The Important Contribution of Non-Saccharomyces Yeasts to the Aroma Complexity of Wine: A Review

Non-Saccharomyces yeast plays an important role in the initial stages of a wild ferment, as they are found in higher abundance in the vineyard than Saccharomyces cerevisiae. As such, there has been a focus in recent years to isolate these yeast species and characterize their effect on wine fermentation and subsequent aroma. This effect on wine aroma is often species and strain dependent, as the enzymatic profile of each yeast will determine which aroma compounds are formed as secondary metabolites. Semi-fermentative yeast, such as Hanseniaspora spp., Candida spp. and Metschnikowia pulcherrima, are commonly in high abundance in fresh grape must and have diverse enzymatic profiles, however they show a weak tolerance to ethanol, limiting their impact to the initial stages of fermentation. Fully fermentative non-Saccharomyces yeast, characterized by high ethanol tolerance, are often found at low abundance in fresh grape must, similar to Saccharomyces cerevisiae. Their ability to influence the aroma profile of wine remains high, however, due to their presence into the final stages of fermentation. Some fermentative yeasts also have unique oenological properties, such as Lanchancea thermotolerans and Schizosaccharomyces pombe, highlighting the potential of these yeast as inoculants for specific wine styles.

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Selection Process of a Mixed Inoculum of Non-Saccharomyces Yeasts Isolated in the D.O.Ca. Rioja

Fermentation ◽

10.3390/fermentation7030148 ◽

2021 ◽

Vol 7 (3) ◽

pp. 148

Author(s):

Rocío Escribano-Viana ◽

Lucía González-Arenzana ◽

Patrocinio Garijo ◽

Rosa López ◽

Pilar Santamaría ◽

...

Keyword(s):

Selection Process ◽

Debaryomyces Hansenii ◽

Aroma Compound ◽

Wine Aroma ◽

Candida Spp ◽

Torulaspora Delbrueckii ◽

Mixed Inoculum ◽

Organoleptic Characteristics ◽

Saccharomyces Yeasts ◽

Lachancea Thermotolerans

The use of non-Saccharomyces yeasts in sequential fermentations with S. cerevisiae has been proposed to improve the organoleptic characteristics involved in the quality of wine. The present study set out to select a non-Saccharomyces inoculum from the D.O.Ca. Rioja for use in winemaking. Strains included in the study belonged to Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, Zygosaccharomyces bailii, Williopsis pratensis, Debaryomyces hansenii, Pichia kluyveri, Sporidiobolus salmonicolor, Candida spp., Cryptococcus spp. and two mixed inocula of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio. In the first stage of the process, SO2 resistance and presence of enzymatic activities related to wine aroma and wine color and fining (esterase, esterase-lipase, lipase, leucine arylamidase, valine arylamidase, cystine arylamidase, β-glucosidase, pectinase, cellulose, xylanase and glucanase) were studied. In the later stages, selection criteria such as fermentative behavior, aroma compound production or influence on phenolic compounds were studied in laboratory scale vinifications. Taking into account the results obtained in the different stages of the process, a mixed inoculum of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio was finally selected. This inoculum stood out for its high implantation capacity, the production of compounds of interest such as glycerol and lactic acid and the consequent modulation of wine acidity. Given these characteristics, the selected inoculum is suitable for the production of quality wines.

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Strain-Specific Responses by Saccharomyces cerevisiae to Competition by Non-Saccharomyces Yeasts

Fermentation ◽

10.3390/fermentation7030165 ◽

2021 ◽

Vol 7 (3) ◽

pp. 165

Author(s):

Cristobal A. Onetto ◽

Anthony R. Borneman ◽

Simon A. Schmidt

Keyword(s):

Saccharomyces Cerevisiae ◽

Yeast Species ◽

Microbial Interactions ◽

Fermentation Performance ◽

Competitive Fitness ◽

Sequential Fermentation ◽

Saccharomyces Yeast ◽

Negative Impacts ◽

The Impact ◽

Saccharomyces Yeasts

The use of non-Saccharomyces yeast species generally involves sequential or co-inoculation of a Saccharomyces cerevisiae strain to complete fermentation. While most studies have focused on characterising the impact that S. cerevisiae has on the growth and metabolic activity of these non-Saccharomyces species, microbial interactions work reciprocally. Antagonism or competition of non-Saccharomyces species against S. cerevisiae has been shown to impact subsequent fermentation performance. To date, it remains unclear whether these negative interactions are strain specific. Hence, characterisation of strain-specific responses to co-inoculation would enable the identification of specific S. cerevisiae strain/non-Saccharomyces combinations that minimise the negative impacts of sequential fermentation on fermentation performance. The competitive fitness response of 93 S. cerevisiae strains to several non-Saccharomyces species was simultaneously investigated using a barcoded library to address this knowledge gap. Strain-specific fitness differences were observed across non-Saccharomyces treatments. Results obtained from experiments using selected S. cerevisiae strains sequentially inoculated after Metschnikowia pulcherrima and Torulaspora delbrueckii were consistent with the competitive barcoded library observations. The results presented in this study indicate that strain selection will influence fermentation performance when using non-Saccharomyces species, therefore, appropriate strain/yeast combinations are required to optimise fermentation.

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Effect of Sequential Inoculation with Non-Saccharomyces and Saccharomyces Yeasts on Riesling Wine Chemical Composition

Fermentation ◽

10.3390/fermentation5030079 ◽

2019 ◽

Vol 5 (3) ◽

pp. 79 ◽

Cited By ~ 16

Author(s):

Ophélie Dutraive ◽

Santiago Benito ◽

Stefanie Fritsch ◽

Beata Beisert ◽

Claus-Dieter Patz ◽

...

Keyword(s):

Volatile Compounds ◽

Positive Influence ◽

Wine Industry ◽

Wine Aroma ◽

Wine Quality ◽

Sequential Inoculation ◽

Saccharomyces Yeast ◽

Pichia Kluyveri ◽

Grape Varieties ◽

Saccharomyces Yeasts

In recent years, studies have reported the positive influence of non-Saccharomyces yeast on wine quality. Many grape varieties under mixed or sequential inoculation show an overall positive effect on aroma enhancement. A potential impact by non-Saccharomyces yeast on volatile and non-volatile compounds should benefit the flavor of Riesling wines. Following this trend, four separate sequential fermentations (using the non-Saccharomyces yeasts Torulaspora delbrueckii, Metschnikowia pulcherrima, Pichia kluyveri, and Lachancea thermotolerans with Saccharomyces cerevisiae) were carried out on Riesling must and compared to a pure culture of S. cerevisiae. Sequential fermentations influenced the final wine aroma. Significant differences were found in esters, acetates, higher alcohols, fatty acids, and low volatile sulfur compounds between the different trials. Other parameters, including the production of non-volatile compounds, showed significant differences. This fermentation process not only allows the modulation of wine aroma but also chemical parameters such as glycerol, ethanol, alcohol, acidity, or fermentation by-products. These potential benefits of wine diversity should be beneficial to the wine industry.

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Assessing the Oenological Potential of Nakazawaea ishiwadae, Candida railenensis and Debaryomyces hansenii Strains in Mixed-Culture Grape Must Fermentation with Saccharomyces cerevisiae

Fermentation ◽

10.3390/fermentation6020049 ◽

2020 ◽

Vol 6 (2) ◽

pp. 49 ◽

Cited By ~ 1

Author(s):

Niel van Wyk ◽

Isak S. Pretorius ◽

Christian von Wallbrunn

Keyword(s):

Saccharomyces Cerevisiae ◽

Acetic Acid ◽

Mixed Culture ◽

Debaryomyces Hansenii ◽

Fermentation Performance ◽

Acetate Esters ◽

Grape Must ◽

Saccharomyces Yeast ◽

Grape Must Fermentation

Recently, there has been a growing interest in the role of non-Saccharomyces yeast (NSY) as a coculturing partner with Saccharomyces cerevisiae during grape must fermentation. We investigated three new strains, namely Nakazawaea ishiwadae, Candida railenensis and Debaryomyces hansenii, for their oenological potential in mixed-culture micro-vinifications with S. cerevisiae Vin13 using Muscaris grape must. None of the NSY strains impeded the fermentation performance as all the mixed-culture experiments finished at the same time. Coculturing with N. ishiwadae yielded significantly higher concentrations of ethyl and acetate esters in the final wine product. Apart from higher acetic acid levels, wines produced with C. railenensis and D. hansenii yielded much lower esters concentrations. The concentrations of certain terpenes and norisoprenoids were also significantly modulated in the mixed-culture fermentations. This study reveals the rarely reported species of N. ishiwadae as a promising coculturing partner for increasing aroma-active compounds in a wine.

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Effect of Myclobutanil Pesticide on the Physiological Behavior of Two Newly Isolated Saccharomyces cerevisiae Strains during Very-High-Gravity Alcoholic Fermentation

Microorganisms ◽

10.3390/microorganisms7120666 ◽

2019 ◽

Vol 7 (12) ◽

pp. 666 ◽

Cited By ~ 3

Author(s):

Antonia Terpou ◽

Maria Dimopoulou ◽

Aikaterini Belka ◽

Stamatina Kallithraka ◽

George-John E. Nychas ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Ethanol Production ◽

Substrate Uptake ◽

High Gravity ◽

Protective Agent ◽

Grape Must ◽

Very High Gravity ◽

High Ethanol ◽

Very High ◽

Grape Musts

Yeasts are able to act as biosorbents, as their cell wall includes several components capable of binding organic xenobiotic compounds that can potentially be removed during various fermentation processes. In the present investigation, two novel Saccharomyces cerevisiae strains (LMBF-Y 16 and LMBF-Y-18), previously isolated from grapes, were studied regarding their physiological behavior (dry cell weight—DCW production, substrate uptake, and ethanol and glycerol biosynthesis) during fermentations of grape must, in some cases enriched with commercial glucose and fructose (initial total sugar concentration approximately 150 and 250 g/L, respectively). Myclobutanil (a chiral triazole fungicide broadly used as a protective agent of vine) was also added to the culture media at various concentrations in order to assess the ability of the yeasts to simultaneously perform alcoholic fermentations and detoxify the medium (i.e., to remove the fungicide). In the first set of experiments and for both tested strains, trials were carried out in either 250 mL or 2.0 L agitated shake flasks in either synthetic glucose-based experiments or grape musts. Since the results obtained in the trials where the cultures were placed in 2.0 L flasks with grape musts as substrates were superior in terms of both DCW and ethanol production, these experimental conditions were selected for the subsequent studies. Both strains showed high fermentative efficiency, producing high amounts of DCW (9.5–10.5 g/L) in parallel with high ethanol production, which in some cases achieved values very close to the maximum theoretical ethanol production yield (≈0.49 g of ethanol per g of sugar). When using grape must with initial total sugars at approximately 250 g/L (very high gravity fermentation media, close to winemaking conditions), significantly high ethanol quantities (i.e., ranging between 105 and 123 g/L) were produced. Myclobutanil addition slightly negatively affected sugar conversion into ethanol; however, in all cases, ethanol production was very satisfactory. A non-negligible myclobutanil removal during fermentation, which ranged between 5%–27%, as a result of the adsorptive or degradative capacity of the yeast was also reported. The presence of myclobutanil had no effect on DCW production and resulted in no significant differences in the biosynthesis of glycerol. Therefore, these newly isolated yeast strains could be excellent candidates for simultaneous high ethanol production and parallel pesticide removal in a general biorefinery concept demonstrating many environmental benefits.

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Reciprocal hemizygosity analysis reveals that the Saccharomyces cerevisiae CGI121 gene affects lag time duration in synthetic grape must

G3 Genes|Genome|Genetics ◽

10.1093/g3journal/jkab061 ◽

2021 ◽

Author(s):

Runze Li ◽

Rebecca C Deed

Keyword(s):

Saccharomyces Cerevisiae ◽

Low Temperatures ◽

Lag Time ◽

Lag Phase ◽

Phenotypic Traits ◽

Time Duration ◽

Phase Duration ◽

Grape Must ◽

The Impact ◽

Reciprocal Hemizygosity Analysis

Abstract It is standard practice to ferment white wines at low temperatures (10-18 °C). However, low temperatures increase fermentation duration and risk of problem ferments, leading to significant costs. The lag duration at fermentation initiation is heavily impacted by temperature; therefore, identification of Saccharomyces cerevisiae genes influencing fermentation kinetics is of interest for winemaking. We selected 28 S. cerevisiae BY4743 single deletants, from a prior list of open reading frames (ORFs) mapped to quantitative trait loci (QTLs) on chromosomes VII and XIII, influencing the duration of fermentative lag time. Five BY4743 deletants, Δapt1, Δcgi121, Δclb6, Δrps17a, and Δvma21, differed significantly in their fermentative lag duration compared to BY4743 in synthetic grape must (SGM) at 15 °C, over 72 h. Fermentation at 12.5 °C for 528 h confirmed the longer lag times of BY4743 Δcgi121, Δrps17a, and Δvma21. These three candidate ORFs were deleted in S. cerevisiae RM11-1a and S288C to perform single reciprocal hemizygosity analysis (RHA). RHA hybrids and single deletants of RM11-1a and S288C were fermented at 12.5 °C in SGM and lag time measurements confirmed that the S288C allele of CGI121 on chromosome XIII, encoding a component of the EKC/KEOPS complex, increased fermentative lag phase duration. Nucleotide sequences of RM11-1a and S288C CGI121 alleles differed by only one synonymous nucleotide, suggesting that intron splicing, codon bias, or positional effects might be responsible for the impact on lag phase duration. This research demonstrates a new role of CGI121 and highlights the applicability of QTL analysis for investigating complex phenotypic traits in yeast.

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Application of Autochthonous Yeast Saccharomyces cerevisiae XG3 in Treixadura Wines from D.O. Ribeiro (NW Spain): Effect on Wine Aroma

Fermentation ◽

10.3390/fermentation7010031 ◽

2021 ◽

Vol 7 (1) ◽

pp. 31

Author(s):

Pilar Blanco ◽

María Vázquez-Alén ◽

Teresa Garde-Cerdán ◽

Mar Vilanova

Keyword(s):

Saccharomyces Cerevisiae ◽

Pilot Scale ◽

Sensory Level ◽

Wine Aroma ◽

Nw Spain ◽

Volatile Phenols ◽

Yeast Saccharomyces Cerevisiae ◽

Spontaneous Fermentation ◽

Autochthonous Yeast ◽

Active Dry Yeast

Yeast plays an essential role in winemaking. Saccharomyces cerevisiae strains involved in fermentation determine the chemical and sensory characteristics of wines. S. cerevisiae XG3, isolated in Galicia (NW Spain), has desirable oenological potential, which has been proved at a pilot scale to produce quality wines. This study applies XG3 as active dry yeast at an industrial scale for Treixadura wine elaboration, and compares it with commercial yeast and spontaneous fermentation within three wineries included in Denomination of Origin Ribeiro over two vintages. Fermentations are monitored using conventional methods, and microbiological implantation controls are carried out by mtDNA-RFLPs analysis. Wine basic chemical parameters are determined using OIV official methodology, and volatile aroma compounds are determined by GC-MS. Finally, wine sensory analysis is also performed. S. cerevisiae XG3 shows an acceptable implantation ability—as compared to commercial control strains. The wines from XG3 have a higher total acidity and lower alcohol content. Their volatile composition differs from control wines, since XG3 produces significantly higher concentrations of acetates, volatile acids, esters and volatile phenols, depending on the vintage and winery. However, lower differences are perceived at the sensory level, where fruity and floral descriptors are perceived by the panellists in XG3 wines. Therefore, XG3 constitutes an alternative to differentiate Treixadura wines.

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Learning from 80 years of studies: a comprehensive catalogue of non-Saccharomyces yeasts associated with viticulture and winemaking

FEMS Yeast Research ◽

10.1093/femsyr/foab017 ◽

2021 ◽

Vol 21 (3) ◽

Author(s):

João Drumonde-Neves ◽

Ticiana Fernandes ◽

Teresa Lima ◽

Célia Pais ◽

Ricardo Franco-Duarte

Keyword(s):

Yeast Species ◽

Starter Cultures ◽

Biotechnological Applications ◽

Major Focus ◽

Wine Production ◽

Sampling Studies ◽

Volatile Acidity ◽

Wine Sector ◽

Saccharomyces Yeast ◽

Saccharomyces Yeasts

ABSTRACT Non-Saccharomyces yeast species are nowadays recognized for their impact on wine´s chemical composition and sensorial properties. In addition, new interest has been given to the commercial exploitation of non-Saccharomyces starter cultures in the wine sector. However, over many years, these yeast species were considered sources of contamination in wine production and conservation, mainly due to the high levels of volatile acidity obtained. The present manuscript systematizes 80 years of literature describing non-Saccharomyces yeast species isolated from grapes and/or grape musts. A link between each reference, the accepted taxonomic name of each species and their geographical occurrence is presented, compiling information for 293 species, in a total of 231 citations. One major focus of this work relates to the isolation of non-Saccharomyces yeasts from grapevines usually ignored in most sampling studies, also as isolation from damaged grapes. These particular niches are sources of specific yeast species, which are not identified in most other explored environments. These yeasts have high potential to be explored for important and diversified biotechnological applications.

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High Potential of Pichia kluyveri and Other Pichia Species in Wine Technology

International Journal of Molecular Sciences ◽

10.3390/ijms22031196 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1196

Author(s):

Javier Vicente ◽

Fernando Calderón ◽

Antonio Santos ◽

Domingo Marquina ◽

Santiago Benito

Keyword(s):

Ethanol Yield ◽

Color Stability ◽

Quality Parameters ◽

Wine Industry ◽

Wine Aroma ◽

Wine Quality ◽

Promising Tool ◽

Pichia Kluyveri ◽

The Impact ◽

Fermentative Yeast

The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non-Saccharomyces yeasts such as Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, and Pichia kluyveri have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non-Saccharomyces species. P. kluyveri stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as Saccharomyces cerevisiae is indispensable to achieve a proper fermented alcohol. The impact of P. kluyveri is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no Pichia species other than P. kluyveri is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future.

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Influence of Microencapsulation on Fermentative Behavior of Hanseniaspora osmophila in Wine Mixed Starter Fermentation

Fermentation ◽

10.3390/fermentation7030112 ◽

2021 ◽

Vol 7 (3) ◽

pp. 112

Author(s):

Grazia Alberico ◽

Angela Capece ◽

Gianluigi Mauriello ◽

Rocchina Pietrafesa ◽

Gabriella Siesto ◽

...

Keyword(s):

Yeast Species ◽

Starter Culture ◽

Cell Immobilization ◽

Mixed Fermentation ◽

Wine Composition ◽

Saccharomyces Yeast ◽

Suitable System ◽

Saccharomyces Yeasts ◽

Fermentative Process

In recent years, as a consequence of the re-evaluation of the role of non-Saccharomyces yeasts, several studies have been conducted on the use of controlled mixed fermentations with Saccharomyces and different non-Saccharomyces yeast species from the winemaking environment. To benefit from the metabolic particularities of some non-Saccharomyces yeasts, the management of a non-Saccharomyces strain in mixed fermentation is a crucial step, in particular the use of procedures addressed to increase the persistence of non-Saccharomyces strains during the fermentative process. The use of microencapsulation for cell immobilization might represent a strategy for enhancing the competitiveness of non-Saccharomyces yeasts during mixed fermentation. This study was aimed to assess the fermentative performance of a mixed starter culture, composed by a wild Hanseniaspora osmophila strain (ND1) and a commercial Saccharomyces cerevisiae strain (EC1118). For this purpose, free and microencapsulated cells of ND1 strain were tested in co-culture with EC1118 during mixed fermentations in order to evaluate the effect of the microencapsulation on fermentative behavior of mixed starter and final wine composition. The data have shown that H. osmophila cell formulation affects the persistence of both ND1 and EC1118 strains during fermentations and microencapsulation resulted in a suitable system to increase the fermentative efficiency of ND1 strain during mixed starter fermentation.

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