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. 2024 May 24:22:101508.
doi: 10.1016/j.fochx.2024.101508. eCollection 2024 Jun 30.

Comprehensive analysis of spatial heterogeneity reveals the important role of the upper-layer fermented grains in the fermentation and flavor formation of Qingxiangxing baijiu

Lei Tian  1   2 , Pei Xu  1   2 , Junyu Chen  1   2 , Hang Chen  3 , Ji Qin  4 , Xiaotian Wu  1   2 , Chengzhe Liu  5 , Zongjun He  5 , Ying Liu  1   2 , Tongwei Guan  1   2
Affiliations

Comprehensive analysis of spatial heterogeneity reveals the important role of the upper-layer fermented grains in the fermentation and flavor formation of Qingxiangxing baijiu

Lei Tian et al. Food Chem X. .

Abstract

Different spatial positions lead to inconsistent fermentation effects and flavors, however, the spatial heterogeneity of Qingxiangxing (QXX) Baijiu remains unknown. We investigated the microbes, flavors, and physicochemical properties of different layers in fermented grains of QXX Baijiu using Illumina HiSeq sequencing, two-dimensional gas chromatography-mass spectrometry (GC × GC-MS) and ultra-high performance liquid chromatography-mass (UHPLC-MS). A total of 79 volatiles, 1596 metabolites, 50 bacterial genera, and 52 fungal genera were identified. The contents distribution followed the order: upper layer > bottom layer > middle layer. Organic acids and derivatives were the main differential metabolites across the three layers. Starch, pH, and reducing sugar levels increased from the upper to bottom layer. Saccharomyces and Lactobacillus were dominant microbes. Pediococcus, the biomarker of upper layer, showed positive correlations with formic acid, ethyl lactate, acetic acid, ethyl linoleate, and ethyl oleate. These findings deepen our understanding of the fermentation and flavor formation mechanisms of QXX Baijiu.

Keywords: Different layers of fermented grains; Metabolites; Microbial composition; Physicochemical properties; Qingxiangxing baijiu; Volatile compounds.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this paper. The following are the supplementary data related to this article. Supplementary Fig. S1The richness, diversity, Lefse analysis of microbial communities in different FG layers. Shannon (A), Chao (B), Simpson (C), and Sobs (D) indices of bacteria. Shannon (E), Chao (F), Simpson (G), and Sobs (H) indices of fungi. Venn diagram of bacteria (I) and fungi (J) communities at genus level. Lefse analysis of bacteria (K) and fungi (L).Supplementary Fig. S1 Supplementary Fig. S2Microbial differences analysis among groups. (A) bacteria and (B) fungi (* P < 0.05) evaluated by Kruskal–Wallis H test.Supplementary Fig. S2 Supplementary Fig. S3Analysis of group differences in some characteristic microorganisms and metabolites. (A) The differences of genus Pediococcus among three layers. (B) The differences of Lactobacillus among four groups. (C) The differences of norleucine between UL and BL groups. (D) The differences of norleucine between AF and BF groups.Supplementary Fig. S3 Supplementary Fig. S4The top 50 significantly different metabolites with VIP > 1.0 and P < 0.05 were shown in the heatmap.Supplementary Fig. S4 Supplementary Fig. S5Functional annotation and classification of differential metabolites identified in the comparison between different layers. Classification of differential metabolites between (A) AF and BF, (B) UL and BL, (C) UL and ML, (D) BL and ML in the superclass. Classification of differential metabolites between (E) AF and BF, (F) UL and BL, (G) UL and ML, (H) BL and ML in the subclass.Supplementary Fig. S5 Supplementary Fig. S6The correlation between the bacterial genus and non-VFCs by heatmap analysis.Supplementary Fig. S6 Supplementary Fig. S7The correlation between the fungal genus and non-VFCs by heatmap analysis.Supplementary Fig. S7 Supplementary material 1Image 1 Supplementary data to this article can be found online at https://doi.org/10.1016/j.fochx.2024.101508.

Figures

Fig. 1
Fig. 1
Microbial community characteristics analysis of different FG. Beta diversity of the (A) bacterial and (B) fungal communities assessed by PCA analysis. The circos plots of the (C) bacterial and (D) fungal communities at the phylum level. The bar plots of the (E) bacterial and (F) fungal communities at the genus level.
Fig. 2
Fig. 2
Physicochemical factors in different FG. (A) Moisture content; (B) Acidity; (C) pH value; (D) Starch content; (E) Reducing sugar content.
Fig. 3
Fig. 3
The correlation network analysis between the microbial communities and physicochemical properties as well as flavor compounds of the FG. The CCA analysis of the correlation between the microbial communities (bacteria A, fungi B) and physicochemical properties at the genus level (P < 0.05). (C) The correlation between the microbial communities and VFCs by network correlation analysis. The line thickness indicates the correlation strength, with a yellow line denoting a positive correlation and a gray line signifying a negative correlation. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 4
Fig. 4
VFCs analysis of FG in the different layers. (A) VFC types and quantities; (B) VFC relative content in different layers; (C) Venn diagram of types of VFC; (D) The heatmap of the VFCs; (E) VFCs displaying distinct differences between the groups are identified via multivariate VIP analysis; (F) Analysis of the main characteristic VFC of QXX Baijiu.
Fig. 5
Fig. 5
Multivariate statistical analysis of non-volatile metabolites in the fermentation grains of different layers. The PCA analysis of (A) negative and (B) positive ions. Topological analysis of KEGG pathway between (C) AF and BF, (D) ML and BL, (E) UL and BL, (F) UL and ML.
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