Stuck Fermentations Research Articles

Enhancing the fermentation rate and aroma compounds of kiwi wine via the addition of nitrogen during the stationary phase

Nitrogen addition is a viable strategy for preventing sluggish or stuck fermentation and enhancing aroma profiles in the winemaking industry. Herein, nitrogen, in the form of amino acids mixture or diammonium hydrogen phosphate (DAP), was added at different fermentation periods to investigate its effects on the fermentation characteristics, non-volatile compounds and aroma profiles of kiwi wines. Both the amino acids mixture and DAP shortened fermentation time by 1–2 days and balanced organic acid proportion by reducing malic acid content and increasing citric acid content. Adding DAP significantly decreased higher alcohols content, whereas higher alcohols content was remarkably increased with amino acids mixture supplementation. Furthermore, nitrogen addition during the stationary phase was most conducive to synthesizing other beneficial aromas, especially acetate esters and medium-chain fatty acid ethyl esters (MCFAEEs), and improving the perception of tropical fruity and cream aromas in kiwi wines. Partial least-squares regression (PLSR) results showed there was a tight correlation between amino acids and volatile compounds. This study provides an insight into the development of high-quality kiwi wine and should serve as the theoretical foundation to support the application of nitrogen addition in the wine industry.

Metabolome and Microbiome Analysis to Study the Flavor of Summer Black Tea Improved by Stuck Fermentation.

Tea is the most popular and widely consumed beverage worldwide, especially black tea. Summer tea has a bitter and astringent taste and low aroma compared to spring tea due to the higher content of polyphenols and lower content of amino acids. Microbial fermentation is routinely used to improve the flavor of various foods. This study analyzed the relationship between the quality of black tea, metabolic characteristics, and microbial communities after microbial stuck fermentation in summer black tea. Stuck fermentation decreased the bitterness, astringency sourness, and freshness, and increased the sweetness, mellowness, and smoothness of summer black tea. The aroma also changed from sweet and floral to fungal, with a significant improvement in overall quality. Metabolomics analysis revealed significant changes in 551 non-volatile and 345 volatile metabolites after fermentation. The contents of compounds with bitter and astringent taste were decreased. Sweet flavor saccharides and aromatic lipids, and acetophenone and isophorone that impart fungal aroma showed a marked increase. These changes are the result of microbial activities, especially the secretion of extracellular enzymes. Aspergillus, Pullululanibacillus, and Bacillus contribute to the reduction of bitterness and astringency in summer black teas after stuck fermentation, and Paenibacillus and Basidiomycota_gen_Incertae_sedis contribute positively to sweetness. In addition, Aspergillus was associated with the formation of fungal aroma. In summary, our research will provide a suitable method for the improvement of tea quality and utilization of summer tea, as well as provide a reference for innovation and improvement in the food industry.

Single nucleotide polymorphisms associated with wine fermentation and adaptation to nitrogen limitation in wild and domesticated yeast strains

For more than 20 years, Saccharomyces cerevisiae has served as a model organism for genetic studies and molecular biology, as well as a platform for biotechnology (e.g., wine production). One of the important ecological niches of this yeast that has been extensively studied is wine fermentation, a complex microbiological process in which S. cerevisiae faces various stresses such as limited availability of nitrogen. Nitrogen deficiencies in grape juice impair fermentation rate and yeast biomass production, leading to sluggish or stuck fermentations, resulting in considerable economic losses for the wine industry. In the present work, we took advantage of the “1002 Yeast Genomes Project” population, the most complete catalogue of the genetic variation in the species and a powerful resource for genotype-phenotype correlations, to study the adaptation to nitrogen limitation in wild and domesticated yeast strains in the context of wine fermentation. We found that wild and domesticated yeast strains have different adaptations to nitrogen limitation, corroborating their different evolutionary trajectories. Using a combination of state-of-the-art bioinformatic (GWAS) and molecular biology (CRISPR-Cas9) methodologies, we validated that PNP1, RRT5 and PDR12 are implicated in wine fermentation, where RRT5 and PDR12 are also involved in yeast adaptation to nitrogen limitation. In addition, we validated SNPs in these genes leading to differences in fermentative capacities and adaptation to nitrogen limitation. Altogether, the mapped genetic variants have potential applications for the genetic improvement of industrial yeast strains.

Continuous automated microbial monitoring in laboratory-scale winemaking experiments

The management of microbial flora is a key element of the winemaking process. It impacts process time, fermentation quality (e.g., production of alcohol, development of aroma compounds, absence of undesirable microorganisms) and the overall quality of the final wine product. In microbial flora management, real-time microbial monitoring is crucial. When applying currently used classical methods (e.g., cultivation on plates) or more recent technologies (e.g., flow cytometry), the flora sampling frequency is not optimal and taking around-the-clock measurements involves high labour costs. The objective of this study was to evaluate the feasibility of automated microbial monitoring based on an online flow cytometry system for different laboratory-scale applications in the field of winemaking. Initially, a protocol for automated sampling, double-staining and analysis was validated on yeast and bacterial populations after rehydration of the starter cultures. The system was then tested on a variety of increasingly complex biological systems, simulating its applications in the winemaking process. First, a yeast starter culture preparation for “prise de mousse” was tested. Then, a mixed-culture alcoholic fermentation was monitored. Finally, a microbial-focused observation of wine aging was emulated. By overcoming specific constraints linked to the must medium (e.g., sugars, alcohol contents, production of gas, fermentation duration), the results show the potential of this technology for (1) automated yeast or bacterial monitoring in a wide range of laboratory-scale wine environments, (2) simultaneous monitoring of both total and intact populations of multiple microorganisms, (3) long observation periods, and (4) high sampling frequencies for high-resolution data. It could be particularly useful for facilitating and improving control of potential contaminants or stuck fermentations, as well as better piloting starter preparations, alcoholic fermentations, or malolactic fermentations.

Men Who Brew: Masculinity and the Production of Drink in Medieval Icelandic Literature

Men Who Brew: Masculinity and the Production of Drink in Medieval Icelandic Literature

Impact of rare yeasts in Saccharomyces cerevisiae wine fermentation performance: Population prevalence and growth phenotype of Cyberlindnera fabianii, Kazachstania unispora, and Naganishia globosa

Saccharomyces cerevisiae is a highly fermentative species able to complete the wine fermentation. However, the interaction with other non-Saccharomyces yeasts can determine the fermentation performance of S. cerevisiae. We have characterised three rare non-Saccharomyces yeasts (Cyberlindnera fabianii, Kazachstania unispora and Naganishia globosa), studying their impact on S. cerevisiae fitness and wine fermentation performance. Using a wide meta-taxonomic dataset of wine samples, analysed through ITS amplicon sequencing, we show that about a 65.07% of wine samples contains Naganishia spp., a 27.21% contains Kazachstania spp., and only a 4.41% contains Cyberlindnera spp; in all cases with average relative abundances lower than 1% of total fungal populations. Although the studied N. globosa strain showed a limited growth capacity in wine, both K. unispora and C. fabianii showed a similar growth phenotype to that of S. cerevisiae in different fermentation conditions, highlighting the outstanding growth rate values of K. unispora. In mixed fermentations with S. cerevisiae, the three yeast species affected co-culture growth parameters and wine chemical profile (volatile compounds, polysaccharides and proteins). K. unispora DN201 strain presents an outstanding capacity to compete with S. cerevisiae strains during the first stage of wine fermentation, causing stuck fermentations in both synthetic and natural grape musts.

Influence of ergosterol and phytosterols on wine alcoholic fermentation with Saccharomyces cerevisiae strains

Sterols are a fraction of the eukaryotic lipidome that is essential for the maintenance of cell membrane integrity and its good functionality. During alcoholic fermentation, they enhance yeast growth, metabolism and viability, as well as resistance to high sugar content and ethanol stress. Grape musts clarified in excess lead to the loss of solid particles rich in sterols, resulting in sluggish and stuck fermentations. Two sterol sources can help Saccharomyces cerevisiae yeasts to adapt to fermentation stress conditions: ergosterol (synthesized by yeast under aerobic conditions) and phytosterols (plant sterols imported by yeast cells from grape musts under anaerobiosis). Little is known about the physiological impact of phytosterols assimilation in comparison with ergosterol and the influence of sterol type on fermentation kinetics parameters. Moreover, studies to date have analyzed a limited number of yeast strains. Thus, the aim of this work was to compare the performances of a set of Saccharomyces cerevisiae wine strains that represent the diversity of industrial wine yeast, fermenting with phytosterols or ergosterol under two conditions: sterol limitation (sterol starvation) and high sugar content (the most common stress during fermentation). Results indicated that yeast cell viability was negatively impacted by both stressful conditions, resulting in sluggish and stuck fermentations. This study revealed the huge phenotype diversity of the S. cerevisiae strains tested, in particular in terms of cell viability. Indeed, strains with better viability maintenance completed fermentation earlier. Interestingly, we showed for the first time that sterol type differently affects a wide variety of phenotype, such as viability, biomass, fermentation kinetics parameters and biosynthesis of carbon central metabolism (CCM) metabolites. Ergosterol allowed preserving more viable cells at the end of fermentation and, as a consequence, a better completion of fermentation in both conditions tested, even if phytosterols also enabled the completion of alcoholic fermentation for almost all strains. These results highlighted the essential role of sterols during wine alcoholic fermentation to ensure yeast growth and avoid sluggish or stuck fermentations. Finally, this study emphasizes the importance of taking into account sterol types available during wine fermentation.

Importance of micronutrients and organic nitrogen in fermentations with Torulaspora delbrueckii and Saccharomyces cerevisiae

The current use of non-Saccharomyces yeasts in mixed fermentations increases the relevance of the interactions between yeast species. In this work, the interactions between Saccharomyces cerevisiae and Torulaspora delbrueckii were analyzed. For this purpose, fermentations with and without contact between strains of those yeast species were performed in synthetic must. Fermentation kinetics, yeast growth and dynamics were measured over time. Additionally, the effects of nitrogen and other nutrient supplementations on the mixed fermentations were determined. Our results showed that S. cerevisiae did not always dominate the sequential fermentations, and experiments without yeast contact (in which T. delbrueckii cells were removed from the medium before inoculating S. cerevisiae at 48 h) resulted in stuck fermentations except when the inoculum size was increased (from 2 × 106 to 108 cells/mL) or there was a supplementation of thiamine, zinc and amino acids at the same concentration as initially found in the synthetic must. Our findings highlight the importance of inoculum size and ensuring the availability of enough micronutrients for all yeast species, especially in sequential fermentations.

Characterization and Role of Sterols in Saccharomyces cerevisiae during White Wine Alcoholic Fermentation

Responsible for plasma membrane structure maintenance in eukaryotic organisms, sterols are essential for yeast development. The role of two sterol sources in Saccharomyces cerevisiae during wine fermentation is highlighted in this review: ergosterol (yeast sterol produced by yeast cells under aerobic conditions) and phytosterols (plant sterols imported by yeast cells from grape musts in the absence of oxygen). These compounds are responsible for the maintenance of yeast cell viability during white wine fermentation under stress conditions, such as ethanol stress and sterol starvation, to avoid sluggish and stuck fermentations.

Timing of inoculation of Oenococcus oeni and Lactobacillus plantarum in mixed malo-lactic culture along with compatible native yeast influences the polyphenolic, volatile and sensory profile of the Shiraz wines

Malolactic fermentation (MLF) is a secondary wine fermentation resulting from lactic acid bacteria (Oenococcus oeni or Lactobacillus plantarum), especially in cool climates. MLF reduces acidity, improves body, mouthfeel, aroma complexity, and stabilizes the wine. Nevertheless, the traditional MLF using single culture is often associated with stuck fermentation, increased volatile acidity and color loss in wine. Thus, studies on mixed blend of L. plantarum and O. oeni to improve chemical and sensory profiles are gaining importance. The first study on different timing of inoculation of dual malo-lactic culture for the vinification of Shiraz wine aims to understand interactions between the wine cultures (Saccharomyces cerevisiae AAV2, L. plantarum Lp 1 and O. oeni Oo 1). The wines were compared based on chemical properties, phenolic and volatile profiles, and sensory analysis. The study highlighted that early or mid-inoculated MLF wines have higher anthocyanins, flavonoids, syringol, esters, vanillate derivatives, benzaldehyde, and free terpenes compared to traditional MLF wines. The wines were rated high for purple red colour, tannin, body, and overall acceptability. Hence, it was concluded that the inoculation of Lp 1 and Oo 1, either after 7 or 14 days of alcoholic fermentation by yeast AAV2, yield the desired quality Shiraz wine.

In-situ muconic acid extraction reveals sugar consumption bottleneck in a xylose-utilizing Saccharomyces cerevisiae strain

BackgroundThe current shift from a fossil-resource based economy to a more sustainable, bio-based economy requires development of alternative production routes based on utilization of biomass for the many chemicals that are currently produced from petroleum. Muconic acid is an attractive platform chemical for the bio-based economy because it can be converted in chemicals with wide industrial applicability, such as adipic and terephthalic acid, and because its two double bonds offer great versatility for chemical modification.ResultsWe have constructed a yeast cell factory converting glucose and xylose into muconic acid without formation of ethanol. We consecutively eliminated feedback inhibition in the shikimate pathway, inserted the heterologous pathway for muconic acid biosynthesis from 3-dehydroshikimate (DHS) by co-expression of DHS dehydratase from P. anserina, protocatechuic acid (PCA) decarboxylase (PCAD) from K. pneumoniae and oxygen-consuming catechol 1,2-dioxygenase (CDO) from C. albicans, eliminated ethanol production by deletion of the three PDC genes and minimized PCA production by enhancing PCAD overexpression and production of its co-factor. The yeast pitching rate was increased to lower high biomass formation caused by the compulsory aerobic conditions. Maximal titers of 4 g/L, 4.5 g/L and 3.8 g/L muconic acid were reached with glucose, xylose, and a mixture, respectively. The use of an elevated initial sugar level, resulting in muconic acid titers above 2.5 g/L, caused stuck fermentations with incomplete utilization of the sugar. Application of polypropylene glycol 4000 (PPG) as solvent for in situ product removal during the fermentation shows that this is not due to toxicity by the muconic acid produced.ConclusionsThis work has developed an industrial yeast strain able to produce muconic acid from glucose and also with great efficiency from xylose, without any ethanol production, minimal production of PCA and reaching the highest titers in batch fermentation reported up to now. Utilization of higher sugar levels remained conspicuously incomplete. Since this was not due to product inhibition by muconic acid or to loss of viability, an unknown, possibly metabolic bottleneck apparently arises during muconic acid fermentation with high sugar levels and blocks further sugar utilization.

Impact of Nitrogen Addition on Wine Fermentation by S. cerevisiae Strains with Different Nitrogen Requirements.

In modern oenology, supplementation of nitrogen sources is an important strategy to prevent sluggish or stuck fermentation. The present study thoroughly determined the effect of nitrogen addition timing and nitrogen source type on fermentation kinetics and aroma production, carried out by yeast strains with low and high nitrogen requirements. The results revealed that yeast strains with different nitrogen requirements have divergent reactions to nitrogen addition. Nitrogen addition clearly shortened the fermentation duration, especially for the high-nitrogen-demanding yeast strain. Nitrogen addition at 1/3 fermentation was the most effective in terms of fermentation activity, nitrogen assimilation, and production of acetate esters. Interestingly enough, yeast cells preferentially took up amino acids related to fermentative aroma synthesis with the addition at 2/3 fermentation. The addition of nitrogen sources also largely affected the production of important metabolites. Generally speaking, acetic acid, glycerol, and succinic acid reduced with the supplementation of nitrogen sources. The results revealed significant application importance for the winemaking industry.

Influence of nitrogen, minerals and vitamins supplementation on honey wine production using response surface methodology

Mead or honey wine is an alcoholic beverage traditionally produced by the fermentation of a mixture of honey and water. Mead fermentation faces several problems that include sluggish and stuck fermentations, mainly caused by nutrient disequilibrium. In this context, the present study aimed at optimizing the mead fermentation process based on three nutritional factors: nitrogen, minerals, and vitamins, evaluating their effect on several fermentation parameters. The experiments included a response surface methodology (RSM) experiment, and the evaluation of specific minerals on mead parameters: fermentation rate, biomass, final residual sugars, ethanol, glycerol/ethanol ratio, volatile acidity, total acidity, and residual nitrogen. The results showed that the models established by the RSM were adequate for predicting mead fermentation and physic-chemical parameters. Mead´s fermentation rate, ethanol, glycerol/ethanol ratio, total acidity, and residual nitrogen were significantly influenced by nitrogen and minerals supplementation, while fermentation rate and volatile acidity were influenced by nitrogen and vitamins supplementation. An “all minus one” experiment showed that potassium addition is essential from healthy mead fermentation. Considering all the variables analyzed, the optimal conditions for mead fermentation include nitrogen supplementation to achieve 150 mg.L−1 YAN, the addition of minerals, in order to obtain at least 600 mg.L−1 potassium, and supplementation with a vitamin complex.

Selection of low nitrogen demand yeast strains and their impact on the physicochemical and volatile composition of mead.

Mead is an ancient alcoholic beverage produced through the fermentation of a diluted solution of honey. Due to the peculiar and varied composition of honey, mead production faces several problems, such as slow or stuck fermentations mainly due to the low nitrogen concentration, lack of uniformity of the final product and the production of unpleasant aromas. In this context, this work aimed to select low nitrogen-demand yeast strains and evaluate their potential for the production of mead. Therefore, among 21 commercial wine yeast strains, 5 were selected based on their fermentative behavior at low assimilable nitrogen concentrations. The selected strains were further evaluated for their contributions in meads produced with limited nitrogen availability, and the results showed significant differences on some physicochemical parameters like biomass production, residual sugars, glycerol concentration, and fermentative rate. Moreover, meads obtained with selected strains differed in the concentration of several volatile compounds. The volatile compounds concentration and the principal component analysis based on odor activity values allowed separating strains into three groups. In general, S. cerevisiae var bayanus strains (QA23, Spark, and AWRI-R2) were the largest producers of aromatic compounds, particularly those with floral and fruity descriptors. The selection of yeast strains with low nitrogen-demand and different volatile compounds production can be explored by mead makers to limit fermentation problems and obtain characteristic products.

Natural allelic variations of Saccharomyces cerevisiae impact stuck fermentation due to the combined effect of ethanol and temperature; a QTL-mapping study

BackgroundFermentation completion is a major prerequisite in many industrial processes involving the bakery yeast Saccharomyces cerevisiae. Stuck fermentations can be due to the combination of many environmental stresses. Among them, high temperature and ethanol content are particularly deleterious especially in bioethanol and red wine production. Although the genetic causes of temperature and/or ethanol tolerance were widely investigated in laboratory conditions, few studies investigated natural genetic variations related to stuck fermentations in high gravity matrixes.ResultsIn this study, three QTLs linked to stuck fermentation in winemaking conditions were identified by using a selective genotyping strategy carried out on a backcrossed population. The precision of mapping allows the identification of two causative genes VHS1 and OYE2 characterized by stop-codon insertion. The phenotypic effect of these allelic variations was validated by Reciprocal Hemyzygous Assay in high gravity fermentations (> 240 g/L of sugar) carried out at high temperatures (> 28 °C). Phenotypes impacted were mostly related to the late stage of alcoholic fermentation during the stationary growth phase of yeast.ConclusionsOur findings illustrate the complex genetic determinism of stuck fermentation and open new avenues for better understanding yeast resistance mechanisms involved in high gravity fermentations.

Occurrence of yeast cell death associated with micronutrient starvation during wine fermentation varies with nitrogen sources

Aim: Nitrogen availability is an essential parameter for wine alcoholic fermentation. Moreover, recent results have shown that it plays a key role in yeast cell death in interaction with micronutrients limitations such as lipids or vitamins. We found that yeast cell death was triggered by starvation for a set of micronutrients, including oleic acid, ergosterol, pantothenic acid and nicotinic acid whenever the level of nitrogen was high, but not in low nitrogen conditions. We examined here the impact of the nature of the nitrogen source supplementation in the light of these previous results.Methods and results: 19 amino acids or NH4+ were added, in amounts corresponding to 354 mg/L assimilable nitrogen, to an oenological medium that was low in nitrogen and oleic acid. Yeast viability in function of the fermentation progress was assessed and showed differences in cell death during the alcoholic fermentation in function of the amino acid added. The addition of NH4+ was also tested at two different times during wine fermentation. The results obtained show that various nitrogen sources (amino acids, ammonium) can trigger cell death but with different intensities.Conclusion: It appears that some amino acids are preferable to others in alcoholic fermentation because they do not trigger cell death. We also provide evidence that the timing of nitrogen addition has a strong impact on cell death in musts with micronutrient limitations: an early nitrogen addition is more likely to trigger cell death than a late addition.Significance and impact of the study: Our results provide a novel frame for managing nitrogen supplementation of grape musts and to avoid stuck fermentation.

Taxonomic and Functional Characterization of the Microbial Community During Spontaneous in vitro Fermentation of Riesling Must.

Although there is an extensive tradition of research into the microbes that underlie the winemaking process, much remains to be learnt. We combined the high-throughput sequencing (HTS) tools of metabarcoding and metagenomics, to characterize how microbial communities of Riesling musts sampled at four different vineyards, and their subsequent spontaneously fermented derivatives, vary. We specifically explored community variation relating to three points: (i) how microbial communities vary by vineyard; (ii) how community biodiversity changes during alcoholic fermentation; and (iii) how microbial community varies between musts that successfully complete alcoholic fermentation and those that become ‘stuck’ in the process. Our metabarcoding data showed a general influence of microbial composition at the vineyard level. Two of the vineyards (4 and 5) had strikingly a change in the differential abundance of Metschnikowia. We therefore additionally performed shotgun metagenomic sequencing on a subset of the samples to provide preliminary insights into the potential relevance of this observation, and used the data to both investigate functional potential and reconstruct draft genomes (bins). At these two vineyards, we also observed an increase in non-Saccharomycetaceae fungal functions, and a decrease in bacterial functions during the early fermentation stage. The binning results yielded 11 coherent bins, with both vineyards sharing the yeast bins Hanseniaspora and Saccharomyces. Read recruitment and functional analysis of this data revealed that during fermentation, a high abundance of Metschnikowia might serve as a biocontrol agent against bacteria, via a putative iron depletion pathway, and this in turn could help Saccharomyces dominate the fermentation. During alcoholic fermentation, we observed a general decrease in biodiversity in both the metabarcoding and metagenomic data. Unexpected Micrococcus behavior was observed in vineyard 4 according to metagenomic analyses based on reference-based read mapping. Analysis of open reading frames using these data showed an increase of functions assigned to class Actinobacteria in the end of fermentation. Therefore, we hypothesize that bacteria might sit-and-wait until Saccharomyces activity slows down. Complementary approaches to annotation instead of relying a single database provide more coherent information true species. Lastly, our metabarcoding data enabled us to identify a relationship between stuck fermentations and Starmerella abundance. Given that robust chemical analysis indicated that although the stuck samples contained residual glucose, all fructose had been consumed, we hypothesize that this was because fructophilic Starmerella, rather than Saccharomyces, dominated these fermentations. Overall, our results showcase the different ways in which metagenomic analyses can improve our understanding of the wine alcoholic fermentation process.

Low Phenotypic Penetrance and Technological Impact of Yeast [GAR +] Prion-Like Elements on Winemaking.

[GAR+] prion-like elements partially relieve carbon catabolite repression in Saccharomyces cerevisiae. They have been hypothesized to contribute to wine yeast survival and alcohol level reduction, as well as communication with bacteria and stuck fermentation. In this work, we selected [GAR+] derivatives from several genetic backgrounds. They were characterized for phenotypic penetrance, heritability and confirmed as prion-like through curing by desiccation. In terms of fermentation kinetics, the impact of the prion on anaerobic wine fermentation (natural grape juice) was either neutral or negative, depending on the genetic background. Likewise, residual sugars were higher or similar for [GAR+] as compared to the cognate [gar-] strains. The prions had little or no impact on glycerol and ethanol yields; while acetic acid yields experienced the highest variations between [GAR+] and [gar-] strains. Strains analyzed under aerobic conditions followed the same pattern, with either little or no impact on fermentation kinetics, ethanol or glycerol yield; and a clearer influence on volatile acidity. Although no clear winemaking advantages were found for [GAR+] strains in this work, they might eventually show interest for some combinations of genetic background or winemaking conditions, e.g., for reducing acetic acid yield under aerated fermentation.

Selection of Indigenous Saccharomyces cerevisiae Strains for Winemaking in Northwest China

To avoid the risk for sluggish or stuck fermentations and quality deterioration caused by increasing sugar contents of grape grown in northwest China, a selection of indigenous Saccharomyces cerevisiae strains with excellent enological characteristics could be of great value for the local wine industry. For this purpose, 52 indigenous S. cerevisiae strains, isolated from five wine-producing regions of northwest China, were evaluated for several important enological properties, including stress tolerance, killer phenotype, hydrogen sulfide and foam production levels, and flocculation and fermentation rates. Microfermentations with eight preselected yeast strains were performed on Cabernet Sauvignon. All selected strains could complete fermentations, and the enochemical parameters were within the acceptable ranges of the wine industry. Four strains (XJ3, SAX2, SAX5, and SAX6) produced higher amounts of esters than the control commercial strains, and the corresponding wines were highly regarded by wine judges as having the best sensory quality. The fermentation performance of these strains at high initial sugar concentrations was also analyzed. They could be used as starter cultures, and the selected indigenous strains could potentially improve the regional character of the wine. Moreover, these strains may have the potential to produce wines from grapes with high initial sugar content in the Xinjiang and Ningxia regions of northwest China.

Quantitative assessment of DNA damage in the industrial ethanol production strain Saccharomyces cerevisiae PE-2

Lignocellulosic hydrolysates remain one of the most abundantly used substrates for the sustainable production of second generation fuels and chemicals with Saccharomyces cerevisiae. Nevertheless, fermentation inhibitors such as acetic acid, furfural and hydroxymethylfurfural are formed during the process and can lead to slow or stuck fermentations and/or act as genotoxic agents leading to production strain genetic instability. We have developed a novel dominant deletion (DEL) cassette assay for quantification of DNA damage in both wild-type and industrial yeast strains. Using this assay, the ethanol production strain S. cerevisiae PE-2 was shown to be more resistant to hydrogen peroxide and furfural than the laboratory DEL strain RS112. Indeed, the PE-2 strain also showed a lower tendency for recombination, consistent with a more efficient DNA protection. The dominant DEL assay presented herein should prove to be a useful tool in the selection of robust yeast strains and process conditions for second generation feedstock fermentations.