Design of a minimal starter culture for controlled fermentation of maguey sap: Microbial and sensory comparison with traditional pulque

Pulque is a traditional Mexican beverage produced by the spontaneous fermentation of Agave (maguey) sap (aguamiel). Although of great cultural and economic importance, artisanal pulque production relies on empirical practices that lead to variability and limited scalability. In this study, we designed a defined minimal starter culture to achieve controlled fermentation of microfiltered aguamiel, preserving the key characteristics of traditional pulque. Microfiltration provided a sterile substrate, allowing functional assessment of three representative species from the pulque core microbiota: Saccharomyces cerevisiae, Zymomonas mobilis, and Lactiplantibacillus plantarum . Laboratory-scale fermentations were conducted to evaluate growth dynamics, sugar consumption, metabolite production, and sensory profiles. Comparing monocultures and co-cultures of these three core microorganisms revealed functional complementarity, resulting in efficient sugar conversion and balanced ethanol and lactic acid production. The resulting beverage maintained the characteristic aroma, acidity, and mouthfeel of artisanal pulque. Sensory evaluation using modified Flash profiling and Generalized Procrustes Analysis confirmed that this formulation converged toward the traditional reference while reducing variability. These findings demonstrate that spontaneous fermentation can be replaced by a reproducible, defined minimal inoculum without compromising the beverage's authenticity and illustrate how translating microbial ecology into minimal starter consortia can bridge tradition with biotechnology and innovation in food fermentation. • A defined starter culture was developed using core pulque microorganisms • Microfiltered aguamiel served as a sterile fermentation substrate • Fermented product matched traditional pulque in key metabolites • Sensory profiling showed similarity to traditional fermentation • S. cerevisiae , Z. mobilis , and L. plantarum were essential for functionality