Compositions and methods are provided for enhanced production of ethanol from fermentation of tobacco biomass. Nicotine resistant microorganisms are provided, as well as methods for making these nicotine resistant microorganisms. A biologically pure culture is provided of a nicotine resistant Saccharomyces cerevisiae strain or a mutant thereof having all the identifying characteristics thereof. Methods are provided for producing ethanol from fermentation of tobacco biomass in which the nicotine resistant microorganisms are used in the fermentation of the tobacco biomass, wherein a higher amount of ethanol can be produced from the fermentation. The nicotine resistant yeast strains of the present disclosure can improve ethanol production in tobacco biomass extract fermentations and shorten the fermentation time.