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This study addresses winter fermentation instability in high-temperature Daqu by establishing a closed-loop monitoring system that links microenvironmental conditions with microbiome and metabolome data. The research identifies vertical CO2 gradients as the primary driver of microbial stratification, which restricts aerobic communities like Bacillus species and blocks key metabolic pathways.

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Abstract

High-temperature Daqu (HTD)’s quality determines the characteristics and yield of the Chinese sauce-aroma baijiu. However, winter production frequently encounters challenges such as fermentation instability and metabolic fluctuations, primarily stemming from complex, unmonitored microenvironmental changes within the HTD pile. This study established a closed-loop system linking the microenvironment, HTD quality, microbiome, and metabolome. Through continuous monitoring of the winter fermentation pile’s microenvironmental conditions and integrating multi-omics analyses, we revealed that CO2 concentration within fermentation piles is the core factor causing quality variations in HTD. By breaking the respiratory bottleneck formed by carbon dioxide (CO2) accumulation through the turning anaerobic stress can be alleviated, thereby driving metabolic succession. The study found that vertical CO2 concentration heterogeneity severely restricts the enrichment of aerobic core functional microbial communities such as the Bacillus species. This directly blocks key metabolic pathways including amino acid metabolism and energy supply via ABC transporters. Moreover, the specific accumulation of Amadori products further confirms that this low-temperature environment under CO2 stress causes the Maillard reaction to stall at intermediate stages. Consequently, this study proposes a steady-state control strategy centered on oxygen and CO2 gas characteristics. By actively regulating the gaseous microenvironment to eliminate metabolic heterogeneity, it provides theoretical support for standardizing traditional fermentation processes.

Key findings

  • Vertical CO2 concentration heterogeneity severely limits the enrichment of aerobic core functional microbial communities, specifically Bacillus species.
  • High CO2 accumulation creates a respiratory bottleneck that blocks amino acid metabolism and energy supply via ABC transporters.
  • Turning-induced anaerobic stress alleviates this bottleneck, driving metabolic succession and enabling Amadori product accumulation.

Keywords

FermentationCarbon dioxideMicrobial metabolismFood scienceHeterotrophMetabolic pathway

Identifiers

DOI
10.3390/foods15050799
Journal
Foods
Year
2026