Selection and Production-Scale Evaluation of an Autochthonous Saccharomyces cerevisiae A5GM Strain for Ancestral Mezcal

Spontaneous fermentations contribute to the identity of ancestral mezcal, although duration and yield may vary. This study selected an indigenous yeast and evaluated it as a starter candidate. Four native isolates (Torulaspora delbrueckii A1GM and A6GM, Kluyveromyces marxianus A2GM, and Saccharomyces cerevisiae A5GM) and a spontaneous control were compared in Agave cupreata microfermentations. Selection considered distillate alcohol content and preliminary ethanol-tolerance assays. A5GM was subsequently evaluated in three production batches containing A. angustifolia + A. karwinskii, A. angustifolia, or A. cupreata + A. angustifolia, with three independent vats per treatment. Fermentation profiles, distillate composition, yield, and sensory acceptance were assessed. A5GM produced the highest microfermentation distillate alcohol content (43.91 ± 0.89% Alc. Vol.) and grew at 16% ethanol (v/v). In the first two production batches, inoculated vats showed higher volumes standardized to 45% Alc. Vol. and lower calculated agave requirements than the controls; these were batch-specific observations under the operating conditions studied. Sensory evaluation used pooled vat-derived distillate composites, and the A5GM composites received the highest mean hedonic scores. Because strain-level establishment and persistence of A5GM were not confirmed, the observed outcomes cannot be attributed exclusively to this strain. Sensory interpretation is limited by fixed presentation order. Further independent batches, standardized management, and strain-level monitoring are needed to assess reproducibility.