AbstractEmerging low-emission production technologies make ethanol an interesting substrate for yeast biotechnology, but information on growth rates and biomass yields of yeasts on ethanol is scarce. Strains of 52 Saccharomycotina yeasts were screened for growth on ethanol. The 21 fastest strains, among which representatives of the Phaffomycetales order were overrepresented, showed specific growth rates in ethanol-grown shake-flask cultures between 0.12 and 0.46 h-1. Seven strains were studied in aerobic, ethanol-limited chemostats (dilution rate 0.10 h-1).Saccharomyces cerevisiaeandKluyveromyces lactis, whose genomes do not encode Complex-I-type NADH dehydrogenases, showed biomass yields of 0.59 and 0.56 gbiomassg-1, respectively. Different biomass yields were observed among species whose genomes do harbour Complex-I genes:Phaffomyces thermotolerans(0.58 g g-1),Pichia ethanolica(0.59 g g-1),Saturnispora dispora(0.66 g g-1),Ogataea parapolymorpha(0.67 g g-1), andCyberlindnera jadinii(0.73 g g-1). The biomass yield ofC. jadinii, which also showed the highest biomass protein content of these yeasts, corresponded to 88% of the theoretical maximum achieved when growth is limited by assimilation rather than by energy availability. This study indicates that energy coupling of mitochondrial respiration and its regulation are key factors for selecting and improving yeast strains for ethanol-based processes.