Multi-Omics Insights into Microbial Community Dynamics and Functional Shifts During Double-Round Bottom Fermentation of Strong-Flavor Baijiu

Double-round bottom fermentation (DRBF) represents an important technological innovation in strong-flavor Baijiu production, yet the microbial succession and metabolic mechanisms underlying this process remain insufficiently understood. In this study, physicochemical analyses combined with multi-omics approaches were employed to elucidate the dynamic variations in physicochemical parameters, volatile compounds, and microbial community structure and function during DRBF, as well as to reconstruct key metabolic pathways involved in fermentation. A total of 153 volatile compounds were identified, with esters, alcohols, and acids as the major components showing distinct accumulation patterns across fermentation stages. High-throughput sequencing detected 505 bacterial and 175 fungal genera, dominated by <i>Lactobacillus</i>, <i>Aspergillus</i>, and <i>Saccharomyces</i>. Functional annotation revealed that metabolic pathways predominated, shifting from energy- and growth-related processes in the early stage to amino acid, fatty acid, and secondary metabolite biosynthesis in the later stage. Reconstruction of metabolic pathways identified 57 key enzymes linking starch degradation, pyruvate metabolism, the tricarboxylic acid (TCA) cycle, and ester biosynthesis, indicating cooperative metabolism among bacteria, yeasts, and molds. These findings elucidate the synergistic metabolic mechanisms of flavor formation during DRBF and provide a scientific basis for optimizing fermentation control and improving Baijiu quality.