Traditional Chinese fermented foods are diverse and loved by people for their rich nutrition and unique flavors. In the fermentation processes of these foods, the microorganisms in the fermentation systems play a crucial role in determining the flavor and quality. Currently, some microorganisms in the fermentation systems of traditional Chinese fermented foods are in a state of being unculturable or difficult to culture, which hinders the comprehensive analysis and resource development of the microbial communities in the fermentation systems. This article provides an overview of the uncultured microorganisms in the natural environment, in the fermentation systems of traditional Chinese fermented foods, and the research methods for studying such microorganisms. It also discusses the prospects of utilizing the uncultured microorganisms in the fermentation systems of traditional Chinese fermented foods. The aim is to gain a comprehensive understanding of the microbial diversity and uncultured microorganisms in the fermentation systems of traditional Chinese fermented foods in order to better exploit and utilize these microorganisms and promote the development of traditional Chinese fermented foods.

Broomcorn millet Huangjiu brewing is usually divided into primary fermentation and post-fermentation. Microbial succession is the major factor influencing the development of the typical Huangjiu flavor. Here, we report the changes in flavor substances and microbial community during the primary fermentation of broomcorn millet Huangjiu. Results indicated that a total of 161 volatile flavor compounds were measured during primary fermentation, and estragole was detected for the first time in broomcorn millet Huangjiu. A total of 82 bacteria genera were identified. Pediococcus, Pantoea, and Weissella were the dominant genera. Saccharomyces and Rhizopus were dominant among the 30 fungal genera. Correlation analysis showed that 102 microorganisms were involved in major flavor substance production during primary fermentation, Lactobacillus, Photobacterium, Hyphodontia, Aquicella, Erysipelothrix, Idiomarina, Paraphaeosphaeria, and Sulfuritalea were most associated with flavoring substances. Four bacteria, Lactobacillus (R1), Photobacterium (R2), Idiomarina (R3), and Pediococcus (R4), were isolated and identified from wheat Qu, which were added to wine Qu to prepare four kinds of fortified Qu (QR1, QR2, QR3, QR4). QR1 and QR2 fermentation can enhance the quality of Huangjiu. This work reveals the correlation between microorganisms and volatile flavor compounds and is beneficial for regulating the micro-ecosystem and flavor of the broomcorn millet Huangjiu.

Baijiu is a traditional distilled liquor in China. The unique flavor developed during the fermentation process of Baijiu is closely related to the microorganisms in the fermented grains of the Baijiu. Fermented grain is a solid material that has not been distilled after the fermentation of Baijiu. It is of great significance to study the bacterial diversity in fermented grains and to isolate and culture them to reveal the formation mechanism of the flavor substances in Baijiu. In this study, the diversity of bacteria in fermented grains was studied by the combination of amplicon sequencing and culturomics, and a pure culture of culturable strains was obtained. The results of amplicon sequencing showed that the bacteria detected in the fermented grains were classified into 5 phyla, 8 classes, 24 orders, 34 families, and 45 genera, of which Lactobacillus (66.95%) and Acetobacter (32.36%) were the dominant genera. By using the method of culturomics, five pH gradients (pH = 3, 4, 5, 6, and 7) of enrichment culture medium and solid medium with different components were designed to enrich and isolate the culturable bacteria in the fermented grains. A total of 323 strains of bacteria were obtained, belonging to 53 genera and 118 species, and 14 suspected new species were obtained. Among the bacteria isolated by the culturomics, 14 genera were detected by amplicon sequencing, and 39 genera were not detected, indicating that the culturomics method can isolate and culture bacteria with low abundance in fermented grains. Compared with the traditional culture method, culturomics expanded the culturable bacteria in the fermented grains to 188%. The diversity of the bacteria in the fermented grains of Baijiu was analyzed by amplicon sequencing and culturomics, and the microbial community composition and relative abundance of fermented grains were comprehensively revealed. At the same time, the acquisition of more pure cultures of culturable bacteria provides more choices for the liquor-making microbial resource library.

Tartary buckwheat is a healthy grain rich in nutrients and medicinal ingredients and consequently is commonly used for Huangjiu brewing. In order to reveal the correlation between microbial succession and higher alcohols production, in this study, Huangjiu fermentation was conducted using Tartary buckwheat as the raw material and wheat Qu as the starter culture. Microbial community dynamics analysis indicated that the bacterial diversity initially decreased rapidly to a lower level and then increased and maintained at a higher level during fermentation. Lactococcus was the dominant bacteria and Ralstonia, Acinetobacter, Cyanobacteria, and Oxalobacteraceae were the bacterial genera with higher abundances. In sharp contrast, only 13 fungal genera were detected during fermentation, and Saccharomyces showed the dominant abundance. Moreover, 18 higher alcohol compounds were detected by GC-MS during fermentation. Four compounds (2-phenylethanol, isopentanol, 1-hexadecanol, and 2-phenoxyethanol) were stably detected with high concentrations during fermentation. The compound 2-ethyl-2-methyl-tridecanol was detected to be of the highest concentration in the later period of fermentation. Correlation analysis revealed that the generation of 2-phenylethanol, isopentanol, 1-hexadecanol, and 2-phenoxyethanol were positively correlated with Granulicatella and Pelomonas, Bacteroides, Pseudonocardia and Pedomicrobium, and Corynebacterium, respectively. The verification fermentation experiments indicated that the improved wheat Qu QT3 and QT4 inoculated with Granulicatella T3 and Acidothermus T4 led to significant increases in the contents of 2-phenylethanol and pentanol, as well as isobutanol and isopentanol, respectively, in the Tartary buckwheat Huangjiu. The findings benefit understanding of higher alcohols production and flavor formation mechanisms in Huangjiu fermentation.