Search

Aging is a time-consuming step in the manufacturing of fermented alcoholic beverages. Natural-aging huangjiu sealed in pottery jars was taken as an example to investigate the changes of physiochemical indexes during aging and to quantify intercorrelations between aging-related factors and metabolites through machine learning methods. Machine learning models provided significant predictions for 86% of metabolites. Physiochemical indexes well reflected the metabolic profile, and total acid was the most important index that needed to be controlled. For aging-related factors, several aging biomarkers of huangjiu were also well predicted. Feature attribution analysis showed aging year was the most powerful predictive factor, and several microbial species were significantly associated with aging biomarkers. Some of the correlations, mostly connected to environmental microorganisms, were newly found, showing considerable microbial influence on aging. Overall, our results reveal the potential determinants that affect the metabolic profile of aged huangjiu, paving the way for a systematical understanding of changes in metabolites of fermented alcoholic beverages.

Daqu and wheat Qu are saccharification and fermenting agents in Chinese huangjiu and baijiu production. This study aimed to investigate the difference between Daqu and wheat Qu in physicochemical indices, microbial communities, functional genes, and the metabolic network of key microbes responsible for flavor synthesis by whole-metagenome sequencing and metabolite analysis. Herein, physicochemical indices indicated that compared with wheat Qu, Daqu exhibited higher protease and cellulase activity and acidity, and lower glucoamylase and amylase enzyme activity. Metagenomic sequencing reveals that although Daqu and wheat Qu community composition have significant differences at species level, they have similar functional genes. Daqu were enriched in Pediococcus pentosaceus, Weissella paramesenteroides, Rasamsonia emersonii and Byssochlamys spectabilis (22.48% of the total abundance), while wheat Qu harbored greater abundances of Saccharopolyspora (54.78%, Saccharopolyspora rectivirgula, Saccharopolyspora shandongensis, Saccharopolyspora hirsuta, Saccharopolyspora spinose, and Saccharopolyspora erythraea). From a functional perspective, the important functions of Daqu and wheat Qu are both amino acid metabolism and carbohydrate metabolism. Meanwhile, a combined analysis among microbiota, functional genes, and dominant flavors indicated S. shandongensis, S. rectivirgula, and S. spinose might be the main contributor to the synthesis of flavor compounds in wheat Qu, while R. emersonii, W. paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Weissella cibaria and P. pentosaceus may make the greatest contribution to flavor compounds synthesis in Daqu. This study reveals the microbial and functional dissimilarities of Daqu and wheat Qu, and helps elucidating different metabolic roles of microbes during flavor formation.

Aromatic alcohols (including β-phenylethanol, tyrosol and tryptophol) could contribute positively to the aroma flavor of huangjiu at an appropriate amount. In this study, it was found that the aromatic alcohols in huangjiu were predominantly synthesized by Saccharomyces cerevisiae HJ01. The Ehrlich pathway and shikimate pathway, including 31 genes, are responsible for aromatic alcohol biosynthesis in S. cerevisiae. During the first 120 h of huangjiu fermentation, the genes GAP1HJ01 (permease), BAT2HJ01 (transaminase), PDC1HJ01 and PDC5HJ01 (pyruvate decarboxylase), and ADH1HJ01 (alcohol dehydrogenase) formed integrated Ehrlich pathway are responsible for aromatic alcohol biosynthesis. In S. cerevisiae HJ01, the mutations in the promoter region of genes GAP1HJ01, BAT2HJ01, PDC1HJ01, and PDC5HJ01 involved in the Ehrlich pathway increased the promoter strength and transcriptional levels of related genes. The addition of 20 U/g acid protease could substantially increase the expression levels of genes involved in the Ehrlich pathway, and therefore there were 34.7% increasing for the concentration of total aromatic alcohols.

Inoculated raw wheat Qu (IRWQ), which can be produced with high efficiency and low cost while maintaining stable quality, is a new Qu that has been applied to Huangjiu brewing. In this study, single molecule real-time DNA sequencing technology and culture-dependent methods were combined for the first time to study the microbiota and the function of the principle microorganisms in IRWQ. The glucoamylase, amylase and protease contents of IRWQ were 1.17, 1.55 and 2.87-times greater, respectively, than those of traditional wheat Qu. Like traditional wheat Qu, the main volatile flavor compounds in IRWQ were alcohols; however, the esters content was much higher and the acids content was much lower. Single molecule real-time DNA sequencing technology identified 18 fungal species and 59 bacterial species in IRWQ. Then, 30 species were isolated by culture-dependent methods. These species represented about 80% of the total fungal microbiota and 35% of the total bacteria microbiota. Molds were the main contributors of glucoamylase, amylase and protease activities. Aspergillus flavus had the highest glucoamylase and protease activity. Bacillus subtilis and Bacillus amyloliquefaciens also produced high hydrolytic enzyme activities. Saccharomyces cerevisiae produced the largest amounts of aromatics compounds, alcohols, esters and acids. Aldehydes and ketones are mainly produced by molds.