素含量在发酵过程中逐渐降低,后期趋于稳定。     

葡萄汁有孢汉逊酵母与酿酒酵母混合发酵对嘎啦苹果酒品质的影响

以嘎啦苹果为实验材料,探究了葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）（云-268）与酿酒酵母（Saccharomyces cerevisiae）混合发酵对苹果酒基本理化指标、酚类物质、香气物质及感官特性的影响。结果表明,相比于酿酒酵母单一发酵,混合发酵苹果酒中乙醇和总酸含量显著降低（P<0.05）,降幅分别为8.72%和9.02%,总酚含量显著增加（P<0.05）,增幅为23.01%;其香气物质种类（36种）明显多于单一发酵苹果酒（31种）,其中香气活度值OAV>1的物质有12种;进一步分析发现,混合发酵苹果酒中具果香味的乙酸酯类和花香味的萜烯类等物质含量显著提高（P<0.05）,具酸腐味的脂肪酸类物质含量显著降低（P<0.05）,对具青草味的C6醇类物质含量影响不大;感官品评结果表明混合发酵苹果酒酒液澄清、禾杆黄色、入口酸度中等偏低,甜度适中,口感平衡,具有丰富的果香味。因此,葡萄汁有孢汉逊酵母与酿酒酵母混合发酵（1:1）能够明显提高苹果酒的品质,增加其发酵香,为低醇果香味的苹果酒酿造奠定了一定的研究基础。     

非酿酒酵母与酿酒酵母混合发酵对葡萄酒香气的影响

为研究自选非酿酒酵母对葡萄酒香气物质的影响,选取毕赤克鲁维酵母（Pichia kluyveri）HSX-5、长孢洛德酵母（Lodderomyces elongisporus）MNS-6和戴尔有孢圆酵母（Torulaspora debrueckii）YQX-8与酿酒酵母（Saccharomyces cerevisiae）AWRI 796混合发酵赤霞珠葡萄酒,采用顶空固相萃取-气相色谱质谱联用（HS-SPME-GC-MS）技术检测其挥发性香气物质,并对结果进行主成分分析（PCA）。结果表明,相较于酿酒酵母单菌发酵酒样,菌株HSX-5和YQX-8混合发酵酒样中己酸乙酯提升了2倍,菌株HSX-5、YQX-8、MNS-6混合发酵酒样中的辛酸乙酯分别提升76.6%、35.5%、5.5%,乙酸乙酯含量增加均为10%,混合发酵增强了葡萄酒花果类香气及醇香;同时混合发酵各组均产生了新物质,增加了葡萄酒风味体系的丰度。PCA结果表明,菌株HSX-5和YQX-8混合发酵的发酵酒样主要贡献香气种类较多且气味活度值（OAV）较高,对葡萄酒香气的影响显著。     

Modulation of Glycerol and Ethanol Yields During Alcoholic Fermentation in Saccharomyces cerevisiae Strains Overexpressed or Disrupted for GPD1 Encoding Glycerol 3-Phosphate Dehydrogenase

The possibility of the diversion of carbon flux from ethanol towards glycerol in Saccharomyces cerevisiae during alcoholic fermentation was investigated. Variations in the glycerol 3-phosphate dehydrogenase (GPDH) level and similar trends for alcohol dehydrogenase (ADH), pyruvate decarboxylase and glycerol-3-phosphatase were found when low and high glycerol-forming wine yeast strains were compared. GPDH is thus a limiting enzyme for glycerol production. Wine yeast strains with modulated GPD1 (encoding one of the two GPDH isoenzymes) expression were constructed and characterized during fermentation on glucose-rich medium. Engineered strains fermented glucose with a strongly modified [glycerol] : [ethanol] ratio. gpd1Δ mutants exhibited a 50% decrease in glycerol production and increased ethanol yield. Overexpression of GPD1 on synthetic must (200 g/l glucose) resulted in a substantial increase in glycerol production (×4) at the expense of ethanol. Acetaldehyde accumulated through the competitive regeneration of NADH via GPDH. Accumulation of by-products such as pyruvate, acetate, acetoin, 2,3 butane-diol and succinate was observed, with a marked increase in acetoin production. © 1997 John Wiley & Sons, Ltd.     

酒精发酵过程中酿酒酵母海藻糖代谢的研究

研究了酿酒酵母在酒精发酵过程中酵母细胞内海藻糖的代谢。结果表明海藻糖的代谢受几种因素如底物浓度、发酵温度以及其他条件的调节与控制。在试验中对酿酒酵母细胞内海藻糖在整个酒精发酵过程中的生物功能作了简要的评价。