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Although Saccharomyces cerevisiae is the yeast species predominantly used for alcoholic fermentation, non-Saccharomyces yeast species are also important because they produce secondary metabolites that can contribute to the final flavour and taste of wines. In this study, 37 strains representing seven nonSaccharomyces species were characterised and evaluated for potential use in wine production, as well as for their effects on malolactic fermentation (MLF). Contour-clamped homogeneous electric field (CHEF) gel electrophoresis and matrix-assisted laser desorption ionisation using a time-of-flight mass spectrometer (MALDI-TOF MS) were used to verify species identity and to determine intra-species variation. Extracellular enzyme production, malic acid degradation and the fermentation kinetics of the yeasts were also investigated. CHEF karyotyping and MALDI-TOF MS were useful for identifying and typing Hanseniaspora uvarum, Lachancea thermotolerans, Candida zemplinina (synonym: Starmerella bacillaris) and Torulaspora delbrueckii strains. Only H. uvarum and Metschnikowia pulcherrima strains were found to have β-glucosidase activity. M. pulcherrima strains also had protease activity. Most of the strains showed limited malic acid degradation, and only Schizosaccharomyces pombe and the C. zemplinina strains showed mentionable degradation. In synthetic wine fermentations, C. stellata, C. zemplinina, H. uvarum, M. pulcherrima and Sc. pombe strains were shown to be slow to medium fermenters, whereas L. thermotolerans and T. delbrueckii strains were found to be medium to strong fermenters. The effect of the yeasts on MLF varied, but inhibition was strain dependent.

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Biological antimicrobial compounds from yeast can be used to address the critical need for safer preservatives in food, fruit and beverages. The inhibition of Candida guilliermondii, a common fermented beverage spoilage organism, was achieved using antimicrobial compounds produced by Candida pyralidae KU736785. The antimicrobial production system was modelled and optimised using response surface methodology, with 22.5 ℃ and pH of 5.0 being the optimum conditions. A new concept for quantifying spoilage organism inhibition was developed. The inhibition activity of the antimicrobial compounds was observed to be at a maximum after 17-23 h of fermentation, with C. pyralidae concentration being between 0.40 and 1.25 × 10 CFU ml, while its maximum specific growth rate was 0.31-0.54 h. The maximum inhibitory activity was between 0.19 and 1.08 l contaminated solidified media per millilitre of antimicrobial compound used. Furthermore, the antimicrobial compound formation rate was 0.037-0.086 l VZI ml ACU h, respectively. The response surface methodology analysis showed that the model developed sufficiently described the antimicrobial compound formation rate 1.08 l VZI ml ACU, as 1.17 l VZI ml ACU, predicted under the optimum production conditions.

The use of non-Saccharomyces yeasts to improve complexity and diversify wine style is increasing; however, the interactions between non-Saccharomyces yeasts and lactic acid bacteria (LAB) have not received much attention. This study investigated the interactions of seven non-Saccharomyces yeast strains of the genera Candida, Hanseniaspora, Lachancea, Metschnikowia and Torulaspora in combination with S. cerevisiae and three malolactic fermentation (MLF) strategies in a Shiraz winemaking trial. Standard oenological parameters, volatile composition and sensory profiles of wines were investigated. Wines produced with non-Saccharomyces yeasts had lower alcohol and glycerol levels than wines produced with S. cerevisiae only. Malolactic fermentation also completed faster in these wines. Wines produced with non-Saccharomyces yeasts differed chemically and sensorially from wines produced with S. cerevisiae only. The Candida zemplinina and the one L. thermotolerans isolate slightly inhibited LAB growth in wines that underwent simultaneous MLF. Malolactic fermentation strategy had a greater impact on sensory profiles than yeast treatment. Both yeast selection and MLF strategy had a significant effect on berry aroma, but MLF strategy also had a significant effect on acid balance and astringency of wines. Winemakers should apply the optimal yeast combination and MLF strategy to ensure fast completion of MLF and improve wine complexity.