We aimed to identify the best yeast or mixture of native yeasts for initiating fermentation during artisan production of mezcal, in an effort to standardize thefinal product. Three native yeasts were identified as good candidates individually. The mixture composed of 75%S. cerevisiaeITD-00185 and 25% T. delbrueckiiITD-00014a showed the best performance overall. This mixture produced mezcal with a high yield and richness of volatile compounds, particularly esters and terpenes, as reflected in its better acceptability in the sensory tests.S. cerevisiae provides the ability to produce ethanol rapidly and in large quantities. However, in terms of compounds that provideflavor and aroma, the mezcal composition was improved by the presence ofT. delbrueckii. This mixture of yeasts can potentially be used as a basis to develop a supply product for the industrial production of mezcal and, perhaps, other spirits from different agave species.