Synergistic effects of simultaneous and sequential fermentation by non-Saccharomyces yeasts and lactic acid bacteria on nutritional and aroma profiles of low-alcohol pomegranate beverages.

Abstract

Background: Mixed-culture fermentation improves polyphenol accumulation and aroma biosynthesis in fruit beverages through microbial synergism. The strain-specific and inoculation time-dependent effects of such synergism, however, remain unexplored in pomegranate beverage fermentation.

Results: Two indigenous non-Saccharomyces yeasts were combined with two lactic acid bacteria (LAB) strains under simultaneous and sequential inoculation schemes. Physicochemical properties, organic acids, polyphenols, antioxidant capacity, and volatile aroma compounds were analyzed comprehensively. Sequential inoculation significantly increased total phenolic content, improved 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, and enriched aroma-active volatiles, particularly esters and higher alcohols. Sequential fermentation with LAB strain 21805 produced the most diverse aroma profile and intensified tropical fruit and floral sensory notes.

Conclusion: This study provides the first systematic evaluation of these specific non-Saccharomyces and LAB combinations in pomegranate fermentation, demonstrating clear effects of inoculation timing, bioactive compound accumulation, and aroma complexity. The observed strain- and timing-dependent synergism offers a practical strategy to improve the nutritional and sensory qualities of low-alcohol fruit beverages. © 2026 Society of Chemical Industry.