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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.

Soaking is an important process in Chinese rice wine brewing. In this study, the influence of vacuum soaking on Chinese rice wine production was investigated. Rice subjected to a 1-h vacuum soaking process or a traditional 2-days soaking process was steamed and fermented. Our results showed that vacuum soaking led to similar absorbed water but less leached solids compared with traditional soaking and showed limited influence on the physiochemical characteristics of steamed rice. Monitoring of the fermentation process suggested that the content of amino acid nitrogen in the vacuum-soaked group was significantly higher than that of the traditional-soaked group, while the other indexes were similar. The detection of flavor substances in the rice wine indicated that the contents of organic acids and free amino acids were higher in the vacuum-soaked group, and the main kinds of volatile flavor compounds from the two groups were similar. Additionally, sensory evaluation reflected that the rice wine brewed with rice subjected to either of the two different soaking treatments had similar sensory performances. Our research indicated that vacuum soaking could effectively shorten the soaking time of rice in Chinese rice wine production, thus shortening the brewing cycle without sacrificing the quality of the rice wine., (Copyright © 2020 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The use of selected Saccharomyces and non-Saccharomyces strains as mixed starters in winemaking would have advantages over the traditional spontaneous fermentation, producing wines with predictable and desirable characteristics. The aim of this study was to evaluate the impact of metabolic interactions between Patagonian indigenous Saccharomyces cerevisiae MMf9 and beta-glucosidase producer Candida pulcherrima V(6) strains on alcoholic fermentation behaviour and wine aroma Three inoculation strategies, simultaneous, sequential and final, were assayed at laboratory-scale fermentations using Muscat d'Alexandrie grape juice as substrate. The fermentation and yeast growth kinetics as well as the physicochemical and the sensory quality of wine were evaluated. Results evidenced that the sequential inoculation is the most adequate strategy of strains combination. The kinetic behaviour of sequential fermentation was similar to a successful spontaneous fermentation and its wine showed differential aromatic quality as evidenced through PC analysis using physicochemical and aromatic composition data. This wine presented the highest total concentration of higher alcohol, esters and terpenols and the strongest fruity and floral aroma., (Copyright 2010 Elsevier B.V. All rights reserved.)

The aim of this work was to evaluate the fate of ochratoxin A (OTA) content from must to wine during the red wine making process in a pilot scale vinification. The study was done using musts obtained from two red grape varieties (Bonarda and Tempranillo) artificially contaminated with two OTA levels. A duplicate set of tanks of 100 I each was established for each must (Bonarda and Tempranillo). The fermentations were initiated by inoculation of two Saccharomyces spp. strains having different fermentation performance. The must from the Tempranillo variety was spiked with 6 microg/I of OTA while that from the Bonarda variety with 0.3 microg/I of the toxin. Samples were collected at different stages of the process. Performance of the alcoholic and malolactic fermentations was monitored. Titratable and volatile acidity, pH, ethanol, sugar and SO2 concentrations were determined following standard methods proposed by the Office International de la Vigne et du Vin (OIV). OTA analysis was done by HPLC. Detection and quantification limits were 0.01 and 0.1 ng/ml, respectively. The OTA levels during the vinification trials dropped to an average of about 86.5%. The type of Saccharomyces strains used showed no effect on toxin reduction.

The impact of mixed cultures of Hanseniaspora osmophila and Saccharomyces cerevisiae with different initial yeast ratios on wine composition has been examined. The mixed culture significantly affected sugar consumption, the main enological parameters and ester concentrations, with the exception of glycerol, isoamyl acetate and diethyl succinate levels. Remarkably, in wines obtained with mixed cultures the concentration of 2-phenylethyl acetate was approximately 3- to 9-fold greater than that produced by S. cerevisiae pure culture. Moreover sensory evaluation revealed a stronger fruity character in wines fermented with mixed cultures than in control wines. Independently of the mixed culture used, all wines showed concentrations of acetic acid and ethyl acetate within the ranges described for wines. Our data suggest that a mixed culture of H. osmophila and S. cerevisiae can be used as a tool to increase 2-phenylethyl acetate in wine and that its concentration can be controlled by modulating the initial yeast ratio in the culture.

The main objective of this study was to investigate the effect of different Saccharomyces cerevisiae wine yeast strains on the concentration of aroma-enhancing volatile thiols and fermentation metabolites in Sauvignon Blanc wine. Seven commercial wine yeast strains were selected based on their putative ability to modulate the concentrations of the fruity volatile thiols, 4-mercapto-4-methylpentan-2-one (4MMP), 3-mercapto-hexanol (3MH) and 3-mercapto-hexylacetate (3MHA). Each of these yeasts was used to produce Sauvignon Blanc wines under controlled conditions, in triplicate, in 20-L quantities. The levels of 4MMP, 3MH and 3MHA in these wines were quantified using the p-hydroxymercuribenzoate method. In addition, a total of 24 volatile yeast-derived fermentation aroma compounds were also quantified using headspace solid-phase microextraction coupled with gas chromatography mass spectrometry (HS-SPME-GC-MS). Formal sensory analysis was conducted by 12 trained assessors and, additionally, a panel of 24 experienced wine industry professionals assessed the wines and ranked them in order of preference. The results indicated that the yeast strains varied significantly in terms of their capabilities to (i) produce volatile thiols and fermentation metabolites; and (ii) to modulate the varietal characters of Sauvignon Blanc wine. Yeast strains that produced the highest levels of volatile thiols were responsible for wines with the highest perceived intensity of fruitiness, and these wines were preferred by the tasting panels. While the 'green' characters in Sauvignon Blanc wines can be manipulated through vineyard management, the 'tropical fruity' characters appear to be largely dependent on the wine yeast strain used during fermentation. Therefore, the choice of yeast strain offers great potential to modulate wine aroma profiles to definable styles and predetermined consumer market specifications.

In winemaking, spontaneous grape must fermentations have been replaced by inoculation of commercial active dry wine yeast (ADWY). Yeast rehydration is the key to avoiding stuck and sluggish fermentations. Despite the importance of this step, not enough is known about what this process implies for winemaking as a whole or about what kind of practices could help to improve it. The main aim of this study is to determine the best yeast rehydration conditions for ensuring good cell viability and vitality before inoculation into the must. The experimental rehydration media in this study can be divided into four groups: carbon and nitrogen compounds, metallic ions, oxidant and antioxidant agents, and membrane fluidity agents. We studied the biochemical and biophysical behaviour of ADWY after rehydration in the various media under oenological conditions, i.e. incubation at 37 degrees C for 30 min. The viability of rehydrated yeast cells was evaluated by plating, and assessed by fluorescence microscopy and flow cytometry. The vitality of rehydrated cells was estimated by indirect impedance. The rehydrating solution complemented with magnesium provided the best vitality rate because the time taken to reach the activity threshold was cut by two thirds. This improvement was also illustrated by the less time needed to stop the leakage of intracellular compounds during the rehydration process.

Evolution of the microbial composition during the production of Vinsanto wine was investigated under different fermentation conditions to determine their impact on the yeast population and the wine sensorial characteristics. Fermentations were carried out according to the traditional process in 50-l barrels. Different fermentation conditions were applied (yeast inoculum, "mother sediment" addition and temperature) to standardise the Vinsanto production process. Fermentations and products were monitored over the aging period by chemical, microbial and sensory evaluation. The low temperature at the beginning of the fermentations under traditional cellar conditions results in prolonged survival of the non-Saccharomyces yeast. In contrast, Saccharomyces yeast populations dominated throughout the fermentation when the temperature of the cellar was maintained at a constant 16-18 degrees C. Results indicate that inoculation with a commercial yeast strain and fermentation temperature strongly influence the evolution of Vinsanto wine. The "mother sediment" seems to have no direct role as a microbiological starter in Vinsanto production but, despite this, it does have a strong influence on the sensory attributes of the Vinsanto wine. Our work highlights the importance of managing the fermenting microflora to improve the sensorial characteristics of Vinsanto wine.

Interspecific hybrids obtained from flocculent Saccharomyces cerevisiae and non-flocculent Saccharomyces uvarum (S. bayanus var. uvarum) are flocculent to a higher degree and have the capacity to ferment well at both low and high temperatures, from 6 to 36 degrees C. These interspecific hybrids generate minor compounds of fermentation in intermediate quantities with respect to the parent strains and produce high-quality sparkling wines, which are not inferior to those of the parent strains. For these reasons, interspecific strains are particularly adapted to production of sparkling wines that are re-fermented in bottles.

To analyse the yeast population diversity during wine fermentations, specific fluorescein-labelled oligonucleotide probes targeted to the D1/D2 region of the 26S rRNA of different yeast species known to occur frequently in this environment were designed and tested with reference strains. The probes were then used to identify wine must isolates and to follow, in combination with plate counts, the evolution of yeast populations in two winery fermentations of white and red grape musts. In both cases, a high diversity of non-Saccharomyces yeast species was detected, including Candida stellata, Hanseniaspora uvarum, H. guilliermondii, Kluyveromyces marxianus, K. thermotolerans and Torulaspora delbrueckii. Some of these species (e.g., K. marxianus, K. thermotolerans and T. delbrueckii) were present in significant amounts during the tumultuous fermentation stage, despite the predominance of Saccharomyces cerevisiae cells following the inoculation of the wine musts with a starter strain. To further clarify the yeast population dynamics at the late phase of the fermentations, and because winery conditions do not allow a reliable control of experimental variables, strains isolated from the industrial musts were used to conduct two laboratory microvinifications in synthetic grape juice, using different ratios of S. cerevisiae/non-Saccharomyces in the inocula. Under these conditions, the results were similar to those obtained in the winery, showing a yeast profile with mixed species throughout the first fermentation stage, i.e. until about 40-50% of the total sugar was consumed. Non-Saccharomyces yeasts were outgrown by S. cerevisiae only after ethanol reached concentrations around 4-5% (v/v), which argues in favour of a potential important role of non-Saccharomyces in the final organoleptic characteristics of the wine.

Penicillium was used to immobilize Saccharomyces cerevisiae, without using physico-chemical external supports, to form yeast biocapsules. The biocapsules, once the Penicillium was killed by the ethanol produced, were used in a grape must fermentation. Must fermentation was carried out for 160 h with the biocapsules and for 300 h with free yeast cells. Acetaldehyde (84 vs. 63 mg/l), isobutanol (217 vs. 194 mg/l), L: -proline (7.7 vs. 6.5 mM: ) and aspartic acid (0.42 vs. 0 mM: ) in final wine were higher with the biocapsules than with free cells.

Methods for immobilizing champagne yeasts, physiological and biochemical characteristics of the immobilized cells, and problems of their utilization in the production of quality champagne wines are reviewed. Studies aimed at the development of efficient biocatalysts for champaignizing wines using bottle fermentation (method champenoise) and tank processing (bulk, or Charmat process), based on the use of immobilized yeast cells, are described. Data on the industrial use of such biocatalysts in countries manufacturing champagne wines are presented. Problems and prospects of further research in this field are discussed.