Screening of Tobacco Leaf Medium for Enriching Functional Microorganisms and Its Effect on Improving Flue-cured Tobacco Quality

doi:10.13560/j.cnki.biotech.bull.1985.2025-1252

Abstract

Abstract:

Objective To investigate the effects of targeted enrichment using a tobacco leaf-derived liquid medium on the metabolic characteristics of functional strain fermentation broths and their impact on improving the quality of flue-cured tobacco. Method Two functional strains were isolated from aged tobacco leaves and identified as Microbacterium lateritum M2 and Bacillus subtilis M24 based on physiological, biochemical, and 16S rDNA sequence analyses. The strains were enriched in LB liquid medium and tobacco leaf liquid medium, respectively. Non-targeted metabolomics was employed to analyze differences in metabolite composition in the fermentation broths. Systematic evaluations were conducted on changes in chemical components, macromolecular substance content, and sensory quality of tobacco leaves after spraying treatment with the fermentation broths. Result Both strains M2 and M24 were capable of producing amylase, protease, and cellulase, with strain M2 additionally producing hemicellulase. Non-targeted metabolomics analysis revealed significant differences in the metabolite profiles of fermentation broths enriched in different media. The content of polyphenols, such as chlorogenic acid and quinic acid, was significantly higher in the tobacco leaf medium group compared to the LB medium group. After spraying treatment, the fermentation broths obtained from tobacco leaf medium enrichment (T2, T4) demonstrated superior tobacco quality improvement effects. Compared to the LB medium group, tobacco leaves treated with these broths showed a reduction in cellulose content by 5.32%–14.63%, a significant increase in total sugar and reducing sugar content, an improved sugar-to-alkaloid ratio, a decrease in total nitrogen and nitrogen-to-alkaloid ratio, and notable enhancements in sensory quality, including reduced off-flavors and irritancy, as well as improved aroma quality, aroma quantity, and sweetness. Conclusion Tobacco leaf medium optimizes the metabolic characteristics of functional strains, promoting the enrichment of more polyphenols beneficial for tobacco quality in the fermentation broth, thereby more effectively enhancing aroma and reducing harshness in flue-cured tobacco. This study provides a new theoretical basis for the targeted application of microbial fermentation technology in tobacco processing.

Key words: functional microorganisms, tobacco medium, enzyme activity, chemical quality, sensory quality

HU Yi-chao, QIN Tian, SUN Jian-sheng, SU Zan, ZHOU Yi, LIU Jin-cang, LIU Zheng-qin, ZHANG Tong-zhuo, GUAN Ming-xin, MA Ting-ting, LI Juan. Screening of Tobacco Leaf Medium for Enriching Functional Microorganisms and Its Effect on Improving Flue-cured Tobacco Quality[J]. Biotechnology Bulletin, 2026, 42(5): 113-123.

Figures/Tables 10

References 42

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处理Treatment	总氮 Nitrogen （%）	全钾 Potassium （%）	氯离子 Chloride ion （%）	总糖 Total sugar （%）	还原糖 Reduced sugar （%）	烟碱Nicotine （%）	糖碱比 Sugar-nicotine ratio	氮碱比Nitrogen-nicotine ratio	钾氯比Potassium-chloride ratio
CK	1.96±0.03a	2.94±0.07a	0.42±0.07a	32.60±1.59ab	22.37±0.79c	2.74±0.02a	11.88±0.55b	0.71±0.01bc	6.99±0.42a
T1	1.97±0.11a	2.96±0.12a	0.42±0.12a	30.61±0.97b	23.65±1.25ab	2.62±0.07a	11.69±0.23b	0.75±0.01a	7.32±1.73a
T2	1.88±0.08cd	2.91±0.15a	0.39±0.15a	34.13±0.47a	24.66±1.04a	2.62±0.16a	13.04±0.43a	0.72±0.02bc	7.50±1.07a
T3	1.92±0.05bc	3.03±0.26a	0.35±0.26a	31.75±1.00ab	23.23±0.67ab	2.62±0.04a	12.11±0.32b	0.73±0.03ab	8.86±2.00a
T4	1.87±0.08d	3.03±0.09a	0.43±0.09a	32.44±2.04ab	25.19±0.86a	2.69±0.12a	12.04±0.57b	0.69±0.02c	7.13±1.47a

处理Treatment	总氮 Nitrogen （%）	全钾 Potassium （%）	氯离子 Chloride ion （%）	总糖 Total sugar （%）	还原糖 Reduced sugar （%）	烟碱Nicotine （%）	糖碱比 Sugar-nicotine ratio	氮碱比Nitrogen-nicotine ratio	钾氯比Potassium-chloride ratio
CK	1.96±0.03a	2.94±0.07a	0.42±0.07a	32.60±1.59ab	22.37±0.79c	2.74±0.02a	11.88±0.55b	0.71±0.01bc	6.99±0.42a
T1	1.97±0.11a	2.96±0.12a	0.42±0.12a	30.61±0.97b	23.65±1.25ab	2.62±0.07a	11.69±0.23b	0.75±0.01a	7.32±1.73a
T2	1.88±0.08cd	2.91±0.15a	0.39±0.15a	34.13±0.47a	24.66±1.04a	2.62±0.16a	13.04±0.43a	0.72±0.02bc	7.50±1.07a
T3	1.92±0.05bc	3.03±0.26a	0.35±0.26a	31.75±1.00ab	23.23±0.67ab	2.62±0.04a	12.11±0.32b	0.73±0.03ab	8.86±2.00a
T4	1.87±0.08d	3.03±0.09a	0.43±0.09a	32.44±2.04ab	25.19±0.86a	2.69±0.12a	12.04±0.57b	0.69±0.02c	7.13±1.47a

处理Treatment	蛋白质Protein （%）	淀粉Starch （%）	纤维素Cellulose （%）
CK	12.22±0.20a	6.47±0.21a	13.30±0.98a
T1	11.97±0.30ab	6.20±0.54ab	13.17±1.70a
T2	11.77±0.19b	5.75±0.09b	12.47±1.27bc
T3	11.86±0.24ab	6.45±0.41a	13.47±1.86a
T4	11.73±0.10b	5.99±0.18ab	11.50±0.75c

处理Treatment	蛋白质Protein （%）	淀粉Starch （%）	纤维素Cellulose （%）
CK	12.22±0.20a	6.47±0.21a	13.30±0.98a
T1	11.97±0.30ab	6.20±0.54ab	13.17±1.70a
T2	11.77±0.19b	5.75±0.09b	12.47±1.27bc
T3	11.86±0.24ab	6.45±0.41a	13.47±1.86a
T4	11.73±0.10b	5.99±0.18ab	11.50±0.75c

处理 Treatment	评吸单项得分 Score of single evaluation （0-9.0）								综合得分Total score
处理 Treatment	香气质Aroma quality	香气量Aroma quantity	刺激性Irritation	杂气Offensive flavor	余味 Aftertaste	甜度Sweetness	燃烧性Burning property	灰色 Ash color	综合得分Total score
CK	7	6.3	6.5	6.5	6	6.3	3	4	45.6
T1	7.3	6.5	6.5	6.3	6	6.3	3	4	45.9
T2	7.5	6.8	6.8	7	6.5	7	3	4	48.6
T3	7	6.5	6.3	6.3	6.4	6.5	3	4	46.0
T4	7.5	6.7	7	6.8	6.5	6.8	3	4	48.3