Search Results (136)

This study aimed to provide novel information on the impact of indigenous non-Saccharomyces yeasts, including Metschnikowia chrysoperlae, Metschnikowia sinensis/shanxiensis, Metschnikowia pulcherrima, Lachancea thermotolerans, Hanseniaspora uvarum, Hanseniaspora guilliermondii, and Pichia kluyveri, on metabolites related to the metabolism of tryptophan, phenylalanine, and tyrosine. The experiment included two fermentation practices: monoculture and sequential fermentation with commercial Saccharomyces cerevisiae, using sterile Maraština grape juice. A targeted approach through ultrahigh-resolution liquid chromatography associated with mass spectrometry was used to quantify 38 metabolites. All the indigenous yeasts demonstrated better consumption of tryptophan in monoculture than in interaction with S. cerevisiae. M. sinensis/shanxiensis was the only producer of indole-3-carboxylic acid, while its ethyl ester was detected in monoculture fermentation with H. guilliermondii. H. guilliermondii consumed the most phenylalanine among the other isolates. 5-hydroxy-L-tryptophan was detected in fermentations with M. pulcherrima and M. sinensis/shanxiensis. M. pulcherrima significantly increased tryptophol content and utilised tyrosine in monoculture fermentations. Sequential fermentation with M. sinensis/shanxiensis and S. cerevisiae produced higher amounts of N-acetyl derivatives of tryptophan and phenylalanine, while H. guilliermondii-S. cerevisiae fermentation resulted in wines with the highest concentrations of L-kynurenine and 3-hydroxyanthranilic acid. P. kluyveri produced the highest concentration of N-acetyl-L-tyrosine in monoculture fermentations. These findings highlight the different yeast metabolic pathways. Full article

The semi-dry fermentation processing (SFP) of Coffea arabica is a traditional primary processing method in the coffee industry, which crucially impacts the coffee’s flavor. To further obtain useful information on microbial communities and chemical compounds during the SFP of C. arabica from Yunnan, China, the microbial community structures and the differentially changed non-volatile compounds (DCnVCs) were comprehensively analyzed. The results showed that Tatumella, Staphylococcus, Klebsiella, Brevundimonas, and Gluconobacter were the most prevalent bacteria genera, and Candida, Hannaella, Hanseniaspora, Pichia, and Lachancea were the most abundant fungal genera. Furthermore, 117 DCnVCs were found in the fermentation-finished samples compared to the raw materials. Therefore, this study can provide useful information for understanding the SFP of coffee beans, and its impact on coffee’s quality parameters. Full article

This study evaluates the fermentation performances of non-Saccharomyces strains in fermenting cherry must from Italian cherries unsuitable for selling and not intended to be consumed fresh, and their effects on the chemical composition of the resulting wine. Fermentation trials in 100 and 500 mL of must were carried out to select 21 strains belonging to 11 non-Saccharomyces species. Cherry wines obtained by six select strains were chemically analyzed for fixed and volatile compounds. Quantitative data were statistically analyzed by agglomerative hierarchical clustering, partial least squared discriminant analysis, and principal component analysis. Wines revealed significant differences in their composition. Lactic acid and phenylethyl acetate levels were very high in wines produced by Lachancea and Hanseniaspora, respectively. Compared to S. cerevisiae wine, non-Saccharomyces wines had a lower content of fatty acid ethyl esters 4-vinyl guaiacol and 4-vinyl phenol. The multivariate analysis discriminated between wines, demonstrating the different contributions of each strain to aroma components. Specifically, all wines from non-Saccharomyces strains were kept strictly separate from the control wine. This study provided comprehensive characterization traits for non-conventional strains that enhance the aroma complexity of cherry-based wine. The use of these yeasts in cherry wine production appears promising. Further investigation is required to ascertain their suitability for larger-scale fermentation. Full article

The culturable yeast communities in temperate forest soils under the ornithogenic influence were studied in a seasonal dynamic. To investigate the intense ornithogenic influence, conventional and “live” feeders were used, which were attached to trees in the forest and constantly replenished throughout the year. It was found that the yeast abundance in the soil under strong ornithogenic influence reached the highest values in winter compared to the other seasons and amounted to 4.8 lg (cfu/g). This was almost an order of magnitude higher than the minimum value of yeast abundance in ornithogenic soils determined for summer. A total of 44 yeast species, 21 ascomycetes and 23 basidiomycetes, were detected in ornithogenic soil samples during the year. These included soil-related species (Barnettozyma californica, Cyberlindnera misumaiensis, Cutaneotrichosporon moniliiforme, Goffeauzyma gastrica, Holtermanniella festucosa, Leucosporidium creatinivorum, L. yakuticum, Naganishia adeliensis, N. albidosimilis, N. globosa, Tausonia pullulans, and Vanrija albida), eurybionts (yeast-like fungus Aureobasidium pullulans, Debaryomyces hansenii, and Rhodotorula mucilaginosa), inhabitants of plant substrates and litter (Cystofilobasidium capitatum, Cys. infirmominiatum, Cys. macerans, Filobasidium magnum, Hanseniaspora uvarum, Metschnikowia pulcherrima, and Rh. babjevae) as well as a group of pathogenic and opportunistic yeast species (Arxiozyma bovina, Candida albicans, C. parapsilosis, C. tropicalis, Clavispora lusitaniae, and Nakaseomyces glabratus). Under an ornithogenic influence, the diversity of soil yeasts was higher compared to the control, confirming the uneven distribution of yeasts in temperate forest soils and their dependence on natural hosts and vectors. Interestingly, the absolute dominant species in ornithogenic soils in winter (when the topsoil temperature was below zero) was the basidiomycetous psychrotolerant yeast T. pullulans. It is regularly observed in various soils in different geographical regions. Screening of the hydrolytic activity of 50 strains of this species at different temperatures (2, 4, 10, 15 and 20 °C) showed that the activity of esterases, lipases and proteases was significantly higher at the cultivation temperature. Ornithogenic soils could be a source for the relatively easy isolation of a large number of strains of the psychrotolerant yeast T. pullulans to test, study and optimize their potential for the production of cold-adapted enzymes for industry. Full article

Grape skin serves as a reservoir for many bacteria and fungi, which can affect grape health, quality, and safety. However, grape skin microbiota and mycobiota of table grapes remain largely understudied. This study investigated bacterial and fungal communities residing on different table grapevine cultivars (‘Summer Black’ and ‘Kyoho’) as well as the impact of potential contributors thereby, via culture-dependent and culture-independent (analysis of 16S rRNA gene and internal transcribed spacer sequences) methods with different purposes. Microbiota of both grapevine cultivars were dominated by Cladosporium, Alternaria, Aspergillus, Thauera, and Pantoea. In addition, yeast strains belonging to Hanseniaspora opuntiae, Pichia terricola, Rhodotorula mucilaginosa, Candida stellimalicola, and Kodamaea ohmeri were enriched from the studied grapes, while some strains were considered as health-threatening pathogens. Differences in grapevine cultivars did not significantly affect their mycobiota and microbiota profiles. Nevertheless, their mycobiota exhibited significant variations across different grape-sampling sites in Shanghai, indicating the contribution of the grape-growing environment to grape skin mycobiota. Altogether, the current study demonstrated the contribution of the grape-growing environment to table grape skin mycobiota, and highlighted the importance of microbiota management in the production and consumption of table grapes. Full article

This research investigates how different fermentation techniques using non-Saccharomyces yeast (Candida ethanolica Ce, Hanseniaspora guilliermondii Hg, Hanseniaspora thailandica Ht) and Saccharomyces cerevisiae (Sc) affect the synthesis of hesperidin, nobiletin, and other flavonoid and aromatic substances, which play a vital role in improving the overall quality of fruit wines due to their various biological properties. The combination of Sc:(Ce.Ht)-1:100 (Ce 0.5 × 10⁷ CFU/mL, Ht 0.5 × 10⁷ CFU/mL, Sc 1 × 10⁵ CFU/mL) yielded the highest hesperidin content at 4.12 ± 0.08 mg/L, followed by the Sc:(Ce.Hg)-1:1 (Ce 0.5 × 10⁷ CFU/mL, Hg 0.5 × 10⁷ CFU/mL, Sc 1 × 10⁷ CFU/mL) combination at 4.08 ± 0.06 mg/L. The highest nobiletin content was achieved by the (Hg.Ht)-10-Sc (Hg 0.5 × 10⁷ CFU/mL, Ht 0.5 × 10⁷ CFU/mL, Sc 1 × 10⁷ CFU/mL) combination, reaching 1.04 ± 0.05 mg/L, which was significantly higher than other multi-strain combinations. Additionally, the hesperidin content produced by the (Hg.Ht)-10-Sc combination was relatively high at 4.04 ± 0.02 mg/L, demonstrating a richness and complexity of aroma superior to that of fermentation with commercial yeast strains alone. The findings suggest that the (Hg.Ht)-10-Sc combination is the most effective multi-strain combination for increasing the levels of nobiletin and hesperidin in citrus wine, thereby enhancing the overall quality of the wine. These experimental results offer a promising approach for enhancing the quality of citrus wines and other fruit wines. Full article

This study evaluated the biocontrol effect of isolated epiphytic yeasts (Papiliotrema terrestris, Hanseniaspora uvarum, and Rhodosporidium glutinis) against Botrytis cinerea and Alternaria alternata in blueberry fruits and its possible mechanisms. Our findings indicated that the three tested yeasts exerted a good biocontrol effect on postharvest diseases in blueberry, and that H. uvarum was the most effective. In addition, the three tested yeasts could improve the postharvest storage quality of blueberry fruits to some extent. H. uvarum demonstrated the strongest direct inhibitory effect on pathogens by suppressing spore germination, mycelial growth, and antifungal volatile organic compound (VOC) production. P. terrestris showed the highest extracellular lytic enzymes activities. It also had better adaptation to low temperature in fruit wounds at 4 °C. The biofilm formation capacity was suggested to be the main action mechanism of R. glutinis, which rapidly colonized fruit wounds at 20 °C. Several action mechanisms are employed by the superb biocontrol yeasts, while yeast strains possess distinctive characteristics and have substantially different action mechanisms. Full article

An amplicon metagenomic approach based on the ITS2 region of fungal rDNA was used to investigate the diversity of fungi associated with mature strawberries collected from a volcanic orchard and open-air market stands. Based on the Kruskal–Wallis test, no statistically significant differences were observed in both non-phylogenetic and phylogenetic alpha diversity indices. According to beta diversity analyses, significant differences in fungal communities were found between groups (orchard vs. market). Taxonomic assignment of amplicon sequence variables (ASVs) revealed 7 phyla and 31 classes. The prevalent fungal phyla were Basidiomycota (29.59–84.58%), Ascomycota (15.33–70.40%), and Fungi-phy-Insertae-sedis (0.45–2.89%). The most predominant classes among the groups were Saccharomycetes in the market group, and Microbotryomycetes and Tremellomycetes in the orchard group. Based on the analysis of microbiome composition (ANCOM), we found that the most differentially fungal genera were Hanseniaspora, Kurtzmaniella, and Phyllozyma. Endophytic yeasts Curvibasidium cygneicollum were prevalent in both groups, while Candida railenensis was detected in fruits originating only from the market. In addition, Rhodotorula graminis (relative abundance varying from 1.7% to 21.18%) and Papiliotrema flavescens (relative abundance varying from 1.58% to 16.55%) were detected in all samples regardless of origin, while Debaryomyces prosopidis was detected in samples from the market only, their relative abundance varying with the sample (from 0.80% to 19.23%). Their role in fruit quality and safety has not been yet documented. Moreover, several clinically related yeasts, such as Meyerozyma guilliermondii and Candida parapsilosis, were detected in samples only from the market. Understanding the variety and makeup of the mycobiome in ripe fruits during the transition from the orchard to the market is crucial for fruit safety after harvest. Full article

Hanseniaspora vineae (Hv) is a non-Saccharomyces yeast with unique metabolic features, making it appealing for wine production. However, Hv presents high nutritional requirements that may lead to slow fermentation. This study investigated the impact of sequential inoculation of Saccharomyces cerevisiae (Sc) in white winemaking at different time points (24, 48, 74, 100 and 200 h) during Hv fermentation and compared them to simultaneous inoculations. The 200 h protocol extended fermentation by an average of 13 days compared to pure Sc, decreasing with earlier sequential inoculation. Sc wines were richer in isoamyl acetate and ethyl hexanoate than Hv wines, with no significant differences among inoculation protocols. β-phenylethyl acetate was increased in Hv wines, particularly in the 24 h protocol. The 2-phenylethanol concentration was negatively correlated with the S. cerevisiae inoculation delay. Hv altered the wine aroma features, enhancing the compounds associated with rose-like scents. Reducing the Sc inoculation delay aligned Hv with industrial standards while maintaining increased β-phenylethyl acetate production. However, co-inoculation with Sc seems to better meet the Hv requirement without sacrificing the main aromatic features of Hv, demonstrating faster sugar depletion and higher acetate and ethyl ester contents, suggesting that co-inoculation yields a more modulable wine aroma profile. Full article

In view of climate change and the increasingly antagonistic wine market, the exploitation of native genetic resources is revisited in relation to sustainable wine production. ‘Sideritis’ is a late-ripening Greek grape variety, which is quite promising for counteracting wine quality issues associated with the annual temperature rise. The aim of this study was to improve the quality and to enhance the aroma of ‘Sideritis’ wine through the use of native yeasts. To improve vinification, Hanseniaspora opuntiae L1 was used along with Saccharomyces cerevisiae W7 in mixed fermentations (SQ). The addition of H. οpuntiae significantly altered the chemical profile of the wine compared to the single-inoculated fermentations with W7 (IS). H. opuntiae increased all the acetate esters, except for hexyl acetate and (Z)-3-hexen-1-ol acetate. The concentration of 2-phenylethyl acetate, which imparts flowery and sweet notes, exhibited a 2.6-fold increase in SQ as compared to IS wines. SQ also showed higher levels in several ethyl esters, including ethyl butyrate, ethyl heptanoate and ethyl 7-octenoate, which are associated with fruity notes compared to IS. H. opuntiae produced citronellol, a terpene associated with rose and green notes, and increased the overall acceptance of the wine. Present results are thus quite promising for improving ‘Sideritis’ wine quality towards a sustainable wine production in Greece in view of global warming. Full article

Pure, co-, and sequential fermentations of Hanseniaspora uvarum, H. guilliermondii, and Saccharomyces cerevisiae strains were evaluated to improve the aromatic quality of ciders. In sequential fermentations, Hanseniaspora strains were used as starter, followed by S. cerevisiae inoculation succeeding one, two, and three days of fermentation. Kinetics, physicochemical parameters, and volatile compounds were assessed during 10 days of fermentation. The headspace technique was used to capture the volatile compounds from the ciders obtained in each experiment and analyzed by gas chromatography. Fermentations with pure strains of Hansenisaspora sp. showed a high population (>10¹⁰ CFU/mL) but had a low fermentation rate (2.3–3.8 CO₂ g/L/d), low consumption of amino acids (20–40 mg/L) with a high residual content, high sugar consumption (80–90 g/L), and low alcohol content (<2.0% v/v). The H. uvarum strain produced a notably high ester content (245 mg/L). In the co-fermentations, H. guilliermondii with S. cerevisiae highlighted a significant production of higher alcohols, similar to that produced by S. cerevisiae alone (152–165 mg/L). In general, the maximum fermentation rate of the sequential inoculations was lower than co-fermentations but showed low residual nitrogen content (<69 mg/L) and good conversion of sugars into ethanol (4.3–5.7% v/v). The highest concentrations of volatile compounds were observed in treatments involving the two non-conventional strains: H. uvarum with S. cerevisiae inoculation after three days (564 mg/L) and H. guilliermondii after just one day (531 mg/L) of fermentation. These differences stemmed from the metabolic activity of the strains. H. uvarum was influenced by the presence of Saccharomyces, whereas H. guilliermondii did not exhibit this effect. Thus, a pure H. uvarum inoculum has the potential to produce a demi-sec cider with low alcohol content and high content of esters, contributing to a fruity aroma. In addition, ciders with sequential inoculation were the most promising for dry cider processing concerning fermentation parameters and bioaroma enrichment. Full article

The metataxonomic diversity and microbial composition of microorganisms during the coffee fermentation process as well as their relationship with coffee quality were determined across 20 farms in the department of Cesar, Colombia, by sampling coffee fruits from Coffea arabica; Var. Castillo General^®, Var. Colombia, and Var. Cenicafé 1. In each farm, the fruits were processed and the fermentation process took place between 10 and 42 h following this. Three samples of mucilage and washed coffee seeds were collected per farm during the fermentation process. The microorganisms present in the mucilage were identified using metataxonomic methods by amplifying the 16S rRNA gene for bacteria and ITS for fungi. The microorganisms’ morphotypes were isolated and identified. The analysis of bacteria allowed for the identification of the following genera: Gluconobacter, Leuconostoc, Acetobacter, Frateuria, Pantoea, Pseudomonas, Tatumella, and Weisella, as well as unclassified enterobacteria; the Lactobacillacea and Secundilactobacillus families were only identified in the Var. Cenicafé 1. For fungi, the top 11 genera and families found included Hanseniaspora, Candida, Meyerozyma, Wickerhamomyces, Pichia, f-Saccharomycodaceae, f-Nectriciae, unclassified fungi, and Saccharomycetaceae, which were only found in Cenicafé 1. A total of 92% of the coffee samples obtained scored between 80.1 and 84.9, indicating “Very Good” coffee (Specialty Coffee Association (SCA) scale). Farms with the longest fermentation times showed better coffee attributes related to acidity, fragrance, and aroma. During coffee fermentation, there is a central microbiome. The differences between the microorganisms’ genera could be influenced by the coffee variety, while the specific conditions of each farm (i.e., altitude and temperature) and its fermentation processes could determine the proportions of and interactions between the microbial groups that favor the sensory characteristics responsible for coffee cup quality. Full article

Annual surveys of Irish soil samples identified three isolates, CBS 16921 (UCD88), CBS 18246 (UCD443), and CBS 18247 (UCD483), of an apiculate yeast species within the Hanseniaspora genus. The internal transcribed spacer (ITS) and D1/D2 region of the large subunit (LSU) rRNA sequences showed that these are isolates of the recently described species Hanseniaspora menglaensis, first isolated from Southwest China. No genome sequence for H. menglaensis is currently available. The genome sequences of the three Irish isolates were determined using short-read (Illumina) sequencing, and the sequence of one isolate (CBS 16921) was assembled to chromosome level using long-read sequencing (Oxford Nanopore Technologies). Phylogenomic analysis shows that H. menglaensis belongs to the fast-evolving lineage (FEL) of Hanseniaspora. Only one MAT idiomorph (encoding MATα1) was identified in all three sequenced H. menglaensis isolates, consistent with one mating type of a heterothallic species. Genome comparisons showed that there has been a rearrangement near MATα of FEL species compared to isolates from the slowly evolving lineage (SEL). Full article

As a biological alternative to the antimicrobial action of SO₂, bioprotection has been proposed to winemakers as a means to limit or prevent grape musts microbial alteration. Competition for nitrogenous nutrients and for oxygen are often cited as potential explanations for the effectiveness of bioprotection. This study analyses the effect of a bioprotective M. pulcherrima strain on the growth of one H. valbyensis strain and one H. uvarum strain. Bioprotection efficiency was observed only against H. valbyensis inoculated at the two lowest concentrations. These results indicate a potential species-dependent efficiency of the bioprotective strain and a strong impact of the initial ratio between bioprotective and apiculate yeasts. The analysis of the consumption of nitrogen compounds revealed that leucine, isoleucine, lysine and tryptophan were consumed preferentially by all three strains. The weaker assimilation percentages of these amino acids observed in H. valbyensis at 24 h growth suggest competition with M. pulcherrima that could negatively affects the growth of the apiculate yeast in co-cultures. The slowest rate of O₂ consumption of H. valbyensis strain, in comparison with M. pulcherrima, was probably not involved in the bioprotective effect. Non-targeted metabolomic analyses of M. pulcherrima and H. valbyensis co-culture indicate that the interaction between both strains particularly impact lysin and tryptophan metabolisms. Full article

Hanseniaspora uvarum is the predominant yeast species in the majority of wine fermentations, which has only recently become amenable to directed genetic manipulation. The genetics and metabolism of H. uvarum have been poorly studied as compared to other yeasts of biotechnological importance. This work describes the construction and characterization of homozygous deletion mutants in the HuZWF1 gene, encoding glucose-6-phosphate dehydrogenase (G6PDH), which provides the entrance into the oxidative part of the pentose phosphate pathway (PPP) and serves as a major source of NADPH for anabolic reactions and oxidative stress response. Huzwf1 deletion mutants grow more slowly on glucose medium than wild-type and are hypersensitive both to hydrogen peroxide and potassium bisulfite, indicating that G6PDH activity is required to cope with these stresses. The mutant also requires methionine for growth. Enzyme activity can be restored by the expression of heterologous G6PDH genes from other yeasts and humans under the control of a strong endogenous promoter. These findings provide the basis for a better adaptation of H. uvarum to conditions used in wine fermentations, as well as its use for other biotechnological purposes and as an expression organism for studying G6PDH functions in patients with hemolytic anemia. Full article