Editorial

3 pages, 180 KiB

Open AccessEditorial

by Rosanna Tofalo

Microorganisms 2021, 9(4), 700; https://doi.org/10.3390/microorganisms9040700 - 28 Mar 2021

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The Special Issue “Microbial dynamics in wine production” collects nine original research articles and a review concerning wine production, a complex process where microbes have a decisive role [...] Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

Research

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12 pages, 878 KiB

Open AccessArticle

Adhesion Properties, Biofilm Forming Potential, and Susceptibility to Disinfectants of Contaminant Wine Yeasts

by Giorgia Perpetuini, Alessio Pio Rossetti, Noemi Battistelli, Giuseppe Arfelli and Rosanna Tofalo

Microorganisms 2021, 9(3), 654; https://doi.org/10.3390/microorganisms9030654 - 22 Mar 2021

Cited by 16 | Viewed by 2779

Abstract

In this study, yeasts isolated from filter membranes used for the quality control of bottled wines were identified and tested for their resistance to some cleaning agents and potassium metabisulphite, adhesion to polystyrene and stainless-steel surfaces, and formation of a thin round biofilm, [...] Read more.

In this study, yeasts isolated from filter membranes used for the quality control of bottled wines were identified and tested for their resistance to some cleaning agents and potassium metabisulphite, adhesion to polystyrene and stainless-steel surfaces, and formation of a thin round biofilm, referred to as a MAT. A total of 40 strains were identified by rRNA internal transcribed spacer (ITS) restriction analysis and sequence analysis of D1/D2 domain of 26S rRNA gene. Strains belong to Pichia manshurica (12), Pichia kudriavzevii (9), Pichia membranifaciens (1), Candida sojae (6), Candida parapsilosis (3), Candida sonorensis (1), Lodderomyces elongisporus (2), Sporopachydermia lactativora (3), and Clavispora lusitaniae (3) species. Regarding the adhesion properties, differences were observed among species. Yeasts preferred planktonic state when tested on polystyrene plates. On stainless-steel supports, adhered cells reached values of about 6 log CFU/mL. MAT structures were formed only by yeasts belonging to the Pichia genus. Yeast species showed different resistance to sanitizers, with peracetic acid being the most effective and active at low concentrations, with minimum inhibitory concentration (MIC) values ranging from 0.08% (v/v) to 1% (v/v). C. parapsilosis was the most sensible species. Data could be exploited to develop sustainable strategies to reduce wine contamination and establish tailored sanitizing procedures. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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22 pages, 2075 KiB

Open AccessEditor’s ChoiceArticle

Effect of Co-Inoculation of Candida zemplinina, Saccharomyces cerevisiae and Lactobacillus plantarum for the Industrial Production of Negroamaro Wine in Apulia (Southern Italy)

by Maria Tufariello, Vittorio Capozzi, Giuseppe Spano, Giovanni Cantele, Pasquale Venerito, Giovanni Mita and Francesco Grieco

Microorganisms 2020, 8(5), 726; https://doi.org/10.3390/microorganisms8050726 - 13 May 2020

Cited by 29 | Viewed by 2882

Abstract

The employment of multi-species starter cultures has growing importance in modern winemaking for improving the complexity and wine attributes. The assessment of compatibility for selected species/strains at the industrial-scale is crucial to assure the quality and the safety associated with fermentations. An aspect [...] Read more.

The employment of multi-species starter cultures has growing importance in modern winemaking for improving the complexity and wine attributes. The assessment of compatibility for selected species/strains at the industrial-scale is crucial to assure the quality and the safety associated with fermentations. An aspect particularly relevant when the species belong to non-Saccharomyces, Saccharomyces spp. and malolactic bacteria, three categories with different biological characteristics and oenological significance. To the best of our knowledge, the present report is the first study regarding the utilization of a combined starter culture composed of three strains of non-Saccharomyces, Saccharomyces cerevisiae and Lactobacillus plantarum for production of wine at the industrial scale. More in-depth, this work investigated the oenological potential of the autochthonous characterized strains from the Apulian region (Southern Italy), Candida zemplinina (syn. Starmerella bacillaris) 35NC1, S. cerevisiae (NP103), and L. plantarum (LP44), in co-inoculation following a complete scale-up scheme. Microbial dynamics, fermentative profiles and production of volatile secondary compounds were assessed in lab-scale micro-vinification tests and then the performances of the mixed starter formulation were further evaluated by pilot-scale wine production. The above results were finally validated by performing an industrial-scale vinification on 100HL of Negroamaro cultivar grape must. The multi-starter formulation was able to rule the different stages of the fermentation processes effectively, and the different microbial combinations enhanced the organoleptic wine features to different extents. The findings indicated that the simultaneous inoculation of the three species affect the quality and quantity of several volatile compounds, confirming that the complexity of the wine can reflect the complexity of the starter cultures. Moreover, the results underlined that the same mixed culture could differently influence wine quality when tested at the lab-, pilot- and industrial-scale. Finally, we highlighted the significance of employment non-Saccharomyces and L. plantarum, together with S. cerevisiae, autochthonous strains in the design of custom-made starter culture formulation for typical regional wine production with pronounced unique quality. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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11 pages, 2780 KiB

Open AccessArticle

Monitoring the Functionality and Stress Response of Yeast Cells Using Flow Cytometry

by Stephan Sommer

Microorganisms 2020, 8(4), 619; https://doi.org/10.3390/microorganisms8040619 - 24 Apr 2020

Cited by 5 | Viewed by 5435

Abstract

Throughout fermentation, yeast faces continuously changing medium conditions and reacts by adapting its metabolism. The adaptation is a critical process and is dependent on the accurate functioning of the cell. A stable membrane potential, which is, among other roles, responsible for protecting the [...] Read more.

Throughout fermentation, yeast faces continuously changing medium conditions and reacts by adapting its metabolism. The adaptation is a critical process and is dependent on the accurate functioning of the cell. A stable membrane potential, which is, among other roles, responsible for protecting the yeast from low pH, is an important attribute for evaluating functionality. Other factors are storage products such as glycogen, trehalose, and neutral lipids, as well as mitochondrial activity and the integrity of the DNA. These parameters can be complemented by the analysis of viability, cell cycle, intracellular pH, and reactive oxygen species in the cell. The correlation of all these factors provides valuable information for evaluating the performance of a yeast population during fermentation. In order to demonstrate the analytical capabilities of flow cytometry, a Saccharomyces cerevisiae yeast strain was observed in a modified growth medium for 384 h (16 days). The results confirm observations made with other methods and reports from the literature. However, with flow cytometry, it is possible to gain deeper insight into stress response and adaptation behavior of yeast at a cellular level. The causality from the formation of oxygen-radicals to cell death, for example, can be shown, as well as the dependency of the intracellular pH on the stability of the membrane. The proposed bio-monitoring system has the potential to provide applicable information as a process control tool for wineries. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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14 pages, 1317 KiB

Open AccessArticle

Inoculum Strategies and Performances of Malolactic Starter Lactobacillus plantarum M10: Impact on Chemical and Sensorial Characteristics of Fiano Wine

by Silvia Jane Lombardi, Gianfranco Pannella, Massimo Iorizzo, Bruno Testa, Mariantonietta Succi, Patrizio Tremonte, Elena Sorrentino, Massimo Di Renzo, Daniela Strollo and Raffaele Coppola

Microorganisms 2020, 8(4), 516; https://doi.org/10.3390/microorganisms8040516 - 04 Apr 2020

Cited by 28 | Viewed by 2891

Abstract

Malolactic fermentation (MLF) is a biological process that, in addition to deacidifying, also improves biological stability and changes the chemical and sensorial characteristics of wines. However, multiple biotic and abiotic factors, present in must and wine, make the onset and completion of MLF [...] Read more.

Malolactic fermentation (MLF) is a biological process that, in addition to deacidifying, also improves biological stability and changes the chemical and sensorial characteristics of wines. However, multiple biotic and abiotic factors, present in must and wine, make the onset and completion of MLF by indigenous malolactic bacteria or added commercial starters difficult. This work illustrates the metabolic and fermentative dynamics in winemaking Fiano wine, using a commercial starter of Saccharomyces cerevisiae and the selected strain Lactobacillus plantarum M10. In particular, an inoculum of malolactic starter was assessed at the beginning of alcoholic fermentation (early co-inoculum), at half alcoholic fermentation (late co-inoculum), and post alcoholic fermentation (sequential inoculum). The malolactic starter, before its use, was pre-adapted in sub-optimal growth conditions (pH 5.0). In sequential inoculum of the Lb. plantarum M10, even in a wine with high acidity, has confirmed its good technological and enzymatic characteristics, completing the MLF and enriching the wine with desirable volatile compounds. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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19 pages, 2944 KiB

Open AccessArticle

Nitrogen Preferences during Alcoholic Fermentation of Different Non-Saccharomyces Yeasts of Oenological Interest

by Helena Roca-Mesa, Sonia Sendra, Albert Mas, Gemma Beltran and María-Jesús Torija

Microorganisms 2020, 8(2), 157; https://doi.org/10.3390/microorganisms8020157 - 22 Jan 2020

Cited by 61 | Viewed by 6191

Abstract

Non-Saccharomyces yeasts have long been considered spoilage microorganisms. Currently, oenological interest in those species is increasing, mostly due to their positive contribution to wine quality. In this work, the fermentative capacity and nitrogen consumption of several non-Saccharomyces wine yeast (Torulaspora [...] Read more.

Non-Saccharomyces yeasts have long been considered spoilage microorganisms. Currently, oenological interest in those species is increasing, mostly due to their positive contribution to wine quality. In this work, the fermentative capacity and nitrogen consumption of several non-Saccharomyces wine yeast (Torulaspora delbrueckii, Lachancea thermotolerans, Starmerella bacillaris, Hanseniaspora uvarum, and Metschnikowia pulcherrima) were analyzed. For this purpose, synthetic must with three different nitrogen compositions was used: a mixture of amino acids and ammonium, only organic or inorganic nitrogen. The fermentation kinetics, nitrogen consumption, and yeast growth were measured over time. Our results showed that the good fermentative strains, T. delbrueckii and L. thermotolerans, had high similarities with Saccharomyces cerevisiae in terms of growth, fermentation profile, and nitrogen assimilation preferences, although L. thermotolerans presented an impaired behavior when only amino acids or ammonia were used, being strain-specific. M. pulcherrima was the non-Saccharomyces strain least affected by the nitrogen composition of the medium. The other two poor fermentative strains, H. uvarum and S. bacillaris, behaved similarly regarding amino acid uptake, which occurred earlier than that of the good fermentative species in the absence of ammonia. The results obtained in single non-Saccharomyces fermentations highlighted the importance of controlling nitrogen requirements of the wine yeasts, mainly in sequential fermentations, in order to manage a proper nitrogen supplementation, when needed. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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29 pages, 17202 KiB

Open AccessArticle

Fungal Diversity of Deteriorated Sparkling Wine and Cork Stoppers in Catalonia, Spain

by Ernesto Rodríguez-Andrade, Alberto M. Stchigel, Josep Guarro and José F. Cano-Lira

Microorganisms 2020, 8(1), 12; https://doi.org/10.3390/microorganisms8010012 - 19 Dec 2019

Cited by 16 | Viewed by 5524

Abstract

Filamentous fungi are rarely reported as responsible for spoiling wine. Cork taint was detected in sparkling wine; therefore, we investigated fungal contamination as a possible cause of organoleptic alteration. Spoiled wine was filtered and membranes were plated onto potato dextrose agar (PDA). The [...] Read more.

Filamentous fungi are rarely reported as responsible for spoiling wine. Cork taint was detected in sparkling wine; therefore, we investigated fungal contamination as a possible cause of organoleptic alteration. Spoiled wine was filtered and membranes were plated onto potato dextrose agar (PDA). The cork stoppers used for sealing bottles were cut and also plated onto PDA. Fungal strains were phenotypically characterized and molecularly identified by sequencing of a fragment of the 28S nrRNA gene (LSU) and (occasionally) by other additional molecular markers. Twenty-seven strains were isolated and sixteen species were identified, all of them belonging to the phylum Ascomycota. The fungi isolated from wine were three species of Aspergillus section Nidulantes, a species of Penicillium section Exicaulis and Beauveria bassiana. Candida patagonica was isolated from both sort of samples, and the fungi isolated from cork stoppers were Altenaria alternata and Cladosporium cladosporioides. Surprisingly, most of the taxa recovered from the cork stoppers and/or wine were new to the science: a new genus (Dactylodendron) and seven new species belonging to the genera Cladophialophora, Dactylodendron, Kirschsteiniothelia, Rasamsonia, and Talaromyces. Future studies could let us know if these fungi would be able to produce compounds responsible for cork taint. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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14 pages, 1654 KiB

Open AccessArticle

Geographical Origin Has a Greater Impact on Grape Berry Fungal Community than Grape Variety and Maturation State

by Dimitrios Kioroglou, Elena Kraeva-Deloire, Leigh M. Schmidtke, Albert Mas and Maria C. Portillo

Microorganisms 2019, 7(12), 669; https://doi.org/10.3390/microorganisms7120669 - 10 Dec 2019

Cited by 25 | Viewed by 3226

Abstract

We used barcoded sequencing to analyze the eukaryotic population in the grape berries at different ripening states in four Australian vineyards. Furthermore, we used an innovative compositional data analysis for assessing the diversity of microbiome communities. The novelty was the introduction of log-ratio [...] Read more.

We used barcoded sequencing to analyze the eukaryotic population in the grape berries at different ripening states in four Australian vineyards. Furthermore, we used an innovative compositional data analysis for assessing the diversity of microbiome communities. The novelty was the introduction of log-ratio balances between the detected genera. Altogether, our results suggest that fungal communities were more impacted by the geographical origin of the Australian vineyards than grape variety and harvest time. Even if the most abundant genera were Aureobasidium and Mycosphaerella, they were ubiquitous to all samples and were not discriminative. In fact, the balances and the fungal community structure seemed to be greatly affected by changes of the genera Penicillium, Colletotrichum, Aspergillus, Rhodotorula, and Botrytis. These results were not evident from the comparison of relative abundance based on OTU counts alone, remarking the importance of the balance analysis for microbiome studies. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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16 pages, 2742 KiB

Open AccessArticle

Identification of Down-Regulated Proteome in Saccharomyces cerevisiae with the Deletion of Yeast Cathepsin D in Response to Nitrogen Stress

by Jingjin Hu, Lingxiao Yu, Qin Shu and Qihe Chen

Microorganisms 2019, 7(8), 214; https://doi.org/10.3390/microorganisms7080214 - 24 Jul 2019

Cited by 6 | Viewed by 3296

Abstract

Vacuolar proteinase A (Pep4p) is required for the post-translational precursor maturation of vacuolar proteinases in Saccharomyces cerevisiae, and important for protein turnover after oxidative damage. The presence of proteinase A in brewing yeast leads to the decline of beer foam stability, thus [...] Read more.

Vacuolar proteinase A (Pep4p) is required for the post-translational precursor maturation of vacuolar proteinases in Saccharomyces cerevisiae, and important for protein turnover after oxidative damage. The presence of proteinase A in brewing yeast leads to the decline of beer foam stability, thus the deletion or inhibition of Pep4p is generally used. However, the influence of Pep4p deletion on cell metabolism in Saccharomyces cerevisiae is still unclear. Herein, we report the identification of differentially down-regulated metabolic proteins in the absence of Pep4p by a comparative proteomics approach. 2D-PAGE (two-dimensional polyacrylamide gel electrophoresis) presented that the number of significantly up-regulated spots (the Pep4p-deficient species versus the wild type) was 183, whereas the down-regulated spots numbered 111. Among them, 35 identified proteins were differentially down-regulated more than 10-fold in the Pep4p-deficient compared to the wild-type species. The data revealed that Pep4p was required for the synthesis and maturation of several glycolytic enzymes and stress proteins, including Eno2p, Fba1p, Pdc1p, Tpi1p, Ssa1, Hsp82p, and Trr1p. The transcription and post-translational modifications of glycolytic enzymes like Eno2p and Fba1p were sensitive to the absence of Pep4p; whereas the depletion of the pep4 gene had a negative impact on mitochondrial and other physiological functions. The finding of this study provides a systematic understanding that Pep4p may serve as a regulating factor for cell physiology and metabolic processes in S. cerevisiae under a nitrogen stress environment. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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15 pages, 1865 KiB

Open AccessArticle

Engineering Synthetic Microbial Communities through a Selective Biofilm Cultivation Device for the Production of Fermented Beverages

by Sokny Ly, F. Bajoul Kakahi, Hasika Mith, Chanvorleak Phat, Barbara Fifani, Tierry Kenne, Marie-Laure Fauconnier and Frank Delvigne

Microorganisms 2019, 7(7), 206; https://doi.org/10.3390/microorganisms7070206 - 20 Jul 2019

Cited by 10 | Viewed by 4333

Abstract

Production of Cambodian rice wine involves complex microbial consortia. Indeed, previous studies focused on traditional microbial starters used for this product revealed that three microbial strains with complementary metabolic activities are required for an effective fermentation, i.e., filamentous fungi (Rhizopus oryzae), yeast [...] Read more.

Production of Cambodian rice wine involves complex microbial consortia. Indeed, previous studies focused on traditional microbial starters used for this product revealed that three microbial strains with complementary metabolic activities are required for an effective fermentation, i.e., filamentous fungi (Rhizopus oryzae), yeast (Saccharomyces cerevisiae), and lactic acid bacteria (Lactobacillus plantarum). Modulating the ratio between these three key players led to significant differences, not only in terms of ethanol and organic acid production, but also on the profile of volatile compounds, in comparison with natural communities. However, we observed that using an equal ratio of spores/cells of the three microbial strains during inoculation led to flavor profile and ethanol yield close to that obtained through the use of natural communities. Compartmentalization of metabolic tasks through the use of a biofilm cultivation device allows further improvement of the whole fermentation process, notably by increasing the amount of key components of the aroma profile of the fermented beverage (i.e., mainly phenylethyl alcohol, isobutyl alcohol, isoamyl alcohol, and 2-methyl-butanol) and reducing the amount of off-flavor compounds. This study is a step forward in our understanding of interkingdom microbial interactions with strong application potential in food biotechnology. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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Review

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21 pages, 629 KiB

Open AccessReview

Microbial Resources as a Tool for Enhancing Sustainability in Winemaking

by Tiziana Nardi

Microorganisms 2020, 8(4), 507; https://doi.org/10.3390/microorganisms8040507 - 02 Apr 2020

Cited by 31 | Viewed by 5414

Abstract

In agriculture, the wine sector is one of the industries most affected by the sustainability issue. It is responsible for about 0.3% of annual global greenhouse gas emissions from anthropogenic activities. Sustainability in vitiviniculture was firstly linked to vineyard management, where the use [...] Read more.

In agriculture, the wine sector is one of the industries most affected by the sustainability issue. It is responsible for about 0.3% of annual global greenhouse gas emissions from anthropogenic activities. Sustainability in vitiviniculture was firstly linked to vineyard management, where the use of fertilizers, pesticides and heavy metals is a major concern. More recently, the contribution of winemaking, from grape harvest to bottling, has also been considered. Several cellar processes could be improved for reducing the environmental impact of the whole chain, including microbe-driven transformations. This paper reviews the potential of microorganisms and interactions thereof as a natural, environmentally friendly tool to improve the sustainability aspects of winemaking, all along the production chain. The main phases identified as potentially interesting for exploiting microbial activities to lower inputs are: (i) pre-fermentative stages, (ii) alcoholic fermentation, (iii) stage between alcoholic and malolactic fermentation, (iv) malolactic fermentation, (v) stabilization and spoilage risk management, and (vi) by-products and wastewater treatment. The presence of proper yeast or bacterial strains, the management and timing of inoculation of starter cultures, and some appropriate technological modifications that favor selected microbial activities can lead to several positive effects, including (among other) energy savings, reduction of chemical additives such as sulfites, and reuse of certain residues. Full article

(This article belongs to the Special Issue Microbial Dynamics in Wine Production)

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