RESUMO
In this study we investigated the effect of the single strain in stabilization of type I sourdough microbial associations by crossing six different Fructilactobacillus sanfranciscensis with five Kazachstania humilis strains. Furthermore, we compared three predictive models, Zwitwering based on Gompertz's equation, Baranyi and Roberts' function and Schiraldi's function to evaluate which one best fitted the experimental data in determining the behaviour of co-cultivated microorganisms. Specific growth rates (µm) and lag time (λ) values for each mixed population were assessed. Results showed that the different F. sanfranciscensis strains significantly steer the growth kinetics within the pair and affect the ratio bacterial/yeast cells, as data analysis confirmed, whereas K. humilis accommodates to the bacterial strain. To compare the growth models, Root Mean Square (RMS) values were calculated for each predicted curve by implementing an algorithm based on an iterative process to minimize the deviation among observed and calculated data. Schiraldi's function performed better than the others, revealing, on average, the smallest RMS values and providing the best fitting for over 70% of co-cultivation experiments. Models prove to be consistent in predicting growth kinetics of microbial consortia too.
Assuntos
Bactérias/química , Bactérias/crescimento & desenvolvimento , Pão/microbiologia , Consórcios Microbianos , Bactérias/isolamento & purificação , Bactérias/metabolismo , Fermentação , Farinha/microbiologia , Microbiologia de Alimentos , Cinética , Triticum/microbiologiaRESUMO
Given the pharmacological properti es and the potential role of kynurenic acid (KYNA) in human physiology and the pleiotropic activity of the neurohormone melatonin (MEL) involved in physiological and immunological functions and as regulator of antioxidant enzymes, this study aimed at evaluating the capability of Saccharomyces cerevisiae EC1118 to release tryptophan derivatives (dTRPs) from the kynurenine (KYN) and melatonin pathways. The setting up of the spectroscopic and chromatographic conditions for the quantification of the dTRPs in LC-MS/MS system, the optimization of dTRPs' production in fermentative and whole-cell biotransformation approaches and the production of dTRPs in a soybean-based cultural medium naturally enriched in tryptophan, as a case of study, were included in the experimental plan. Variable amounts of dTRPs, with a prevalence of metabolites of the KYN pathway, were detected. The LC-MS/MS analysis showed that the compound synthesized at highest concentration is KYNA that reached 9.146 ± 0.585 mg/L in fermentation trials in a chemically defined medium at 400 mg/L TRP. Further experiments in a soybean-based medium confirm KYNA as the main dTRPs, whereas the other dTRPs reached very lower concentrations. While detectable quantities of melatonin were never observed, two MEL isomers were successfully measured in laboratory media.
Assuntos
Meios de Cultura/metabolismo , Glycine max/metabolismo , Saccharomyces cerevisiae/metabolismo , Triptofano/metabolismo , Cromatografia Líquida/métodos , Fermentação/fisiologia , Humanos , Ácido Cinurênico/metabolismo , Cinurenina/metabolismo , Melatonina/metabolismo , Neurotransmissores/metabolismo , Transdução de Sinais/fisiologia , Espectrometria de Massas em Tandem/métodosRESUMO
Malolactic fermentation (MLF) in Valtellina Superiore DOCG red wine was monitored in 4 cellars and the final products were analysed to determine the content of melatonin (MEL) and other tryptophan (TRP) derivatives, including tryptophan ethyl ester (TEE) and MEL isomers (MISs), and to isolate predominant O. oeni strains. MEL and TEE significantly increased in wines after MLF from two cellars out of four. Six strains were isolated during the MLF of red wines and under laboratory scale, in rich and synthetic wine cultural media, together with other four O. oeni strains able to trigger the MLF. Results showed that the presence of stressful growth factors, like ethanol and acid pH, has a pivotal role in triggering the release of TEE by oenococci. Indeed, all the strains became capable to produce also MEL and MISs, together with TEE. under harsh growth conditions, as in a synthetic wine medium. The production of these compounds was strain-dependent and a maximum amount of 0.0078⯱â¯0.0023 ngT/mL (UMB472) and 619.85⯱â¯196.16 ngT/mL (UMB436) of MEL and TEE was obtained, respectively. In particular, different MISs were detected under oenological and laboratory scale suggesting that other factors (i.e. technological and/or physico-chemical) could affect the synthesis of TRP derivatives.
Assuntos
Melatonina/biossíntese , Oenococcus/metabolismo , Triptofano/biossíntese , Etanol/metabolismo , Fermentação , Concentração de Íons de Hidrogênio , Microbiologia Industrial , Malatos/metabolismo , Oenococcus/química , Triptofano/análogos & derivados , Triptofano/análise , Triptofano/química , Triptofano/metabolismo , Vinho/análise , Vinho/microbiologiaRESUMO
Sulfur dioxide is generally used as an antimicrobial in wine to counteract the activity of spoilage yeasts, including Brettanomyces bruxellensis. However, this chemical does not exert the same effectiveness on different B. bruxellensis yeasts since some strains can proliferate in the final product leading to a negative sensory profile due to 4-ethylguaiacol and 4-ethylphenol. Thus, the capability of deciphering the general molecular mechanisms characterizing this yeast species' response in presence of SO2 stress could be considered strategic for a better management of SO2 in winemaking. A RNA-Seq approach was used to investigate the gene expression of two strains of B. bruxellensis, AWRI 1499 and CBS 2499 having different genetic backgrounds, when exposed to a SO2 pulse. Results revealed that sulphites affected yeast culturability and metabolism, but not volatile phenol production suggesting that a phenotypical heterogeneity could be involved for the SO2 cell adaptation. The transcriptomics variation in response to SO2 stress confirmed the strain-related response in B. bruxellensis and the GO analysis of common differentially expressed genes showed that the detoxification process carried out by SSU1 gene can be considered as the principal specific adaptive response to counteract the SO2 presence. However, nonspecific mechanisms can be exploited by cells to assist the SO2 tolerance; namely, the metabolisms related to sugar alcohol (polyols) and oxidative stress, and structural compounds.
Assuntos
Brettanomyces/genética , Brettanomyces/metabolismo , Fermentação , Estresse Fisiológico , Dióxido de Enxofre/metabolismo , Vinho/microbiologia , Microbiologia de Alimentos , Perfilação da Expressão Gênica , RNA-Seq , TranscriptomaRESUMO
Food plants contain hundreds of bioactive phytochemicals arising from different secondary metabolic pathways. Among these, the metabolic route of the amino acid Tryptophan yields a large number of plant natural products with chemically and pharmacologically diverse properties. We propose the identifier "indolome" to collect all metabolites in the Tryptophan pathway. In addition, Tryptophan-rich plant sources can be used as substrates for the fermentation by yeast strains to produce pharmacologically active metabolites, such as Melatonin. To pursue this technological development, we have developed a UHPLC-MS/MS method to monitor 14 Tryptophan, Tryptamine, amino-benzoic, and pyridine metabolites. In addition, different extraction procedures to improve the recovery of Tryptophan and its derivatives from the vegetal matrix were tested. We investigated soybeans, pumpkin seeds, sesame seeds, and spirulina because of their botanical diversity and documented healthy effects. Four different extractions with different solvents and temperatures were tested, and water extraction at room temperature was chosen as the most suitable procedure to extract the whole Tryptophan metabolites pattern (called by us "indolome") in terms of ease, high efficiency, short time, low cost, and sustainability. In all plant matrices, Tryptophan was the most abundant indole compound, while the pattern of its metabolites was different in the diverse plants extracts. Overall, 5-OH Tryptamine and Kynurenine were the most abundant compounds, despite being 100-1000-fold lower than Tryptophan. Melatonin was undetected in all extracts, but sesame showed the presence of a Melatonin isomer. The results of this study highlight the variability in the occurrence of indole compounds among diverse food plants. The knowledge of Tryptophan metabolism in plants represents a relevant issue for human health and nutrition.
Assuntos
Cucurbita/química , Análise de Alimentos , Glycine max/química , Espectrometria de Massas , Sementes/química , Sesamum/química , Triptofano/análise , Cromatografia LíquidaRESUMO
Vinylphenol reductase of Dekkera bruxellensis, the characteristic enzyme liable for "Brett" sensory modification of wine, has been recently recognized to belong to the short chain dehydrogenases/reductases family. Indeed, a preliminary biochemical characterisation has conferred to the purified protein a dual significance acting as superoxide dismutase and as a NADH-dependent reductase. The present study aimed for providing a certain identification of the enzyme by cloning the VPR gene in S. cerevisiae, a species not producing ethyl phenols. Transformed clones of S. cerevisiae resulted capable of expressing a biologically active form of the heterologous protein, proving its role in the conversion of 4-vinyl guaiacol to 4-ethyl guaiacol. A VPR specific protein activity of 9 ± 0.6 mU/mg was found in crude extracts of S. cerevisiae recombinant strain. This result was confirmed in activity trials carried out with the protein purified from transformant cells of S. cerevisiae by a his-tag purification approach; in particular, VPR-enriched fractions showed a specific activity of 1.83 ± 0.03 U/mg at pH 6.0. Furthermore, in agreement with literature, the purified protein behaves like a SOD, with a calculated specific activity of approximatively 3.41 U/mg. The comparative genetic analysis of the partial VPR gene sequences from 17 different D. bruxellesis strains suggested that the observed polymorphism (2.3%) and the allelic heterozygosity state of the gene do not justify the well described strain-dependent character in producing volatile phenols of this species. Actually, no correlation exists between genotype membership of the analysed strains and their capability to release off-flavours. This work adds valuable knowledge to the study of D. bruxellensis wine spoilage and prepare the ground for interesting future industrial applications.
Assuntos
Dekkera/genética , Oxirredutases/genética , Saccharomyces cerevisiae/genética , Clonagem Molecular , Dekkera/enzimologia , Fermentação , Microbiologia de Alimentos , Genótipo , Oxirredutases/química , Oxirredutases/metabolismo , Fenóis/metabolismo , Polimorfismo Genético , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/metabolismo , Vinho/análiseRESUMO
A three year survey on the dominant yeast populations in samples of air, must and wine in different vineyards and cellars of two northern Italian vine-growing territories (six sites in Franciacorta and eight sites in Oltrepò Pavese areas) was carried out. A total of 505 isolates were ascribed to 31 different species by RFLP analysis of the ITS1-5.8SrRNA-ITS2 region and partial sequence analysis of the 26S rRNA gene. The most commonly found species were Saccharomyces cerevisiae (frequency, F'â=â58.7%; incidence, I'â=â53.5%), Hanseniaspora uvarum (F'â=â14.3%; I'â=â5.3%), Metschnikowia fructicola (F'â=â11.1%; I'â=â5.0%) and Torulaspora delbrueckii (F'â=â10.3%; I'â=â3.8%). Among 270 S. cerevisiae new isolates, 156 (57.8%) revealed a different genetic pattern through polymorphism analysis of the interdelta regions by capillary electrophoresis, while 47 isolates (17.4â%) were clones of starter cultures. By considering the Shannon-Wiener index and results of principal component analysis (PCA) analyses, the year of isolation (vintage) proved to be a factor that significantly affected the biodiversity of the yeast species, whereas the geographical site (terroir) was not. Seventy-five per cent of S. cerevisiae isolates gathered in a unique cluster at a similarity level of 82%, while the remaining 25% were separated into minor groups without any evident relationship between δ-PCR profile and territory, year or source of isolation. However, in six cases a similar strain appeared at the harvesting time both in Franciacorta and Oltrepò Pavese areas, whereas surprisingly no strain was reisolated in the same vineyard or cellar for consecutive years.
Assuntos
Biodiversidade , Vitis/microbiologia , Vinho/microbiologia , Leveduras/isolamento & purificação , Microbiologia do Ar , DNA Fúngico/genética , Fermentação , Microbiologia Industrial , Itália , Dados de Sequência Molecular , Filogenia , Vinho/análise , Leveduras/classificação , Leveduras/genéticaRESUMO
Dekkera bruxellensis and Saccharomyces cerevisiae are considered two phylogenetically distant relatives, but they share several industrial relevant traits such as the ability to produce ethanol under aerobic conditions (Crabtree effect), high tolerance towards ethanol and acids, and ability to grow without oxygen. Beside a huge adaptability, D. bruxellensis exhibits a broader spectrum in utilization of carbon and nitrogen sources in comparison to S. cerevisiae. With the aim to better characterize its carbon source metabolism and regulation, the usage of galactose and the role that glucose plays on sugar metabolism were investigated in D. bruxellensis CBS 2499. The results indicate that in this yeast galactose is a non-fermentable carbon source, in contrast to S. cerevisiae that can ferment it. In particular, its metabolism is affected by the nitrogen source. Interestingly, D. bruxellensis CBS 2499 exhibits the 'short-term Crabtree effect', and the expression of genes involved in galactose utilization and in respiratory metabolism is repressed by glucose, similarly to what occurs in S. cerevisiae.
Assuntos
Brettanomyces/genética , Brettanomyces/metabolismo , Galactose/metabolismo , Redes e Vias Metabólicas/genética , Ácido Acético/metabolismo , Carbono/metabolismo , Etanol/metabolismo , Regulação Fúngica da Expressão Gênica , Glucose/metabolismo , Nitrogênio/metabolismoRESUMO
Melatonin (MEL) has been found in some medicinal and food plants, including grapevine, a commodity of particular interest for the production of wine, a beverage of economic relevance. It has also been suggested that MEL in wine may, at least in part, contribute to the health-promoting properties attributed to this beverage and, possibly, to other traditional Mediterranean foodstuffs. After a preliminary screening of 9 yeast strains in laboratory medium, three selected strains (Saccharomyces cerevisiae EC1118, Torulaspora delbrueckii CBS1146(T) and Zygosaccharomyces bailii ATCC36947(T) ) were inoculated in experimental musts obtained from 2 white (Moscato and Chardonnay) and 2 red (Croatina and Merlot) grape varieties. The production of MEL, melatonin isomers (MIs) and tryptophan ethyl ester (TEE) was monitored during the alcoholic fermentation. The screening showed that the three investigated strains produced the highest concentrations of MEL and two MIs in optimal growth conditions. However, MEL and MIs were not produced in oenological conditions, but the three strains synthesized high concentrations of a new MI and TEE in musts.
Assuntos
Melatonina/metabolismo , Vitis/química , Vitis/metabolismo , Leveduras/metabolismo , Fermentação/fisiologiaRESUMO
Dekkera bruxellensis is a yeast known to affect the quality of wine and beer. This species, due to its high ethanol and acid tolerance, has been reported also to compete with Saccharomyces cerevisiae in distilleries producing fuel ethanol. In order to understand how this species responds when exposed to low temperatures, some mechanisms like synthesis and accumulation of intracellular metabolites, changes in lipid composition and activation of the HOG-MAPK pathway were investigated in the genome sequenced strain CBS 2499. We show that cold stress caused intracellular accumulation of glycogen, but did not induce accumulation of trehalose and glycerol. The cellular fatty acid composition changed after the temperature downshift, and a significant increase of palmitoleic acid was observed. RT-PCR analysis revealed that OLE1 encoding for Δ9-fatty acid desaturase was up-regulated, whereas TPS1 and INO1 didn't show changes in their expression. In D. bruxellensis Hog1p was activated by phosphorylation, as described in S. cerevisiae, highlighting a conserved role of the HOG-MAP kinase signaling pathway in cold stress response.
Assuntos
Metabolismo dos Carboidratos , Dekkera/metabolismo , Proteínas Fúngicas/metabolismo , Metabolismo dos Lipídeos , Temperatura Baixa , Dekkera/genética , Dekkera/crescimento & desenvolvimento , Etanol/metabolismo , Fermentação , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , FosforilaçãoRESUMO
Candida milleri, together with Candida humilis, is the most representative yeast species found in type I sourdough ecosystems. In this work, comparison of the ITS region and the D1/D2 domain of 26S rDNA gene partial sequences, karyotyping, mtDNA-RFLP analysis, Intron Splice Site dispersion (ISS-PCR) and (GTG)5 microsatellite analyses, assimilation test of different carbohydrates, and metabolome assessment by FT-IR analysis, were investigated in seventeen strains isolated from four different companies as well as in type strains CBS6897(T) and CBS5658(T). Most isolates were ascribed to C. milleri, even if a strong relatedness was confirmed with C. humilis as well, particularly for three strains. Genetic characterization showed a high degree of intraspecific polymorphism since 12 different genotypes were discriminated. The number of chromosomes varied from 9 to 13 and their size ranged from less than 0.3 to over 2 Mbp. Phenotypic traits let to recognize 9 different profiles of carbon sources assimilation. FT-IR spectra from yeast cells cultivated in different media and collected at different growth phases revealed a diversity of behaviour among strains in accordance with the results of PCR-based fingerprinting. A clear evidence of the polymorphic status of C. milleri species is provided thus representing an important feature for the development of technological applications in bakery industries.
Assuntos
Pão/microbiologia , Candida/genética , Candida/metabolismo , Polimorfismo Genético , Candida/classificação , Candida/isolamento & purificação , DNA Fúngico/genética , DNA Ribossômico/genética , Genótipo , Dados de Sequência Molecular , Técnicas de Tipagem Micológica , Fenótipo , Filogenia , Polimorfismo de Fragmento de RestriçãoRESUMO
There is a growing interest in various types of kiwifruits, such as the "yellow" kiwifruit (Actinidia chinensis var. chinensis) and the "baby" kiwifruit of Actinidia arguta. These fruits are rich in bioactive compounds, which contribute to their nutraceutical properties, but they nevertheless have a shorter shelf life, resulting in economic losses. This study aims to chemically characterize kiwifruit juices from SunGold and baby kiwifruit varieties (Hortgem Rua and Hortgem Tahi) to improve knowledge and explore the suitability of these products for producing low-alcohol beverages using non-Saccharomyces strains, with the purpose of reducing waste and generating value-added processing. Total soluble solids, acidity, hardness, dry matter, total phenolic content, and antioxidant capacity were used as indicators of fruit quality. Chemical characterization of fresh kiwifruit juices revealed distinct profiles among varieties, with SunGold juice exhibiting higher sugar content and acidity. Citric acid was the predominant organic acid, while, as expected, tartaric was not detected. Kiwifruit juice fermentations by T. delbrueckii UMY196 were always completed regardless of the type of juice used, producing beverages with ethanol content ranging from 6.46 to 8.85% (v/v). The analysis of volatile organic compounds highlighted the presence of several molecules, contributing to aroma profiles with relevant differences among the three kiwifruit-based drinks. In particular, the total concentration of esters in the beverages reached 8.2 mg/L, 9.2 mg/L, and 8.6 mg/L in the Tahi, Rua, and SunGold beverages, respectively. The qualitative and quantitative profiles of the kiwifruit drinks revealed a pronounced perception of acidity and fruity traits, with significant differences observed by Principal Component Analysis (PCA) in aroma descriptors among the proposed beverages. The possibility of using unsold, overripe, or waste fruits to prepare new products with an increase in the sustainability of this supply chain is set.
RESUMO
Brettanomyces bruxellensis displays a high degree of genotypic and phenotypic polymorphism and is the main yeast species involved in wine spoilage. The innate resistance of 108 B. bruxellensis strains to the antimicrobial agent SO2 used in winemaking was investigated. Nineteen strains (17.6%) were sensitive to SO2 , failing to grow at the lowest concentration tested (0.1 mg L(-1) molecular SO2). Twenty-nine strains (26.8%) grew at 0.1 mg L(-1), 42 strains (38.9%) grew at 0.2 mg L(-1) , and 16 strains (14.8%) were able to grow as high as 0.4 mg L(-1) mol. SO2. Two strains able to grow in the presence of 0.6 mg L(-1) mol. SO2 were further studied by GCMS-TOF analysis to define the metabolic response to SO2 treatment. Two hundred and fifty-three intracellular metabolites were detected. The main effect observed was a decrease in cytoplasmic levels of polyols and an increase in levels of some amino acids, alanine, glutamic acid, glycine, proline, 5-oxoproline, serine and valine, which were significantly accumulated in the presence of SO2. No alteration in the pentose phosphate pathway was observed, suggesting NADPH usage could be diverted to other pathways. Finally, a change in metabolites involved in the glycerophospholipid pathway (glycerol-3-phosphate and myo-inositol) was also found.
Assuntos
Antifúngicos/metabolismo , Brettanomyces/efeitos dos fármacos , Brettanomyces/metabolismo , Metaboloma , Dióxido de Enxofre/metabolismo , Antifúngicos/toxicidade , Brettanomyces/química , Farmacorresistência Fúngica , Cromatografia Gasosa-Espectrometria de Massas , Dióxido de Enxofre/toxicidadeRESUMO
Dekkera bruxellensis is mainly associated with lambic beer fermentation and wine production and may contribute in a positive or negative manner to the flavor development. This yeast is able to produce phenolic compounds, such as 4-ethylguaiacol and 4-ethylphenol which could spoil the wine, depending on their concentration. In this work we have investigated how this yeast responds when exposed to conditions causing osmotic stress, as high sorbitol or salt concentrations. We observed that osmotic stress determined the production and accumulation of intracellular glycerol, and the expression of NADH-dependent glycerol-3-phosphate dehydrogenase (GPD) activity was elevated. The involvement of the HOG MAPK pathway in response to this stress condition was also investigated. We show that in D. bruxellensis Hog1 protein is activated by phosphorylation under hyperosmotic conditions, highlighting the conserved role of HOG MAP kinase signaling pathway in the osmotic stress response. Gene Accession numbers in GenBank: DbHOG1: JX65361, DbSTL1: JX965362.
Assuntos
Dekkera/metabolismo , Vinho/microbiologia , Dekkera/enzimologia , Dekkera/genética , Dekkera/crescimento & desenvolvimento , Fermentação , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Glicerol/metabolismo , Glicerolfosfato Desidrogenase/genética , Glicerolfosfato Desidrogenase/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Dados de Sequência Molecular , Osmose , Sais/metabolismo , Sorbitol/metabolismo , Vinho/análiseRESUMO
Riboflavin (RF), or vitamin B2, is an essential compound for yeast growth and a precursor of the flavin coenzymes, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), involved in redox and non-redox processes. RF is a photosensitive compound involved in the light-struck taste (LST), a fault causing the formation of off-flavors that can develop when the wine is exposed to light in the presence of methionine (Met), as well. As both RF and Met can be associated with detrimental changes in wines, a better comprehension of its yeast-mediated production is relevant to predict the maintenance of the desired character of the wine. This study aims at assessing the production of flavin derivatives (FDs) and Met by S. cerevisiae oenological starters under laboratory conditions. The results showed the presence of extra- and intracellular FDs, and Met is a strain-dependent characteristic being also affected by the initial content of RF in the medium. This finding was confirmed when the winemaking was carried out in a relevant environment. Our results evidenced the important impact of the yeast strain on the content of RF and its derivatives.
RESUMO
The current environmental challenge is pushing food systems towards more sustainable models of production that require reorganizing of processes by re-using side products still containing nutrients. This work aimed at valorising a mix of bovine sweet whey and sunflower press cake, through targeted fermentation. After preliminary screening based on growth rate, final pH, lactose/galactose assimilation, phytase activity, six Lactic Acid Bacteria strains (Lacticaseibacillus casei, L. paracasei (2), Lactococcus lactis, Lentilactobacillus parakefiri and Leuconostoc pseudomesenteroides) and three yeasts (Kluyveromyces lactis, K. marxianus and Torulaspora delbrueckii) were co-cultivated in pairs in microcosms (1-part ground press cake: 4-parts whey). All tested microorganisms were able to grow and acidify the blend: the LAB counts increased during the incubation (26 °C for 48 h) of +2.80 log CFU/g, whereas yeasts counts were of +1.98 log CFU/g, with significant differences among the different associations (p < 0.01). Mould counts were always <3 log CFU/g. Interestingly, the bacterial contaminants count significantly varied in samples with different pairs of strains (p < 0.001). Acidification level, acetic acid and ethanol contents were the limiting factors affecting the growth of spoilage micro-organisms. Best performances were attained in microcosms inoculated with L. lactis or L. paracasei and K. lactis or K. marxianus.
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Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi is recognized as the dioecious parental generation of today's cultivars. Climatic change and the arrival of pathogens and pests in Europe led it to be included on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species in 1997. The present work focused on the study of culturable yeast occurrence and diversity of grape berries collected from wild vines. Sampling was performed in 29 locations of Azerbaijan, Georgia, Italy, Romania, and Spain. In total, 3431 yeast colonies were isolated and identified as belonging to 49 species, including Saccharomyces cerevisiae, by 26S rDNA D1/D2 domains and ITS region sequencing. Isolates of S. cerevisiae were also analyzed by SSR-PCR obtaining 185 different genotypes. Classical ecology indices were used to obtain the richness (S), the biodiversity (H'), and the dominance (D) of the species studied. This study highlights the biodiversity potential of natural environments that still represent a fascinating source of solutions to common problems in winemaking.
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Microbial diversity in vineyards and in grapes has generated significant scientific interest. From a biotechnological perspective, vineyard and grape biodiversity has been shown to impact soil, vine, and grape health and to determine the fermentation microbiome and the final character of wine. Thus, an understanding of the drivers that are responsible for the differences in vineyard and grape microbiota is required. The impact of soil and climate, as well as of viticultural practices in geographically delimited areas, have been reported. However, the limited scale makes the identification of generally applicable drivers of microbial biodiversity and of specific microbial fingerprints challenging. The comparison and meta-analysis of different datasets is furthermore complicated by differences in sampling and in methodology. Here we present data from a wide-ranging coordinated approach, using standardized sampling and data generation and analysis, involving four countries with different climates and viticultural traditions. The data confirm the existence of a grape core microbial consortium, but also provide evidence for country-specific microbiota and suggest the existence of a cultivar-specific microbial fingerprint for Cabernet Sauvignon grape. This study puts in evidence new insight of the grape microbial community in two continents and the importance of both location and cultivar for the definition of the grape microbiome.
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This paper reports on a common experiment performed by 17 Research Units of the Italian Group of Microbiology of Vine and Wine (GMVV), which belongs to the Scientific Society SIMTREA, with the aim to validate a protocol for the characterization of wine strains of Saccharomyces cerevisiae. For this purpose, two commercial S. cerevisiae strains (EC 1118 and AWRI796) were used to carry out inter-laboratory-scale comparative fermentations using both synthetic medium and grape musts and applying the same protocol to obtain reproducible, replicable, and statistically valid results. Ethanol yield, production of acetic acid, glycerol, higher alcohols, and other volatile compounds were assessed. Moreover, the Fourier transform infrared spectroscopy was also applied to define the metabolomic fingerprint of yeast cells from each experimental trial. Data were standardized as unit of compounds or yield per gram of sugar (glucose and fructose) consumed throughout fermentation, and analyzed through parametric and non-parametric tests, and multivariate approaches (cluster analysis, two-way joining, and principal component analysis). The results of experiments carried out by using synthetic must showed that it was possible to gain comparable results from three different laboratories by using the same strains. Then, the use of the standardized protocol on different grape musts allowed pointing out the goodness and the reproducibility of the method; it showed the main traits of the two yeast strains and allowed reducing variability amongst independent batches (biological replicates) to acceptable levels. In conclusion, the findings of this collaborative study contributed to the validation of a protocol in a specific synthetic medium and in grape must and showed how data should be treated to gain reproducible and robust results, which could allow direct comparison of the experimental data obtained during the characterization of wine yeasts carried out by different research laboratories.
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The consumption of traditional wine has decreased in Europe during the last fifteen years. In parallel, new wine alternatives obtained by blending wines and fruit juices or by flavoring wines with artificial or natural flavors have appeared on the market. Recently, an innovative fruit wine obtained by co-fermentation of grape must and kiwi juice has been proposed and its potential of attraction for consumers should be exploited. To assess the potential consumer acceptability and expectations towards this new product, an online choice experiment has been conducted involving a consumer group of young adults (18-35 years old; n = 373). After the data collection, participants were divided into two groups according to whether they had already tasted a fruit wine (neophiles) or had never tasted it (new entries). For each group, the individual's responses (on wine consumption habits, expectations and willingness to consume and pay a fruit wine) were analyzed through Principal Component Analysis. Different consumption styles and expectation patterns were defined in the two groups. However, in general, neophiles showed consumption patterns based on the evaluation of fruit quality, sales format, alcoholic content and the presence or not of bubbles, not giving importance to the brand. In contrast, new entries' responses identified consumption patterns driven by the willingness to pay for a new product, the product value for money and packaging features. Differences between the two groups in expectations about the product sensory characteristics also emerged. These findings should contribute to this area of study by integrating environmental, economic and social dimensions and addressing food innovation and sustainability in the fruit and wine chains.