Abstract

BackgroundTo explore the optimum fermentation conditions for tobacco leaves and also screen the microbiota and metabolites that are beneficial for fermentation.MethodsTobacco leaves were fermented at 25 °C, 35 °C, and 45 °C for 2, 4, and 6 weeks, respectively. For identification of the best fermentation temperature, physicochemical properties and sensory quality of fermented tobacco were investigated. Subsequently, based on the appropriate temperature, 16 s rRNA sequencing and metabolomics analysis of tobacco were performed to monitor the change of microbes and metabolites during fermentation process (from 2 to 6 weeks).ResultsSensory quality analysis indicated that fermentation at 45 °C for 6 weeks represented the optimum condition. Metabolomics analysis showed that a total of 415 metabolites were annotated. The increase of fermentation period led to significant changes of metabolites. Results revealed an increase in concentration of L-phenylalanine and sphingosine as well as decreased concentration of betaine and phytosphingosine with the prolongation of fermentation period (2 to 6 weeks). Distinct changes in the microbiota were also observed with prolongation of the fermentation time. Results revealed that Pseudomonas, Pantoea, and Burkholderia were dominant bacteria in fermentation at 45 °C for 6 weeks. With the extension of the fermentation time, the abundance of Pseudomonas increased, while that of Sphingomonas and Methylobacterium decreased. Furthermore, microbiota profiles were tightly relevant to the altered metabolites, especially compounds involved in the sphingolipid metabolism.ConclusionSuitable fermentation conditions were 45 °C for 6 weeks; phytosphingosine and sphingosine might affect tobacco fermentation via the sphingolipid metabolism pathway. This study provides a theoretical basis for guiding tobacco fermentation and gives insights into reducing harmful substances during tobacco fermentation.

Highlights

  • To explore the optimum fermentation conditions for tobacco leaves and screen the microbiota and metabolites that are beneficial for fermentation

  • The results showed that the levels of major neutral aroma components, such as solanone, geranylacetone, dihydroactinidiolide, and megastigmatrienone, were elevated with the increase in fermenting temperature

  • Beta-diversity analysis We further evaluated the impact of fermentation time on microbial tobacco structure by principal component analysis (PCA), principal coordinate analysis (PCoA), and non-metric multidimensional scaling (NMDS) analysis (Supplementary Figure 5)

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Summary

Introduction

To explore the optimum fermentation conditions for tobacco leaves and screen the microbiota and metabolites that are beneficial for fermentation. Relying on the heat generated by the self-heating action of tobacco, it promotes the biochemical changes in leaves and improves the quality and processing characteristics of tobacco During this process, heat is not dissipated due to the poor thermal conductivity of the tobacco bulk itself, and the temperature in the stack can rise to 55– 65 °C. Undesirable fermentation by-products are produced during the fermentation process, such as tobacco-specific nitrosamines (TSNAs), whose increase in concentration parallels that of nitrite [13]. Another unpleasant by-product of organic acid metabolism is oxalic acid, which can negatively affect the taste of tobacco [14]. Taken together, minimizing the production of TSNAs and nitrite is a major goal of tobacco fermentation technology [16]

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