A method for stacking fermentation of cigar tobacco leaves based on compound enzymes

Abstract

This invention discloses a method for cigar tobacco leaf stacking fermentation based on a compound enzyme, belonging to the field of cigar processing technology. The method includes: S1, mixing a compound enzyme with deionized water to prepare an enzyme treatment solution; S2, spraying the enzyme treatment solution onto the surface of cigar tobacco leaves; S3, stacking the sprayed cigar tobacco leaves into natural tobacco stacks for stacking fermentation; S4, aging the stacked fermented cigar tobacco leaves in a constant temperature and humidity environment; wherein the compound enzyme includes aroma-producing enzymes. This invention, through the synergistic effect of a specific compound enzyme system combined with precise fermentation and aging processes, can directionally increase the content and diversity of aroma substances in cigar tobacco leaves, while effectively reducing the irritation of the smoke, achieving a synergistic effect of "high aroma and low harshness".

Description

Technical Field

[0001] This invention belongs to the field of cigar processing technology, specifically relating to a method for cigar tobacco leaf stacking fermentation based on compound enzymes. Background Technology

[0002] As a special tobacco product, the quality and style of cigars are highly dependent on the fermentation and aging process of the tobacco leaves. The sensory qualities of cigar tobacco leaves, especially those used for the filler, such as the richness of aroma, the smoothness of the smoke, the irritation, and the comfort of the finish, are core elements determining the grade of a cigar and its acceptance by consumers. High-quality cigar tobacco leaves should possess characteristics such as a rich and full-bodied aroma, strong character, smooth and delicate smoke, low irritation, and a clean and comfortable finish.

[0003] In recent years, with the advancement of the "Chinese-style cigar" development strategy, the cultivation and industrial application of domestically produced cigar tobacco have made significant progress. However, compared with high-quality foreign cigar tobacco, domestically produced cigar tobacco generally suffers from some common problems: First, it lacks sufficient aroma, with poor aroma burst and richness, and lacks the complex aroma required for high-end cigars; second, the smoke is more irritating and strong, affecting the smoothness and comfort of the smoke; and third, it has a heavier off-flavor and an impure aftertaste. These problems seriously restrict the proportion of domestically produced cigar tobacco used in high-quality cigar products and its industrial availability, becoming a technical bottleneck restricting the high-quality development of Chinese-style cigars.

[0004] To improve tobacco leaf quality, existing technologies have explored the use of bio-enzyme treatment. The basic idea is to utilize one or a few enzymes to degrade specific macromolecular components in tobacco leaves (such as pectin, cellulose, protein, and starch), aiming to loosen the cell wall structure and release some of the encapsulated aroma substances, thereby increasing the aroma content. However, these existing technologies still have the following inherent drawbacks: First, the technological fields differ, and the root causes of the problems are different. Existing technologies are mainly applied to flue-cured tobacco, which differs fundamentally from cigar tobacco in terms of raw material characteristics, processing techniques (such as the degree of fermentation and combustion state), and quality evaluation standards. Cigar tobacco leaves require a richer smoky aroma, nutty aroma, and lower nasal irritation. Directly applying the enzyme treatment methods used for flue-cured tobacco is not targeted enough and cannot solve the unique sensory defects of cigar tobacco leaves.

[0005] Second, the enzyme system combination does not match the target. Existing enzyme system designs primarily target the release of glycoside aroma precursors in flue-cured tobacco, without fully considering the specific pathways of nitrogenous compounds such as proteins and alkaloids in cigar tobacco transforming into aroma components, nor do they address the regulation of nicotine transfer behavior in smoke. Therefore, their aroma-enhancing effect is limited, and they cannot effectively reduce the irritation of cigar tobacco.

[0006] Third, the effects are uneven and it is difficult to achieve synergistic improvement. Although existing technologies can enhance aroma to a certain extent, they are often accompanied by an increase in irritation, or the improvement in irritation is not significant, and it is impossible to achieve the synergistic effect of "significantly enhancing aroma" and "significantly reducing irritation". This bottleneck of "enhancing aroma increases irritation" has not been overcome for a long time, which limits its application value in the improvement of high-end cigar raw materials.

[0007] Fourth, the process control is crude and the stability is poor. The processing technology in the current technology (such as parameters such as enzyme concentration, temperature and humidity, fermentation cycle, etc.) is relatively crude and has not formed a refined control that matches the characteristics of cigar tobacco leaves, resulting in unstable processing effects that are difficult to reproduce in industrial production.

[0008] In summary, there is an urgent need in this field for a compound enzyme preparation specifically designed for cigar tobacco leaves, capable of synergistically achieving "high aroma and low harshness," along with a corresponding industrialized stacking fermentation method. This method should be able to directionally increase the content and diversity of aroma substances in cigar tobacco leaves, while effectively reducing the harshness of the smoke, achieving a comprehensive improvement in sensory quality. This would solve the long-standing technical bottlenecks in domestically produced cigar tobacco leaves and enhance their industrial usability and application value. Summary of the Invention

[0009] The purpose of this invention is to target and enhance the content and diversity of aroma substances in cigar tobacco leaves, while effectively reducing the irritation of the smoke, thereby achieving a comprehensive improvement in sensory quality and enhancing its industrial usability and application value.

[0010] To solve the above-mentioned technical problems, the present invention provides the following technical solution: On one hand, a method for cigar tobacco leaf stacking fermentation based on compound enzymes includes the following steps: S1. Mix the compound enzyme with deionized water to prepare an enzyme treatment solution; S2. Spray the enzyme treatment solution onto the surface of the cigar tobacco leaves; S3. After spraying, the cigar tobacco leaves are piled into natural tobacco stacks for fermentation. S4. The fermented cigar tobacco leaves are aged in a constant temperature and humidity environment. Among them, the compound enzymes include aroma-producing enzymes.

[0011] Furthermore, the compound enzyme contains cellulase, pectinase, protease, saccharifying enzyme, xylanase and aroma-producing enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme and xylanase; the concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

[0012] Furthermore, the compound enzyme contains cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

[0013] Furthermore, the compound enzyme contains hemicellulase, amylase, and aroma-producing enzyme, or is composed of hemicellulase, amylase, and aroma-producing enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.05%~0.15%, or 0.075%~0.125%, or 0.1% by mass-volume concentration.

[0014] Furthermore, the compound enzyme contains hemicellulase, amylase, and aroma-producing enzyme, or is composed of hemicellulase, amylase, and aroma-producing enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

[0015] Furthermore, the amount of enzyme-treated solution applied is 1% to 20% of the weight of the cigar tobacco leaves, preferably 3% to 15%, and more preferably 5% to 10%, by mass percentage.

[0016] Furthermore, cigar tobacco leaves are one or more of the following: cigar filler tobacco leaves, cigar binder tobacco leaves, or cigar wrapper tobacco leaves.

[0017] Furthermore, the equilibration process is carried out under the following conditions: temperature of 20℃~30℃, or 23℃~27℃, or 25℃; relative humidity of 60%~70%, or 63%~67%, or 65%; and time of 48 hours~96 hours, or 60 hours~84 hours, or 72 hours.

[0018] Furthermore, the fermentation cycle of the stack is 30 to 50 days, or 35 to 45 days, or 40 days; the turning conditions are: turning the stack once every 8 to 12 days, or turning the stack once when the core temperature reaches 40℃ to 50℃, and adding moisture during the turning process.

[0019] In a second aspect, an enzyme treatment solution for use in the method of the first aspect is provided, the enzyme treatment solution comprising a complex enzyme.

[0020] Compared with the prior art, the technical solution provided by the present invention has the following beneficial technical effects: Achieving a synergistic effect of "high aroma and low harshness": This invention breaks through the bottleneck of existing technologies where "increasing aroma increases harshness." By screening specific compound enzyme systems including aroma-producing enzymes, it achieves the targeted and synergistic transformation of aroma precursors and irritating components in cigar tobacco leaves. Experimental data show that, taking 0.3% enzyme A treatment as an example, the relative content of total aroma substances in the mainstream smoke increases significantly, while successfully inducing or enhancing the core aromas of cigars such as smokiness and nutty roasted notes; furthermore, the nicotine content in the mainstream smoke is significantly reduced, significantly decreasing the irritation of the smoke.

[0021] The cigar tobacco leaves processed by the method of this invention exhibit a comprehensive improvement in sensory quality. Specifically, the aroma is more mellow, the aroma intensity is richer, off-flavors are significantly reduced, and the aftertaste is more pleasant. The overall product style and quality are greatly enhanced, effectively improving the industrial usability of domestically produced cigar tobacco leaves.

[0022] This invention discloses various compound enzyme systems and their applicable concentrations, which can be selected according to different quality improvement goals. For example, 0.1% enzyme C is most effective in promoting the release of aroma substances; 0.3% enzyme C is outstanding in reducing nicotine; and 0.3% enzyme B performs excellently in enhancing the aroma quality and sophistication. This provides a flexible and precise technical means for industrial production tailored to different raw material characteristics and product style requirements.

[0023] This invention establishes a refined control process tailored to the characteristics of cigar tobacco leaves, specifying the types, concentrations, and application rates of the compound enzymes, as well as key parameters such as the fermentation cycle, turning conditions, and temperature, humidity, and time for equilibration. This process exhibits good stability, is easily reproducible and applicable in industrial production, aligns with the development strategy of "Chinese-style cigars" and the industry direction of "cost reduction and efficiency improvement," and is of great significance for increasing the proportion of domestically produced cigar raw materials used. Attached Figure Description

[0024] The above description of the present invention and the following detailed embodiments will be better understood when read in conjunction with the accompanying drawings. It should be noted that the drawings are merely examples of the claimed technical solutions.

[0025] Figure 1 This is a schematic flowchart of a cigar tobacco stacking fermentation method based on a complex enzyme according to an embodiment of the present invention. Detailed Implementation

[0026] The detailed features and advantages of this application are described below in the specific embodiments. The content of this description is sufficient to enable any person skilled in the art to understand the technical content of this application and implement it accordingly. Based on the specification, claims and drawings disclosed in this specification, a person skilled in the art can easily understand the related objectives and advantages of this application.

[0027] In this specification and claims, several terms will be used, and unless otherwise indicated, these terms will be defined to have the following meanings: All figures used to represent component amounts, properties (e.g., weight-average molecular weight), reaction conditions, etc., should be considered in all cases modified by the terms "within the unavoidable margin of error" or "about". Therefore, the numerical values ​​set forth herein are approximate and may vary depending on the desired properties sought to be obtained by the present invention. The principle of equivalents, which is applied to a minimum and not intended to limit the scope of the claims, should be applied, for example, each value should be interpreted at least according to the reported significant digits and by applying conventional rounding techniques.

[0028] All other terms used herein for special definition are intended to have the general meaning understood by one of ordinary skill in the art, in particular meaning that one of ordinary skill in the art can directly and without doubt determine how the technical solutions of the invention can be implemented after reading the claims, specification and drawings.

[0029] Even if there are incomplete descriptions, omissions, or ambiguities in the grammar, words, punctuation, graphics, symbols, etc. of the claims, specification, and drawings of this invention, those skilled in the art can still arrive at the only correct understanding by reading the claims, specification, and drawings as a whole without extensive reasoning or experimentation, and effectively exclude various incorrect interpretations that are not aimed at achieving the purpose of this invention.

[0030] The "range" disclosed herein is defined by a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, which define the boundaries of a particular range. Ranges defined in this way can include or exclude endpoints and can be arbitrarily combined; that is, any lower limit can be combined with any upper limit to form a range. For example, if ranges of 60-120 and 80-110 are listed for a specific parameter, it is expected that ranges of 60-110 and 80-120 are also expected. Furthermore, if minimum range values ​​of 1 and 2 are listed, and if maximum range values ​​of 3, 4, and 5 are listed, then the following ranges are all expected: 1-3, 1-4, 1-5, 2-3, 2-4, and 2-5. In this application, unless otherwise stated, the numerical range "ab" represents a shortened representation of any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0-5" indicates that all real numbers between "0-5" have been listed in this article; "0-5" is simply a shortened representation of these numerical combinations. Furthermore, when a parameter is stated as an integer ≥2, it is equivalent to disclosing that the parameter is, for example, an integer such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0031] Unless otherwise specified, all embodiments and preferred embodiments mentioned herein can be combined to form new technical solutions.

[0032] Unless otherwise specified, all the technical features and preferred features mentioned herein can be combined to form new technical solutions.

[0033] Unless otherwise specified, all steps mentioned herein may be performed sequentially or randomly, but are preferably performed sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the mention that the method may also include step (c) indicates that step (c) may be added to the method in any order; for example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0034] Unless otherwise specified, the terms "comprising" and "including" as used herein can be open-ended or closed-ended. For example, "comprising" and "including" may mean that other components not listed may also be included, or that only the listed components may be included.

[0035] In the description of this article, it should be noted that, unless otherwise stated, "above" and "below" include the number itself, and "several" in "one or more" means two or more.

[0036] In this description, unless otherwise stated, the term "or" is inclusive. For example, the phrase "A or B" means "A, B, or both A and B". More specifically, the condition "A or B" is satisfied by any of the following conditions: A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); or both A and B are true (or exist).

[0037] Unless otherwise specified, percentages (%) in this document refer to percentages by mass relative to the composition.

[0038] Unless otherwise stated herein, the sum of the contents of the components in the composition is 100%.

[0039] In this document, unless otherwise stated, “combination of” means a multi-component mixture of the elements, such as two, three, four, and up to the maximum possible multi-component mixture.

[0040] Unless otherwise specified, the term "a" as used in this specification means "at least one".

[0041] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0043] To make the objectives, technical solutions, and advantages of the present invention clearer, the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods, and the materials and reagents used in the following embodiments are commercially available unless otherwise specified.

[0044] One object of the present invention is to provide a method for stacking fermentation of cigar tobacco leaves based on compound enzymes, comprising the following steps: S1. Mix the compound enzyme with deionized water to prepare an enzyme treatment solution; S2. Spray the enzyme treatment solution onto the surface of the cigar tobacco leaves; S3. After spraying, the cigar tobacco leaves are piled into natural tobacco stacks for fermentation. S4. The fermented cigar tobacco leaves are aged in a constant temperature and humidity environment. Among them, the compound enzymes include aroma-producing enzymes.

[0045] As used in this article, "flavor-enhancing enzymes" are the core biocatalysts for food fermentation and flavor regulation. They are essentially hydrolases, transferases, oxidases, etc., and achieve product aroma enhancement, aroma improvement, aroma addition, and flavor enhancement through three major pathways: ester synthesis, release of bound aroma, and degradation of macromolecules.

[0046] As used in this article, "pile fermentation" refers to the process of piling cigar tobacco leaves into natural stacks after they have been sprayed with enzyme treatment solution. Under natural ventilation or controlled conditions, the process utilizes the microorganisms and enzyme systems carried by the tobacco leaves themselves, combined with the synergistic effect of added compound enzymes. By controlling the fermentation cycle and turning conditions, a series of complex biochemical changes occur inside the tobacco leaves to degrade macromolecules, transform aroma precursors, and reduce irritation, thereby improving the sensory quality of the tobacco leaves.

[0047] As used in this article, "balanced aging" refers to the post-processing of placing cigar tobacco leaves after stacking fermentation in a constant temperature and humidity environment. By controlling the temperature, relative humidity, and time, the internal moisture distribution, enzyme reaction products, and aroma substances of the tobacco leaves are further made more uniform, stable, and coordinated, so as to facilitate subsequent processing and quality consistency.

[0048] In some embodiments, the compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, xylanase and aroma-producing enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme and xylanase; the concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

[0049] As used in this article, "w / v" refers to the percentage of solute mass to solution volume, such as 1 g / 100 mL, which is expressed as 1%.

[0050] In some embodiments, the compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1% to 0.5%, or 0.2% to 0.4%, or 0.3% by mass volume.

[0051] In some embodiments, the compound enzyme comprises hemicellulase, amylase, and aroma-producing enzyme, or is composed of hemicellulase, amylase, and aroma-producing enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.05%~0.15%, or 0.075%~0.125%, or 0.1% by mass-volume concentration.

[0052] In some embodiments, the compound enzyme comprises hemicellulase, amylase and flavoring enzyme, or is composed of hemicellulase, amylase and flavoring enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1% to 0.5%, or 0.2% to 0.4%, or 0.3% by mass-volume concentration.

[0053] In some embodiments, the amount of enzyme-treated solution applied is 1% to 20% of the weight of the cigar tobacco leaves, preferably 3% to 15%, and more preferably 5% to 10%, by mass percentage.

[0054] In some embodiments, the cigar tobacco is one or more of the following: filler tobacco, binder tobacco, or wrapper tobacco.

[0055] In some implementations, the equilibration process is carried out under the following conditions: temperature of 20°C to 30°C, or 23°C to 27°C, or 25°C; relative humidity of 60% to 70%, or 63% to 67%, or 65%; and time of 48 hours to 96 hours, or 60 hours to 84 hours, or 72 hours.

[0056] In some implementations, the fermentation cycle of the stack is 30 to 50 days, or 35 to 45 days, or 40 days; the turning conditions are: turning the stack once every 8 to 12 days, or turning the stack once when the core temperature reaches 40°C to 50°C, and adding moisture during the turning process.

[0057] Another object of the present invention is to provide an enzyme treatment solution for use in the method of the first aspect, the enzyme treatment solution comprising a complex enzyme.

[0058] In some embodiments, the compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, xylanase and aroma-producing enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme and xylanase; the concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

[0059] In some embodiments, the compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme, amylase and flavoring enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1% to 0.5%, or 0.2% to 0.4%, or 0.3% by mass volume.

[0060] In some embodiments, the compound enzyme comprises hemicellulase, amylase, and aroma-producing enzyme, or is composed of hemicellulase, amylase, and aroma-producing enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.05%~0.15%, or 0.075%~0.125%, or 0.1% by mass-volume concentration.

[0061] In some embodiments, the compound enzyme comprises hemicellulase, amylase and flavoring enzyme, or is composed of hemicellulase, amylase and flavoring enzyme; the concentration of the compound enzyme in the enzyme treatment solution is 0.1% to 0.5%, or 0.2% to 0.4%, or 0.3% by mass-volume concentration.

[0062] Example

[0063] Experimental materials and instruments

[0064] The tobacco leaves used for the test cigars were selected from the middle section of the "Chu Xue 14" variety produced in Danjiangkou, Shiyan, Hubei Province in 2024, and were provided by Hubei China Tobacco Industry Co., Ltd. The tobacco leaves were grade three in appearance, and the initial moisture content was controlled between 13% and 15%.

[0065] Enzyme sources: cellulase (purchased from Shandong Longket Enzyme Preparation Co., Ltd.), amylase (purchased from Shandong Longket Enzyme Preparation Co., Ltd.), pectinase (purchased from Shandong Longket Enzyme Preparation Co., Ltd.), protease (purchased from Shandong Longket Enzyme Preparation Co., Ltd.), saccharifying enzyme (purchased from Shandong Longket Enzyme Preparation Co., Ltd.), xylanase (purchased from Ningxia Xiasheng Industrial Group Co., Ltd.), and aroma-producing enzyme (purchased from Shandong Longket Enzyme Preparation Co., Ltd.).

[0066] Experimental equipment: Electronic balance (ME303), Mettler Toledo Instruments (Shanghai) Co., Ltd.; Gas chromatography-mass spectrometry (GCMS 2400), PerkinElmer; Linear cigar smoker (SML1000C), Hefei Zhongwo Instrument Technology Co., Ltd.; Constant temperature and humidity chamber KBF240, Xiamen Southeast Chuangsheng Instrument Equipment Co., Ltd.; Drying oven DHG-9070A electric heating forced-air drying oven, Shanghai Yiheng Scientific Instrument Co., Ltd.

[0067] Preparation Example

[0068] Four compound enzyme preparations were prepared according to Table 1 below: enzyme A, enzyme B, enzyme C and enzyme D.

[0069] Table 1 Composition of Compound Enzyme Preparations

[0070] Prepare the enzyme treatment solutions for each group according to Table 2 below. All concentrations are volume-to-mass ratios.

[0071] Table 2 Composition and concentration of enzyme treatment solutions in each group

[0072] The enzyme treatment solutions for each group were evenly sprayed onto the surface of the cigar filler tobacco leaves, with a spraying amount of 10% of the tobacco leaf mass (i.e., approximately 100g of solution per kilogram of tobacco leaves). The sprayed cigar filler tobacco leaves were then piled into natural stacks; the fermentation period was 40 days, with the stacks turned every 10 days or when the core temperature reached 45℃, and simultaneously moistened. After fermentation, the stacks were equilibrated at 25℃ and RH=65% for 72 hours. The treated tobacco leaves were then used for rolling cigars and subjected to sensory and physicochemical evaluation.

[0073] Test Example 1

[0074] This experiment determined the content of petroleum ether extract in cigar tobacco leaves to reflect changes in their aroma compounds. The testing procedure is as follows: Sample preparation followed the method in standard YC / T31—1996. After curing, the tobacco leaf samples were dried in an oven at 40℃ for 3–4 h until they could be crushed by fingers. After cooling and pulverizing, the samples were passed through a 40-mesh sieve and stored in a vacuum bag for later use. The petroleum ether extraction rate was determined according to standard YC / T 176—2003, "Determination of Petroleum Ether Extracts in Tobacco and Tobacco Products". The extracted petroleum ether extract was concentrated to 1 mL using a rotary evaporator and stored for later use.

[0075] The test results are shown in the table below: Table 3. Analytical results of petroleum ether extract

[0076] Test results of petroleum ether extracts in cigar tobacco leaves showed that, compared with Comparative Example 1 (blank control), the extract content of all enzyme-treated groups was significantly increased, with increases ranging from 22% to 120%. Among them, the 0.1% enzyme C group showed the most significant effect, with a content as high as 13.2%. Different enzymes exhibit different modes of action: enzymes C and D may rapidly release encapsulated fat-soluble aroma substances at low concentrations through efficient degradation of cell walls (such as pectin and cellulose), but their effects decrease with increasing concentration; the effects of enzymes A and B are relatively mild and gradual, reaching peak values ​​at 0.3% and 0.1%, respectively.

[0077] Test Example 2

[0078] This experiment determined the nicotine content in cigar tobacco leaves to reflect the alkaloids and aging progress within the leaves. The testing procedure was as follows: Cigar tobacco leaf samples were dried in an oven at 40℃ for 3–4 hours, cooled, pulverized, and passed through a 40-mesh sieve. Acetic acid extraction solution was added, and the samples were shaken and filtered through qualitative filter paper. The filtrate was then prepared for analysis. The samples were tested using a continuous flow analyzer method, referring to the standard "Determination of Nicotine, Nornicotinic Acid, Neonicotinic Acid, Maxamine and Equisetine in Tobacco and Tobacco Products by Gas Chromatography-Mass Spectrometry" (YC / T 383-2010).

[0079] The test results are shown in Table 4: Table 4. Results of Nicotine Content Detection in Treated Cigar Tobacco Leaves

[0080] Nicotine affects the strength of cigar smoke during smoking. Analysis of the nicotine content in treated cigar tobacco leaves, as shown in Table 1, revealed a significant decrease in nicotine content compared to the blank control group (31.69 mg / g). Other experimental groups showed no significant effect. The 0.3% enzyme C treatment showed the most significant effect. Enzyme-promoted metabolic processes may utilize and convert nicotine as a nitrogen source, leading to a decrease in its content. Enzyme degradation of cellular structures or macromolecules bound to nicotine may also cause a temporary increase in free nicotine content during measurement.

[0081] Test Example 3

[0082] This experiment measured the nicotine release from mainstream cigarette smoke to reflect the nicotine content in cigarette samples treated with different methods. The testing procedure was as follows: A cigarette was smoked on a cigar smoking machine, and total particulate matter was collected using a Cambridge filter smoke trap. The mass of the total particulate matter was weighed. The extracted total particulate matter was used for gas chromatography to determine the moisture and nicotine content.

[0083] The test results are shown in Table 5: Table 5 Analysis of Nicotine Content in Mainstream Smoke Gas

[0084] By capturing particulate matter in the flue gas using Cambridge filters and analyzing the nicotine content, it was found that, compared with the blank control, the nicotine content in the mainstream flue gas of the exogenously added compound enzymes was reduced in all groups except for 0.1% enzyme A, 0.3% enzyme B, and 0.3% D, with 0.3% enzyme C having the lowest nicotine content.

[0085] Test Example 4

[0086] This experiment aims to reflect the aroma richness, aroma characteristics, and sensory quality differences of cigar tobacco leaves by determining their aroma compound content. The testing procedure is as follows: Processed filler and binder tobacco leaves are rolled into 15mm × 90mm cigars using standard techniques. A linear cigar smoking machine is used in "solid puff mode" to collect the mainstream smoke. A Cambridge filter is used to capture the total particulate matter in the smoke. After smoking, the filter is extracted with dichloromethane solution and then analyzed. GC-MS qualitative analysis is employed, using GC-MS 2400s to detect aroma compounds in the mainstream smoke and calculate the relative content of each component.

[0087] The test results are shown in the table below: Table 6. Relative content analysis of aroma compounds

[0088]

[0089] Table 7 Aroma Classification and Description

[0090] All enzyme treatment groups significantly enhanced nitrogen-containing heterocyclic compounds and advanced flavoring components, promoted the conversion of precursor substances in tobacco leaves, and allowed for targeted regulation of the aroma profile of cigar smoke. 0.3% enzyme A exhibited high levels of phenols, nitrogen-containing heterocyclic compounds, terpenes, and advanced flavoring components, resulting in a rich and complex aroma. 0.3% enzyme B had the highest content of advanced flavoring components among all groups, demonstrating outstanding aroma quality. Enzymes C and D were substrate-specific, suitable for fine-tuning the aroma. In terms of concentration effects, 0.3% treatment of enzymes A, C, and D was superior to 0.2%, while enzyme B exhibited an optimal concentration window. Compound enzymes can precisely regulate the aroma composition of cigar smoke, pursuing both richness and complexity, with 0.3% enzyme A being the optimal choice.

[0091] Test Example 5

[0092] Comparative smoking evaluation was used to assess the aroma quality, aroma intensity, irritation, and off-flavors of the cigarettes, following the evaluation rules of GB15269.4—2011 "Cigars Part 4: Sensory Technical Requirements". A smoking evaluation team of seven technicians from the Technical Center of Hubei China Tobacco Industry Co., Ltd. was assembled. Evaluation indicators included: aroma quality, aroma intensity, off-flavors (higher scores indicate less off-flavors), irritation (higher scores indicate less irritation), aftertaste, sweetness, combustibility, and ash. Each indicator had three sub-categories, and the average score was used as the final score. Evaluation indicators also included: strength, power, style intensity, and quality grade. Each indicator had a maximum score of 5, and the average score was used as the final score. The evaluation results are shown in the table below. Table 8 Sensory Evaluation Form

[0093] Most enzyme-treated groups showed improved sensory scores compared to the control group, with the compound enzyme treatment demonstrating significant effects. The 0.2% and 0.3% enzyme A groups achieved total scores of 82.5 and 83 respectively, far exceeding the project's expected score improvement targets. The 0.3% enzyme A group had the highest sensory score, highly consistent with its physicochemical indicators: aroma quality (15.5) and aroma quantity (12.5) were both optimal, corresponding to a total aroma substance content of 12.9% in the smoke. Nutty, roasted, smoky, and grassy aromas collectively enhanced the aroma's smoothness and fullness; the irritation score was 9 (compared to 7 in the control group), matching an 87% reduction in nicotine, validating the high-aroma, low-irritation technology approach. This group exhibited reduced off-flavors, achieving a style level and quality grade of 3.5, degrading large off-flavor molecules, and integrating the sweetness and rich aroma of Hubei raw materials with a smooth taste, making it the optimal treatment group for both physicochemical properties and smoking experience.

[0094] The 0.3% enzyme B group has the same style and quality level as the best. The enzyme A group has slightly lower aroma quantity and aroma quality, and its core advantage is to improve quality rather than increase intensity. Components such as succinyl ol and geraniol significantly enhance the sophistication and elegant recognizability of the aroma.

[0095] In summary, treatment with 0.3% enzyme A increased the relative content of total aroma compounds in mainstream smoke from 3.1% to 12.9%, a significant increase of 316%. The aroma of the tobacco leaves became richer, successfully inducing or enhancing the core aromas of cigars, such as smokiness (phenols), nutty roasted aroma (nitrogenous heterocyclic compounds), and vegetal freshness (terpenes). The 0.3% enzyme A treatment significantly reduced nicotine in mainstream smoke. The key mechanism lies in altering the nicotine combustion transfer rate, rather than destructive degradation, thus reducing irritation while perfectly preserving the rich character of the tobacco leaves. The 0.3% enzyme A treatment group scored 83 points, an improvement of 8 points compared to the control group (75 points), far exceeding the expected target. Specifically, this manifested as a more mellow aroma, a richer aroma volume, reduced off-flavors, and a more comfortable aftertaste, resulting in a significant improvement in product style and quality. Based on a comprehensive analysis of physicochemical indicators, aroma components, and sensory evaluation, the 0.3% enzyme A treatment group demonstrated the best technical effect.

[0096] In addition, 0.1% enzyme C has the strongest aroma release. 0.3% enzyme C has an outstanding alkali-reducing effect. 0.3% enzyme B has the best style and quality grade, and is preferred for enhancing the aroma quality of high-end product lines.

[0097] This technology can effectively improve the sensory quality and industrial usability of domestically produced cigar raw materials, help increase the proportion of domestic raw materials used, and is in line with the national and industry strategic directions of "developing Chinese-style cigars" and "reducing costs and increasing efficiency".

[0098] In the foregoing description of exemplary embodiments / specific implementations of the present invention, various features of the invention are sometimes combined in a single embodiment / specific implementation or its figures and description, with the aim of simplifying the disclosure and aiding in the understanding of one or more of the various aspects of the invention. However, the method of description of the invention should not be construed as reflecting an intention that the claimed features are more than expressly stated in each claim, except where explicitly stated otherwise or in obvious technical contradiction or exclusion. Rather, the inventive aspect reflected in the claims lies in not all the features of a single foregoing disclosed embodiment / specific implementation. Therefore, the claims following the detailed description are expressly incorporated herein by reference, each claim existing independently as a separate embodiment / specific implementation of the invention.

[0099] The terminology and expressions used in this specification are for illustrative purposes and not for limitation. Their use is not intended to exclude any equivalents of the shown and described features or portions thereof, but rather to facilitate the understanding that various modifications may be possible within the scope of the invention. Therefore, it should be understood that while the invention has been specifically disclosed through preferred embodiments, exemplary embodiments, and optional features, variations or modifications of the concepts disclosed herein may be adopted by those skilled in the art, and such variations and modifications are therefore considered to be within the scope of the invention as defined by the appended claims. The specific embodiments given in this specification are examples of useful embodiments of the invention, and it will be apparent to those skilled in the art that the invention can be practiced using many variations of the devices, device components, and method steps disclosed herein.

[0100] The foregoing description of specific embodiments has fully disclosed the general features of the invention, enabling others to easily modify and / or adapt such embodiments for various applications by applying knowledge within the scope of the art, without excessive experimentation and without departing from the general conception of the invention. Therefore, based on the teachings and guidance provided herein, it is intended that such modifications and alterations be included within the meaning and scope of equivalents of the disclosed embodiments. It should be understood that the wording or terminology used herein is for descriptive purposes and is not intended to be limiting; thus, the wording or terminology in this specification will be interpreted by those skilled in the art based on the foregoing teachings and guidance.

[0101] Furthermore, the scope of the invention should not be limited to any of the exemplary embodiments described above, but only to the appended claims and their equivalents.

Claims

1. A method for stacking fermentation of cigar tobacco leaves based on compound enzymes, characterized in that, Includes the following steps: S1. Mix the compound enzyme with deionized water to prepare an enzyme treatment solution; S2. Spray the enzyme treatment solution onto the surface of cigar tobacco leaves; S3. The sprayed cigar tobacco leaves are piled into natural tobacco stacks for fermentation. S4. The cigar tobacco leaves after stacking and fermentation are aged in a constant temperature and humidity environment. The compound enzyme includes aroma-producing enzyme.

2. The method according to claim 1, characterized in that, The compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, xylanase and aroma-producing enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme and xylanase; The concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

3. The method according to claim 1, characterized in that, The compound enzyme comprises cellulase, pectinase, protease, saccharifying enzyme, amylase and flavor-producing enzyme, or is composed of cellulase, pectinase, protease, saccharifying enzyme, amylase and flavor-producing enzyme; The concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

4. The method according to claim 1, characterized in that, The compound enzyme contains hemicellulase, amylase and flavoring enzyme, or is composed of hemicellulase, amylase and flavoring enzyme; The concentration of the compound enzyme in the enzyme treatment solution is 0.05%~0.15%, or 0.075%~0.125%, or 0.1% by mass-volume concentration.

5. The method according to claim 1, characterized in that, The compound enzyme contains hemicellulase, amylase and flavoring enzyme, or is composed of hemicellulase, amylase and flavoring enzyme; The concentration of the compound enzyme in the enzyme treatment solution is 0.1%~0.5%, or 0.2%~0.4%, or 0.3% by mass-volume concentration.

6. The method according to claim 1, characterized in that, The amount of the enzyme-treated solution applied is 1% to 20% of the mass of the cigar tobacco leaves, preferably 3% to 15%, and more preferably 5% to 10%, by mass percentage.

7. The method according to claim 1, characterized in that, The cigar tobacco leaves are one or more of the following: cigar filler tobacco leaves, cigar binder tobacco leaves, or cigar wrapper tobacco leaves.

8. The method according to claim 1, characterized in that, The equilibration process is carried out under the following conditions: temperature of 20℃~30℃, or 23℃~27℃, or 25℃; relative humidity of 60%~70%, or 63%~67%, or 65%; and time of 48 hours~96 hours, or 60 hours~84 hours, or 72 hours.

9. The method according to claim 1, characterized in that, The fermentation cycle of the stack is 30 to 50 days, or 35 to 45 days, or 40 days; the turning conditions are: turning the stack once every 8 to 12 days, or turning the stack once when the core temperature reaches 40℃ to 50℃, and adding moisture during the turning process.

10. An enzyme treatment solution for use in the method of any one of claims 1 to 9, said enzyme treatment solution comprising the compound enzyme.

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