The selection of new yeast strains could lead to improvements in bioethanol production. Here, we have studied the fermentative capacity of different auxotrophic mutants of Saccharomyces cerevisiae, which are routinely used as hosts for the production of heterologous proteins. It has recently been found that these strains exhibit physiological alterations and peculiar sensitivities with respect to the parental prototrophic strains from which they derive. In this work the performance of auxotrophic S. cerevisiae CEN.PK strains was compared to the corresponding prototrophic strain, to S. cerevisiae T5bV, a strain isolated from grape must and to another auxotrophic strain, S. cerevisiae BY4741. The results indicate that the fermentative capacity of strains grown in 2% glucose was similar in all the strains tested. However, in 15% initial glucose, the auxotrophic strains exhibited a more than doubled ethanol yield on biomass (10 g g^-1_dw) compared to the prototrophic strains (less than 5 g g^-1_dw). Other tests have also evidenced that in medium depletion conditions, ethanol production continues after growth arrest. Our results highlight the need of specific applied research on the use of auxotrophic strains of S. cerevisiae in the production of ethanol in both homogeneous and heterogeneous phases (immobilized systems). The higher ethanol yield relative to biomass would be specially advantageous in immobilized cell systems, as a reduced yeast biomass could greatly reduce the mass transfer limitations through the immobilization matrix.