Recently, non-Saccharomyces yeast have become very popular in wine and beer fermentation. Their interesting abilities introduce novel aromatic profiles to the fermented product. In this study, screening of eight non-Saccharomyces yeast (Starmerella bombicola, Lindnera saturnus, Lindnera jadinii, Zygosaccharomyces rouxii, Torulaspora delbrueckii, Pichia kluyveri, Candida pulcherrima, and Saccharomycodes ludwigii) revealed their potential in non-alcoholic beer production. Conditions for non-alcoholic beer production were optimised for all strains tested (except T. delbrueckii) with the best results obtained at temperature 10 to 15 °C for maximum of 10 days. Starmerella bombicola, an important industrial producer of biosurfactants, was used for beer production for the first time and was able to produce non-alcoholic beer even at 20°C after 10 days of fermentation. Aromatic profile of the beer fermented with S. bombicola was neutral with no negative impact on organoleptic properties of the beer. The most interesting organoleptic properties were evaluated in beers fermented with L. jadinii and L. saturnus, which produced banana-flavoured beers with low alcohol content. This work confirmed the suitability of mentioned yeast to produce non-alcoholic beers and could serve as a steppingstone for further investigation.

The popularity of non-alcoholic beers has been increasing over the past few years. Maltose-negative strains of different genera are frequently used to obtain beers of low alcohol content. S. cerevisiae hybrids with other Saccharomyces species offer interesting inherited flavour characteristics; however, their use in non-alcoholic beer production is rare. In this work, we constructed six hybrids of maltose-negative S. cerevisiae parental strains (modified to produce higher amounts of organic acids) and S. mikatae (wild-type). Growth behaviour, osmotolerance and fermentation features of the offspring were compared with parental strains. One hybrid with mitochondrial DNA inherited from both parents was used to produce non-alcoholic beer in which organic metabolites were evaluated by HPLC and HS-SPME-GC-MS. This hybrid produced non-alcoholic beer (≤0.05% (v/v)) with an increased organic acid content, just as its parent S. cerevisiae, but without producing increased amounts of acetic acid. The beer had a neutral aromatic profile with no negative off-flavours, similar to the beer produced by the parent S. mikatae, which was used for the first time to produce non-alcoholic beer. Overall, both parents and hybrid yeast produced non-alcoholic beers with increased amounts of higher alcohols compared with esters.

The development of non-alcoholic beer (NAB) with health benefits, using non-conventional potential probiotic yeasts, offers an interesting alternative to standard NAB brewing strategies. In this study, potential probiotic non-Saccharomyces yeasts Pichia manshurica, Kluyveromyces lactis, and Kluyveromyces marxianus, along with commercial probiotic yeast Saccharomyces boulardii, were characterised and tested for functional NAB production, whereas P. manshurica was used in NAB production for the first time. Growth and viability were assessed across a range of temperatures, pH, and iso-α-bitter acids. The tested yeasts withstood conditions typical of the beer matrix and human digestive tract and had a positive phenolic off-flavour phenotype. Two strains, K. lactis and K. marxianus, showed strong β-glucosidase activity, which may enhance beverage aroma complexity. Ethanol levels in beers fermented with non-Saccharomyces yeasts remained below the NAB limit (≤0.5% v/v). An analysis of volatile organic compound profiles revealed the potential of these yeasts to produce higher alcohols and esters valuable from a brewer’s perspective. This study provides valuable insight into novel probiotic fermentations and the potential application of unconventional yeasts in functional, aromatic, and health-oriented non-alcoholic beverages.

The development of innovative non-alcoholic beer (NAB) with health benefits, with the use of non-conventional potential probiotic yeasts offers an interesting alternative to standard NAB brewing strategies. In this study, potential probiotic non-Saccharomyces yeasts Pichia manshurica, Kluyveromyces lactis, and Kluyveromyces marxianus, along with commercial probiotic yeast Saccharomyces boulardii, were characterized and tested for functional NAB production, whereas P. manshurica was used in NAB production for the first time. Growth and viability were assessed across a range of temperatures, pH, and iso-α-bitter acids. The tested yeasts withstood conditions typical of the beer matrix and human digestive tract and had positive phenolic off-flavour phenotype. Two strains, K. lactis and K. marxianus, showed strong ß-glucosidase activity, which may enhance beverage aroma complexity. Ethanol levels in beers fermented with non-Saccharomyces yeasts remained below the NAB limit (≤0.5% v/v). Analysis of volatile organic compound profiles revealed the potential of these yeasts to produce higher alcohols and esters valuable from a brewer’s perspective. Sensorial evaluation of prepared NAB produced with potential probiotic non-Saccharomyces yeasts revealed floral and clove-like aroma traces. This study provides valuable insight into novel probiotic fermentations and the potential application of unconventional yeasts in functional, aromatic, and health-oriented non-alcoholic beverages.