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1.
Food Res Int ; 161: 111891, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36192917

RESUMO

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.


Assuntos
Hanseniaspora , Vinho , Etanol/análise , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Nitrogênio/análise , Nutrientes/análise , Odorantes/análise , Saccharomyces cerevisiae , Saccharomycetales , Vinho/análise
2.
Food Microbiol ; 107: 104064, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35953174

RESUMO

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.


Assuntos
Hanseniaspora , Mel , Vinho , Fermentação , Mel/análise , Saccharomyces cerevisiae , Sicília , Vinho/análise
3.
Int J Food Microbiol ; 379: 109868, 2022 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-35961159

RESUMO

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.


Assuntos
Eriobotrya , Hanseniaspora , Vinho , Cerveja , Etanol/análise , Fermentação , Saccharomyces cerevisiae , Açúcares , Vinho/análise
4.
J Econ Entomol ; 115(4): 999-1007, 2022 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-35385117

RESUMO

Since the early phase of the intercontinental dispersal of Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), fermentation baits have been used for monitoring. Self-made lures and commercial products are often based on wine and vinegar. From an ecological perspective, the formulation of these baits is expected to target especially vinegar flies associated with overripe fruit, such as Drosophila melanogaster (Meigen) (Diptera: Drosophilidae). Hanseniaspora uvarum (Niehaus) (Ascomycota: Saccharomyceta) is a yeast closely associated with D. suzukii and fruit, and furthermore attractive to the flies. Based on this relation, H. uvarum might represent a suitable substrate for the development of lures that are more specific than vinegar and wine. In the field, we therefore, compared H. uvarum to a commercial bait that was based on vinegar and wine with respect to the number of trapped D. suzukii relative to other drosophilids and arthropods. Trap captures were higher with the commercial bait but specificity for D. suzukii was greater with H. uvarum. Moreover, H. uvarum headspace extracts, as well as a synthetic blend of H. uvarum volatiles, were assayed for attraction of D suzukii in a wind tunnel and in the field. Headspace extracts and the synthetic blend induced strong upwind flight in the wind tunnel and confirmed attraction to H. uvarum volatiles. Furthermore, baited with H. uvarum headspace extract and a drowning solution of aqueous acetic acid and ethanol, 74% of field captured arthropods were D. suzukii. Our findings suggest that synthetic yeast headspace formulations might advance the development of more selective monitoring traps with reduced by-catch.


Assuntos
Drosophila , Hanseniaspora , Controle de Insetos , Ácido Acético/farmacologia , Animais , Drosophila melanogaster , Frutas , Controle de Insetos/métodos , Leveduras
5.
J Food Sci ; 87(3): 886-894, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35142373

RESUMO

ß-Glucosidase is a key enzyme that hydrolyzes nonvolatile glycosylated precursors of aroma compounds and enhances the organoleptic quality of wines. In this study, a novel ß-glucosidase from Hanseniaspora uvarum Yun268 was localized, purified, and characterized. Results indicated that ß-glucosidase activity was mainly distributed within the cells. After purification via ammonium sulfate precipitation combined with chromatography, ß-glucosidase specific activity increased 8.36 times, and the activity recovery was 56.90%. The enzyme had a molecular mass of 74.22 kDa. It has a Michaelis constant (Km ) of 0.65 mmol/L, and a maximum velocity (Vmax ) of 5.1 nmol/min under optimum conditions; and Km of 0.94 mmol/L, and Vmax of 2.8 nmol/min under typical winemaking conditions. It exhibited the highest activity at 50°C and pH 5.0 and was stable at a temperature range of 20-80°C and pH range of 3.0-8.0. The enzyme has good tolerance to Fe3+ , especially maintaining 93.68% of its activity with 10 mmol/L of Fe3+ . Ethanol (<20%) and glucose (<150 g/L) inhibited its activity only slightly. Therefore, ß-glucosidase from H. uvarum Yun268 has excellent biochemical properties and a good application potential in winemaking. PRACTICAL APPLICATION: Winemaking is a biotechnological process in which exogenous ß-glucosidase is used to overcome the deficiency of endogenous ß-glucosidase activity in grapes. By localizing, purifying, and characterizing of ß-glucosidase from Hanseniaspora uvarum Yun268, it is expected to reveal its physical and chemical characteristics to evaluate its oenological properties in winemaking. The results may provide the basis for promoting the release of varietal aroma and improving wine sensory quality in the wine industry.


Assuntos
Hanseniaspora , Vinho , Fermentação , Odorantes/análise , Vinho/análise , beta-Glucosidase/metabolismo
6.
Food Microbiol ; 101: 103896, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34579856

RESUMO

Cocoa fermentation is the key and most relevant process in the synthesis of aroma and flavor precursor molecules in dry beans or raw material for producing chocolate. Because this process occurs in an uncontrolled manner, the chemical and sensory quality of beans can vary and be negatively affected. One of the strategies for the standardization and improvement of the sensory quality of chocolate is the introduction of microbial starter cultures. Among these, yeasts involved in fermentation have been studied because of their pectinolytic and metabolic potential in the production of volatile compounds. This study was aimed at isolating and characterizing, both sensory and chemically, yeasts involved in cocoa fermentation that could be used as starter cultures from two agro-ecological regions for the cultivation of cocoa in Colombia. The microbiological analyses identified 22 species represented mostly by Saccharomyces cerevisiae, Wickerhamomyces anomalus and Pichia sp. The preliminary sensory analysis of eight of these species showed that Hanseniaspora thailandica and Pichia kluyveri presented sensory profiles characterized by high intensity levels of fruity notes, which could be ascribed to the production of ethyl acetate, isoamyl acetate, and 2-phenylethyl acetate.


Assuntos
Bioprospecção , Cacau , Chocolate , Fermentação , Leveduras , Chocolate/microbiologia , Hanseniaspora , Pichia , Saccharomyces cerevisiae , Saccharomycetales
7.
Food Chem ; 368: 130807, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34411859

RESUMO

The combined use of selected Saccharomyces cerevisiae and non-Saccharomyces strains is becoming an effective way to achieve wine products with distinctive aromas. The purpose of this study was to further improve the wine aroma complexity through optimizing inoculation protocols of multi-starters. The three indigenous non-Saccharomyces strains (Torulaspora delbrueckii, Hanseniaspora vineae, and Lachancea thermotolerans) and their pairwise combinations (co-inoculation) were sequentially inoculated with S. cerevisiae in Petit Manseng grape must, respectively. Results evidenced a higher divergence in aroma compounds produced by two different non-Saccharomyces species compared to single species. Especially for the combination of T. delbrueckii and L. thermotolerans, the concentrations of most ethyl esters were further increased, contributing to a higher score of 'pineapple' note in agreement with sensory analysis. Our results highlighted that the inoculation of more than one non-Saccharomyces species is a potential strategy to improve the aroma diversity and quality of industrial wines.


Assuntos
Compostos Orgânicos Voláteis , Vinho , Fermentação , Hanseniaspora , Saccharomyces cerevisiae , Saccharomycetales , Compostos Orgânicos Voláteis/análise , Vinho/análise
8.
Pest Manag Sci ; 78(3): 1287-1295, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34854220

RESUMO

BACKGROUND: The invasive insect Drosophila suzukii (Matsumura) is an important pest of several red grape varieties. The yeast Hanseniaspora uvarum (Niehaus), which is associated with D. suzukii, strongly attracts flies and stimulates them to feed on yeast-laden food. In the present study, a formulation based on H. uvarum culture with spinosad insecticide was applied to the foliage of vineyards and control of D. suzukii was compared to applying spinosad to the whole plant. After successful H. uvarum and insecticide application in the vineyard, we tested additional H. uvarum-based formulations with spinosad in a greenhouse to determine their capacity to control D. suzukii. RESULTS: Application of the H. uvarum-spinosad formulation at 36.4 g of spinosad per hectare reduced the D. suzukii field infestation at the same rate as applying 120 g of spinosad per hectare and prevented spinosad residues on grapes. Leaves treated with H. uvarum and spinosad in the field and transferred to a laboratory assay caused high mortality to flies and reduced the number of eggs laid on fruits. Formulations with spinosad applied in the greenhouse showed that both H. uvarum culture and the yeast cell-free supernatant of a centrifuged culture increased fly mortality and reduced the number of eggs laid compared to the unsprayed control. CONCLUSION: In comparison to typical spinosad spray applications, the use of H. uvarum in combination with spinosad as an attract-and-kill formulation against D. suzukii reduces pesticide residues on the fruits by targeting the treatment to the canopy and decreasing the amount of insecticide per hectare without compromising control efficacy.


Assuntos
Inseticidas , Vitis , Animais , Drosophila , Combinação de Medicamentos , Frutas , Hanseniaspora , Controle de Insetos , Inseticidas/farmacologia , Macrolídeos
9.
FEMS Microbiol Lett ; 368(20)2021 11 27.
Artigo em Inglês | MEDLINE | ID: mdl-34755861

RESUMO

Non-Saccharomyces yeasts are important players during winemaking and may come from grapes grown in vineyards. To study the diversity of non-Saccharomyces yeasts on grape berry surfaces, 433 strains were isolated from different Cabernet Sauvignon vineyards grown in Henan Province. Our results demonstrated that these strains were classified into 16 morphotypes according to their growth morphology on Wallerstein Laboratory agar medium, and were identified as seven species from four genera-Hanseniaspora opuntiae, Hanseniaspora vineae, Hanseniaspora uvarum, Pichia occidentalis, Pichia kluyveri, Issatchenkia terricola and Saturnispora diversa-based on a series of molecular biological experiments. Hanseniaspora opuntiae was obtained from all sampling sites except Changyuan County, while Pichia kluyveri and Saturnispora diversa were only found in sites of Zhengzhou Grape Resource Garden and Minquan County, respectively. The site Minquan was home of the greatest species richness, while only one single species (Hanseniaspora opuntiae) was detected at NAPA winery from Zhengzhou or at Anyang County. Finally, this study suggested that the geographic distribution and diversity of non-Saccharomyces yeast populations on Cabernet Sauvignon grape berries were likely to be determined by a combination of grape varieties and environmental factors.


Assuntos
Biodiversidade , Frutas , Vitis , Leveduras , China , Fazendas , Fermentação , Frutas/microbiologia , Hanseniaspora/classificação , Hanseniaspora/isolamento & purificação , Pichia/classificação , Pichia/isolamento & purificação , Saccharomycetales/classificação , Saccharomycetales/isolamento & purificação , Vitis/microbiologia , Vinho/microbiologia , Leveduras/classificação , Leveduras/isolamento & purificação
10.
Food Res Int ; 147: 110549, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34399526

RESUMO

Kombucha is a very distinct naturally fermented sweetened tea that has been produced for thousands of years. Fermentation relies on metabolic activities of the complex autochthonous symbiotic microbiota embedded in a floating biofilm and used as a backslop for successive fermentations. Here, we designed a tailor-made microbial consortium representative of the core Kombucha microbiota to drive this fermentation. Microbial (counts, metagenetics), physico-chemical (pH, density) and biochemical (organic acids, volatile compounds) parameters were monitored as well as biofilm formation by confocal laser scanning microscopy and scanning electron microscopy. While nine species were co-inoculated, four (Dekkera bruxellensis, Hanseniaspora uvarum, Acetobacter okinawensis and Liquorilactobacillus nagelii) largely dominated. Microbial activities led to acetic, lactic, succinic and oxalic acids being produced right from the start of fermentation while gluconic and glucuronic acids progressively increased. A distinct shift in volatile profile was also observed with mainly aldehydes identified early on, then high abundances of fatty acids, ketones and esters at the end. Correlation analyses, combining metabolomic and microbial data also showed a shift in species abundances during fermentation. We also determined distinct bacteria-yeast co-occurence patterns in biofilms by microscopy. Our study provides clear evidence that a tailor-made consortium can be successfully used to drive Kombucha fermentations.


Assuntos
Consórcios Microbianos , Microbiota , Acetobacter , Biofilmes , Brettanomyces , Fermentação , Hanseniaspora
11.
mBio ; 12(5): e0234521, 2021 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-34607457

RESUMO

During fermentation, Saccharomyces cerevisiae metabolizes sugars and other nutrients to obtain energy for growth and survival, while also modulating these activities in response to cell-environment interactions. Here, differences in S. cerevisiae gene expression were explored over a time course of fermentation and used to differentiate fermentations, using Pinot noir grapes from 15 unique sites. Data analysis was complicated by the fact that the fermentations proceeded at different rates, making a direct comparison of time series gene expression data difficult with conventional differential expression tools. This led to the development of a novel approach combining diffusion mapping with continuous differential expression analysis (termed DMap-DE). Using this method, site-specific deviations in gene expression were identified, including changes in gene expression correlated with the non-Saccharomyces yeast Hanseniaspora uvarum, as well as initial nitrogen concentrations in grape musts. These results highlight novel relationships between site-specific variables and Saccharomyces cerevisiae gene expression that are linked to repeated fermentation outcomes. It was also demonstrated that DMap-DE can extract biologically relevant gene expression patterns from other contexts (e.g., hypoxic response of Saccharomyces cerevisiae) and offers advantages over other data dimensionality reduction approaches, indicating that DMap-DE offers a robust method for investigating asynchronous time series gene expression data. IMPORTANCE In this work, Saccharomyces cerevisiae gene expression was used as a biosensor to capture differences across and between fermentations of Pinot noir grapes from 15 unique sites representing eight American Viticultural Areas. This required development of a novel analysis method, DMap-DE, for investigation of asynchronous gene expression data. It was demonstrated that DMap-DE reveals biologically relevant shifts in gene expression related to cell-environment interactions in the context of hypoxia and fermentation. Using these data, it was discovered that gene expression by non-Saccharomyces yeasts and initial nitrogen content in grape musts are correlated with differences in gene expression among fermentations. These findings highlight important relationships between site-specific variables and gene expression that may be used to understand why foods and beverages, including wine, possess sensory characteristics associated with or derived from their place of origin.


Assuntos
Biologia Computacional/métodos , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Fermentação , Regulação Fúngica da Expressão Gênica , Hanseniaspora/genética , Hanseniaspora/crescimento & desenvolvimento , Hanseniaspora/metabolismo , RNA-Seq , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Vitis/microbiologia
12.
Food Microbiol ; 99: 103806, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34119099

RESUMO

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.


Assuntos
Cerveja/microbiologia , Hanseniaspora/metabolismo , Mel/microbiologia , Ácido Acético/análise , Ácido Acético/metabolismo , Cerveja/análise , Etanol/metabolismo , Fermentação , Microbiologia de Alimentos , Hanseniaspora/crescimento & desenvolvimento , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Resíduos/análise , Leveduras/crescimento & desenvolvimento , Leveduras/metabolismo
13.
Food Res Int ; 143: 110311, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33992330

RESUMO

The aim of the present study was to assess the cultivable microbiota of "mothers" of Vino cotto collected from production of different years 1890, 1895, 1920, 1975, 2008. A total of 73 yeasts and 81 bacteria were isolated. Starmerella lactis-condensi, Starmerella bacillaris, Hanseniaspora uvarum, Saccharomyces cerevisiae, Hanseniaspora guillermondi and Metschnikowia pulcherrima were identified. Bacteria isolates belonged to lactic acid bacteria (Lactiplantibacillus plantarum and Pediococcus pentosaceus) and acetic acid bacteria (Gluconobacter oxydans). Remarkable biodiversity was observed for Starm. bacillaris, as well as L. plantarum and G. oxydans. Organic acids and volatile compounds were also determined. Malic and succinic acids were the main ones with values ranging from 8.49 g/L to 11.76 g/L and from 4.15 g/L to 7.73 g/L respectively, while citric acid was present at low concentrations (<0.2 g/L) in all samples. Esters and higher alcohols were the main volatile compounds detected followed by alkanes. This study permits to better understand the microbial communities associated to this product and could be considered a starting point for the definition of tailored starter cultures to improve the quality of Vino cotto preserving its typical traits.


Assuntos
Vinho , Fermentação , Hanseniaspora , Metschnikowia , Saccharomycetales , Vinho/análise
14.
J Food Sci ; 86(6): 2327-2345, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33929752

RESUMO

ß-glucosidase is a pivotal enzyme that hydrolyzes bound volatile aromatic compounds. However, the activity of ß-glucosidase in winemaking and the mechanism by which it affects the flavor and taste of wines have not been fully investigated. In this study, we profiled the characteristics of ß-glucosidase derived from wine-related yeasts isolated from different wine-making regions in China, and analyzed the enzyme activity from different parts of the cells under aerobic and anaerobic conditions. A total of 56 strains of wine-related yeasts producing ß-glucosidases were screened using the YNB-C medium (YNB 6.7 g L-1 , cellobiose 5 g L-1 , pH 5.0). We found that strain Clavispora lusitaniae C117 produced the highest enzyme activity (152.39 µmol pNP ml-1 h-1 ). In most strains, ß-glucosidase were located in whole cells (periplasmic space) and permeabilized cells (intracellular). The non-Saccharomyces species had the highest enzymatic activity in a strain-dependent manner. Under aerobic conditions, C. lusitaniae C117, Hanseniaspora guilliermondii A27-3-4, Metschnikowia pulcherrima F-1-6, and Pichia anomala C84 had the highest ß-glucosidase activity. We further investigated the ß-glucosidase activity during the wine fermentation and the effects of sugar, pH, temperature, and ethanol on the enzyme activities of P. anomala C84 and commercial Saccharomyces yeast strains RC212 and VL1. The presence of fructose, glucose, and sucrose strongly inhibited enzyme activity. Similarly, low pH and low temperature inhibited the activity of ß-glucosidase, whereas ethanol promoted enzyme activity. Our findings provide a theoretical basis on understanding the different yeast characteristics of ß-glucosidase and their potential application for further improving wine aroma complexity.


Assuntos
Hanseniaspora/enzimologia , Metschnikowia/enzimologia , Odorantes/análise , Saccharomycetales/enzimologia , Vinho/análise , beta-Glucosidase/metabolismo
15.
J Insect Sci ; 21(2)2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33693806

RESUMO

Background odors produced by plants in the environment can interfere with the response of insects to a point-releasing attractant, especially when their compositions overlap. In this study, a series of binary choice tests was conducted in a wind tunnel to investigate whether background odors emitted from cherry, blueberry, blackberry, or raspberry fruits would affect the level of Drosophila suzukii (Matsumura) attraction to its symbiotic yeast, Hanseniaspora uvarum (Niehaus) (Saccharomycetales: Saccharomycetaceae). Whether an increase in the intensity of background odors would affect the attractiveness of H. uvarum to D. suzukii was also investigated, either by increasing the number of cherry or raspberry fruit per cup or by increasing the number of fruit cups surrounding the cup baited with the yeast. In wind tunnel assays, background fruit odors interfering with D. suzukii attraction to the yeast varied among fruit types. Raspberry odor inhibited the attractiveness of H. uvarum to the fly the most, followed by blackberry odor, whereas cherry and blueberry odors had no significant impact on the attraction. An increase in the intensity of odors by adding more cherry or raspberry fruit per cup did not increase the impact of fruit odor on the attraction; however, adding more raspberry cups around H. uvarum linearly decreased its attractiveness, suggesting that background host fruit abundance and likely increase in host odor may influence D. suzukii attraction to yeast odor depending on host species.


Assuntos
Drosophila , Frutas/fisiologia , Hanseniaspora , Odorantes , Animais , Bioensaio/métodos , Mirtilos Azuis (Planta)/fisiologia , Drosophila/microbiologia , Drosophila/fisiologia , Prunus avium/fisiologia , Rubus/fisiologia , Saccharomycetales , Simbiose
16.
Int J Mol Sci ; 22(4)2021 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-33672220

RESUMO

Hanseniaspora vineae is a non-Saccharomyces yeast that has a powerful impact on the sensory profile of wines. Its effect on the aromatic profile of non-aromatic grape varieties, such as Albillo Mayor (Vitis vinifera, L), during vinification is a useful biotechnology to improve sensory complexity. Fermentation in steel barrels using Hanseniaspora vineae and sequential inoculation with Saccharomyces cerevisiae have been used to study the formation of terpenes and cell lysis in the production of Albillo white wines. The GC-MS analysis profile shows a significant effect of H. vineae fermentation on the contents of terpenes (≈×3), mainly in linalool (>×3), ß-citronellol (>×4), geraniol (>×2) and α-terpineol (≈×2). The contents of several polyoxygenated terpenes and some volatile phenols with a spicy aroma were increased during fermentation. In summary, Hanseniaspora vineae releases a large number of cell wall polysaccharides during fermentation that affect wine palatability and structure. Hanseniaspora vineae is a powerful bio-tool to enhance the fruitiness, floral notes and freshness in non-aromatic white varieties.


Assuntos
Hanseniaspora/fisiologia , Terpenos/análise , Vinho/microbiologia , Fermentação , Microbiologia de Alimentos , Cromatografia Gasosa-Espectrometria de Massas , Hanseniaspora/genética , Odorantes/análise , Polifenóis/análise , Polifenóis/metabolismo , Polissacarídeos/análise , Polissacarídeos/metabolismo , Saccharomyces cerevisiae , Terpenos/metabolismo , Compostos Orgânicos Voláteis/análise , Vinho/análise
17.
Int J Mol Sci ; 22(4)2021 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-33669299

RESUMO

Apiculate yeasts belonging to the genus Hanseniaspora are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of Hanseniaspora uvarum. This was employed for the disruption of the HuATF1 gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the HuTEF1 promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative HuATF1 gene were deleted in a diploid H. uvarum strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast.


Assuntos
Deleção de Genes , Engenharia Genética/métodos , Hanseniaspora/enzimologia , Hanseniaspora/genética , Microrganismos Geneticamente Modificados/genética , Proteínas/genética , Acetatos/metabolismo , Alelos , Fermentação/genética , Genes Fúngicos , Fases de Leitura Aberta , Fenótipo , Regiões Promotoras Genéticas , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Vitis/metabolismo , Vinho
18.
Molecules ; 26(3)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530641

RESUMO

Wine fermentation processes are driven by complex microbial systems, which comprise eukaryotic and prokaryotic microorganisms that participate in several biochemical interactions with the must and wine chemicals and modulate the organoleptic properties of wine. Among these, yeasts play a fundamental role, since they carry out the alcoholic fermentation (AF), converting sugars to ethanol and CO2 together with a wide range of volatile organic compounds. The contribution of Saccharomyces cerevisiae, the reference organism associated with AF, has been extensively studied. However, in the last decade, selected non-Saccharomyces strains received considerable commercial and oenological interest due to their specific pro-technological aptitudes and the positive influence on sensory quality. This review aims to highlight the inter-specific variability within the heterogeneous class of non-Saccharomyces in terms of synthesis and release of volatile organic compounds during controlled AF in wine. In particular, we reported findings on the presence of model non-Saccharomyces organisms, including Torulaspora delbrueckii, Hanseniaspora spp,Lachancea thermotolerans, Metschnikowia pulcherrima, Pichia spp. and Candida zemplinina, in combination with S. cerevisiae. The evidence is discussed from both basic and applicative scientific perspective. In particular, the oenological significance in different kind of wines has been underlined.


Assuntos
Odorantes/análise , Saccharomycetales/fisiologia , Vinho/microbiologia , Fermentação , Hanseniaspora/fisiologia , Metschnikowia/fisiologia , Pichia/fisiologia , Torulaspora/fisiologia , Compostos Orgânicos Voláteis/química , Vinho/análise
19.
BMC Genomics ; 22(1): 131, 2021 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-33622260

RESUMO

BACKGROUND: Saccharomycodes ludwigii belongs to the poorly characterized Saccharomycodeacea family and is known by its ability to spoil wines, a trait mostly attributable to its high tolerance to sulfur dioxide (SO2). To improve knowledge about Saccharomycodeacea our group determined whole-genome sequences of Hanseniaspora guilliermondii (UTAD222) and S. ludwigii (UTAD17), two members of this family. While in the case of H. guilliermondii the genomic information elucidated crucial aspects concerning the physiology of this species in the context of wine fermentation, the draft sequence obtained for S. ludwigii was distributed by more than 1000 contigs complicating extraction of biologically relevant information. In this work we describe the results obtained upon resequencing of S. ludwigii UTAD17 genome using PacBio as well as the insights gathered from the exploration of the annotation performed over the assembled genome. RESULTS: Resequencing of S. ludwigii UTAD17 genome with PacBio resulted in 20 contigs totaling 13 Mb of assembled DNA and corresponding to 95% of the DNA harbored by this strain. Annotation of the assembled UTAD17 genome predicts 4644 protein-encoding genes. Comparative analysis of the predicted S. ludwigii ORFeome with those encoded by other Saccharomycodeacea led to the identification of 213 proteins only found in this species. Among these were six enzymes required for catabolism of N-acetylglucosamine, four cell wall ß-mannosyltransferases, several flocculins and three acetoin reductases. Different from its sister Hanseniaspora species, neoglucogenesis, glyoxylate cycle and thiamine biosynthetic pathways are functional in S. ludwigii. Four efflux pumps similar to the Ssu1 sulfite exporter, as well as robust orthologues for 65% of the S. cerevisiae SO2-tolerance genes, were identified in S. ludwigii genome. CONCLUSIONS: This work provides the first genome-wide picture of a S. ludwigii strain representing a step forward for a better understanding of the physiology and genetics of this species and of the Saccharomycodeacea family. The release of this genomic sequence and of the information extracted from it can contribute to guide the design of better wine preservation strategies to counteract spoilage prompted by S. ludwigii. It will also accelerate the exploration of this species as a cell factory, specially in production of fermented beverages where the use of Non-Saccharomyces species (including spoilage species) is booming.


Assuntos
Hanseniaspora , Vinho , Fermentação , Saccharomyces cerevisiae , Saccharomycetales
20.
ACS Synth Biol ; 10(3): 495-504, 2021 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-33576609

RESUMO

Ethyl acetate can be synthesized from acetyl-CoA and ethanol via a reaction by alcohol acetyltransferases (AATase) in yeast. In order to increase the yield of acetyl-CoA, different terminators were used to optimize the expressions of acetyl-CoA synthetase (ACS1/2) and aldehyde dehydrogenase (ALD6) to increase the contents of acetyl-CoA in Saccharomyces cerevisiae. ATF1 coding AATase was coexpressed in expression cassettes of ACS1/ACS2 and ALD6 to promote the carbon flux toward ethyl acetate from acetyl-CoA. Further to improve ethyl acetate production, four heterologous AATase including HuvEAT1 (Hanseniaspora uvarum), KamEAT1 (Kluyveromyces marxianus), VAAT (wild strawberry), and AeAT9 (kiwifruit) were introduced. Subsequently mitochondrial transport and utilization of pyruvate and acetyl-CoA were impeded to increase the ethyl acetate accumulation in cytoplasm. Under the optimal fermentation conditions, the engineered strain of PGAeΔPOR2 produced 1.69 g/L ethyl acetate, which was the highest value reported to date by metabolic engineering methods.


Assuntos
Acetatos/metabolismo , Engenharia Metabólica , Saccharomyces cerevisiae/metabolismo , Acetatos/química , Acetilcoenzima A/biossíntese , Aciltransferases/genética , Aciltransferases/metabolismo , Coenzima A Ligases/genética , Coenzima A Ligases/metabolismo , Hanseniaspora/enzimologia , Kluyveromyces/enzimologia , Ácido Pirúvico/metabolismo
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