Impact of phenylalanine on Hanseniaspora vineae aroma metabolism during wine fermentation.

Hanseniaspora vineae exhibits extraordinary positive oenological characteristics contributing to the aroma and texture of wines, especially by its ability to produce great concentrations of benzenoid and phenylpropanoid compounds compared with conventional Saccharomyces yeasts. Consequently, in practice, sequential inoculation of H. vineae and Saccharomyces cerevisiae allows to improve the aromatic quality of wines. In this work, we evaluated the impact on wine aroma produced by increasing the concentration of phenylalanine, the main amino acid precursor of phenylpropanoids and benzenoids. Fermentations were carried out using a Chardonnay grape juice containing 150 mg N/L yeast assimilable nitrogen. Fermentations were performed adding 60 mg/L of phenylalanine without any supplementary addition to the juice. Musts were inoculated sequentially using three different H. vineae strains isolated from Uruguayan vineyards and, after 96 h, S. cerevisiae was inoculated to complete the process. At the end of the fermentation, wine aromas were analysed by both gas chromatography-mass spectrometry and sensory evaluation through a panel of experts. Aromas derived from aromatic amino acids were differentially produced depending on the treatments. Sensory analysis revealed more floral character and greater aromatic complexity when compared with control fermentations without phenylalanine added. Moreover, fermentations performed in synthetic must with pure H. vineae revealed that even tyrosine can be used in absence of phenylalanine, and phenylalanine is not used by this yeast for the synthesis of tyrosine derivatives.

Isolation of local strains of the yeast Metschnikowia for biocontrol and lipid production purposes.

The bioprospection of indigenous microorganism strains with biotechnological potential represents a prominent trend. Metschnikowia yeasts exhibit diverse capabilities, such as ethanol reduction in winemaking, biocontrol potential, and lipid production. In this work, local Metschnikowia strains were isolated from different fruits by their ability to produce pulcherrimic acid, a molecule that has been linked to biocontrol activity and that binds iron giving colored colonies. Five strains were selected, each from one of five distinct sources. All of them were identified as M. pulcherrima. All five were able inhibit other yeasts and one M. pulcherrima, called M7, inhibited the growth of Aspergillus nidulans. The selected strains accumulated lipid bodies in stationary phase. Certain non-conventional yeasts like Hanseniaspora vineae are very sensitive to biomass drying, but cell extracts from M. pulcherrima added to the growth media as a source of antioxidant lipids increased their tolerance to drying. All strains isolated showed good stress tolerance (particularly to heat) and have nutrient requirements similar to a commercial M. pulcherrima strain. In addition, the M7 strain had a good growth in sugarcane and beet molasses and behaved like Saccharomyces cerevisiae in a growth medium derived from agricultural waste, a persimmon hydrolysate. Therefore, the isolation of local strains of Metschnikowia able to grow in a variety of substrates is a good source of biocontrol agents.

Spontaneous fermentation of Marastina wines: The correlation between autochthonous mycobiota and phenolic compounds.

Understanding fungal community dynamics during fermentation is important for assessing their influence on wine's phenolic content. The present study represents the first effort to explore the correlation between the autochthonous mycobiota of Marastina grapes collected from Dalmatian winegrowing sub-regions in Croatia and the phenolic composition, as well as the physicochemical parameters of wines produced through spontaneous fermentation. The metataxonomic approach revealed Metschnikowia pulcherrima, Metschnikowia fructicola and Hanseniaspora uvarum as the core mycobiota detected at the initial phase of fermentation. By contrast, Saccharomyces cerevisiae took over the dominance starting from the middle stage of fermentation. The wine's phenolic compounds were revealed by high-performance liquid chromatography, with tyrosol being the most abundant. Rhodotorula babjevae and Botrytis cinerea showed a positive correlation with p-hydroxybenzoic acid, gentisic acid, caffeic acid and cinnamic acid, while demonstrating a negative correlation with protocatechuic acid and chlorogenic acid. Heterophoma novae-verbascicola exhibited the opposite behaviour regarding the same phenolic compounds. The concentration of lactic acid was positively correlated with B. cinerea and negatively correlated with Het. novae-verbascicola. These findings serve as a foundation for in-depth investigations into the role of autochthonous grape mycobiota in phenolic transformation during spontaneous fermentation, potentially leading to the production of high-quality wines with unique terroir characteristics. Future studies should aim to explore the specific role played by individual yeast isolates in the formation of phenolic compounds.

Exploring future applications of the apiculate yeast Hanseniaspora.

As a metaphor, lemons get a bad rap; however the proverb 'if life gives you lemons, make lemonade' is often used in a motivational context. The same could be said of Hanseniaspora in winemaking. Despite its predominance in vineyards and grape must, this lemon-shaped yeast is underappreciated in terms of its contribution to the overall sensory profile of fine wine. Species belonging to this apiculate yeast are known for being common isolates not just on grape berries, but on many other fruits. They play a critical role in the early stages of a fermentation and can influence the quality of the final product. Their deliberate addition within mixed-culture fermentations shows promise in adding to the complexity of a wine and thus provide sensorial benefits. Hanseniaspora species are also key participants in the fermentations of a variety of other foodstuffs ranging from chocolate to apple cider. Outside of their role in fermentation, Hanseniaspora species have attractive biotechnological possibilities as revealed through studies on biocontrol potential, use as a whole-cell biocatalyst and important interactions with Drosophila flies. The growing amount of 'omics data on Hanseniaspora is revealing interesting features of the genus that sets it apart from the other Ascomycetes. This review collates the fields of research conducted on this apiculate yeast genus.

Genome-scale metabolic models reveal determinants of phenotypic differences in non-Saccharomyces yeasts.

BACKGROUND: Use of alternative non-Saccharomyces yeasts in wine and beer brewing has gained more attention the recent years. This is both due to the desire to obtain a wider variety of flavours in the product and to reduce the final alcohol content. Given the metabolic differences between the yeast species, we wanted to account for some of the differences by using in silico models. RESULTS: We created and studied genome-scale metabolic models of five different non-Saccharomyces species using an automated processes. These were: Metschnikowia pulcherrima, Lachancea thermotolerans, Hanseniaspora osmophila, Torulaspora delbrueckii and Kluyveromyces lactis. Using the models, we predicted that M. pulcherrima, when compared to the other species, conducts more respiration and thus produces less fermentation products, a finding which agrees with experimental data. Complex I of the electron transport chain was to be present in M. pulcherrima, but absent in the others. The predicted importance of Complex I was diminished when we incorporated constraints on the amount of enzymatic protein, as this shifts the metabolism towards fermentation. CONCLUSIONS: Our results suggest that Complex I in the electron transport chain is a key differentiator between Metschnikowia pulcherrima and the other yeasts considered. Yet, more annotations and experimental data have the potential to improve model quality in order to increase fidelity and confidence in these results. Further experiments should be conducted to confirm the in vivo effect of Complex I in M. pulcherrima and its respiratory metabolism.

Comparing the hierarchy of inter- and intra-species interactions with population dynamics of wine yeast cocultures.

In winemaking, the development of new fermentation strategies, such as the use of mixed starter cultures with Saccharomyces cerevisiae (Sc) yeast and non-Saccharomyces (NS) species, requires a better understanding of how yeasts interact, especially at the beginning of fermentation. Despite the growing knowledge on interactions between Sc and NS, few data are available on the interactions between different species of NS. It is furthermore still unclear whether interactions are primarily driven by generic differences between yeast species or whether individual strains are the evolutionarily relevant unit for biotic interactions. This study aimed at acquiring knowledge of the relevance of species and strain in the population dynamics of cocultures between five yeast species: Hanseniaspora uvarum, Lachancea thermotolerans, Starmerella bacillaris, Torulaspora delbrueckii and Sc. We performed cocultures between 15 strains in synthetic grape must and monitored growth in microplates. Both positive and negative interactions were identified. Based on an interaction index, our results showed that the population dynamics seemed mainly driven by the two species involved. Strain level was more relevant in modulating the strength of the interactions. This study provides fundamental insights into the microbial dynamics in early fermentation and contribute to the understanding of more complex consortia encompassing multiple yeasts trains.

Exploring the effects of the fermentation method on the quality of Lycium barbarum and Polygonatum cyrtonema compound wine based on LC-MS metabolomics.

This study aimed to examine the effect of fermentation methods on the quality of Lycium barbarum and Polygonatum cyrtonema compound wine (LPW) by combining non-targeted metabolomic approaches with chemometrics and path profiling to determine the chemical and metabolic properties of LPW. The results demonstrated that SRA had higher leaching rates of total phenols and flavonoids, reaching 4.20 ± 0.10 v/v ethanol concentration. According to LC-MS non-targeting genomics, the metabolic profiles of LPW prepared by different mixtures of fermentation methods (Saccharomyces cerevisiae RW; Debaryomyces hansenii AS2.45) of yeast differed significantly. Amino acids, phenylpropanoids, flavonols, etc., were identified as the differential metabolites between different comparison groups. The pathways of tyrosine metabolism, biosynthesis of phenylpropanoids, and metabolism of 2-oxocarboxylic acids enriched 17 distinct metabolites. SRA stimulated the production of tyrosine and imparted a distinctive saucy aroma to the wine samples, providing a novel research concept for the microbial fermentation-based production of tyrosine.

Yeast diversity during the spontaneous fermentation of wine with only the microbiota on grapes cultivated in Japan.

Making wine via spontaneous fermentation without sulfur dioxide and commercial yeast (spontaneous winemaking) is increasing in recent year, but there is scant research regarding microbial communities present in Japan during spontaneous winemaking using culture-independent molecular methods. We analyzed fungal communities and populations during laboratory-scale spontaneous winemaking using sterilized labware to avoid winery-resident microbes. In the spontaneous fermentation of four grape varieties (Pinot Noir, Riesling, Koshu, and Koshusanjaku) grown in the same Japanese vineyard, our analysis of yeast and other fungal species by next-generation sequencing based on the ITS1 region demonstrated that Saccharomyces cerevisiae was eventually dominant in seven of 12 fermentation batches (three replications for each grape variety), whereas non-Saccharomyces species (e.g., Schizosaccharomyces japonicus, Lachancea dasiensis, and Hanseniaspora valbyensis) became dominant in four batches at the end of fermentation. In another batch, lactic acid bacteria (LAB) became dominant and the fermentation remained incomplete. Diverse microbes were involved in the spontaneous fermentation (particularly in Koshusanjaku), indicating that residual sugar remained and lactic and acetic acid largely increased. Compared to the control wine made with SO2 and commercial yeast, the concentration of lactic acid was 47-fold higher in the must dominated by L. dasiensis, and the concentrations of acetic acid and lactic acid were 10-fold and 20-fold higher in the must dominated by LAB, respectively. Even when indigenous S. cerevisiae became dominant, the finished wines obtained high sensory-analysis scores for complexity but low scores for varietal typicality, indicating the risk of fermentation with unselected wild yeast on the grapes grown in Japan.

Fermentation Characteristics and Aromatic Profiles of Plum Wines Produced with Hanseniaspora thailandica Zal1 and Common Wine Yeasts.

Plum has long been cultivated in northern Thailand and evolved into products having long shelf lives. In this study, plum processing was analyzed by comparing the production of plum wine using three types of yeast, Saccharomyces cerevisiae var. burgundy, Hanseniaspora thailandica Zal1, and S. cerevisiae Lalvin EC1118. EC1118 exhibited the highest alcohol content (9.31%), similar to that of burgundy (9.21%), and H. thailandica Zal1 had the lowest alcohol content (8.07%) after 14 days of fermentation. Plum wine fermented by S. cerevisiae var. burgundy had the highest total phenolic (TP) content and antioxidant activity of 469.84 ± 6.95 mg GAE/L and 304.36 ± 6.24 µg TE/g, respectively, similar to that fermented by EC1118 (418.27 ± 3.40 mg GAE/L 288.2 ± 7.9 µg TE/g). H. thailandica Zal1 exhibited the least amount of TP content and antioxidant activity; however, the volatility produced by H. thailandica Zal1 resulted in a plum wine with a distinct aroma.

Insights into the transcriptional regulation of poorly characterized alcohol acetyltransferase-encoding genes (HgAATs) shed light into the production of acetate esters in the wine yeast Hanseniaspora guilliermondii.

Hanseniaspora guilliermondii is a well-recognized producer of acetate esters associated with fruity and floral aromas. The molecular mechanisms underneath this production or the environmental factors modulating it remain unknown. Herein, we found that, unlike Saccharomyces cerevisiae, H. guilliermondii over-produces acetate esters and higher alcohols at low carbon-to-assimilable nitrogen (C:N) ratios, with the highest titers being obtained in the amino acid-enriched medium YPD. The evidences gathered support a model in which the strict preference of H. guilliermondii for amino acids as nitrogen sources results in a channeling of keto-acids obtained after transamination to higher alcohols and acetate esters. This higher production was accompanied by higher expression of the four HgAATs, genes, recently proposed to encode alcohol acetyl transferases. In silico analyses of these HgAat's reveal that they harbor conserved AATs motifs, albeit radical substitutions were identified that might result in different kinetic properties. Close homologues of HgAat2, HgAat3, and HgAat4 were only found in members of Hanseniaspora genus and phylogenetic reconstruction shows that these constitute a distinct family of Aat's. These results advance the exploration of H. guilliermondii as a bio-flavoring agent providing important insights to guide future strategies for strain engineering and media manipulation that can enhance production of aromatic volatiles.

Biology and physiology of Hanseniaspora vineae: metabolic diversity and increase flavour complexity for food fermentation.

Apiculate yeasts belonging to the genus Hanseniaspora are predominant on grapes and other fruits. While some species, such as Hanseniaspora uvarum, are well known for their abundant presence in fruits, they are generally characterized by their detrimental effect on fermentation quality because the excessive production of acetic acid. However, the species Hanseniaspora vineae is adapted to fermentation and currently is considered as an enhancer of positive flavour and sensory complexity in foods. Since 2002, we have been isolating strains from this species and conducting winemaking processes with them. In parallel, we also characterized this species from genes to metabolites. In 2013, we sequenced the genomes of two H. vineae strains, being these the first apiculate yeast genomes determined. In the last 10 years, it has become possible to understand its biology, discovering very peculiar features compared to the conventional Saccharomyces yeasts, such as a natural and unique G2 cell cycle arrest or the elucidation of the mandelate pathway for benzenoids synthesis. All these characteristics contribute to phenotypes with proved interest from the biotechnological point of view for winemaking and the production of other foods.

Dynamic analysis of microbial communities and flavor properties in Merlot wines produced from inoculation and spontaneous fermentation.

The microbiota is of great importance in forming flavor compounds and improving sensory characteristics during wine fermentation. Understanding microbial succession is critical for controlling its contribution to wine flavor with predictable sensory quality. In this study, microbial community composition and characteristic flavor compounds were identified during the inoculation fermentation (IF) and spontaneous fermentation (SF) to provide a basis for exploring the relationship between these microorganisms and volatile components. The results demonstrated that SF had higher fungal community diversity and lower bacterial community diversity than IF. Eleven (11) fungal and 10 bacterial genera (relative abundance > 0.1 %) were considered beneficial microbiota. Saccharomyces, Hanseniaspora, and Alternaria were the leading fungal genera in SF. Massilia, Nesterenkonia, and Halomonas were the predominant bacteria in IF, while Tatumella and Ochrobactrum were mainly from SF. In addition, the microbial community composition was reshaped via correlational analysis between microbiota succession and physicochemical properties, mainly attributed to the changes in environmental factors during fermentation. The SF wines had more aromatic higher alcohols, acetate esters, and terpenes. Also, the sensory evaluation showed that the SF wines were characterized by more fruity, floral, intense, and typical aromas. The associations between the microbial community and the volatile components indicated that the dominant species largely determined the characteristic flavor compounds during fermentation.

Wine yeast selection in the Iberian Peninsula: Saccharomyces and non-Saccharomyces as drivers of innovation in Spanish and Portuguese wine industries.

Yeast selection for the wine industry in Spain started in 1950 for the understanding of the microbial ecology, and for the selection of optimal strains to improve the performance of alcoholic fermentation and the overall wine quality. This process has been strongly developed over the last 30 years, firstly on Saccharomyces cerevisiae, and, lately, with intense activity on non-Saccharomyces. Several thousand yeast strains have been isolated, identified and tested to select those with better performance and/or specific technological properties. The present review proposes a global survey of this massive ex-situ preservation of eukaryotic microorganisms, a reservoir of biotechnological solutions for the wine sector, overviewing relevant screenings that led to the selection of strains from 12 genera and 22 species of oenological significance. In the first part, the attention goes to the selection programmes related to relevant wine-producing areas (i.e. Douro, Extremadura, Galicia, La Mancha and Uclés, Ribera del Duero, Rioja, Sherry area, and Valencia). In the second part, the focus shifted on specific non-Saccharomyces genera/species selected from different Spanish and Portuguese regions, exploited to enhance particular attributes of the wines. A fil rouge of the dissertation is the design of tailored biotechnological solutions for wines typical of given geographic areas.

Effect of fermentation methods on the quality and in vitro antioxidant properties of Lycium barbarum and Polygonatum cyrtonema compound wine.

Lycium barbarum and Polygonatum cyrtonema are known for their medicinal, edible, and ornamental properties. The sensory indices of the novel high-quality L. barbarum and P. cyrtonema compound wine (LPCW) fermented by Saccharomyces cerevisiae RW and Debaryomyces hansenii AS2.45 under different inoculation methods were analyzed. The alcohol content of the LPCW ranged from 3.88 to 4.75 % under three mixed inoculations. The total saponin and total polysaccharide contents in LPCW inoculated with D. hansenii first and S. cerevisiae after 24 h were 4.39 mg/mL and 0.21 mg/mL, respectively. Ethyl butyrate, citronellol, and 3-(methylthio) propanol were unique metabolites of D. hansenii. 4-Methoxybenzoic acid was the core product of brewing of by S. cerevisiae. Except for wine inoculated with S. cerevisiae only, the acceptability scores of all the LPCW samples were higher than 7.3. Our data provided the foundation for the development and application of medicinal and food homologous substances in food fermentation.

Effect of Hanseniaspora uvarum-Saccharomyces cerevisiae Mixed Fermentation on Aroma Characteristics of Rosa roxburghii Tratt, Blueberry, and Plum Wines.

Hanseniaspora uvarum, a non-Saccharomyces cerevisiae species, has a crucial effect on the aroma characteristics of fruit wines, thus, attracting significant research interest in recent years. In this study, H. uvarum-Saccharomyces cerevisiae mixed fermentation was used to ferment Rosa roxburghii Tratt, blueberry fruit wine, and plum fruit wines using either a co-inoculated or a sequentially inoculated approach. The three fruit wines' volatile aroma characteristics were analyzed by headspace-solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). The results showed that the mixed inoculation of H. uvarum and S. cerevisiae reduced the alcoholic content of Kongxinli fruit wine. Moreover, H. uvarum-S. cerevisiae fermented Rosa roxburghii Tratt, blueberry, and plum fruit wines and further enriched their flavor compounds. The overall flavor characteristics of sequentially inoculated fruit wines differed significantly from those fermented with S. cerevisiae alone, although several similarities were also observed. Sequential inoculation of H. uvarum and S. cerevisiae positively affected the mellowness of the wine and achieved a better harmony of the overall wine flavors. Therefore, H. uvarum-Saccharomyces cerevisiae mixed fermentation can improve the complexity of the wines' aromatic composition and empower them with a unique identity. In particular, H. uvarum-Saccharomyces cerevisiae blueberry wine produced by mixed fermentation had the widest variety and content of aroma compounds among the fermented wines. Therefore, H. uvarum-Saccharomyces cerevisiae mixed-fermentation inoculation in the three fermented fruit wines significantly increased the aroma compound variety and content, thus, enriching their aroma richness and complexity. This study is the first comparative evaluation of the aroma characteristics of different fruit wines fermented with a mixed inoculation of H. uvarum and S. cerevisiae and provides a preliminary guide for these fruit wines produced with non-Saccharomyces yeast.

Co-inoculations of Lachancea thermotolerans with different Hanseniaspora spp.: Acidification, aroma, biocompatibility, and effects of nutrients in wine.

The use of non-Saccharomyces yeast in the winemaking industry and even more their co-inoculations to maximize their growth and to express phenotypic characteristic is gaining more and more relevance. This study aimed to shed light on the biocompatibilities between Lachancea thermotolerans and Hanseniaspora spp., using different types of nutrients and considering the effect on Yeast Assimilable Nitrogen (YAN), at low temperature (16 °C) and medium SO2 (50 mg/L), in white must. L. thermotolerans has been used for its positive effect on pH reduction and Hanseniaspora spp. for improving the sensory profile. The behaviour of these yeasts was evaluated in co-inoculation, always finishing the fermentation with the sequential inoculation of S. cerevisiae. Significant results were obtained on the population count (CFU/mL) in CHROMagar™, with higher populations of Hanseniaspora spp. with respect to L. thermotolerans. Fermentations with L. thermotolerans/H. vineae, showed inhibition of acidification, generating up to 0.41 g/L of lactic acid. On the contrary, a synergistic effect when L. thermotolerans/H. opuntiae was used, achieved 2.44 g/L of lactic acid and a pH reduction of up to 0.16 and always more significant with Nutrient Vit BlancTM. At the same time ethanol concentration decreased by 3.4 % and volatile acidity never exceeded 0.5 g/L. Aromatic composition was analysed and it was found that all fermentations retained more aromatic esters and that on day 7 the amount of 2-phenylethyl acetate was at least 3 times higher in all fermentations compared to the control (Sc + Nutrient Vit BlancTM) which had 5.96 mg/L. Less yellow intensity (-17.3 %) typical of oxidation were observed in all fermentations in which Nutrient Vit BlancTM had been used and in the sensory analysis the co-inoculations with H. vineae generated better scores.

Use of sequentially inoculation of Saccharomyces cerevisiae and Hanseniaspora uvarum strains isolated from honey by-products to improve and stabilize the quality of mead produced in Sicily.

Mead is a beverage produced by alcoholic fermentation of honey-must. The starter yeasts that are commonly used for the alcoholic fermentation of honey-must are oenological Saccharomyces cerevisiae strains. The objective of the present work was, for the first time, to apply yeasts of honey by-products origin to evaluate the influences the taste-olfactory attributes of mead. For this purpose, three experimental productions were set up, which included: (i) single inoculation of S. cerevisiae; (ii) single inoculation of Hanseniaspora uvarum; (iii) sequential inoculation of H. uvarum/S. cerevisiae. Two control trials were performed, using a commercial strain of S. cerevisiae of oenological origin and a spontaneous fermentation. The results of the chemical parameters showed differences between the trials in terms of residual sugars, acetic acid, glycerol, ethanol and volatile organic compounds. Sensorial analysis also showed a high heterogeneity among trials. The attributes of sweetness, honey and floral were found in mead fermented with H. uvarum, whereas all meads obtained with S. cerevisiae were dry, balanced and without off-odors and off-flavours. The results obtained showed that the controlled application of conventional and non-conventional yeast strains isolated from honey by-products origin could be a promising approach to improve the quality of meads.

Influence of indigenous Hanseniaspora uvarum and Saccharomyces cerevisiae from sugar-rich substrates on the aromatic composition of loquat beer.

The demand for unique and exclusive food products and beverages is constantly on the increase. One of the products that mostly evolved to encounter market dynamics in the last decade is craft beer. For a long time, craft breweries have included fruit in beer production to enrich flavour and aroma profile of different beer styles. In this study, for the first time, the use of Saccharomyces and non-Saccharomyces yeast strains isolated from high-sugar matrices (manna and fermented honey by-products) were investigated to diversify fruit craft beer production, in order to improve the fermentation process and highlight the complexity of aroma profiles generated during alcoholic fermentation. Two yeast strains, Hanseniaspora uvarum YGA34 and Saccharomyces cerevisiae MN113, were tested as co-starters and starters for their beer production capacity. Commercial yeast strain US-05 was used as control. Loquat juice was added at the end of primary alcoholic fermentation in all trials. Interestingly, S. cerevisiae MN113 consumed sugars faster than control strain S. cerevisiae US-05, including maltose, even in the case of sequential inoculation. This strain showed an excellent ability to consume rapidly sugars present. All strains showed their concentrations ranged between 5 and 8 Log cycles during fermentation. The absence of off-odours and the improvement of aromatic perception were observed in experimental trials involving the use of S. cerevisiae MN113 as a monoculture and in sequential combination with H. uvarum YGA34. Esters and alcohols were the most abundant compounds emitted from the beers. The beers produced with sequential inoculation of H. uvarum YGA34 and S. cerevisiae MN113 or US-05 are characterised by a higher ester and lower alcohol concentration. These two unconventional yeast strains from high sugar matrices showed great technological properties, representing promising co-starters and starter during craft fruit beer production.

Impact of vineyard management on grape fungal community and Montepulciano d'Abruzzo wine quality.

In this study the effect of farming practices on the diversity of grape fungal community of Vitis vinifera L. cultivar Montepulciano and the microbial metabolic activity was investigated. Small-scale vinifications were performed and wines were characterized. Agronomic management system affected the fungal community composition. Hanseniaspora, Areobasidium and Botrytis genera represented 30%, 20% and 10% of the total reads in all samples. A. pullulans, and Cladosprium cladosporioides mainly occurred on organic and biodynamic grapes. Saccharomyces and Pseudopithomyces genera were present only on organic or biodynamic grapes, respectively. The agronomic managements also influenced the potential functionality of microbial community. In fact, the metabolic function was increased in organic and biodynamic grapes. Polymers were used only by organic and biodynamic microbial communities, which also showed the highest values of Shannon's diversity index, and substrate richness. However, no significative differences were observed for the oenological parameters analyzed, with the only exception of the content of sugars which were higher on conventional grapes. The fermentation profiles showed that conventional wines had a lower residual sugars content, and a higher amount of alcohol. The differences observed in the volatile composition of the wines were both quantitative and qualitative. Conventional wines showed a lower content of esters, and a higher concentration of alcohols than organic and biodynamic ones. Biodynamic wines were characterized by the highest content of organic acids. The obtained results revealed that farming practices shape the fungal community influencing wine traits linking the wine with the viti-vinicultural area of origin.

Growth kinetics and fermentation properties of autochthonous yeasts in pineapple juice fermentation for starter culture development.

Autochthonous yeasts associated with pineapple fermentation were isolated and their fermentation behaviours were investigated for development of specific culture in pineapple winemaking. Autochthonous yeast isolates, Saccharomycodes ludwigii and Hanseniaspora uvarum, were selected due to their generated products of alcohol and 2-phenylethyl acetate, respectively. The fermentation kinetic parameters of selected autochthonous yeasts as single and co-cultures in chaptalized pineapple juice were investigated comparing to commercial Saccharomyces cerevisiae. The ethanol production rate of S'codes ludwigii (0.104%(v/v)/h) during the initial stage of fermentation was relatively slower compared to those of S. cerevisiae (0.129%(v/v)/h) but increased during middle through final stages with similar ethanol content to the commercial S. cerevisiae (~12%(v/v)). In pineapple juice, fructose was firstly assimilated, S'codes ludwigii (K = 0.405) and S. cerevisiae (K = 0.552), while glucose was secondly used, S'codes ludwigii (K = 0.281) and S. cerevisiae (K = 0.217) for first-order kinetic model. In co-cultures, the two isolated strains displayed synergistic behaviours during fermentation. S'codes ludwigii supported the growth of H. uvarum so that it generated more desirable volatile organic compounds (VOCs) at an early stage. Interestingly, the VOCs could not be produced in co-cultures of H. uvarum with the commercial strains. Then, S'codes ludwigii further completed the alcoholic fermentation through final stage. The fermentation performances of co-cultured autochthonous yeasts demonstrated a new approach for successful pineapple winemaking over S. cerevisiae. In addition, growth kinetics and fermentation behaviour, as observed in this study, could be a key information in development of potential substrates and strains for future alcoholic fermentation.