The solid-state fermentation of Baijiu is complicated by the co-fermentation of many microorganisms. The instability of the composition and abundance of the microorganisms in the fermentation process leads to fluctuations of product quality, which is one of the bottleneck problems faced by the Strong-flavor Baijiu industry. In this study, we established a combination of functional microorganisms for the stable fermentation of the main flavor compounds of Baijiu, including medium and long-chain fatty acid ethyl esters such as hexanoic acid, ethyl ester; butanoic acid, ethyl ester; octanoic acid, ethyl ester; acetic acid, ethyl ester; 9,12-octadecadienoic acid, ethyl ester; and decanoic acid, ethyl ester in the fermented grains. Our study investigated the effects of microbial combinations on the fermentation from three aspects: microbial composition, microbial interactions, and microbial association with flavor compounds. The results showed that the added functional microorganisms (Lactobacillus, Clostridium, Caproiciproducens, Saccharomyces, and Aspergillus) became the dominant species in the fermentation system and formed positive interactions with other microorganisms, while the negative interactions between microorganisms were significantly reduced in the fermentation systems that contained both Daqu and functional microorganisms. The redundancy analysis showed that the functional microorganisms (Lactobacillus, Saccharomyces, Clostridium, Cloacibacterium, Chaenothecopsis, Anaerosporobacter, and Sporolactobacillus) showed strong positive correlations with the main flavor compounds (hexanoic acid, ethyl ester; lactic acid, ethyl ester; butanoic acid, ethyl ester; acetic acid, ethyl ester; and octanoic acid, ethyl ester). These results indicated that it was feasible to produce Baijiu with a functional microbial combination, and that this could promote stable Baijiu production.

The compositions of the microbial community in fermented grains of Sauce-flavor baijiu produced in different regions have diverse characteristics; however, the reasons for this remain unclear. The present study investigated the contributions of environmental microorganisms to the microbial community as well as the volatile compounds in the fermented grains of Sauce-flavor baijiu produced in the Beijing region using high-throughput sequencing combined with sourcetracker analysis, and compared the differences of environmental microorganism and their roles in the production process of Sauce-flavor baijiu from different regions.The results showed that the environmental microorganisms in the tools were the main contributors of the bacterial and fungal communities in fermented grains during heap fermentation and at the beginning of pit fermentation. At the end of pit fermentation, pit mud was the main environmental source of bacterial community in fermented grains, while tools and Daqu were the main environmental sources of fungal community in fermented grains.Environmental microorganisms thrived on the functional microorganisms in the fermented grains of Sauce-flavor baijiu produced in the Beijing region and thus shaped the profiles of volatile compounds. Environmental microorganisms of Sauce-flavor baijiu in the Guizhou province and the Beijing region differed significantly, which is partially responsible for the distinctive characteristics in the microbial community structure of Sauce-flavor baijiu-fermented grains from different regions.

Daqu is a microbial-rich baijiu fermentation starter. The high-temperature Daqu plays an essential role in the manufacturing of sauce-flavored baijiu. However, few studies have focused on three kinds of high-temperature Daqu (white, yellow, and black Daqu) in northern China. In this study, the physicochemical indexes, volatile flavor compounds, and microbial characteristics of the three different colors of high-temperature Daqu in northern China were comparatively analyzed to reveal their potential functions. White Daqu (WQ) exhibited the highest liquefying power and starch, and black Daqu (BQ) showed the highest saccharifying and esterifying powers. A total of 96 volatile components were identified in the three types of Daqu, and the contents of the volatile components of yellow Daqu (YQ) were the highest. The microbial community structure analysis showed that Bacillus and Byssochlamys were dominant in BQ, Kroppenstedtia and Thermoascus were dominant in WQ, and Virgibacillus and Thermomyces dominated the YQ. The RDA analysis revealed the correlation between the dominant microorganisms and different physicochemical indexes. The Spearman correlation analysis indicated that Oceanobacillus, Saccharopolyspora, Staphylococcus, Pseudogracilibacillus, Byssochlamys, and Thermomyces showed positive correlations with part of the majority of the key volatile flavor compounds. This work provides a scientific basis for the actual production of different colors of high-temperature Daqu in the northern region of China for sauce-flavored baijiu.

A mesophilic xylanase PjxA from Penicillium janthinellum MA21601 has high specific activity under acidic condition and holds great potential for applications in the animal feed industry. To enhance the thermostability of xylanase PjxA, two mutation strategies in the N-terminal region were examined and then integrated into the xylanase to further improvement. The recombinant xylanase PTxA-DB (The meaning of DB is disulfide-bridge.) was constructed by replacement of five residues in the mutated region in TfxA (T10Y, N11H, N12D, Y15F, N30 L), combined with an additional disulfide bridge in the N-terminal region. The Tm value of mutant PTxA-DB was improved from 21.3 °C to 76.6 °C, and its half-life was found to be 53.6 min at 60 °C, 107-fold higher than the wild type strain. The location of the disulfide bridge (T2C-T29C) was between the irregular loop and the β-strand A2, accounting for most of the improvement in thermostability of PjxA. Further analysis indicated T2C, T29C, N30 L and Y15F lead to increase N-terminal hydrophobicity. Moreover, the specific activity and substrate affinity of PTxA-DB were also enhanced under the acidic pH values. These results indicated PTxA-DB could be a prospective additive to industrial animal feeds.

Sauce-flavor baijiu produced in the Beijing and Guizhou regions has regional characteristic flavors, but the differences in flavor compounds and reasons for their formation remain unclear. The sauce-flavor baijiu brewing process involves several rounds of fermentation. In this study, we investigated the temporal distribution of microbial communities and flavor substances during the third round of sauce-flavor baijiu fermentation in the Beijing region, and we then compared and analyzed the differences of flavor substances and microorganisms in the fermented grains of sauce-flavor baijiu in the Beijing and Guizhou regions. It was found that 10 bacterial genera and 10 fungal genera were dominant in the fermented grains. The acidity of the fermented grains had a significant driving effect on the microbial community succession. A total of 81 volatile compounds were identified and quantified in the fermented grains, of which esters and alcohols were relatively abundant. The differences in 30 microbial community compositions and their resulting differences in terms of the fermentation parameters of fermented grains are responsible for the differences in the profiles of flavor compounds between sauce-flavor baijiu produced in the Beijing and Guizhou regions.