Soy sauce aroma type baijiu (also known as Maotai-flavor baijiu) is one of the most popular types of baijiu in China. Traditionally, it is mainly produced in Southwest China. However, in recent decades, some other regions in China have also been able to produce high-quality soy sauce aroma type baijiu, but their microbial flora characteristics during fermentation are still unclear. Here, the bacterial microbial community structure of fermented grains in different rounds of Lutaichun soy sauce aroma type baijiu produced in North China was studied by high-throughput sequencing technology, and the potential probiotics strains with good characteristics (alcohol tolerance, etc.) were screened. The results showed that lactic acid bacteria were the main bacteria in the process of baijiu fermentation. However, as the number of repeated fermentation rounds increased, the proportion of lactic acid bacteria decreased. Firmicutes (96.81%) were the main bacteria in baijiu fermentation at the phylum level, and Lactobacillus (66.50%) were the main bacteria at the genus level. Finally, two strains with high resistance to alcohol stress, Lactiplantibacillus plantarum LTJ12 and Pediococcus acidilactici LTJ28, were screened from 48 strains of lactic acid bacteria in the fermented grains. The survival rates of L. plantarum LTJ12 and P. acidilactici LTJ28 under the 8% alcohol stress treatment were 59.01% and 55.50%, respectively. To the best of our knowledge, this study is the first to reveal the microbial succession of fermented grains in different rounds of soy sauce aroma type baijiu from North China, and has the benefit of explaining the deep molecular mechanism in the process of baijiu fermentation. In addition, the obtained lactic acid bacteria strains with high alcohol tolerance could be conducive to the development of new products such as active probiotic alcoholic beverages and may have important industrial development prospects also.

Lactic acid bacteria have received a significant amount of attention due to their probiotic characteristics. The species Lactiplantibacillus plantarum and Lactiplantibacillus pentosus are genotypically closely related, and their phenotypes are so similar that they are easily confused and mistaken. In the previous study, an ethanol-resistant strain, LTJ12, isolated from the fermented grains of soy sauce aroma type baijiu in North China, was originally identified as L. plantarum through a 16S rRNA sequence analysis. Here, the genome of strain LTJ12 was further sequenced using PacBio and Illumina sequencing technology to obtain a better understanding of the metabolic pathway underlying its resistance to ethanol stress. The results showed that the genome of strain LTJ12 was composed of one circular chromosome and three circular plasmids. The genome size is 3,512,307 bp with a GC content of 46.37%, and the number of predicted coding genes is 3248. Moreover, by comparing the coding genes with the GO (Gene Ontology), COG (Cluster of Orthologous Groups) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases, the functional annotation of the genome and an assessment of the metabolic pathways were performed, with the results showing that strain LTJ12 has multiple genes that may be related to alcohol metabolism and probiotic-related genes. Antibiotic resistance gene analysis showed that there were few potential safety hazards. Further, after conducting the comparative genomics analysis, it was found that strain LTJ12 is L. pentosus but not L. plantarum, but it has more functional genes than other L. pentosus strains that are mainly related to carbohydrate transport and metabolism, transcription, replication, recombination and repair, signal transduction mechanisms, defense mechanisms and cell wall/membrane/envelope biogenesis. These unique functional genes, such as gene 2754 (encodes alcohol dehydrogenase), gene 3093 (encodes gamma-D-glutamyl-meso-diaminopimelate peptidase) and some others may enhance the ethanol tolerance and alcohol metabolism of the strain. Taken together, L. pentosus LTJ12 might be a potentially safe probiotic with a high ethanol tolerance and alcohol metabolism. The findings of this study will also shed light on the accurate identification and rational application of the Lactiplantibacillus species.