Abstract
To enhance the quality of tobacco leaves and optimize the smoking experience, diverse strains of functional bacteria and their associated metabolites have been used in tobacco aging. Exogenous cellulase additives are frequently employed to facilitate the degradation of cellulose and other macromolecular matrices and enhance the quality of the tobacco product. However, little is known about how microbial metabolites present in exogenous enzyme additives affect tobacco quality. In this study, crude cellulase solutions, produced by a tobacco-originating bacterium Bacillus subtilis FX-1 were employed on flue-cured tobacco. The incorporation of cellulase solutions resulted in the reduction of cellulose crystallinity in tobacco and the enhancement of the overall sensory quality of tobacco. Notably, tobacco treated with cellulase obtained from laboratory flask fermentation demonstrated superior scent and flavor attributes in comparison to tobacco treated with enzymes derived from industrial bioreactor fermentation. The targeted and untargeted metabolomic analysis revealed the presence of diverse flavor-related precursors and components in the cellulase additives, encompassing sugars, alcohols, amino acids, organic acids, and others. The majority of these metabolites exhibited significantly higher levels in the flask group compared to the bioreactor group, probably contributing to a pronounced enhancement in the sensory quality of tobacco. Our findings suggest that the utilization of metabolic products derived from B. subtilis FX-1 as additives in flue-cured tobacco holds promise as a viable approach for enhancing sensory attributes, establishing a solid theoretical foundation for the potential development of innovative tobacco aging additives.
Graphical abstract
Cellulase solutions obtained from Bacillus subtilis FX-1, a bacterium originating from tobacco, were applied to flue-cured tobacco during the aging process. The cellulase structure of tobacco was analyzed using 13C cross-polarization magic angle spinning NMR spectroscopy. Sensory evaluation was utilized to assess the sensory attributes and quality characteristics. Targeted and untargeted metabolomics were utilized to investigate the chemical components present in the cellulase solutions. The inclusion of cellulase additives altered the cellulose structure in tobacco and introduced various flavor-related precursors and components. The utilization of cellulase additives derived from B. subtilis FX-1 substantially improved the overall quality of the tobacco product.
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Acknowledgements
This work was funded by the Xiamen Natural Science Foundation (3502Z202371034), the National Natural Science Foundation of China (42188102), the Opening Foundation of Fujian Key Laboratory of Marine Carbon Sequestration Research Fund (FKLMCS2023005), the Science and Technology Project of China National Tobacco Corporation (110202202022, 110202102017), and the Opening Foundation of Fujian Key Laboratory of Marine Carbon Sequestration Research Fund (FKLMCS2023006). The authors would also like to express their gratitude to the editor and anonymous reviewers for their valuable comments, which greatly contributed to the improvement of this manuscript.
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This work was supported by the Xiamen Natural Science Foundation (3502Z202371034), the National Natural Science Foundation of China (42188102), the Opening Foundation of Fujian Key Laboratory of Marine Carbon Sequestration Research Fund (FKLMCS2023005), the Science and Technology Project of China National Tobacco Corporation (110202202022, 110202102017), and the Opening Foundation of Fujian Key Laboratory of Marine Carbon Sequestration Research Fund (FKLMCS2023006).
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XC and TL performed the experiments equally. XC analyzed data, wrote the first draft of the manuscript, and edited the manuscript. TL analyzed data and edited the manuscript. SH, YC, and ZJ prepared material and performed the experiments. HL analyzed data. CC edited the manuscript. JL, SC, and WH provided technical support for the study. XT and JF designed and supervised the study. All authors read and approved the submitted final manuscript.
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Chen, X., Long, T., Huang, S. et al. Metabolomics-based study of chemical compositions in cellulase additives derived from a tobacco-origin Bacillus subtilis and their impact on tobacco sensory attributes. Arch Microbiol 206, 163 (2024). https://doi.org/10.1007/s00203-024-03876-x
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DOI: https://doi.org/10.1007/s00203-024-03876-x