Metabolic enzyme activity and microbial composition of the air-curing and fermentation processes determine the quality of cigar tobacco leaves (CTLs). In this study, we reveal the evolution of the dominant microorganisms and microbial community structure at different stages of the air-curing and fermentation processes of CTLs. The results showed that the changes in metabolic enzymes occurred mainly during the air-curing phase, with polyphenol oxidase (PPO) being the most active at the browning phase. Pseudomonas, Bacteroides, Vibrio, Monographella, Bipolaris, and Aspergillus were the key microorganisms in the air-curing and fermentation processes. Principal coordinate analysis revealed significant separation of microbial communities between the air-curing and fermentation phases. Redundancy analysis showed that bacteria such as Proteobacteria, Firmicutes, Bacteroidota, and Acidobacteriota and fungi such as Ascomycota and Basidiomycota were correlated with enzyme activity and temperature and humidity. Bacteria mainly act in sugar metabolism, lipid metabolism, and amino acid metabolism, while fungi mainly degrade lignin, cellulose, and pectin through saprophytic action. Spearman correlation network analysis showed that Firmicutes, Proteobacteria, and Actinobacteria were the key bacterial taxa, while Dothideomycetes, Sordariomycetes, and Eurotiomycetes were the key fungal taxa. This research provides the basis for improving the quality of cigars by improving the air-curing and fermentation processes. Key points: • Changes in POD and PPO activity control the color change of CTLs at the air-curing stage. • Monographella, Aspergillus, Pseudomonas, and Vibrio play an important role in air-curing and fermentation. • Environmental temperature and humidity mainly affect the fermentation process, whereas bacteria such as Proteobacteria, Firmicutes, Bacteroidota, and Acidobacteriota and fungi such as Ascomycota and Basidiomycota are associated with enzyme activity and temperature and humidity. Graphical

Changes in volatile metabolites during cigar tobacco leaves fermentation as well as the metabolic pathways of metabolites with significant differences were investigated to determine the influence of cigar tobacco leaves fermentation on its flavor. The volatile substances in cigar tobacco leaves at different stages were detected by headspace-solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS), and the main differences in volatile substances in cigar tobacco leaves at different fermentation stages of Yunxue1 in Yuxi production area were analyzed by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The results show that in the process of cigar tobacco leaves fermentation (YXF0, YXF1, YXF2, YXF3, YXF4, YXF5), a total of 613 volatile metabolites were detected, and a significant difference was found in 263 kinds of metabolites. Among them, the main upregulated differential metabolites were 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, [S-(E,Z,E,E)]-, Benzoic acid, Benzaldehyde, etc. While the main downregulated differential metabolites included beta.-Myrcene, trans-Farnesol, etc. The metabolites with significant differences are mainly concentrated in the biosynthesis of monoterpenes, diterpenes, sesquiterpenes and triterpenes, the degradation metabolism of amino acids, such as valine, leucine and isoleucine, and the biosynthesis of phenylpropyl. There were 8 different metabolites in 5 groups, including 4- (1-methylethyl) -1-cyclohexene-1-formaldehyde、2, 4-dihydroxyacetophenone、2-methylbutyl 3-methylbutyrate and methylpyrazine, all of which showed upregulation trend during fermentation. In the fermentation process, volatile metabolites participate in various synthesis and degradation pathways. The biosynthesis pathway of terpenes and amino acid synthesis and degradation pathway are connected to produce various terpenes, aldehydes and other substances, such as 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, [S-(E,Z,E,E)]-、benzaldehyde and 4-hydroxybenzaldehyde, which are conducive to the overall flavor and quality of cigar tobacco leaves.

The surface color of cigar tobacco leaves (CTLs) is largely determined by the browning response and is one of the most important quality traits affecting consumer preferences. The physiological changes and metabolic network of browning in CTLs after harvest have not been reported. We investigated the molecular mechanism of browning in CTLs by comparing the physiological characteristics and metabolites of CTLs browning at five postharvest periods. Phenotypic traits and browning-related enzyme activity data indicated that the browning reaction was most intense at approximately 12 d when polyphenol oxidase and peroxidase activities reached their maximum. Postharvest CTLs polyphenols, polyacids, and carbon and nitrogen compounds were rapidly degraded during the yellowing phase, and the biosynthesis rate of polyphenols and carotenoids was greater than the degradation rate during the browning phase. Metabolomic analysis discovered 2027 metabolites that were annotated mainly to lipids and lipid-like molecules, organic acids and their derivatives, and organic heterocyclic components. Hierarchical cluster analysis discovered glutamate, serine, threonine, ornithine, and arginine as the major amino acids involved in the browning reaction. KEGG pathway enrichment analysis indicated that the metabolic pathways with important effects on enzymatic browning were phenylpropanoid biosynthesis, flavonoid biosynthesis, isoquinoline alkaloid biosynthesis, phenylalanine metabolism, and linoleic acid metabolism. The metabolic pathway network was mined, and the main polyphenols involved in the browning reaction of CTLs were found to be cyanidin, rutin, caffeoylquinic acid, kaempferol, naringin, and neohesperidin. This study provides a reference for the browning physiology and metabolism network of postharvest CTLs. Graphical Abstract

The purpose of this study was to elucidate how the microbiota affects the metabolic state and characteristic flavor development of cigar tobacco leaves (CTL) in the fermentation process through microbial metabolism and co-metabolism with the host. The results showed that core bacterial communities in the fermentation process were Cyanobacteria , Pseudomonas , and Staphylococcus , and Aspergillus , Penicillium , and Inocybe were the most metabolically active fungi. Pseudomonas fulva , Staphylococcus nepalensis , Bacillus subtilis , Stenotrophomonas rhizophila , Alternaria alternata , and Aspergillus cristatus could degrade carbon and nitrogen compounds, such as protein, starch, lignin, and nicotine. There were 12 common non-volatile metabolites and three common volatile metabolites in DH, LC, PE, and YX before and after fermentation, of which menatetrenone, solanidine, γ-glutamylphenylalanine, carotol, and phenol, 4-(1,1,3,3-tetramethylbutyl) were significantly different before and after fermentation. The synthesis and degradation metabolism of various amino acids, alkaloids, and flavonoids are the key metabolic pathways of characteristic flavor development during CTL fermentation. Co-occurrence and interaction patterns showed that seven bacteria and 12 fungi were strictly linearly positively or negatively correlated with 72 and 55 volatile compounds, respectively. In conclusion, this study preliminarily confirmed that CTL fermentation is a microbially mediated carbon–nitrogen coupling metabolism. The carbohydrates in tobacco leaves were largely decomposed and consumed, providing energy sources for microorganisms and the carbon skeleton required for cell construction. The nitrogen-containing macromolecular compounds were degraded to form volatile compounds or flavor precursors with typical flavor. IMPORTANCE: The development of the metabolic state and characteristic flavor of cigar tobacco during fermentation is the key to process control. Innovative discoveries in the development of the metabolic state and characteristic flavor of cigar tobacco during fermentation are the key to process control. Innovative discoveries of core functional microorganisms and key metabolites were made during fermentation, suggesting potential pathways for carbon and nitrogen metabolisms. We demonstrate for the first time that cigar tobacco leaf fermentation is a microbially mediated carbon–nitrogen coupling metabolism. Many carbohydrates in tobacco leaves are decomposed and consumed to provide energy source and carbon skeleton for the construction of microbial cells, and nitrogen-containing macromolecular compounds are degraded to form volatile compounds or flavor precursors with typical flavors.

Potato and its products have become indispensable foods and snacks for most people. Steroidal glycoalkaloids (SGAs) occur in all tissues of the potato, and consuming potatoes with a high SGA content harms human health. Therefore, the effects of different cooking methods on the SGA content in potato foods were investigated in this study. The results indicated that adding food-grade acetic acid during the manufacturing process did not affect the SGA content in stir-fried shredded potatoes or fresh mashed potatoes. However, the SGA content in potato food after peeling was significantly lower than that in non-peeled food, and the volume ratio of potato skin to flesh decreased with the increase of the potato tuber volume. Therefore, potato breeders and farmers should make the most hard to increase the proportion of commodity potato via corresponding science and technology. In addition, frying significantly reduced the SGA content in potato chips. Further research indicated that SGAs degraded slowly at 150°C, while they degraded rapidly at 190°C within 30 min. The temperature of rapeseed oil in the frying process can be as high above 200°C. Thus, frying significantly decreased the SGA content in potato chips, which could be attributed to temperature. These results will provide a theoretical basis and practical guidance for potato breeding and cooking.