Identification of Dominant Microbes and Their Successions During Solid-State Fermentation of Luzhou-Flavour Liquor Based on High-Throughput Sequencing Following Culture

Microorganisms are crucial for the liquor brewing process and substantially impact liquor flavour and quality; therefore, understanding microbial succession is necessary. Most studies use a single-method approach and fail to provide an in-depth analysis. We aimed to combine traditional culture method with high-throughput sequencing (HTS) to identify the microbial diversity and succession in Luzhou-flavour fermentation. HTS revealed 932 bacterial and 980 fungal operational taxonomic units. 16S rDNA, 26S D1/D2 rDNA, and ITS v4/v5 isolated and identified 256 bacterial and 130 yeast strains. Population succession analysis showed that the dominant populations were yeasts, Lactobacillus, and Bacillus (early stage), and yeasts and Lactobacillus (late stage). Lactobacillus, Pichia, Bacillus, and Candida were abundant among all three layers of fermented grains. However, C. ethanolica, Saccharomycetes sp., and an unidentified Saccharomyces cerevisiae were more abundant in the lower layer than in the middle and upper layers, while L. parabuchneri, Oceanobacillus oncorhynchi, and Thermoactinomyces sp. were present only in the lower layer. Correlations among enzyme activity, volatile production, and dominant microbes during fermentation indicated that P. fermentans, L. suebicus, L. acetotolerans, P. kudriavzevii, P. exigua, and B. tequilensis were significantly affected during brewing. Our results lay a foundation for elucidating the microbial fermentation mechanism of Luzhou-flavour liquor and will assist in improving traditional liquor brewing quality and efficiency.