Brewing and probiotic potential activity of wild yeasts Hanseniaspora uvarum PIT001, Pichia kluyveri LAR001 and Candida intermedia ORQ001

Non-conventional yeasts can be isolated from a wide range of environmental sources and are often found in the beverage industry in mixed fermentations, in which the microbial community is usually not fully known. However, it is important to know the compositions of these starter cultures because in addition to enabling reproducibility during fermentation, other properties can be discovered. Thus, the objective of this work was to identify and characterize non-conventional yeasts isolated from the environment, evaluating their probiotic potential and possible use in beer brewing. Isolates were obtained from flowers, fruits, leaves and mixed-fermentation beers, with the species being identified by PCR. Yeasts with promising activity were evaluated regarding their growth under different pHs, temperature and the presence of organic acids. To explore probiotic potential, in vitro tests were performed for antimicrobial activity and co-aggregation with food-spoiling microorganisms, auto-aggregation and survival in simulated gastrointestinal tract conditions. In this study, Pichia kluyveri (LAR001), Hanseniaspora uvarum (PIT001) and Candida intermedia (ORQ001) were selected among 20 isolates for further study. P. kluyveri was the only strain that tolerated pH 2.5. Lactic acid was not inhibitory, but acetic acid and incubation at 37 °C had partially inhibitory effects on yeast growth. All yeasts tolerated α-acids from hops and up to 1% NaCl. Our results also suggest that these isolates are able to adhere to intestinal cells and positively influence the host to combat pathogens, as they showed auto-aggregation rates > 99% and antagonistic activity to pathogenic bacteria. The yeasts tolerated gastric environment conditions, but were more sensitive to pancreatic conditions. We conclude that these non-conventional yeasts have probiotic potential and promising application in beer fermentation.