Composite biological treatment method for improving quality of cigar-flavored heated cigarettes

Abstract

The invention provides a composite biological treatment method for improving the quality of cigar-flavored heated cigarettes. A composite biological treatment method for improving the quality of cigar-flavored heated cigarettes comprises the following steps: (1) removing tobacco stems from cigar tobacco leaves, and performing physical crushing and sieving to obtain cigar tobacco powder; (2) adding a cellulase solution into the cigar tobacco powder, uniformly stirring, and treating at 35-45 DEG C for 2-6 hours; (3) after the step (2) is completed, adding bacillus subtilis DES-59 bacterial liquid into the cigar tobacco powder, and uniformly stirring; and (4) fermenting the treated cigar tobacco powder for 7-15 days. According to the treatment method disclosed by the invention, an efficient composite biological fermentation system is constructed through cooperative treatment of cellulase and bacillus subtilis DES-59. The system not only obviously accelerates and deepens the fermentation process of cigar tobacco, but also realizes directional regulation and control of chemical components of the tobacco.

Description

Technical Field

[0001] This application relates to the field of tobacco processing technology, and in particular to a composite biological treatment method for improving the flavor and quality of heated cigarettes for cigars. Background Technology

[0002] In the field of heated tobacco products (HNB), higher demands are placed on the efficient aroma release and pleasant smoke characteristics of raw materials under low-temperature heating conditions. Cigar tobacco, with its naturally rich aroma, is considered a high-potential premium raw material. However, its inherent chemical composition—high cellulose and protein content and low soluble sugar content—constitutes a major bottleneck in its application. The former easily generates undesirable off-flavors during pyrolysis, while the latter directly leads to a lack of aroma precursors, ultimately making it difficult to achieve ideal aroma quality and a smooth smoke profile in HNB applications.

[0003] Currently, targeted quality improvement technologies have significant shortcomings: traditional aging or fermentation methods are inefficient; while single-enzyme treatments can degrade specific macromolecules (such as cellulose or proteins), they cannot simultaneously synergistically promote aroma precursors (such as sugars and amino acids) or utilize aroma precursors for aroma production, resulting in limited improvement effects. Therefore, there is an urgent need for an innovative treatment method that can synergistically transform multiple undesirable components in cigar tobacco and simultaneously promote aroma and flavor generation, in order to unlock its full application potential in high-end HNB products. Summary of the Invention

[0004] In view of this, the purpose of this application is to provide a composite biological treatment method for improving the quality of heated cigar cigarettes with cigar flavor. The core of this method is to sequentially treat cigar tobacco raw materials with cellulase, inoculate them with Bacillus subtilis DES-59, and carry out solid-state fermentation to directionally transform macromolecular substances, enrich aroma precursors, and ultimately improve the overall quality of heated cigar cigarettes with cigar flavor.

[0005] This application provides a composite biological treatment method for improving the flavor and quality of heated cigarettes, comprising the following steps: (1) After removing the stems from the cigar tobacco leaves, they are physically crushed, sieved, and cigar tobacco powder is obtained. (2) Add cellulase solution to the cigar tobacco powder at 20-28℃ and stir evenly, then treat at 35-45℃ for 2-6 hours; (3) After completing step (2), add Bacillus subtilis DES-59 bacterial solution to the cigar tobacco powder at 20-28℃ and stir evenly; (4) The treated cigar tobacco powder is fermented for 7-15 days at 20-28℃ and 60-70% relative humidity.

[0006] Preferably, the amount of cellulase used is 180-240 U / g of the cigar tobacco powder.

[0007] Preferably, the inoculum size of Bacillus subtilis DES-59 is 5 × 10⁻⁶. 8 -7×10 8 CFU / g of the cigar tobacco powder.

[0008] Preferably, the total amount of solution and bacterial solution applied in steps (2) and (3) is 0.5-0.7 mL / g of the cigar tobacco powder.

[0009] Preferably, the solution and bacterial solution are applied by spraying in steps (2) and (3).

[0010] Preferably, the processing method further includes deactivating the cigar tobacco powder processed in step (4) and drying it.

[0011] Preferably, the inactivation temperature is 80-100℃, and the treatment time is 20-40 minutes.

[0012] Preferably, the drying temperature is 40-80℃, and the treatment time is 5-10 hours.

[0013] This application also provides a cigar-flavored heated cigarette, using the cigar tobacco powder obtained by any of the above-described processing methods as raw material.

[0014] This application's processing method constructs a highly efficient composite bio-fermentation system through the synergistic treatment of cellulase and Bacillus subtilis DES-59. This system not only significantly accelerates and deepens the fermentation process of cigar tobacco but also achieves targeted regulation of tobacco chemical components: on the one hand, the synergistic effect of cellulase and protease secreted by the strain efficiently degrades high-content macromolecules such as cellulose and protein in tobacco; on the other hand, this composite system actively promotes the generation and accumulation of small-molecule aroma precursors such as soluble sugars and amino acids, and enhances the overall metabolic aroma-producing capacity of the microbial community. Therefore, this application's processing method can significantly improve fermentation efficiency under optimized environmental conditions, effectively shorten the traditional process cycle, and ultimately achieve a comprehensive improvement in the aroma release characteristics and sensory quality of cigar tobacco in heated tobacco products.

[0015] The cigar-flavored heated tobacco produced in this application exhibits significantly improved aroma release and sensory quality under low-temperature heating conditions. The aroma originates from within the tobacco itself, closely resembling the natural aroma of a cigar. The smoke is delicate, without any jarring artificial flavoring, and the aroma and smoke are highly integrated, releasing slowly, continuously, and stably during heating. Attached Figure Description

[0016] Figure 1The results of the chemical composition analysis of cigar tobacco powder are as follows: (A): cellulose; (B): protein; (C): total sugar; (D): reducing sugar; (E): amino acids; (F): total alkaloids.

[0017] Figure 2 The results show the detection of the release of various pyrolysis aroma-related substances. Detailed Implementation

[0018] To better understand the technical solution of this application, the above technical solution will be described in detail below with reference to the accompanying drawings and specific embodiments. Unless otherwise specified, the experimental methods in the following embodiments are conventional methods. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in this field, or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0019] A composite biological treatment method for improving the flavor and quality of heated cigarettes for cigars includes the following steps: (1) After removing the stems from the cigar tobacco leaves, they are physically crushed and sieved to obtain cigar tobacco powder.

[0020] (2) Add cellulase solution to cigar tobacco powder at 20-28℃ and stir evenly. The amount of cellulase is preferably 180-240 U / g cigar tobacco powder. Treat at 35-45℃ for 2-6 hours.

[0021] (3) After completing step (2), add Bacillus subtilis DES-59 bacterial solution to the cigar tobacco powder at 20-28℃ and stir evenly. The preferred inoculum amount of Bacillus subtilis DES-59 is 5×10⁻⁶. 8 -7×10 8 CFU / g cigar tobacco powder. The total amount of solution and bacterial solution applied in steps (2) and (3) is preferably 0.5-0.7 mL / g cigar tobacco powder.

[0022] (4) The treated cigar tobacco powder is fermented at 20-28℃ and 60-70% relative humidity for 7-15 days. The cigar tobacco powder is then inactivated at 80-100℃ for 20-40 minutes, followed by drying at 40-80℃ for 5-10 hours.

[0023] This application's processing method first treats cigar tobacco with cellulase, then inoculates it with Bacillus subtilis DES-59, which has high protease activity and aroma-enhancing functions, and then carries out solid-state fermentation under controlled conditions. This method, through complex biological processes, efficiently degrades cellulose and protein in the tobacco, simultaneously accumulating aroma precursors such as sugars and amino acids, and further develops aroma through microbial fermentation. This comprehensively improves the chemical composition and thermal decomposition aroma characteristics of the cigar tobacco raw materials, ultimately enhancing the sensory quality of heated cigar cigarettes. The aroma substances are endogenously generated and evenly distributed, releasing more stably under heated, non-combustion conditions, resulting in a more harmonious smoke without any abrupt artificial flavoring. In contrast, adding flavorings to heated cigarette tobacco often leads to a strong initial aroma followed by a weak, unharmonious finish. Furthermore, the processing method of this application has a short cycle of 7-14 days.

[0024] The source of biological materials in the embodiments of this application is: Cigar tobacco leaves: Cigar tobacco leaves of the "Yunxue No. 6" variety after drying and preliminary agricultural fermentation, provided by Yunnan Provincial Academy of Tobacco Agricultural Sciences.

[0025] Cellulase preparation: 10000U / g, Shanghai Maclean Biochemical Technology Co., Ltd.

[0026] Bacillus subtilis ( Bacillus subtilis DES-59: A strain screened and preserved by the School of Biological Sciences, South China University of Technology, provided by the Key Laboratory of Fermentation and Enzyme Engineering, School of Biological Sciences and Engineering, South China University of Technology.

[0027] Example 1 A composite biological treatment method for improving the flavor and quality of heated cigarettes for cigars includes the following steps: (1) After removing the stems from the cigar tobacco leaves, they are physically crushed at a temperature range of 20-28℃ and then screened through a 200-mesh wire mesh to obtain cigar tobacco powder.

[0028] (2) Transfer the cigar tobacco powder to a tissue culture bottle. Spray cellulase solution into the bottle at 24°C with constant stirring. The amount of cellulase used is 180 U / g of cigar tobacco powder, and the amount of solution applied is 0.4 mL / g of cigar tobacco powder. Place the tissue culture bottle in a constant temperature and humidity incubator at 35°C, cover it with a cap with a breathable filter membrane, and let it stand for 4 hours.

[0029] (3) After completing step (2), add Bacillus subtilis DES-59 bacterial solution to the cigar tobacco powder at 24℃ with constant stirring. The inoculum amount of Bacillus subtilis DES-59 is 5×10⁻⁶. 8 The bacterial solution was applied at a concentration of CFU / g of cigar tobacco powder, with a dosage of 0.2 mL / g of cigar tobacco powder.

[0030] (4) The treated cigar tobacco powder was placed in a constant temperature and humidity incubator and fermented in a solid state for 7 days at 20°C and 60% relative humidity. Then it was inactivated at 80°C for 20 minutes and then dried at 40°C for 8 hours. The obtained cigar tobacco powder was made into cigar-flavored heated cigarettes according to industry standards.

[0031] Comparative Example 1 Compared with Example 1, the processing method lacks step (3), while the remaining steps are the same as in Example 1.

[0032] Comparative Example 2 Compared with Example 1, the processing method lacks step (2), while the remaining steps are the same as in Example 1.

[0033] Comparative Example 3 Compared with Example 1, the treatment method is the same as that in Example 1, except that steps (2) and (3) are replaced by adding 0.6 mL / g of sterile water.

[0034] I. Detection of chemical components in cigar tobacco powder The chemical composition of the cigar tobacco powder samples from Examples 1 and Comparative Examples 1-3 was determined, including the content of cellulose, protein, total soluble sugars, reducing sugars, total amino acids, and total alkaloids. Figure 1 As shown, cellulase treatment and inoculation with Bacillus subtilis DES-59 to enhance fermentation significantly affected the conventional chemical composition of cigar tobacco powder. Cellulose and protein macromolecules were degraded, while soluble total sugars, reducing sugars, and amino acids accumulated, and the content of total alkaloids also decreased. Compared to before fermentation, after 7 days of fermentation, the cellulose content of all groups decreased significantly, with the largest decreases in Example 1 and Comparative Example 1, with degradation rates of 14.94% and 17.31%, respectively, indicating the effective degradation of cellulose by microorganisms and cellulase during fermentation. After 7 days of fermentation, the protein content decreased in different groups, with significant decreases observed in Comparative Example 2 and Example 1 (p < 0.05), with degradation rates of 10.21% and 9.13%, respectively, indicating that inoculation with Bacillus subtilis DES-59 effectively promoted protein degradation in cigar tobacco powder. After the degradation of cellulose and protein, the content of total sugars, reducing sugars, and amino acids in the cigar tobacco powder also increased.

[0035] II. Testing the quality of heated cigar-flavored cigarettes The cigar-flavored heated cigarettes prepared in Example 1 and Comparative Examples 1-3 were subjected to sensory quality evaluation by several smoking experts, and the results are shown in Table 1.

[0036] Table 1

[0037] Different lowercase letters a, b, c represent significant differences, P≤0.05.

[0038] As shown in Table 1, compared with Comparative Example 3, both Example 1 and Comparative Examples 1-2 significantly improved the overall sensory quality score of heated cigar tobacco powder. Example 1 scored the highest, improving by 7.1 points compared to Comparative Example 3, and significantly higher than Comparative Examples 1-2. Compared with Comparative Example 3, Example 1 and Comparative Example 1 significantly improved the smoke volume score. In addition, Example 1 and Comparative Example 2 also significantly improved the aroma score of heated cigar tobacco powder, with Example 1 showing a 14.67% increase in aroma score. The strength, harmony, irritation, and mouthfeel scores of Example 1 were significantly improved.

[0039] III. Detection of the release of various pyrolysis aroma-related substances The types and amounts of aroma-related substances released by thermal decomposition of cigar tobacco powder after 7 days of solid-state fermentation in Examples 1 and Comparative Examples 1-3 at 350°C were determined by heating release experiments and GC / MS analysis. Figure 2 As shown, inoculation with Bacillus subtilis DES-59 and cellulase treatment enhanced fermentation affected the types, quantities, and release amounts of various important components in the smoke produced by the thermal decomposition of cigar tobacco powder under heating conditions, and altered the proportion of different types of compounds in the smoke release. Compared with Comparative Example 3, in Comparative Example 1, the proportion of aldehydes, ketones, and organic acids released by the thermal decomposition of cigar tobacco powder increased, while the proportion of alcohols and alkenes decreased; in Comparative Example 2, the proportions of aldehydes, ketones, esters, alkenes, phenols, organic acids, and heterocyclic compounds increased, while the proportion of nicotine decreased; in Example 1, the proportions of aldehydes, ketones, esters, alkenes, and organic acids increased, while the proportions of nicotine and phenols decreased.

[0040] In summary, this application constructs a highly efficient composite bio-fermentation system through the synergistic treatment of cellulase and Bacillus subtilis DES-59. This system not only significantly accelerates and deepens the fermentation process of cigar tobacco but also achieves targeted regulation of tobacco chemical components: on the one hand, the synergistic effect of cellulase and protease secreted by the strain efficiently degrades high-content macromolecules such as cellulose and protein in tobacco; on the other hand, this composite system actively promotes the generation and accumulation of small-molecule aroma precursors such as soluble sugars and amino acids, and enhances the overall metabolic aroma-producing capacity of the microbial community. Therefore, the method of this application can significantly improve fermentation efficiency under optimized environmental conditions, effectively shorten the traditional process cycle, and ultimately achieve a comprehensive improvement in the aroma release characteristics and sensory quality of cigar tobacco in heated tobacco products.

[0041] Although this application has been described in detail above with general descriptions, specific embodiments, and experiments, some modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of this application fall within the scope of protection claimed in this application.

Claims

1. A composite biological treatment method for improving the flavor and quality of heated cigarettes, characterized in that, Includes the following steps: (1) After removing the stems from the cigar tobacco leaves, they are physically crushed, sieved, and cigar tobacco powder is obtained. (2) Add cellulase solution to the cigar tobacco powder at 20-28℃ and stir evenly, then treat at 35-45℃ for 2-6 hours; (3) After completing step (2), add Bacillus subtilis DES-59 bacterial solution to the cigar tobacco powder at 20-28℃ and stir evenly; (4) The treated cigar tobacco powder is fermented for 7-15 days at 20-28℃ and 60-70% relative humidity.

2. The processing method according to claim 1, characterized in that, The amount of cellulase used is 180-240 U / g of the cigar tobacco powder.

3. The processing method according to claim 1, characterized in that, The inoculum size of Bacillus subtilis DES-59 was 5 × 10⁻⁶. 8 -7×10 8 CFU / g of the cigar tobacco powder.

4. The processing method according to claim 1, characterized in that, In steps (2) and (3), the total amount of solution and bacterial solution applied is 0.5-0.7 mL / g of the cigar tobacco powder.

5. The processing method according to claim 1, characterized in that, In steps (2) and (3), the solution and bacterial solution are applied by spraying.

6. The processing method according to claim 1, characterized in that, It also includes deactivating and drying the cigar tobacco powder processed in step (4).

7. The processing method according to claim 6, characterized in that, The inactivation temperature is 80-100℃, and the treatment time is 20-40 minutes.

8. The processing method according to claim 6, characterized in that, The drying temperature is 40-80℃, and the treatment time is 5-10 hours.

9. A cigar-flavored heated cigarette, characterized in that, The cigar tobacco powder obtained by any one of the processing methods described in claims 1-8 is used as raw material.

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