Non-Saccharomyces yeasts have aroused interest in brewing science as an innovative and seminal way of creating new beer flavors. A screening system for potential brewing strains of non-Saccharomyces yeasts was set up to investigate the yeast's utilization of wort sugars and to examine the effect of hop acids as well as ethanol on the growth of different yeast strains. Additionally, phenolic off-flavor (POF) and sensory odor tests of fermented wort samples were performed. The promising strains were further investigated for their propagation ability and for following fermentation trials. The produced beers were analyzed for secondary metabolites, ethanol content and judged by trained panelists. Subsequently to the screening, it was discovered that among the 110 screened yeast strains, approx. 10 strains of the species Saccharomycopsis fibuligera, Schizosaccharomyces pombe and Zygosaccharomyces rouxii generate promising fruity flavors during fermentation and were able to metabolize maltose and maltotriose as a prerequisite for the production of alcoholic beers. Consequently, the screening method described in this study makes it possible to investigate a tremendous number of different non-Saccharomyces yeasts and to test their brewing ability in a relatively short period of time.Fermentation 2019, 5, 101 2 of 23 brewer's wort to a desirable beer. Ravasio et al. also evaluated the fermentation and aroma profile of 60 different non-Saccharomyces strains in 2018. The applied yeasts were cultured in a medium based on glucose and the resulting volatile compounds were detected by gas chromatography/mass spectrometry (GC/MS) measurement. Only the promising species were additionally analyzed for their maltose utilization on serial-dilution plate assays [7].The study presented here includes screening the brewing ability of 110 non-Saccharomyces strains with optimized screening conditions as the metabolism of the full range of main wort carbohydrates (glucose, fructose, sucrose, maltose and maltotriose [8]) were taken into account. As maltose and maltotriose represent more than 80% of the total carbohydrates in brewer's wort [9], the ability to ferment these two substances is essential for a fast, complete and predictable fermentation [10]. Melibiose was additionally included into the screening, as a study of Wickerham indicated that bottom-fermenting yeasts are able to metabolize this type of sugar whereas top-fermenting yeasts do not [11]. Furthermore, part of the first screening step looked at the effect of hop iso-α-and β-acids as well as ethanol on the growth of the yeast strains to determine whether there are any existing resistances at certain concentrations that would restrict the production of a conventional beer. As hop acids have antimicrobial properties and β-acids, in particular, were reported to have an even stronger antimicrobial effect than iso-α-acids [12,13], it is necessary to test the yeast's tolerance to these acids. Although a conventional Pils has up to 38 IBU, which is approximately comparable to 38 mg ...