Abstract
The quality of Zhejiang rosy vinegar (ZRV) is defined by its complex chemical profile, yet the biochemical links between microbial metabolism and flavor evolution remain unclear. We employed an integrated multi-omics approach (amplicon sequencing, GC-MS, GC-IMS, and UHPLC-MS/MS) to elucidate the molecular underpinnings of ZRV flavor formation. Results revealed a microbial succession dominated by Acetobacter, Lactobacillus, Saccharomyces, and Monascus. Metabolomics demonstrated a dynamic shift in volatile profiles and the accumulation of non-volatile organic acids, driven largely by branched-chain amino acid metabolism. Integration identified critical biochemical linkages: Saccharomyces and Acetobacter activity shaped the early ester/acid framework, while Monascus and Pseudomonas drove acetoin accumulation. Furthermore, Lactobacillus and Monascus abundances were strongly associated with flavonoid enrichment, underpinning ZRV's antioxidant capacity. These findings provide mechanistic insights into microbial-chemical interactions governing flavor and functional compound synthesis, offering a robust statistical and chemical foundation for the industrial quality control of traditional fermented foods.