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Sherry wines are film wines produced in the Jerez-Xérès-Sherry and Montilla-Moriles regions in southern Spain which require an aging process under flor biofilms, known as "biological aging". The presence of mites in Sherry wine wineries has been reported and associated with improved wine volatile properties. This work analyzes the microbial diversity in flor biofilms and mites in Sherry wine wineries using Matrix-Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) and ITS/gene amplification. Two mite species, Carpoglyphus lactis and Tyrophagus putrescentiae , were spotted in the sampled winery and 32 microorganism species were identified in their exoskeleton or surrounding biofilms. To our knowledge, 26 of these species were never described before in sherry wine environments. We hypothesized that mites feed on the flor biofilms as well as another type of biofilm located in barrel cracks, known by winemakers as "natas" (cream in English). These non-studied biofilms showed the highest microbiome diversity among all samples (followed by C. lactis spotted nearby) thus, representing a niche of microorganisms with potential biotechnological interest. Besides mites, Drosophila flies were spotted in the sampling areas. The role of flies and mites as vectors that transport microorganisms among different niches (i.e., flor biofilms and natas) is discussed. • 32 microorganism species were identified, 26 not described before in Sherry wineries. • "Natas" biofilm formed by multiple microorganisms, is described for the first time. • Mites may act as microorganism vectors between different winery environments. [ABSTRACT FROM AUTHOR]

Abstract In food industry and winemaking, the use of active dehydrated yeast (ADY) Saccharomyces cerevisiae is a frequent practice because of the long-term stability and high efficiency of ADY. Nowadays, there is an increasing interest for new yeasts strains, such as Torulaspora delbrueckii (Td), Metschnikowia pulcherrima (Mp) and Lachancea thermotolerans (Lt). However, the yeasts transformation processes into the solidified form generate several stresses that reduce the cell viability. In this case, understanding the phenomena of yeast cell resistance before, during and after dehydration is of great importance. In this study we analyzed two compounds associated with resistance to stress and produced by cells, glutathione (total, oxidized and reduced) and trehalose, at different stages of the process. The impact of growing and dehydration conditions on cell viability was analyzed by flow cytometry and two-photon laser scanning microscopy. The results showed that cells naturally enriched in glutathione or trehalose acquired resistance to dehydration, preventing the oxidation of glutathione in a growth/dehydration condition dependent manner. This is the first time that simultaneous metabolic and dehydration responses were observed in three non- Saccharomyces strains. These findings represent an opportunity to better understand the yeast's dehydration resistance phenomena and thus to promote the efficient industrial production of new dried yeasts. Highlights • Multiple growing-dehydration responses were compared between yeasts T. delbrueckii , M. pulcherrima and L. thermotolerans. • GSH and TRE accumulation was correlated with high cell survival. • Trehalose rich-cells were partially protected from oxidation. • Cell viability is strongly correlated to dehydration kinetics for non- Saccharomyces. [ABSTRACT FROM AUTHOR]

Abstract Oxidative stress is a common stress in yeasts during the stages of the winemaking process in which aerobic growth occurs, and it can modify the cellular lipid composition. The aim of this study was to evaluate the oxidative stress tolerance of two non-conventional yeasts (Torulaspora delbrueckii and Metschnikowia pulcherrima) compared to Saccharomyces cerevisiae. Therefore, their resistance against H 2 O 2 , the ROS production and the cellular lipid composition were assessed. The results showed that the non- Saccharomyces yeasts used in this study exhibited higher resistance to H 2 O 2 stress and lower ROS accumulation than Saccharomyces. Regarding the cellular lipid composition, the two non- Saccharomyces species studied here displayed a high percentage of polyunsaturated fatty acids, which resulted in more fluid membranes. This result could indicate that these yeasts have been evolutionarily adapted to have better resistance against the oxidative stress. Furthermore, under external oxidative stress, non- Saccharomyces yeasts were better able to adapt their lipid composition as a defense mechanism by decreasing their percentage of polyunsaturated fatty acids and squalene and increasing their monounsaturated fatty acids. Highlights • Non- Saccharomyces yeasts better resist oxidative stress induced by H 2 O 2. • Non- Saccharomyces have higher levels of PUFAs which results in more fluid membranes. • The yeast stress tolerance is associated with higher levels of UFAs. • Higher PC/PE and ergosterol/squalene ratios increase oxidative stress resistance. [ABSTRACT FROM AUTHOR]

While sequentially inoculating non- Saccharomyces yeasts with Saccharomyces cerevisiae can lower the alcohol contents of wine, the abilities of these yeasts to utilize/produce ethanol or generate other byproducts remained unclear. Metschnikowia pulcherrima or Meyerozyma guilliermondii were inoculated into media with or without S. cerevisiae to assess byproduct formation. Both species metabolized ethanol in a yeast-nitrogen-base medium but produced the alcohol in a synthetic grape juice medium. In fact, Mt. pulcherrima and My. guilliermondii generated less ethanol per gram of metabolized sugar (0.372 and 0.301 g/g, respectively) compared to S. cerevisiae (0.422 g/g). Sequentially inoculating each non- Saccharomyces species with S. cerevisiae into grape juice media achieved up to 3.0% v/v alcohol reduction compared to S. cerevisiae alone while producing variable glycerol, succinic acid, and acetic acid concentrations. However, neither non- Saccharomyces yeasts released appreciable CO 2 under fermentative conditions regardless of incubation temperature. Despite equivalent peak populations, S. cerevisiae produced more biomass (2.98 g/L) than the non- Saccharomyces yeasts while sequential inoculations yielded higher biomass with Mt. pulcherrima (3.97 g/L) but not My. guilliermondii (3.03 g/L). To reduce ethanol concentrations, these non- Saccharomyces species may metabolize ethanol and/or produce less from metabolized sugars compared to S. cerevisiae but also divert carbon towards glycerol, succinic acid, and/or biomass. • Inoculating non- Saccharomyces yeasts before S. cerevisiae yielded less ethanol. • Mt. pulcherrima and My. guilliermondii could metabolize or produce ethanol. • Byproducts of these yeasts were glycerol, succinic acid, and biomass but not CO 2. [ABSTRACT FROM AUTHOR]

The use of Torulaspora delbrueckii in the alcoholic fermentation (AF) of grape must is increasingly studied and used in the wine industry. In addition to the organoleptic improvement of wines, the synergy of this yeast species with the lactic acid bacterium Oenococcus oeni is an interesting field of study. In this work, 60 strain combinations were compared: 3 strains of Saccharomyces cerevisiae (Sc) and 4 strains of Torulaspora delbrueckii (Td) in sequential AF, and four strains of O. oeni (Oo) in malolactic fermentation (MLF). The objective was to describe the positive or negative relationships of these strains with the aim of finding the combination that ensures better MLF performance. In addition, a new synthetic grape must has been developed that allows the success of AF and subsequent MLF. Under these conditions, the Sc-K1 strain would be unsuitable for carrying out MLF unless there is prior inoculation with Td-Prelude, Td-Viniferm or Td-Zymaflore always with the Oo-VP41 combination. However, from all the trials performed, it appears that the combinations of sequential AF with Td-Prelude and Sc-QA23 or Sc-CLOS, followed by MLF with Oo-VP41, reflected a positive effect of T. delbrueckii compared to inoculation of Sc alone, such as a reduction in L-malic consumption time. In conclusion, the obtained results highlight the relevance of strain selection and yeast-LAB strain compatibility in wine fermentations. The study also reveals the positive effect on MLF of some T. delbrueckii strains. • Synthetic grape musts fermented by Torulaspora / Saccharomyces combinations enabled us to perform a screening of strains of O. oeni. • Sequential fermentations with T. delbrueckii strains had higher equivalent mannose values. • Oo-VP41 LAB strain has been the best MLF performer of 4 strains in almost all combinations with T. delbrueckii strains. • T. delbrueckii improves malolactic fermentation in wines fermented with a S. cerevisiae not recommended for MLF. • QA23 or CLOS strains in combination with T. delbrueckii Prelude followed by a MLF with O. oeni VP41 is the best strain combination. [ABSTRACT FROM AUTHOR]

Way-a-linah, an alcoholic beverage produced from the fermented sap of Eucalyptus gunnii , and tuba , a fermented drink made from the syrup of Cocos nucifera fructifying bud, are two of several fermented beverages produced by Australian Aboriginal and Torres Strait people. Here we describe the characterisation of yeast isolates from samples associated with the fermentation of way-a-linah and tuba. Microbial isolates were obtained from two different geographical locations in Australia - the Central Plateau in Tasmania, and Erub Island in the Torres Strait. While Hanseniaspora species and Lachancea cidri were the most abundant species in Tasmania, Candida species were the most abundant in Erub Island. Isolates were screened for tolerance to stress conditions found during the production of fermented beverages and for enzyme activities relevant to the appearance, aroma and flavour of these beverages. Based on screening results, eight isolates were evaluated for their volatile profile during the fermentation of wort, apple juice and grape juice. Diverse volatile profiles were observed for beers, ciders and wines fermented with different isolates. These findings reveal the potential of these isolates to produce fermented beverages with unique aroma and flavour profiles and highlight the vast microbial diversity associated with fermented beverages produced by Australia's Indigenous peoples. • Way-a-linah and tuba are two of several fermented beverages produced by Australia's Indigenous peoples. • Yeast isolates associated with the fermentation of way-a-linah and tuba were identified and characterised. • Isolates were evaluated for their volatile profile during the fermentation of wort, apple juice and grape juice. • Diverse volatile profiles were observed for beers, ciders and wines fermented with different isolates. [ABSTRACT FROM AUTHOR]

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. • The prevalence of Cyberlindnera spp. in wine populations is about 4% of samples. • The prevalence of Kazachstania spp. in wine populations is about 27% of samples. • The prevalence of Naganishia spp. in wine populations is about 65% of samples. • Kazachstania unispora DN201 caused stuck fermentations. • K.unispora DN201 has high nitrogen consumption rate and production of acetic acid. [ABSTRACT FROM AUTHOR]

Abstract: Eight non-Saccharomyces wine strains, previously selected for their ability to modulate the final concentrations of various volatile compounds and to persist with Saccharomyces cerevisiae in mixed inocula fermentations of grape juice, have been analyzed in the present work to test their ability to release mannoproteins. The eight strains were members of different genera originally isolated from grape: Hansensiaspora osmophila, Lachancea thermotolerans, Metschnikowia pulcherrima, Pichia fermentans, Saccharomycodes ludwigii, Starmerella bacillaris, Torulaspora delbrueckii and Zygosaccharomyces florentinus. A synthetic polysaccharide-free grape juice, was used to characterize the mannoproteins released during the alcoholic fermentation. Mannoproteins profiles were characterized by gel electrophoresis and carbohydrate composition was analyzed both by HPLC and by mass spectrometry. The eight non-Saccharomyces yeasts demonstrated a higher capacity to release polysaccharides compared to S. cerevisiae. The proteins released by the eight yeast strains showed a wide variety of protein sizes, ranging from 25 kDa to greater than 250 kDa. The mass spectrometric profile of the N-glycans ranged from 1600 to 4000 Da and was characteristic for each strain. Detailed investigation of the degree of polymerization of released N-glycans revealed variable composition from 8 to 15 units of monosaccharides. [Copyright &y& Elsevier]

Abstract: In Mediterranean countries the most diffuse practice to obtain the valorization of grape marc, the main by-product from winemaking, is the production of spirits. During this process, marc storage for sugar fermentation represents a crucial step, since side-fermentations leading to off-flavours production can very easily occur. In this study we evaluated the effect of the addition of two yeast strains, inoculated separately at the beginning of the storage period, into marcs from two Italian grape varieties with the aim to control the development of autochthonous microbiota and to improve spirit quality. The presence of the inoculated strains was monitored by means of PCR-based approaches. A commercial Saccharomyces cerevisiae strain, chosen as this species is notably the best ethanol producer, showed excellent ability to dominate the autochthonous microflora and to reduce off-flavours as demonstrated by chemical analysis and sensory evaluation. A Saccharomycodes ludwigii strain, chosen for increasing varietal compounds thus enhancing spirit aroma, showed a level of implantation not sufficient to assure a clear beneficial effect on quality. The implantation level of this strain was affected by S. cerevisiae competition since the highest level was found in grape marc with lower sugar content, where indigenous S. cerevisiae were less persistent. [Copyright &y& Elsevier]

This study used a double-compartment fermenter to assess yeast growth, fermentation activity, and aroma production in response to cell-cell contact during mixed culture fermentation of Pinot noir grape must with Pichia kluyveri and Saccharomyces cerevisiae. Furthermore, amino acids were analyzed in order to study yeast interactions and possible reasons for aroma modulation as a response to cell-cell contact. Our results show that cell-cell contact between the two yeasts decreased cell viability of each yeast during mixed culture fermentation, and that it increased acetate and ethyl ester production and decreased varietal volatile levels. Moreover, it increased the consumption of glutamic acid and the biosynthesis of some specific amino acids related to cell growth, mainly histidine, glycine and proline, while suppressing the production of higher alcohols through the Ehrlich pathway. These results may contribute to an improved understanding, and thus control, of aroma production in mixed culture wine fermentations. • A cell-cell contact interaction was found between P. kluyveri and S. cerevisiae. • Cell-cell contact suppressed cell growth of each yeast in their mixed fermentation. • Cell-cell contact increased the production of both acetate and ethyl esters. • Cell-cell contact induced the biosynthesis of amino acids related to cell growth. [ABSTRACT FROM AUTHOR]

Abstract: 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. In this specific pairing of yeast strains in mixed fermentations (S. cerevisiae EC1118 and L. thermotolerans 101), this non-Saccharomyces yeast showed a high level of competitiveness. Nevertheless the S. cerevisiae strain dominated the fermentation over the spontaneous S. cerevisiae strains also under the industrial fermentation conditions. The different condition tested (modalities of inoculum, temperature of fermentation, different grape juice) influenced the specific interactions and the fermentation behaviour of the co-culture of S. cerevisiae and L. thermotolerans. However, some metabolic behaviours such as pH reduction and enhancement of 2-phenylethanol and glycerol, were shown here under all of the conditions tested. The specific chemical profiles of these wines were confirmed by the sensory analysis test, which expressed these results at the tasting level as significant increases in the spicy notes and in terms of total acidity increases. [Copyright &y& Elsevier]

Abstract: There has been increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. The main reason is that the multistarter fermentation process is thought to simulate indigenous fermentation, thus increasing wine aroma complexity while avoiding the risks linked to natural fermentation. However, multistarter fermentation is characterised by complex and largely unknown interactions between yeasts. Consequently the resulting wine quality is rather unpredictable. In order to better understand the interactions that take place between non-Saccharomyces and Saccharomyces yeasts during alcoholic fermentation, we analysed the volatile profiles of several mono-culture and co-cultures. Candida zemplinina, Torulaspora delbrueckii and Metschnikowia pulcherrima were used to conduct fermentations either in mono-culture or in co-culture with S. cerevisiae. Up to 48 volatile compounds belonging to different chemical families were quantified. For the first time, we show that C. zemplinina is a strong producer of terpenes and lactones. We demonstrate by means of multivariate analysis that different interactions exist between the co-cultures studied. We observed a synergistic effect on aromatic compound production when M. pulcherrima was in co-culture with S. cerevisiae. However a negative interaction was observed between C. zemplinina and S. cerevisiae, which resulted in a decrease in terpene and lactone content. These interactions are independent of biomass production. The aromatic profiles of T. delbrueckii and S. cerevisiae in mono-culture and in co-culture are very close, and are biomass-dependent, reflecting a neutral interaction. This study reveals that a whole family of compounds could be altered by such interactions. These results suggest that the entire metabolic pathway is affected by these interactions. [Copyright &y& Elsevier]

Abstract: The presence of Saccharomyces cerevisiae in grape berries and fresh musts is usually very low. However, as fermentation progresses, the population levels of this species considerably increase. In this study, we use the concept of fitness advantage to measure how increasing ethanol concentrations (0–25%) and temperature values (4–46 °C) in wine fermentations affects competition between S. cerevisiae and several non-Saccharomyces yeasts (Hanseniaspora uvarum, Torulaspora delbrueckii, Candida zemplinina, Pichia fermentans and Kluyveromyces marxianus). We used a mathematical approach to model the hypothetical time needed for S. cerevisiae to impose itself on a mixed population of the non-Saccharomyces species described above. This approach also took into consideration the influence of environmental factors and the initial population levels of S. cerevisiae (0.1, 1.0 and 10.0%). Our results suggest that Saccharomyces niche construction via ethanol production does not provide a clear ecological advantage (at least not until the ethanol concentration exceeds 9%), whereas a temperature rise (above 15 °C) does give S. cerevisiae a considerable advantage. The initial frequency of S. cerevisiae considerably influences the time it needs to impose itself (until it reaches a final frequency of 99% in the mixed culture), the lowest time values being found at the highest initial frequency. In light of these results, the application of low temperatures in the wine industry could favor the growth and survival of non-Saccharomyces species for a longer period of time. [Copyright &y& Elsevier]

Abstract: Thirty-eight yeast strains belonging to the genera Candida, Hanseniaspora, Pichia, Torulaspora and Zygosaccharomyces were screened for ester formation on synthetic microbiological medium. The genera Hanseniaspora and Pichia stood out as the best acetate ester producers. Based on the ester profile Hanseniaspora guilliermondii 11027 and 11102, Hanseniaspora osmophila 1471 and Pichia membranifaciens 10113 and 10550 were selected for further characterization of enological traits. When growing on must H. osmophila 1471, which displayed a glucophilic nature and was able to consume more than 90% of initial must sugars, produced levels of acetic acid, medium chain fatty acids and ethyl acetate, within the ranges described for wine. On the other hand, it was found to be a strong producer of 2-phenylethyl acetate. Our data suggest that H. osmophila 1471 is a good candidate for mixed starters, although the possible interactions with S. cerevisiae deserve further research. [Copyright &y& Elsevier]

Abstract: The growth dynamics of three non-Saccharomyces strains in combination with Saccharomyces cerevisiae during fermentation of a sterile grape juice have been studied. The influence of the initial concentrations of S. cerevisiae on the whole yeast community was the main purpose of this research. The progression of S. cerevisiae within the first 5 days of fermentation was monitored by enumeration on selective and non-selective media. The population of each species was evaluated by morphological criteria. After 24h, Hanseniaspora uvarum represented more than 50% of the whole yeast community, including ferments with the highest initial concentration of S. cerevisiae. As the population of S. cerevisiae increased, H. uvarum decreased. Metchnikowia pulcherrima was more inhibited by S. cerevisiae than H. uvarum, whereas the growth of Candida stellata was less inhibited. After thirty days, irrespective of the initial concentration of S. cerevisiae, only S. cerevisiae was detected in all ferments. Conversion of sugars to ethanol correlated with the initial population of S. cerevisiae. Glucose was almost completely exhausted in all cases, independent of the initial S. cerevisiae concentration used. Grape juices inoculated with composite inocula of non-S. cerevisiae and S. cerevisiae, with its initial concentration lower than 5cfu/ml did not produce wines according to wine regulations. The concentration of ethanol in the wines did not reach the minimum amount of 9vol%. Samples of wines fermented with a composite inoculum the concentration of S. cerevisiae represented 50cfu/ml of grape juice, were judged to have the best sensorial properties. [Copyright &y& Elsevier]

The use of non- Saccharomyces yeast together with S. cerevisiae in winemaking is a current trend. Apart from the organoleptic modulation of the wine, the composition of the resulting yeast lees is different and may thus impact malolactic fermentation (MLF). Yeasts of Saccharomyces cerevisiae, Torulaspora delbrueckii and Metschnikowia pulcherrima were inactivated and added to a synthetic wine. Three different strains of Oenococcus oeni were inoculated and MLF was monitored. Non- Saccharomyces lees, especially from some strains of T. delbrueckii , showed higher compatibility with some O. oeni strains, with a shorter MLF and a maintained bacterial cell viability. The supplementation of lees increased nitrogen compounds available by O. oeni. A lower mannoprotein consumption was related with longer MLF. Amino acid assimilation by O. oeni was strain specific. There may be many other compounds regulating these yeast lees- O. oeni interactions apart from the well-known mannoproteins and amino acids. This is the first study of MLF with different O. oeni strains in the presence of S. cerevisiae and non- Saccharomyces yeast lees to report a strain-specific interaction between them. • Non- Saccharomyces lees showed higher compatibility with O. oeni. • Lower mannoprotein consumption by O. oeni was related with longer fermentations. • Amino acid contribution of the lees and assimilation by O. oeni are strain specific. [ABSTRACT FROM AUTHOR]

The increasing interest in novel beer productions focused on non- Saccharomyces yeasts in order to pursue their potential in generating groundbreaking sensory profiles. Traditional fermented beverages represent an important source of yeast strains which could express interesting features during brewing. A total of 404 yeasts were isolated from fermented honey by-products and identified as Saccharomyces cerevisiae , Wickerhamomyces anomalus , Zygosaccharomyces bailii , Zygosaccharomyces rouxii and Hanseniaspora uvarum. Five H. uvarum strains were screened for their brewing capability. Interestingly, Hanseniaspora uvarum strains showed growth in presence of ethanol and hop and a more rapid growth than the control strain S. cerevisiae US-05. Even though all strains showed a very low fermentation power, their concentrations ranged between 7 and 8 Log cycles during fermentation. The statistical analyses showed significant differences among the strains and underlined the ability of YGA2 and YGA34 to grow rapidly in presence of ethanol and hop. The strain YGA34 showed the best technological properties and was selected for beer production. Its presence in mixed- and sequential-culture fermentations with US-05 did not influence attenuation and ethanol concentration but had a significant impact on glycerol and acetic acid concentrations, with a higher sensory complexity and intensity, representing promising co-starters during craft beer production. • Νon-S accharomyces strains to be used as co-starter for craft beer production. • Hanseniaspora uvarum strains with high resistance to ethanol and hop. • Use of Hanseniaspora uvarum to improve aromatic profile of craft beer. [ABSTRACT FROM AUTHOR]

Beer production is predominantly carried out by Saccharomyces species, such as S. cerevisiae and S. pastorianus. However, the introduction of non- Saccharomyces yeasts in the brewing process is now seen as a promising strategy to improve and differentiate the organoleptic profile of beer. In this study, 17 non- Saccharomyces strains of 12 distinct species were isolated and submitted to a preliminary sensory evaluation to determine their potential for beer bioflavouring. Hanseniaspora guilliermondii IST315 and H. opuntiae IST408 aroma profiles presented the highest acceptability and were described as having 'fruity' and 'toffee' notes, respectively. Their presence in mixed-culture fermentations with S. cerevisiae US-05 did not influence attenuation and ethanol concentration of beer but had a significant impact in its volatile composition. Notably, while both strains reduced the total amount of ethyl esters, H. guilliermondii IST315 greatly increased the concentration of acetate esters, especially when sequentially inoculated, leading to an 8.2-fold increase in phenylethyl acetate ('rose', 'honey' aroma) in the final beverage. These findings highlight the importance of non- Saccharomyces yeasts in shaping the aroma profile of beer and suggest a role for Hanseniaspora spp. in improving it. • Non- Saccharomyces yeasts able to add organoleptic complexity to beer were isolated. • H. opuntiae IST408 increases abundance of acetate esters in mixed fermentations with S. cerevisiae. • This effect is even more pronounced for H. guilliermondii IST315, which increases phenylethyl acetate abundance by 8.2-fold. • Hanseniaspora species enhance the aroma profile of beer, which was well received by expert panellists. [ABSTRACT FROM AUTHOR]

The use of non- Saccharomyces species as starter cultures together with Saccharomyces cerevisiae is becoming a common practice in the oenological industry to produce wines that respond to new market demands. In this context, microbial interactions with these non- Saccharomyces species must be considered for a rational design of yeast starter combinations. Previously, transcriptional responses of S. cerevisiae to short-term co-cultivation with Torulaspora delbrueckii , Candida sake , or Hanseniaspora uvarum was compared. An activation of sugar consumption and glycolysis, membrane and cell wall biogenesis, and nitrogen utilization was observed, suggesting a metabolic boost of S. cerevisiae in response to competing yeasts. In the present study, the transcription profile of S. cerevisiae was analyzed after 3 h of cell contact with Metschnikowia pulcherrima. Results show an over-expression of the gluco-fermentative pathway much stronger than with the other species. Moreover, a great repression of the respiration pathway has been found in response to Metschnikowia. Our hypothesis is that there is a direct interaction stress response (DISR) between S. cerevisiae and the other yeast species that, under excess sugar conditions, induces transcription of the hexose transporters, triggering glucose flow to fermentation and inhibiting respiration, leading to an increase in both, metabolic flow and population dynamics. • Co-cultivation promotes up-regulation of glycolysis and down-regulation of the TCA. • Co-cultivation with M. pulcherrima also impacts cell-wall remodeling. • The transcriptional response is not dependent on the oxygen availability. • The Direct Interaction Stress Response (DISR) in S. cerevisiae is proposed. [ABSTRACT FROM AUTHOR]

This work was performed to investigate on the yeast ecology of durum wheat to evaluate the interaction between kernel yeasts and the commercial baker's yeast Saccharomyces cerevisiae during dough leavening. Yeast populations were studied in 39 genotypes of durum wheat cultivated in Sicily. The highest level of kernel yeasts was 2.9 Log CFU/g. A total of 413 isolates was collected and subjected to phenotypic and genotypic characterization. Twenty-three yeast species belonging to 11 genera have been identified. Filobasidium oeirense , Sporobolomyces roseus and Aureobasidium pullulans were the species most commonly found in durum wheat kernels. Doughs were co-inoculated with yeasts isolated from wheat kernels and commercial Saccharomyces cerevisiae , in order to evaluate the interactions between yeasts and the leavening performance. Yeast populations of all doughs have been monitored as well as dough volume increase and weight loss (as CO 2) measured after 2 h of fermentation. The doughs whose final volume was higher than control dough (inoculated exclusively with S. cerevisiae) were those inoculated with Naganishia albida , Vishniacozyma dimennae (118 mL each), and Candida parapsilosis (102 mL). The weight losses were variable, depending on the co-culture used with S. cerevisiae and the values were in the range of 0.08–1.00 g CO 2 /100 g. The kernel yeasts species C. parapsilosis , N. albida , P. terrestris , R. mucilaginosa and V. dimennae deserves future attention to be co-inoculated with the commercial starter S. cerevisiae in order to improve the sensory characteristics of bread. • Ecology of yeast populations associated to Triticum turgidum subsp. durum. • Identification of the yeast species isolated to modern genotypes and old lantraces • Interaction between kernel yeast species and Saccharomyces cerevisiae. • Different volume increases in co-inoculated doughs. [ABSTRACT FROM AUTHOR]

Non- Saccharomyces wine yeasts are useful tools for producing wines with complex aromas or low ethanol content. Their use in wine would benefit from their production as active dry yeast (ADY) starters to be used as co-inocula alongside S. cerevisiae. Oxidative stress during biomass propagation and dehydration is a key factor in determining ADY performance, as it affects yeast vitality and viability. Several studies have analysed the response of S. cerevisiae to oxidative stress under dehydration conditions, but not so many deal with non-conventional yeasts. In this work, we analysed eight non- Saccharomyces wine yeasts under biomass production conditions and studied oxidative stress parameters and lipid composition. The results revealed wide variability among species in their technological performance during ADY production. Also, for Metschnikowia pulcherrima and Starmerella bacillaris , better performance correlates with high catalase activity and glutathione levels. Our data suggest that non- Saccharomyces wine yeasts with an enhanced oxidative stress response are better suited to grow under ADY production conditions. • Production of active dry wine yeast is not optimized for non- Saccharomyces yeasts. • High lipid peroxidation in dry crabtree negative yeast cells does not cause low viability. • Total fatty acids composition can be correlated to active dry yeast performance. • Antioxidant protection is relevant for non- Saccharomyces active dry yeast performance. [ABSTRACT FROM AUTHOR]

In the search for the desired oenological features and flavour complexity of wines, there is growing interest in the potential use of non-Saccharomyces yeast that are naturally present in the winemaking environment. Torulaspora delbrueckii is one such yeast that has seen profitable use in mixed fermentations with Saccharomyces cerevisiae and with different grape varieties. T. delbrueckii can have positive and distinctive impacts on the overall aroma of wines, and has also been used at an industrial level. Here, T. delbrueckii was successfully used in pure and mixed secondary fermentations for sparkling wine. The two selected T. delbrueckii strains used completed the secondary fermentation 'prise de mousse' in these pure and mixed fermentations. The sparkling wines obtained with T. delbrueckii showed different aromatic compositions and sensory profiles to those of S. cerevisiae. T. delbrueckii strain DiSVA 130 showed high esters production and significantly high scores for some of the aromatic descriptors that positively influence the sensory profile of sparkling wine. Thus, the use of T. delbrueckii in pure and mixed fermentations is a suitable strategy to further develop the flavour complexity during secondary fermentation of sparkling wines., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The use of non-Saccharomyces strains in aerated conditions has proven effective for alcohol content reduction in wine during lab-scale fermentation. The process has been scaled up to 20 L batches, in order to produce lower alcohol wines amenable to sensory analysis. Sequential instead of simultaneous inoculation was chosen to prevent oxygen exposure of Saccharomyces cerevisiae during fermentation, since previous results indicated that this would result in increased acetic acid production. In addition, an adaptation step was included to facilitate non-Saccharomyces implantation in natural must. Wines elaborated with Torulaspora delbrueckii or Metschnikowia pulcherrima in aerated conditions contained less alcohol than control wine (S. cerevisiae, non-aerated). Sensory and aroma analysis revealed that the quality of mixed fermentations was affected by the high levels of some yeast amino acid related byproducts, which suggests that further progress requires a careful selection of non-Saccharomyces strains and the use of specific N-nutrients., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Today, many non-Saccharomyces strains have been verified can be positive for the development of wine anthocyanin and aroma in different fermentation scenarios. Moreover, oenological tannins are widely used in wine industry to improve the colour profile and aroma complexity. The aim of this work is to analyze the fermentation characters of non-Saccharomyces strains and investigate the effects of pre-fermentative addition of oenological tannins on the wine components as well as sensory properties. For this purpose, five selected non-Saccharomyces strains and grape seed tannin were used to carry out the different fermentation trials. As a result, the grape seed tannin were less likely to influence growth kinetics of non-Saccharomyces strains. Schizosaccharomyces pombe has been proved can be effective to reduce the malic acid content while increase the level of vinylphenolic pyranoanthocyanin, which is positive for wine colour stability. Pre-fermentative use of oenological tannin was verified could be beneficial for the wines fermented with non-Saccharomyces regarding the improvement of wine colour, anthocyanin composition and the complexity of volatile compounds. Nevertheless, sensory analysis showed that oenological tannin could be less effective to modify the aroma impression of non-Saccharomyces wines., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Saccharomyces yeast species are currently the most important yeasts involved in industrial-scale food fermentations. However, there are hundreds of other yeast species poorly studied that are highly promising for flavour development, some of which have also been identified in traditional food fermentations. This work explores natural yeast biodiversity in terms of aroma formation, with a particular focus on aromas relevant for industrial fermentations such as wine and beer. Several non-Saccharomyces species produce important aroma compounds such as fusel alcohols derived from the Ehrlich pathway, acetate esters and ethyl esters in significantly higher quantities than the well-known Saccharomyces species. These species are Starmera caribaea, Hanseniaspora guilliermondii, Galactomyces geotrichum, Saccharomycopsis vini and Ambrosiozyma monospora. Certain species revealed a strain-dependent flavour profile while other species were very homogenous in their flavour profiles. Finally, characterization of a selected number of yeast species using valine or leucine as sole nitrogen sources indicates that the mechanisms of regulation of the expression of the Ehrlich pathway exist amongst non-conventional yeast species., (Copyright © 2016. Published by Elsevier Ltd.)