Composition and function of viruses in sauce-flavor baijiu fermentation.

Viruses are highly abundant in nature, associated with quality and safety of traditional fermented foods. However, the overall viral diversity and function are still poorly understood in food microbiome. Traditional baijiu fermentation is an ideal model system to examine the diversity and function of viruses owing to easy access, stable operation, and domesticated microbial community. Equipped with cutting-edge viral metagenomics, we investigated the viral community in the fermented grain and fermentation environment, as well as their contribution to baijiu fermentation. Viral communities in the fermented grains and fermentation environment are highly similar. The dominant viruses were bacteriophages, mainly including the order Caudovirales and the family Inoviridae. Furtherly, association network analysis showed that viruses and bacteria were significantly negatively correlated (P < 0.01). Viral diversity could significantly influence bacterial and fungal succession (P < 0.05). Moreover, we proved that starter phages could significantly inhibit the growth of Bacillus licheniformis in the logarithmic growth stage (P < 0.05) under culture condition. Based on the functional annotations, viruses and bacteria both showed high distribution of genes related to amino acid and carbohydrate metabolism. In addition, abundant auxiliary carbohydrate-active enzyme (CAZyme) genes were also identified in viruses, indicating that viruses were involved in the decomposition of complex polysaccharides during fermentation. Our results revealed that viruses could crucially affect microbial community and metabolism during traditional fermentation.

Quantitative metaproteomics reveals composition and metabolism characteristics of microbial communities in Chinese liquor fermentation starters.

Introduction: Daqu, the Chinese liquor fermentation starter, contains complex microbial communities that are important for the yield, quality, and unique flavor of produced liquor. However, the composition and metabolism of microbial communities in the different types of high-temperature Daqu (i.e., white, yellow, and black Daqu) have not been well understood. Methods: Herein, we used quantitative metaproteomics based on data-independent acquisition (DIA) mass spectrometry to analyze a total of 90 samples of white, yellow, and black Daqu collected in spring, summer, and autumn, revealing the taxonomic and metabolic profiles of different types of Daqu across seasons. Results: Taxonomic composition differences were explored across types of Daqu and seasons, where the under-fermented white Daqu showed the higher microbial diversity and seasonal stability. It was demonstrated that yellow Daqu had higher abundance of saccharifying enzymes for raw material degradation. In addition, considerable seasonal variation of microbial protein abundance was discovered in the over-fermented black Daqu, suggesting elevated carbohydrate and amino acid metabolism in autumn black Daqu. Discussion: We expect that this study will facilitate the understanding of the key microbes and their metabolism in the traditional fermentation process of Chinese liquor production.

Microbial Community Succession and Its Environment Driving Factors During Initial Fermentation of Maotai-Flavor Baijiu.

The microbial composition and environmental factors can take a great influence on community succession during the solid-state fermentation (SSF) of Maotai-flavor Baijiu. In this paper, high-throughput sequencing was used to reveal the dominant microorganisms and the evolution process of microbial community structure in the initial fermentation of Maotai-flavor Baijiu. The correlation analysis was carried out for the relationship between physicochemical factors and fermented microbes. The results showed that microorganisms were obviously enriched and the diversity of bacteria and fungi showed a downward trend during the heap fermentation process of Maotai-flavor Baijiu. However, the diversity of fungi in the pit fermentation process increased. Generally, Lactobacillus, Pichia, and Saccharomyces were the dominant microorganisms in the initial fermentation of Maotai-flavor Baijiu. According to the redundancy analysis, we found that reducing sugar was the key driving factor for microbial succession in the heap fermentation, while acidity, alcohol, and temperature were the main driving forces in pit fermentation. This study revealed the microbial succession and its related environmental factors in the initial fermentation of Maotai-flavor Baijiu, which will enrich our knowledge of the mechanism of solid-state liquor fermentation.

Effects of initial temperature on microbial community succession rate and volatile flavors during Baijiu fermentation process.

The importance of fermentation temperature has been highlighted as it correlates with biodiversity and microbial metabolism for a microbial community. In this study, microbial community succession and volatile flavors during sauce-flavor Baijiu fermentation at different initial temperatures (LT group: 28 ± 2 °C and HT group: 37 ± 2 °C) were investigated using Illumina Miseq sequencing and gas chromatography-mass spectrometry (GC-MS). First, we found that different initial temperatures had a significant effect on fermentation parameters (P < 0.001); specifically, a higher initial temperature increased the accumulation of acetic acid and decreased the production of ethanol. Second, the microbial communities were characterized by decreased α-diversity and increased ß-diversity (P < 0.05) during heap fermentation. A higher initial temperature accelerated the increase in Lactobacillus and led to a faster microbial succession rate. Lactobacillus could be used as microbial markers of microbial succession rate in sauce-flavor Baijiu fermentation. Next, we found that acetic acid drove microbial succession under a higher fermentation temperature. Molecular ecological network analysis showed that different fermentation temperatures affected microbial interactions. The higher temperature enhanced microbial interactions of Lactobacillus. In addition, 50 volatile flavors were identified in the fermented grains. High temperature increased the content of total acid and reduced total esters, and Lactobacillus and Saccharomyces were the important microbiota related to different flavor compounds between the two groups. Collectively, altering the initial temperature led to differences in microbial succession rates and volatile flavors in the sauce-flavor Baijiu fermentation process. Therefore, these results are valuable for exploring quality control and management strategies in the spontaneous fermentation process.

Effect of Pichia on shaping the fermentation microbial community of sauce-flavor Baijiu.

In spontaneous food fermentation processes, environmental microbiota affects the yield and quality of the fermentation productions. Although the importance of environmental microbiota has been highlighted, the ecological processes that how the environmental microbiota affects the fermentation microbial community are poorly understood. To study the effect of the environmental microbiota on community assembly, the sources of microbiota and the ecological processes of the fermentation were characterized in sauce-flavor Baijiu. Results showed that the process of sauce-flavor Baijiu making could be divided into three phases according to fermentation parameters. Heap fermentation (phase I) was an important period for rapid temperature rise, substrate utilization and production accumulation. The microbial community of heap fermentation was characterized by decrease of diversity and rapid succession of community structure. Virgibacillus, Kroppenstedtia, Bacillus and Oceanobacillus were predominant in the initial heap fermentation, while Lactobacillus was predominant during the later stage. Pichia was the predominant fungal genus during the whole fermentation process. Then, SourceTracker results showed that Daqu provided 95.6% of the bacterial community and 28.10% of the fungal community to heap fermentation, whereas the environments (indoor ground and tools) provided 71.9% of the fungal communities (mainly Pichia) to heap fermentation. Next, the results revealed that the temperature, ethanol and microbial interaction of Pichia synergistically drove the dynamic of the microbial community during the heap fermentation process. Pichia was proved to be the heat-resistant fungi and strong competitor based on growth in different temperature and competition assays in vitro. Finally, the quick succession of heap fermentation microbiota increased the enrichment of volatile flavors such as acids and esters. Our comprehensive methods shows that Pichia, which mainly comes from the environment, can construct the microbial community of Baijiu fermentation, and highlights the importance of environmental microbiota in attempts to control and promote the formation of Baijiu fermentation microbial community. This systematic study of environmental microbiota is valuable for quality control and management during spontaneous fermentation.