Utjecaj kvasaca Saccharomyces cerevisiae i Lachancea thermotolerans na kemijski sastav vina sorte ´Frankovka´

Cilj ovog istraživanja bio je utvrditi učinke primjene različitih sojeva kvasaca Saccharomyces cerevisiae i Lachancea thermotolerans na osnovni kemijski sastav, koncentracije pojedinačnih organskih kiselina, fenolnih spojeva i glicerola u vinu. U istraživanju provedenom 2018. godine, grožđe sorte ´Frankovka´ korišteno je za fermentacije s komercijalnim sojevima kvasca Saccharomyces cerevisiae (Uvaferm BDX™ i Ionys WF™) i sekvencijalnu inokulaciju s Lachancea thermotolerans (Laktia™) u ulozi startera, te Uvaferm BDX™ kvasca, čija je uloga bila provesti fermentaciju do kraja. Rezultati potvrđuju utjecaj kvasca Uvaferm BDX™ na pojačanu ekstrakciju antocijana i tanina, te utjecaj kvasca Ionys™ na povećanu sintezu glicerola u vinu. Iako je došlo do zastoja u fermentaciji, potvrđen je učinak sekvencijalne inokulacije na povećanje ukupne kiselosti te boju i strukturu vina temeljem više koncentracije ukupnih fenola što potencijalno može opravdati njenu primjenu u ekstremno toplim godinama ili toplim klimatima.

An Integrative View of the Role of Lachancea thermotolerans in Wine Technology

The interest in Lachancea thermotolerans, a yeast species with unusual characteristics, has notably increased in all ecological, evolutionary, and industrial aspects. One of the key characteristics of L. thermotolerans is the production of high quantities of lactic acid compared to other yeast species. Its evolution has mainly been driven by the influence of the environment and domestication, allowing several metabolic traits to arise. The molecular regulation of the fermentative process in L. thermotolerans shows interesting routes that play a complementary or protective role against fermentative stresses. One route that is activated under this condition is involved in the production of lactic acid, presenting a complete system for its production, showing the involvement of several enzymes and transporters. In winemaking, the use of L. thermotolerans is nowadays mostly focused in early–medium-maturity grape varieties, in which over-ripening can produce wines lacking acidity and with high concentrations of ethanol. Recent studies have reported new positive influences on quality apart from lactic acid acidification, such as improvements in color, glutathione production, aroma, malic acid, polysaccharides, or specific enzymatic activities that constitute interesting new criteria for selecting better strains. This positive influence on winemaking has increased the availability of commercial strains during recent years, allowing comparisons among some of those products. Initially, the management of L. thermotolerans was thought to be combined with Saccaharomyces cerevisiae to properly end alcoholic fermentation, but new studies are innovating and reporting combinations with other key enological microorganisms such as Schizosaccharomyces pombe, Oenocous oeni, Lactiplantibacillus plantarum, or other non-Saccharomyces.

Use of Lachancea thermotolerans for Biological vs. Chemical Acidification at Pilot-Scale in White Wines from Warm Areas

Climate change is affecting vineyards, resulting in grapes with a low acidity a high pH and sugar at harvest time. The most common procedure so far to improve the acidity and reduce the final pH of wines is to use tartaric acid, but wine can also be acidified microbiologically using Lachancea thermotolerans yeasts, a natural bio-tool that acidifies gradually during the first stage/days of fermentation. Two strains of L. thermotolerans were compared with one Saccharomyces cerevisiae at a pilot-scale under similar fermentation conditions and in duplicate. A sequential inoculation was performed on the third day for the non-Saccharomyces, producing only about 1 g/L of lactic acid, which was suitable for comparison with the Saccharomyces, to which 1.5 g/L of tartaric acid had been added to lower the final pH. The three fermentations ended with a total acidity without significant differences. A significant and normal feature of the L. thermotolerans yeasts is their higher propane-1,2,3-triol production, which was observed in the Laktia yeast, and the acetic acid was <0.3 g/L. The amount of volatile metabolites was generally higher for non-Saccharomyces and the increase was seen in carbonyl compounds, organic acids, lactones, fumaric compounds, and phenols. Finally, the sensory analysis showed that there were hardly any significant differences, even though the non-Saccharomyces had a higher quantity of volatile metabolites, which could lead to a good acceptance of the product, since biological acidification was used, generating a more natural product.

Commercially Available Non-Saccharomyces Yeasts for Winemaking: Current Market, Advantages over Saccharomyces, Biocompatibility, and Safety

About 42 commercial products based on non-Saccharomyces yeasts are estimated as available on the market, being mostly pure cultures (79%), with a predominance of Torulaspora delbrueckii, Lachancea thermotolerans, and Metschnikowia pulcherrima. The others are multi-starter consortia that include non-Saccharomyces/Saccharomyces mixtures or only non-Saccharomyces species. Several commercial yeasts have shown adequate biocompatibility with S. cerevisiae in mixed fermentations, allowing an increased contribution of metabolites of oenological interest, such as glycerol, esters, higher alcohols, acids, thiols, and terpenes, among others, in addition to a lower production of acetic acid, volatile phenols, biogenic amines, or urea. Multi-starter inoculations are also reviewed here, which show adequate biocompatibility and synergy between species. In certain cases, the aromatic profile of wines based on grape varieties considered neutral is improved. In addition, several yeasts show the capacity as biocontrollers against contaminating microorganisms. The studies conducted to date demonstrate the potential of these yeasts to improve the properties of wine as an alternative and complement to the traditional S. cerevisiae.

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

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.

Non-Saccharomyces as Biotools to Control the Production of Off-Flavors in Wines

Off-flavors produced by undesirable microbial spoilage are a major concern in wineries, as they affect wine quality. This situation is worse in warm areas affected by global warming because of the resulting higher pHs in wines. Natural biotechnologies can aid in effectively controlling these processes, while reducing the use of chemical preservatives such as SO2. Bioacidification reduces the development of spoilage yeasts and bacteria, but also increases the amount of molecular SO2, which allows for lower total levels. The use of non-Saccharomyces yeasts, such as Lachancea thermotolerans, results in effective acidification through the production of lactic acid from sugars. Furthermore, high lactic acid contents (>4 g/L) inhibit lactic acid bacteria and have some effect on Brettanomyces. Additionally, the use of yeasts with hydroxycinnamate decarboxylase (HCDC) activity can be useful to promote the fermentative formation of stable vinylphenolic pyranoanthocyanins, reducing the amount of ethylphenol precursors. This biotechnology increases the amount of stable pigments and simultaneously prevents the formation of high contents of ethylphenols, even when the wine is contaminated by Brettanomyces.

Učinak sekvencijalne fermentacije s kvascima Lachancea thermotelerans i Torulaspora delbrueckii na kemijski sastav vina ´Malvazija istarska´

Suočavajući se sa sve drastičnijim utjecajem klimatskih čimbenika na kemijski sastav grožđa, enologija traži i proučava nove metode u tehnologiji proizvodnje vina, posebice bijelih, kako bi se očuvale primarne arome te postigla ravnoteža između alkoholne jakosti i ukupne kiselosti. Kao jedno od rješenja nudi se primjena ne- Saccharomyces kvasaca. U ovom istraživanju analiziran je utjecaj sekvencijalne inokulacije komercijalnih sojeva Torulospora delbrueckii i Lachancea thermotolerans sa sojem kvasca Saccharomycem cerevisiae na vino ´Malvazija istarska´. Istraživanje je obuhvatilo inokulacije mošta s ne-Saccharomyces kvascima, a 48 h kasnije i sa sojem S. cerevisae te kontrolnu varijantu isključivo sa S. cerevisae. Ne-Saccharomyces kvasci utjecali su značajno na koncentraciju alkohola, mliječne kiseline te pH vrijednost. Fermentacija sa S. cerevisiae utjecala je na višu koncentraciju ukupnih aromatskih spojeva u vinu. Intenziteti boje i mirisa najbolje su ocijenjeni u kontrolnom uzorku, a metodom redoslijeda najbolje je rangirana ´Malvazija´ iz tretmana T. delbrueckii/ S. cerevisiae.

Pulsed Electric Fields to Improve the Use of Non-Saccharomyces Starters in Red Wines

New nonthermal technologies, including pulsed electric fields (PEF), open a new way to generate more natural foods while respecting their organoleptic qualities. PEF can reduce wild yeasts to improve the implantation of other yeasts and generate more desired metabolites. Two PEF treatments were applied; one with an intensity of 5 kV/cm was applied continuously to the must for further colour extraction, and a second treatment only to the must (without skins) after a 24-hour maceration of 17.5 kV/cm intensity, reducing its wild yeast load by up to 2 log CFU/mL, thus comparing the implantation and fermentation of inoculated non-Saccharomyces yeasts. In general, those treated with PEF preserved more total esters and formed more anthocyanins, including vitisin A, due to better implantation of the inoculated yeasts. It should be noted that the yeast Lachancea thermotolerans that had received PEF treatment produced four-fold more lactic acid (3.62 ± 0.84 g/L) than the control of the same yeast, and Hanseniaspora vineae with PEF produced almost three-fold more 2-phenylethyl acetate than the rest. On the other hand, 3-ethoxy-1-propanol was not observed at the end of the fermentation with a Torulaspora delbrueckii (Td) control but in the Td PEF, it was observed (3.17 ± 0.58 mg/L).