Light-flavor Baijiu (LFB) fermentation is a representative spontaneous mixed-culture solid-state fermentation process in which sorghum is used as the raw material. Raw materials and microorganisms are crucial to the flavor formation and quality of LFB. However, the microbial and physicochemical dynamics of different sorghum varieties during LFB fermentation, as well as their impact on flavor compounds are still largely unknown. Herein, PacBio single-molecule real-time (SMRT) sequencing and headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) were applied to investigate microbial community succession and volatile flavor formation in glutinous/non-glutinous sorghum-based fermented grains during LFB fermentation. Fermented grains made of glutinous sorghum Liangnuo No. 1 (GLN) had higher bacterial α-diversity and lower fungal α-diversity than those with fermented grains prepared with non-glutinous red sorghum (NRS) (p < 0.05). The dominant microbial species were Saccharomyces cerevisiae, Acetobacter pasteurinus, and Lactobacillus helveticus, the latter two of which were the predominant bacteria observed at the end of fermentation in GLN and NRS, respectively. Moisture content and reducing sugar had a more significant impact on the microorganisms in GLN, while amino acid nitrogen, total free amino acids, and residual starch were the main driving factors driving the microbial community in NRS. The correlation network and discriminant analysis indicated that a relatively high content of 4-vinylguaiacol showed a significant positive association with significant differential microbial species in GLN. These results provided valuable insights for improving the quality of LFB.

To investigate the changes of phenolic metabolite during different grains fermentation stages of Chinse Baijiu, the ultra-performance liquid chromatography-quadrupole time of-flight mass spectrometry (UHPLC-QTOF-MS) was applied to identify and analyze the different phenolic metabolites, combined with principal component analysis and partial least squares discriminant analysis. Results indicated that significant differences in phenolic metabolites during different fermentation stages were found. Among the 231 phenolic metabolites detected, 36, 31, 19, 23, 14, and 50 differential phenolic metabolites were screened between different groups using partial least squares discriminant analysis. Twelve metabolic pathways with high correlation of differential phenolic metabolites and 23 main participating differential metabolites were identified through KEGG metabolic pathway enrichment analysis. The present study preliminarily revealed the differences of phenolic metabolites at different fermentation stages, and providing a theoretical basis for the further improving of the taste and quality of Chinese Baijiu.

The fermentation process of Chinese Baijiu\'s fermented grains involves the intricate succession and metabolism of microbial communities, collectively shaping the Baijiu\'s quality. Understanding the composition and succession of these living microbial communities within fermented grains is crucial for comprehending fermentation and flavor formation mechanisms. However, conducting high-throughput analysis of living microbial communities within the complex microbial system of fermented grains poses significant challenges. Thus, this study addressed this challenge by devising a high-throughput analysis framework using light-flavor Baijiu as a model. This framework combined propidium monoazide (PMA) pretreatment technology with amplicon sequencing techniques. Optimal PMA treatment parameters, including a concentration of 50 μM and incubation in darkness for 5 min followed by an exposure incubation period of 5 min, were identified. Utilizing this protocol, viable microorganism biomass ranging from 8.71 × 106 to 1.47 × 108 copies/μL was successfully detected in fermented grain samples. Subsequent amplicon sequencing analysis revealed distinct microbial community structures between untreated and PMA-treated groups, with notable differences in relative abundance compositions, particularly in dominant species such as Lactobacillus, Bacillus, Pediococcus, Saccharomycopsis, Issatchenkia and Pichia, as identified by LEfSe analysis. The results of this study confirmed the efficacy of PMA-amplicon sequencing technology for analyzing living microbial communities in fermented grains and furnished a methodological framework for investigating living microbial communities in diverse traditional fermented foods. This technical framework holds considerable significance for advancing our understanding of the fermentation mechanisms intrinsic to traditional fermented foods.

Elevating the flavor profile of strong flavors Baijiu has always been a focal point in the industry, and pit mud (PM) serves as a crucial flavor contributor in the fermentation process of the fermented grains (FG). This study investigated the influence of wheat flour and bran (MC and FC) as PM culture enrichment media on the microbiota and metabolites of FG, aiming to inform strategies for improving strong-flavor Baijiu flavor. Results showed that adding PM cultures to FG significantly altered its properties: FC enhanced starch degradation to 51.46% and elevated reducing sugar content to 1.60%, while MC increased acidity to 2.11 mmol/10 g. PM cultures also elevated FG\'s ester content, with increases of 0.36 times for MC-FG60d and 1.48 times for FC-FG60d compared to controls, and ethyl hexanoate rising by 0.91 times and 1.39 times, respectively. Microbial analysis revealed that Lactobacillus constituted over 95% of the Abundant bacteria community, with Kroppenstedtia or Bacillus being predominant among Rare bacteria. Abundant fungi included Rasamsonia, Pichia, and Thermomyces, while Rare fungi consisted of Rhizopus and Malassezia. Metagenomic analysis revealed bacterial dominance, primarily consisting of Lactobacillus and Acetilactobacillus (98.80-99.40%), with metabolic function predictions highlighting genes related to metabolism, especially in MC-FG60d. Predictions from PICRUSt2 suggested control over starch, cellulose degradation, and the TCA cycle by fungal subgroups, while Abundant fungi and bacteria regulated ethanol and lactic acid production. This study highlights the importance of PM cultures in the fermentation process of FG, which is significant for brewing high-quality, strong-flavor Baijiu.

Starch and alcohol serve as pivotal indicators in assessing the quality of lees fermentation. In this paper, two hyperspectral imaging (HSI) techniques (visible-near-infrared (Vis-NIR) and NIR) were utilized to acquire separate HSI data, which were then fused and analyzed toforecast the starch and alcohol contents during the fermentation of lees. Five preprocessing methods were first used to preprocess the Vis-NIR, NIR, and the fused Vis-NIR and NIR data, after which partial least squares regression models were established to determine the best preprocessing method. Following, competitive adaptive reweighted sampling, successive projection algorithm, and principal component analysis algorithms were used to extract the characteristic wavelengths to accurately predict the starch and alcohol levels. Finally, support vector machine (SVM)-AdaBoost and XGBoost models were built based on the low-level fusion (LLF) and intermediate-level fusion (ILF) of single Vis-NIR and NIR as well as the fused data. The results showed that the SVM-AdaBoost model built using the LLF data afterpreprocessing by standard normalized variable was most accurate for predicting the starch content, with an R P 2 $\\ R_P^2$ of 0.9976 and a root mean square error of prediction (RMSEP) of 0.0992. The XGBoost model built using ILF data was most accurate for predicting the alcohol content, with an R P 2 $R_P^2$ of 0.9969 and an RMSEP of 0.0605. In conclusion, the analysis of fused data from distinct HSI technologies facilitates rapid and precise determination of the starch and alcohol contents in fermented grains.

The present study employed Hunan local Tuqu for fermentation and investigated the physicochemical properties, microbial community composition, and volatile flavor compounds of the fermented grains, as well as the correlation between the physicochemical indicators and the microbial community. The findings reveal that the activities of α-amylase and glucoamylase were highest during the initial stages of the fermentation process. The acid protease activity increased to 30.6 U/g on the second day and then decreased. Cellulose and lipase activities both showed an increasing trend. The moisture content increased sharply to 73.41% and then remained relatively stable. The acidity was highest on the eighth day. Fifty genera of bacteria and twenty-two genera of fungi were detected. Lactobacillus was dominant among bacteria, and Saccharomyces was dominant among fungi. A correlation analysis showed that there were positive correlations between moisture, acidity, cellulose, lipase activities and Lactobacillus, and there were positive correlations between moisture content, acidity, cellulase activity, acidic protease activity and Saccharomyces. A total of 46 volatile flavor compounds were detected, of which 6 alcohols and 14 esters constituted the major portion, and 9 key flavor compounds with an ROAV > 1 were identified throughout the fermentation process. Isoamyl acetate had the highest ROAV and made the greatest contribution to the flavor.

UNASSIGNED: While the variation in physicochemical parameters, microbial communities, metabolism, composition, and the proportion of volatile components in fermented grains (FG) affect final Baijiu quality, their complex interactions during the ultra-long fermentation of compound-flavor Baijiu (CFB) are still poorly understood.
UNASSIGNED: In this study, amplicon sequencing was used to analyze the microbial community, and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was used to analyze the volatile components in FG during ultra-long fermentation of CFB. The relationships between the dominant microbial communities, physicochemical parameters, and volatile components were analyzed using redundancy analysis and network analysis.
UNASSIGNED: During ultra-long fermentation, bacterial diversity was initially higher than during the mid and late stages. Fungal diversity in the mid stages was higher than that initially and later in the process. A total of 88 volatile components, including six alcohols, 43 esters, eight aldehydes and ketones, 13 acids, and 18 other compounds were detected in FG. Starch and reducing sugars in FG strongly affected the composition and function of bacterial and fungal communities. However, acidity had little effect on the composition and function of the bacterial flora. Lactobacillus, Bacillus, Weissella, and Pichia were the core microbial genera involved in metabolizing the volatile components of FG.
UNASSIGNED: We provide insights into the relationships and influences among the dominant microbial communities, physicochemical parameters, and volatile components during ultra-long fermentation of CFB. These insights help clarify the fermentation mechanisms of solid-state fermentation Baijiu (SFB) and control and improve the aroma quality of CFB.

Five volatile thiol compounds (methanethiol, ethanethiol, 2-mercapto-1-ethanol, 2-furfurylthiol, and 2-methyl-3-furanethiol) in fermented grains of sauce-aroma baijiu were determined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were pre-treated using a modified QuEChERS method. 4,4\'-Dithiodipyridine (DTDP) derivatization reaction improved the detectability and stability of volatile thiol compounds. From the end of the first round to the end of the seventh round of fermentation and different fermentation states from the fifth round of fermented grains of the sauce-aroma baijiu production process were analyzed. The results showed that the concentrations of methanethiol (67.64-205.37 μg/kg), ethanethiol (1.22-1.76 μg/kg), 2-furfurylthiol (0.51-3.03 μg/kg), and 2-methyl-3-furanthiol (1.70-12.74 μg/kg) were increased with the number of fermentation rounds. Methanethiol, 2-furfurylthiol, and 2-methyl-3-furanthiol increased during fermentation and distillation in the fifth round. Fermentation and distillation were important stages for their widespread production. After distillation, there were still a large number of volatile thiol compounds in the fermented grains. The thermal reaction was of great significance in the formation of these thiols.

Biofilm plays an important role in resisting the adverse environment, improving the taste and texture, and promoting the synthesis of flavor substances. However, to date, the findings on the effect of biofilm and dominating bacteria Bacillus on the ester synthesis in the Baijiu field have been largely lacked. Therefore, the objectives of the present study were to primarily isolate biofilm-producing microbes in the fermented grains, evaluate the stress tolerance capacity, and unveil the effect of biofilm and co-culture with Bacillus on the ester synthesis in the strong flavor Baijiu. Results indicated that after isolation and evaluation of stress-tolerance capacity, bacterial strain BG-5 and yeast strains YM-21 and YL-10 were demonstrated as mediate or strong biofilm-producing microbes and were identified as Bacillus velezensis, Saccharomycopsis fibuligera, and Zygosaccharomyces bailii, respectively. Solid phase microextraction/gas chromatography-mass spectrometer indicated that biofilm could enhance the diversity of esters while reduce the contents of ester. The scanning electron microscopy showed an inhibitory effect of B. velezensis on the growth of S. fibuligera, further restraining the production of esters. Taken together, both biofilm and B. velezensis influence the ester synthesis process. The present study is the first to reveal the biofilm-producing microorganisms in fermented grains and to preliminarily investigate the effect of biofilm on the ester synthesis in the Baijiu field.

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.