Short communicationOccurrence and enological properties of two new non-conventional yeasts (Nakazawaea ishiwadae and Lodderomyces elongisporus) in wine fermentations
Introduction
The use of metagenomics in wine science has revealed a microbial diversity higher than expected both in the vineyard and in the winery (reviewed in Belda et al., 2017a). The microbial complexity of wine fermentations is key to understanding the qualitative determinants when turning a grape must into wine (Bokulich et al., 2016). From spoilage species to wine enhancers, a great diversity of taxa, and thus metabolisms (metabolic routes and regulation patterns), have attracted microbiologists to the field of wine science.
Some decades ago, ‘non-Saccharomyces’ was a term usually associated with wine spoilage; mainly due to increases in volatile acidity (due to the presence of certain apiculate yeasts) or, more particularly, with the production of non-desired compounds such as volatile phenols (i.e. from Brettanomyces species) (Amerine and Cruess, 1960; Le Roux et al., 1973; Rankine, 1972; Van der Walt and Van Kerken, 1958; Van Zyl and Du Plessis, 1961). In the last two decades, a large number of studies reported the metabolic/enzymatic potential of certain non-Saccharomyces yeasts and their potential role for improving some technological and sensorial aspects of wine (reviewed in Balmaseda et al., 2018; Belda et al., 2017b; Ciani et al., 2006; Jolly et al., 2014). Thus, the deliberate use of non-Saccharomyces yeasts as biological tools (inoculum) in wine fermentations has become a trend in modern wine industry. Nowadays, several strains of non-Saccharomyces species are produced commercially (mainly pertaining to Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima and Pichia kluyveri species). These species are used in winemaking with objectives such as: i) increasing the varietal aroma fraction of wines (Belda et al., 2017c; Ruiz et al., 2018; Sadoudi et al., 2012); ii) controlling wine acidity (Gobbi et al., 2013); iii) improving color extraction and mouthfeel properties (Belda et al., 2016a; Lleixà et al., 2016); iv) reducing the ethanol content of wines (Contreras et al., 2014); and, more recently, v) to improve foaming properties of sparkling wines (Medina-Trujillo et al., 2017a).
During the last years, the number of scientific works exploring the use of the above-mentioned species for understanding their enological potential has increased. However, none or at most very few new species has been explored apart from those already reported in the initial works or in classical reviews in the field of more than a decade ago. These include Candida, Debaryomyces, Hanseniaspora/Kloeckera, Hansenula, Metschnikowia, Pichia, Schizosaccharomyces, Torulaspora and Zygosaccharomyces species (Ciani et al., 2006; Esteve-Zarzoso et al., 1998).
The first works using Next Generation Sequencing (NGS) technology to explore the complexity of wine fermentations were carried out by Bokulich et al. (2012) and Bokulich and Mills (2013) for bacterial and fungal communities, respectively. For mycobiome studies, several papers have demonstrated the great diversity of yeast species in wine fermentations and its dependence with vineyard terroir aspects and the enological practices applied in the winery (Bokulich et al., 2014, Bokulich et al., 2016; Grangeteau et al., 2017; Portillo and Mas, 2016). Nevertheless, the microbial diversity detected by NGS analysis is rarely confirmed by culture-dependent methods, and even less by physiological characterization after isolation.
In this work, we reported the incidence in wine samples of two non-Saccharomyces yeast species, pertaining to non-conventional genus for wine environment (Nakazawaea ishiwadae and Lodderomyces elongisporus). Although periodically mentioned in scientific literature associated with wine-related environments, there is no information about their enological properties. One strain of each species was isolated from wine samples, and, after a basic characterization of some technological properties, the enological potential of the mentioned strains was studied in laboratory scale wine fermentations. After this first report, it is necessary to increase the number of N. ishiwadae and L. elongisporus wine strains for further genomic, phenotypic and enological studies for the understanding of their potential intra-specific diversity.
Section snippets
Survey of N. ishiwadae and L. elongisporus occurrence in wine yeast populations
The occurrence and relative abundance of N. ishiwadae and L. elongisporus in wines were determined from an extensive survey of 429 wine samples (coming from different winemaking stages, as is summarized in Table S1, detailed in Table S2). The mycobiome profile of the 429 wines was analyzed by Next Generation Sequencing ITS-amplicon metabarcoding, using WineSeq technology (patent number: Patent WO2017096385), in a MiSeq® Sequencer (Illumina Inc., San Diego, Ca, USA). Considering the total fungal
Occurrence and abundance N. ishiwadae and L. elongisporus in wine
Fig. 1 shows that both N. ishiwadae and L. elongisporus are rare species in wine fermentations. Based on a survey of 429 wine fermentation samples, taken from different fermentation stages, only 5.83% and 20.05% of the samples showed L. elongisporus and N. ishiwadae, respectively, in relative abundance values higher than 0.01% (Table S1). It is notable that both species could be detected throughout the wine production process, from grape juice to wine under maturation. It is especially notable
Conclusions
The growing number of scientific works using NGS technology to explore the microbial complexity of wine fermentations should be the basis for discovering the role of non-conventional species in wine ecosystem and the final flavor properties of wines. In this work, a first enological characterization has been performed for 2 strains of the non-conventional yeasts N. ishiwadae and L. elongisporus species and reporting some interesting features, starting from their ability to grow in winemaking
Declaration of Competing Interest
Alberto Acedo is an employee of Biome Makers and Ignacio Belda developed part of this work as employee of Biome Makers (granted by Spanish Ministry of Economy, Industry and Competitiveness).
Acknowledgements
Funding for the research in this paper was provided by Biome Makers Spain S.L. under the framework of the MINECO-financed project IDI-20180120 (CDTI, Spanish Ministry of Economy, Industry and Competitiveness, Spain). Javier Ruiz acknowledges to the Complutense University of Madrid for his doctoral grant (CT17/17-CT18/17) and Ignacio Belda acknowledges for his postdoctoral grant (PTQ-16-08253) provided by the Spanish Ministry of Economy, Industry and Competitiveness.
References (55)
- et al.
The Technology of Winemaking
(1960) - et al.
Quantification of major grape polysaccharides (Tempranillo V.) released by maceration enzymes during the fermentation process
Anal. Chim. Acta
(2004) - et al.
Non-Saccharomyces in wine: effect upon Oenococcus oeni and malolactic fermentation
Front. Microbiol.
(2018) - et al.
A dynamic approach to predicting bacterial growth in food
Int. J. Food Microbiol.
(1994) - et al.
Dynamic analysis of physiological properties of Torulaspora delbrueckii in wine fermentations and its incidence on wine quality
Appl. Microbiol. Biotechnol.
(2015) - et al.
Selection and use of pectinolytic yeasts for improving clarification and phenolic extraction in winemaking
Int. J. Food Microbiol.
(2016) - et al.
Unraveling the enzymatic basis of wine “flavorome”: a phylo-functional study of wine related yeast species
Front. Microbiol.
(2016) - et al.
From vineyard soil to wine fermentation: microbiome approximations to explain the “terroir” concept
Front. Microbiol.
(2017) - et al.
Microbial contribution to wine aroma and its intended use for wine quality improvement
Molecules
(2017) - et al.
Influence of Torulaspora delbrueckii in varietal thiol (3-SH and 4-MSP) release in wine sequential fermentations
Int. J. Food Microbiol.
(2017)
Impact of mixed Torulaspora delbrueckii-Saccharomyces cerevisiae culture on high-sugar fermentation
Int. J. Food Microbiol.
Physiological features of Schizosaccharomyces pombe of interest in making of white wines
Eur. Food Res. Technol.
Improved selection of internal transcribed spacer-specific primers enables quantitative, ultra-high-throughput profiling of fungal communities
Appl. Environ. Microbiol.
Next-generation sequencing reveals significant bacterial diversity of botrytized wine
PLoS One
Microbial biogeography of wine grapes is conditioned by cultivar, vintage, and climate
Proc. Natl. Acad. Sci. U. S. A.
Associations among wine grape microbiome, metabolome, and fermentation behavior suggest microbial contribution to regional wine characteristics
mBio
Identification and characterization of complex bioactive oligosaccharides in white and red wine by a combination of mass spectrometry and gas chromatography
J. Agric. Food Chem.
Species richness influences wine ecosystem function through a dominant species
Fungal Ecol.
Protein fraction analysis of white wine by FPLC
Am. J. Enol. Vitic.
Divalent cation interactions with oligogalacturonides
J. Agric. Food Chem.
Fermentation behavior and metabolic interactions of multistarter wine yeast fermentations
Int. J. Food Microbiol.
Non-conventional yeast species for lowering ethanol content of wines
Front. Microbiol.
Quantitative and qualitative composition of yeast microbiota in spontaneously fermented grape musts obtained from cool climate grape varieties ‘Rondo’ and ‘Regent’
FEMS Yeast Res.
Evaluation of non–Saccharomyces yeasts for the reduction of alcohol content in wine
Appl. Environ. Microbiol.
Characterization of natural haze protein in Sauvignon white wine
Food Chem.
The role of non-Saccharomyces yeasts in industrial winemaking
Int. Microbiol.
Degradation of malic and tartaric acids by high density cell suspensions of wine yeasts
Food Microbiol.
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These authors contributed equally to this work.