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Activity Funded

SMARTWINE 2.0 - Tailoring 2-in-1 new Yeast starter cultures for production of distinct "greenly-preserved" wines

SMARTWINE 2.0 – Desenvolvimento de Leveduras starter para a produção de vinhos estilísticos sustentavelmente preservados

Reference
2022.06777.PTDC
Project Start Date
2023-03-01
Project End Date
2026-12-31
Principal Investigator
Coinvestigator
Scientific Area
Agricultural sciences
Funding Program
Concurso de Projetos de I&D em Todos os Domínios Científicos - 2022 - ICDT

Abstract

The essential role of the “microbial terroir” (comprised by the microbial fingerprint of a given must, typically composed by multiple bacteria and Non-Saccharomyces yeasts-NSYs) in shaping the properties of wines is becoming increasingly recognized and intimately linked with authenticity and stylistic properties. Indigenous NSYs, formerly seen as spoilage, are now demonstrated powerful bio-flavouring agents, not only for wines, but also for beers and other fermented products. Integrating results from molecular OMICS approaches with extensive characterization of mixed fermentations using NSYs, the consortium leading this proposal has been providing seminal knowledge concerning biology and physiology of these species (specially those belonging to the Saccharomycodeaceae family) and how these interact with Saccharomyces cerevisiae, the leading species in vinification. This proposal roots on the results of SMARTWINE, a previous research project led by this consortium that has produced, among other things, a set of strains capable of being used as starter cultures to improve wine aroma. Leveraging on these results we know aim to develop the concept of “bi-functional” starter cultures, aiming for NSYs with enhanced bio-flavouring potential but also with the ability to control  the activity of spoilage Yeasts (SPYs). Brettanomyces bruxellensis, Saccharomycodes ludwigii and Zygosaccharomyces bailii are prominent wine spoilage species and will be our target SPYs. The fact that NSY species described to have higher bio-flavouring potential coincide with those in which some antagonistic activity against different spoilage microbes (including some SPYs) has been described, reinforces our hypothesis that it will be possible to develop bi-functional strains. To achieve that, an integrated workplan including 5 experimental tasks distributed among the 4 teams, is proposed. In T2 we expect to obtain the portrait of more prevalent SPYs in Portugal by collecting and profiling isolates from different sources (musts, cellars and wines) and obtained across different wine producing regions. The strains will be phenotyped for their spoilage potential and subjected to comparative genomic analyses to establish relevant genotype-ecotype-phenotype associations relevant within the context of wine spoilage. In T3 we will profile two cohorts of available NSY strains (already phenotyped for bio-flavouring potential) to search for those showing higher bio-flavouring potential and higher biocontrol against SPYs. High-throughput screenings coupled with fermentation trials in synthetic medium will be used to select the best performing NSY strains, which will be used for downstream analyses. In T4 we expect to obtain an in-depth molecular view of the NSY/SPY interference resorting to comparative genomic analyses and co-culture transcriptomic profilings coupled with PhenOMIC analyses to identify genes/pathways mediating competitiveness of the two Yeasts while in co-culture and, consequently, map the genetic traits determining biocontrol and bio-flavouring activity, essential for a future legally acceptable design of best-performing strains with dual behavior. In T5 we aim to accelerate the development of dual best-performing NSY strains resorting to adaptive laboratory evolution of strains selected in T3 to obtain non-GMO strains with higher resilience to ethanol and SO2 (two traits that limit NSY performance in wine fermentation) while maintaining anti-SPY activity. T6 is an integration task in which we will use the NSY strains selected in T3 or evolved in T5 as starters, together with S. cerevisiae, to perform fermentations of natural grape musts. The effect of the presence of the selected/improved NSYs in the wine aroma profile (using chemical profiling of volatiles and sensory analysis); in the dynamics of musts’ biodiversity (using metagenomics), including of SPYs; and in microbiological stability of the musts will be monitored in realistic conditions. On the overall the results of the herein proposed project will tailor a set of novel starter cultures that can be used to produce innovative wines with enhanced stylistic properties and exposed to lower levels of chemical preservatives, two aspects that will increase significantly competitiveness of Portuguese winemakers in the international demanding market. Fundamental knowledge concerning multi-microbial ecological interactions involving Yeasts relevant within the context of wine fermentation will be also obtained, including paramount aspects of the biology and physiology of a number of relevant NSY species. This knowledge is expected to advance the exploration of these NSY species in other processes where the value of non-conventional Yeasts is growing prominently such as bio-flavouring or in microbial production of add-value chemicals.

Institutions

Main Institutions

  • Associação do Instituto Superior Técnico para a Investigação e Desenvolvimento (IST-ID)

Other Institutions

  • Universidade Católica Portuguesa (UCP)
  • Universidade de Trás-os-Montes e Alto Douro (UTAD)
  • Universidade de Lisboa Centro de Ecologia Evolução e Alterações Ambientais (CE3C/FC/ULisboa)

Funding 249.370,55 €

Fundação para a Ciência e a Tecnologia (FCT) - Portugal

249.370,55 €