Starch is an essential factor affecting the quality of flue-cured tobacco, and high starch content can affect the sensory quality and safety. Recently, the degradation of macromolecules in tobacco raw materials by using additional microorganisms to improve their intrinsic quality and safety has become a new research hotspot in the tobacco industry. However, the technical maturity and application scale are limited. Our study analyzed the correlation between microbial community composition and volatile components on the surface of tobacco leaves from 14 different grades in Fujian tobacco-producing areas. The PICRUSt software was utilized to predict the function of the microbial community present in tobacco leaves. Furthermore, dominant strains that produced amylase were screened out, and an enzyme solution was prepared to enhance the flue-cured tobacco flavor. Changes in the content of macromolecules and volatile components were determined, and sensory evaluations were conducted to assess the overall quality of the tobacco leaves. The results showed that the dominant bacterial genera on the surface of Fujian tobacco leaves were Variovorax, Sphingomonas, Bacillus, etc. Bacillus was positively correlated with various volatile components, which contributed to the sweet and aromatic flavors of Fujian flue-cured tobacco. The main genetic functions of Fujian flue-cured tobacco surface bacteria were carbohydrate metabolism and amino acid metabolism. After treating flue-cured tobacco with an enzyme preparation prepared by the fermentation of Paenibacillus amylolyticus A17 #, the content of starch, pectin, and cellulose in flue-cured tobacco decreased significantly compared with the control group. Meanwhile, the content of total soluble sugar and reducing sugar was significantly increased, and the volatile aroma components, such as 3-hydroxy--damascone, 2,3-dihydro-3,5-dihydroxy-6-methyl-4 H-Pyran-4-one, ethyl palmitate, ethyl linolenic acid, etc., were significantly increased. The aroma quality and quantity of flue-cured tobacco were enhanced, while impurities were reduced. The smoke characteristics were improved, with increased fineness, concentration, and moderate strength. The taste characteristics were also improved, with reduced irritation and a better aftertaste. In conclusion, Bacillus, as the dominant genus in the abundance of bacterial communities on tobacco surfaces in Fujian, had an essential impact on the flavor of tobacco leaves by participating in carbohydrate metabolism and finally forming the unique flavor style of flue-cured tobacco in Fujian tobacco-producing areas. Paenibacillus amylolyticus A17 #, a target strain with amylase-producing ability, was screened from the surface of Fujian flue-cured tobacco. The enzyme preparation, produced by the fermentation of Paenibacillus amylolyticus A17 #, was utilized to reduce the content of macromolecules, increase the content of water-soluble total sugar and reducing sugar, and produce a variety of crucial volatile aroma components, which had a significant improvement on the quality of tobacco leaves.

To study the relationship between the diversity of the surface microbial community and tobacco flavor, and to improve tobacco quality using microorganisms. The microbial community composition and diversity of 14 samples of flue-cured tobacco from tobacco-producing areas in Yunnan with varying grades were analyzed by high-throughput sequencing. PICRUSt was used for predicting microbial functions. A strain of Bacillus amyloliquefaciens W6-2 with the ability to degrade pectin was screened from the surface of flued-cured tobacco leaves from Yunnan reroasted tobacco leave. The enzyme preparation was prepared through fermentation and then applied for treating flue-cured tobacco. The improvement effect was evaluated by measuring the content of macromolecule and the changes in volatile components, combined with sensory evaluations. The bacterial communities on the surface of flue-cured tobacco exhibited functional diversity, consisting primarily of Variovorax, Pseudomonas, Sphingomonas, Burkholderia, and Bacillus. These bacterial strains played a role in the aging process of flue-cured tobacco leaves by participating in amino acid metabolism and carbohydrate metabolism. These metabolic activity converted complex macromolecules into smaller molecular compounds, ultimately influence the smoking quality and burning characteristics of flue-cured tobacco. The pectinase preparation produced through fermentation using W6-2 has been found to enhance the aroma and sweetness of flue-cured tobacco, leading to improved aroma, reduced impurities, and enhanced smoothness. Additionally, the levels of pectin, cellulose, and hemicellulose decreased, while the levels of water-soluble sugar and reducing sugar increased, and the contents of esters, ketones, and aldehydes increased, and the contents of benzoic acid decreased. The study revealed the correlation between surface microorganisms and volatile components of Yunnan tobacco leaves, and the enzyme produced by the pectin-degrading bacteria W6-2 effectively improved the quality of flue-cured tobacco.

Two strains of Bacillus subtilis designated YY-10 and BY-2, were isolated from the surface of tobacco and found to be capable of significant lignin degradation. The predominant lignin-degrading enzymes produced by these strains were lignin peroxidase (LiP) and manganese peroxidase (MnP), respectively. A notable distinction was observed in the organoleptic evaluation and volatile flavor compounds, as determined by sensory evaluation and GC–MS analysis. The content of volatile flavor compounds, such as geranylacetone, meglumine trienone B, and meglumine trienone C, was found to be significantly increased in roasted tobacco treated with YY-10. This treatment has been shown to reduce the astringent flavor of the roasted tobacco and improve the aroma, which in turn could improve the quality of the roasted tobacco. Conversely, the quality and aroma levels of the roasted tobacco treated with the crude enzyme solution of the BY-2 strain were reduced. Transcriptome analysis revealed that the expression of genes related to amino acid metabolism, genetic material biosynthesis, and protein synthesis was up-regulated in the YY-10 strain compared with the BY-2 strain, which promoted the biosynthesis of LiP. This study provides a preliminary elucidation of the potential mechanism by which YY-10 enhances the quality of tobacco leaves through lignin-degrading enzyme production, thus establishing a research foundation for the subsequent treatment of waste tobacco raw materials and industrial applications.

Aquatic products are highly susceptible to spoilage caused by adherence of spoilage bacteria during storage, so it is crucial to develop new microecological preservatives. The probiotic B. subtilis can achieve potential preservation effects by adhering to the fish gut to regulate the amount of probiotic colonization in the flora. Here, we demonstrated the surface adhesion characteristics of B. subtilis and the mechanism of adhesion-associated gene-mediated adhesion; the regulation of B. subtilis adhesion ability by nitrogen and carbon sources, and the application of tilapia preservation. Our results revealed 10 B. subtilis adhesion-related genes, among which nitrogen sources up-regulate the adhesion genes hag and motA, carbon sources up-regulate sinR and fliG. Based on flagellum-associated high adhesion strain C6 and high adhesion regulation of B. subtilis C6_H allowed the abundance of probiotic bacteria Bacillus, Rhodobacteraceae to increase, attenuate the content of spoilage flavor substances such as 1-Octen-3-ol, 15-Crown-5 and maintain the freshness of tilapia. The results helped to reveal the adhesion mechanism of B. subtilis and to develop and apply microecological preservatives with high quality adhesion ability.