Dynamic Changes and Correlations of Physicochemical Parameters, Flavor Compounds and Microbial Communities During Soy...

Koji production is a critical process that determines the flavor and quality of the final soy sauce product. However, the complex mechanisms underlying microbial metabolism and the evolution of the physicochemical environment still require further analysis. This study focuses on three parallel koji rooms in an industrialized koji fermentation process. This work tracked the dynamics of physicochemical indices, volatile flavor compounds, and microbial communities over a full 40 h cycle. Data integration and correlation analysis elucidated the close linkage between the microbial community, the fermentation environment, and flavor formation. Koji moisture declined gradually, with faster losses at later fermentation stages. This physiological dehydration arose from microbial metabolic heat, forced aeration and structural loosening of koji, not simple physical evaporation. System pH displayed a typical U-shaped trend across fermentation. Values dropped early, most likely driven by accumulating organic acids, before rising from mid to late fermentation. This pH rebound was tentatively attributed to ammonia release from proteolytic breakdown, which may neutralize acidic compounds. These observations cast doubt on the conventional assumption that organic acid levels may be reliably estimated solely from pH measurements. Physicochemical analysis showed continuous accumulation of amino acid nitrogen (0.6-0.9 g/100 g) and total acidity throughout fermentation. By contrast, reducing sugar concentrations differed across individual koji rooms, presumably owing to divergent microbial adaptation in early fermentation. A total of 77 common compounds were identified, among which 13 key odor-active compounds with OAV ≥ 1, such as 4-vinylguaiacol and 3-methylbutyraldehyde, constitute the characteristic flavor profile of soy sauce starter culture. High-throughput sequencing uncovered a distinct ecological pattern: eukaryotic communities, dominated by Aspergillus oryzae, converged under controlled regulation. While prokaryotic communities differentiated dynamically, driven by spatial heterogeneity in the semi-open fermentation environment. Spearman correlation analysis further indicated potential functional partitioning: high-abundance taxa (e.g., Aspergillus oryzae, Weissella) were predominantly associated with macromolecular substrate degradation, whereas rare low-abundance taxa (e.g., Alternaria) displayed significant correlations with the biosynthesis of key characteristic flavor compounds. This study clarifies the synergistic regulatory mechanisms linking physicochemical conditions, microbial metabolism, and flavor precursor formation during industrial koji production. The findings establish a scientific foundation for optimizing process parameters and achieving standardized quality control in soy sauce manufacturing.