A Novel Data Fusion Strategy of GC-MS and 1H NMR Spectra for the Identification of Different Vintages of Maotai-flavor Baijiu.

Counterfeit Baijiu has been emerging because of the price variances of real-aged Chinese Baijiu. Accurate identification of different vintages is of great interest. In this study, the combination of gas chromatography-mass spectrometry (GC-MS) and proton nuclear magnetic resonance (1H NMR) spectroscopy was applied for the comprehensive analysis of chemical constituents for Maotai-flavor Baijiu. Furthermore, a novel data fusion strategy combined with machine learning algorithms has been established. The results showed that the midlevel data fusion combined with the random forest algorithm were the best and successfully applied for classification of different Baijiu vintages. A total of 14 differential compounds (belonging to fatty acid ethyl esters, alcohols, organic acids, and aldehydes) were identified, and used for evaluation of commercial Maotai-flavor Baijiu. Our results indicated that both volatiles and nonvolatiles contributed to the vintage differences. This study demonstrated that GC-MS and 1H NMR spectra combined with a data fusion strategy are practical for the classification of different vintages of Maotai-flavor Baijiu.

Comprehensive genomic and metabolomic analysis revealed the physiological characteristics and pickle like odor compounds metabolic pathways of Bacillus amyloliquefaciens ZZ7 isolated from fermented grains of Maotai-flavor baijiu.

Pickle like odor (PLO) is one of the main defective flavors of Maotai flavor baijiu (MFB). Understanding and controlling the PLO compounds producing strains not only solves the problem of PLO from the source, but also ensures the high-quality production of MFB. However, the relevant research on PLO compounds producing strains has not been reported in MFB. In this study, we identified a Bacillus amyloliquefaciens ZZ7 with high yield of PLO compounds in the fermented grains of MFB, and measured its physiological characteristics. It produces 627 volatile compounds and 1,507 non-volatile compounds. There are 7 volatile sulfur compounds that cause the PLO, the content of dimethyl disulfide, dimethyl trisulfide, and dimethyl sulfur is relatively high, accounting for 89.43% of the total volatile sulfur compounds. The genome size of B. amyloliquefaciens ZZ7 is 3,902,720 bp with a GC content of 46.09%, and a total of 3,948 protein coding genes were predicted. Moreover, the functional annotation of coding genes and an assessment of the metabolic pathways were performed by genome annotation, showing it has strong ability to transport and metabolize amino acids and carbohydrates. Comprehensive genomic and metabolomic analysis, the metabolic pathway of PLO compounds of B. amyloliquefaciens ZZ7 was revealed, which mainly involves 12 enzymes including sulfate adenylyltransferase, cysteine synthase, cystathionine γ-synthase, etc. This work provides biological information support at both genetic and metabolic levels for the mechanism of B. amyloliquefaciens ZZ7 to synthesize PLO compounds, and provides a direction for the subsequent genetic modification of ZZ7 to solve PLO from the source in the MFB.

Spatiotemporal accumulation differences of volatile compounds and bacteria metabolizing pickle like odor compounds during stacking fermentation of Maotai-flavor baijiu.

Pickle like odor (PLO) is undesirable in Maotai-flavor baijiu; however, its formation mechanism is unclear. Furthermore, there is a lack of understanding of the spatiotemporal accumulation of volatile compounds (including PLO compounds, PLOC) and of the microorganisms responsible for the production of PLOC during stacking fermentation. In this study, we analyzed the spatiotemporal distribution differences of 132 volatile compounds in piled fermented grains. PLOC (n = 5) were higher in pile surface than in pile center, reaching their highest levels at 6th and 5th rounds, respectively. The microorganisms in pile center were more conducive to the formation of alcohols, while those in the pile surface more promoted the synthesis of esters. Rhodococcus and Zygosaccharomyces promoted the formation of PLOC. Acetobacter was negatively correlated with the content of sulfur compounds by promoting their conversion into non-volatile sulfur compounds, thereby reducing the content of PLOC. This study provides information on the spatiotemporal differences of volatile compounds (especially PLOC) in piled fermented grains and identified the microorganisms that produce PLOC.

The spatio-temporal diversity and succession of microbial community and its environment driving factors during stacking fermentation of Maotai-flavor baijiu.

Stacking fermentation is an important stage of microbial expansion and enrichment in the brewing process of Maotai-flavor baijiu and has an important impact on quality. However, the structure and succession of microbial communities at different spatial points of fermented grains, as well as the key environmental factors driving community assembly, remain unclear. Here, we analyzed spatio-temporal similarities and differences in the microbial community structure and succession during 1-6 rounds of stacking fermentation of Maotai-flavor baijiu. The microbial diversity and richness in the pile center were higher than those at the pile surface. The dominant bacterial genus changed from Lactobacillus to Acetobacter, while the dominant fungal genus Pichia was gradually replaced by Candida, however, some microorganisms (Acetobacter, Thermoascus) could not occupy community dominance in both the pile surface and the pile center of fermented grains. Most of the biomarkers (Kroppenstedtia, Thermomyces, etc.) of the pile surface showed thermostable or thermophilic characteristics, while most biomarkers (Aspergillus, Hyphopicia, etc.) of the pile center were functional microorganisms. Furthermore, pH and moisture were the main environmental driving factors of community construction at the pile surface and the pile center, respectively, with starch and reducing sugars having a greater impact on the microbial community assembly of the pile center than that of the pile surface. The main differences in the metabolic pathways of the dominant bacterial genera of the pile surface and the pile center were concentrated around cell growth and death, amino acids, leading to enrichment and growth of microbial communities at the pile surface and nitrogen utilization at the pile center, respectively. This study reveals the spatio-temporal differences in microbial community structure, succession and corresponding environmental driving factors during stacking fermentation, which will provide guidance for regulating the microbial community diversity to produce high-quality Maotai-flavor baijiu.

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.