A natural plant aerosol generation matrix for electric heating smoke appliances

Abstract

This invention discloses a natural plant aerosol generating matrix for electric heating tobacco appliances. The natural plant aerosol generating matrix comprises: natural plant raw materials subjected to quantitative cell wall disruption treatment and an exogenous aerosol generating agent loaded within the natural plant raw materials. This invention, through quantitative cell wall disruption treatment to decompose cell wall components, enables quantitative and controllable modification of tobacco raw materials, causing localized disruption of thinner cell walls to form an aerosol loading medium, thereby increasing the aerosol generating agent loading capacity of the tobacco raw materials. Furthermore, by monitoring the loading capacity, the enzymatic cell wall disruption reaction can be adjusted and the overall reaction mechanism controlled. Moreover, while maintaining the original processability, it can absorb more aerosol and release more aroma substances during heating.

Description

Technical Field

[0001] This invention relates to the field of novel tobacco technology, and in particular to a natural plant aerosol generation matrix for use in electrically heated tobacco devices. Background Technology

[0002] In recent years, with the increasingly stringent tobacco control situation and consumers' growing awareness of health, new tobacco products have become increasingly popular. Heated cigarettes and other aerosol-generating products have achieved a good smoking experience after rapid development and optimization in recent years. However, due to the special heating method of heated cigarettes, existing traditional cigarette tobacco cannot be used as the filler material. Therefore, reconstituted tobacco leaves, i.e., tobacco sheets, are mostly used as filler materials. These sheets are mainly composed of tobacco dust, tobacco fragments, tobacco stems, or low-grade tobacco leaves with adhesives and other additives. They are mainly prepared by three methods: papermaking, slurry processing, and rolling. This differs significantly from traditional tobacco and requires the construction of new production lines. There have also been attempts to use traditional cigarette tobacco in heated cigarettes. To release sufficient aroma substances under heating conditions, traditional tobacco needs to undergo high-loss permeability treatments such as steam explosion and freeze-drying. However, direct high-loss permeability treatment severely reduces the processability of the tobacco, resulting in a low yield of whole tobacco leaves. Furthermore, it presents difficulties in rolling and splicing the tobacco leaves on the production line. Summary of the Invention

[0003] To address the shortcomings of existing technologies and practical needs, this invention provides a natural plant aerosol generation matrix for electric heating smoke appliances.

[0004] To achieve this objective, the present invention employs the following technical solution:

[0005] In a first aspect, the present invention provides a natural plant aerosol generating matrix for electric heating smoke appliances, comprising: natural plant raw materials subjected to quantified cell wall disruption treatment and an exogenous aerosol generating agent loaded within the natural plant raw materials.

[0006] Furthermore, the quantitative cell wall breaking treatment includes 1 to 5 enzymatic cell wall breaking steps, each enzymatic cell wall breaking step including: Step A: uniformly spraying the active cell wall breaking enzyme solution onto the natural plant material; Step B: sealing the sprayed natural plant material and carrying out the enzymatic cell wall breaking reaction under constant temperature and humidity conditions.

[0007] Furthermore, in step A, the weight ratio of the active cell wall-breaking enzyme to the natural plant raw material is 0.5-1.0%, 1.0-1.5%, 1.5-2.0%, 2.0-2.5%, or 2.5-3.0%.

[0008] Furthermore, in step A, the active cell wall-breaking enzyme includes any one or a combination of at least two of pectinase, hemicellulase, cellulase, or amylase.

[0009] Furthermore, in step A, the active cell-wall-breaking enzyme is composed of pectinase, hemicellulase, and amylase.

[0010] Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, cellulase, and amylase;

[0011] Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, hemicellulase, cellulase, and amylase, in the following weight ratio:

[0012] Pectinase: 15-25%,

[0013] Total hemicellulase and cellulase: 32-48%,

[0014] Amylase: 32-48%.

[0015] Furthermore, in step B, the conditions for each enzymatic hydrolysis and cell wall breaking reaction are independently as follows: temperature of 25-30℃, 30-35℃, 35-40℃, 40-45℃, 45-50℃, 50-55℃ or 55-60℃, relative humidity of 15-20%, 20-25%, 25-30%, 30-35%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 65-70%, 70-75% or 75-80%, and enzymatic hydrolysis and cell wall breaking reaction time of 0.5-1.0 h, 1.0-2.0 h, 2.0-4.0 h, 4.0 h-6.0 h, 6.0-8.0 h, 8.0-12.0 h or 12.0-24.0 h.

[0016] Furthermore, after step B is completed, step C is performed: the apparent loading and / or micro loading of the natural plant material are detected. When the apparent loading and / or micro loading of the aerosol generating agent of the natural plant material is greater than the target loading, the next enzymatic hydrolysis and cell wall breaking step is not performed.

[0017] Furthermore, based on the difference between the apparent loading and / or microscopic loading of the aerosol generator of the natural plant raw materials and the target loading, the enzymatic hydrolysis reaction temperature, relative humidity, and / or time of the enzymatic hydrolysis reaction in the next enzymatic hydrolysis reaction step are determined.

[0018] Furthermore, the apparent loading test includes weighing the maximum load of the exogenous aerosol generator; the micro loading test includes observing the cell wall and statistically analyzing the cell wall disruption rate.

[0019] Furthermore, after each enzymatic hydrolysis and cell wall breaking reaction is completed, step D is performed: the natural plant raw materials undergo harm reduction treatment.

[0020] Furthermore, the exogenous aerosol generating agent is loaded into the natural plant material that has undergone quantitative cell wall disruption treatment in the following manner: the exogenous aerosol generating agent is sprayed onto the natural plant material that has undergone quantitative cell wall disruption treatment, wherein: the weight of the exogenous aerosol generating agent is less than the weight of the natural plant material × the maximum loading rate × the whole fiber rate; after the exogenous aerosol generating agent has completely penetrated into the natural plant material that has undergone quantitative cell wall disruption treatment, a natural plant aerosol generating matrix is ​​prepared.

[0021] Furthermore, after each enzymatic hydrolysis and cell wall breaking reaction is completed, step E is performed: the whole fiber rate of the natural plant material is detected. If the whole fiber rate of the natural plant material is greater than the whole fiber rate warning value, the next enzymatic hydrolysis and cell wall breaking step will not be performed.

[0022] Furthermore, the warning value for the whole yarn rate is above 60%, above 65%, above 68%, above 70%, above 72%, above 75%, above 78%, or above 80%.

[0023] Secondly, a method for manufacturing the aforementioned aerosol generation matrix is ​​provided.

[0024] Thirdly, the present invention provides an aerosol generating article comprising the natural plant aerosol generating matrix for electric heating smokers as described in the first aspect.

[0025] Fourthly, the present invention provides an aerosol generation system, the aerosol generation system comprising an electrically heated aerosol generation device and the aerosol generation product described in the second aspect.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] (1) In this invention, scanning electron microscopy and thermogravimetric-gas chromatography-mass spectrometry are used to detect and analyze tobacco leaf samples before and after active enzyme treatment, and to explore the influence of active enzyme on the microstructure of tobacco cell wall and heat release pattern. The aim is to enhance the understanding of the properties of tobacco raw materials after active enzyme action and provide technical support for the development of heated cigarette smoke-generating media.

[0028] (2) This invention, through quantitative cell wall disruption treatment to decompose cell wall components, can achieve quantitative and controllable modification of tobacco raw materials, causing local disruption of thinner cell walls to form aerosol loads and increasing the loading capacity of tobacco raw materials for aerosol generating agents. In addition, the enzymatic cell wall disruption reaction can be adjusted and the overall reaction mechanism controlled by monitoring the loading capacity. Moreover, while maintaining the original processability (especially the whole tobacco yield), it can absorb more aerosols and release more aroma substances during heating, enabling traditional cigarette production lines to be reused with heated cigarettes with minor modifications. By selecting specific degradation enzyme compositions, the enzymatic degradation reaction can be achieved in the existing leaf storage environment without the need for additional high-temperature reaction equipment. Since it does not require solvent soaking, microwave heating, or other processes, the loss of aroma substances is avoided, and the natural aroma of tobacco can be fully volatilized. The aerosol addition amount determined by the maximum loading rate and whole tobacco yield can ensure overall processability. Attached Figure Description

[0029] 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.

[0030] Figure 1 This is a morphological image of tobacco leaf cells treated with pectinase under a transmission electron microscope (scale bar: 2 μm).

[0031] Figure 2 This is a morphological image of tobacco leaf cells treated with hemicellulase under a transmission electron microscope (scale bar: 10 μm).

[0032] Figure 3 This is a morphological image of tobacco leaf cells treated with hemicellulase under a transmission electron microscope (scale bar: 2 μm).

[0033] Figure 4 This is a comparative graph showing the effects of different concentrations of pectinase treatment on the pectin content in the cell walls of tobacco leaves.

[0034] Figure 5 This is a comparative graph showing the effects of different concentrations of hemicellulase treatment on the hemicellulose content in the cell walls of tobacco leaves.

[0035] Figure 6 This is a comparative graph showing the effects of different concentrations of amylase treatment on the starch content in the cell walls of tobacco leaves;

[0036] Figure 7 This is a schematic diagram of the maximum atomizing agent load testing device of the present invention, wherein the reference numerals are explained as follows: 1-tobacco shreds, 2-extrusion container, 3-force application probe;

[0037] Figure 8AThis is a graph showing the TG curves of tobacco leaves treated with different concentrations of pectinase.

[0038] Figure 8B These are DSC curves of tobacco leaves treated with different concentrations of pectinase.

[0039] Figure 9A These are TG curves of tobacco leaves treated with different types of enzymes.

[0040] Figure 9B These are DSC curves of tobacco leaves treated with different types of enzymes;

[0041] Figure 10A This is a graph showing the TG curves of tobacco leaves treated with different concentrations of hemicellulase.

[0042] Figure 10B These are DSC curves of tobacco leaves treated with different concentrations of hemicellulase.

[0043] Figure 11A This is a graph showing the TG curves of tobacco leaves treated with different concentrations of amylase.

[0044] Figure 11B These are DSC curves of tobacco leaves treated with different concentrations of amylase.

[0045] Figure 12A These are TG curves of tobacco leaves treated with different enzyme combinations.

[0046] Figure 12B These are DSC curves of tobacco leaves treated with different enzyme combinations;

[0047] Figure 13 This is a graph showing the release of five volatile substances in tobacco leaves after treatment with different concentrations of pectinase;

[0048] Figure 14 This is a graph showing the detection of aroma components in tobacco leaf samples treated under different storage conditions.

[0049] Figure 15 The graph shows the cell wall rupture rate after treatment with different concentrations of pectinase.

[0050] Figure 16A These are TG curves after adding different amounts of atomizing agent;

[0051] Figure 16B These are DSC curves after adding different amounts of atomizing agent.

[0052] Figure 17 This is a schematic diagram of the manufacturing process of the natural plant aerosol generation matrix described in this invention;

[0053] Figure 18 This is a schematic diagram of the main equipment deployment of the production line of the present invention. Detailed Implementation

[0054] To further illustrate the technical means and effects of this invention, the following description, in conjunction with embodiments and accompanying drawings, provides a further explanation of the invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it.

[0055] The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not limited to those elements and may also include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0056] "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event will occur and the possibility that the event will not occur.

[0057] The indefinite articles “a” and “an” preceding an element or component of this invention do not impose any limitation on the quantity (i.e., number of occurrences) of the element or component. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers only to the singular form.

[0058] The terms "one embodiment," "some embodiments," "exemplary," "specific example," or "some examples," etc., used in this invention refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the invention. In this document, the illustrative expressions of the above terms are not necessarily directed at the same embodiment or example.

[0059] I. Raw Materials

[0060] Unless otherwise specified below, the tobacco used is 2021 Henan Sanmenxia Mianchi C3F Qinyan 96 flue-cured tobacco.

[0061] The inventors also used the following flue-cured tobacco leaves in their experiments: Chenzhou Guiyang B2F, C2F, X2F; Yongzhou Lanshan B3F, C2F, X2F; Sanmenxia Mianchi B2F, C3F, X3F; Guizhou Zunyi B2F, C2F, X2F; Bijie Dafang B3F, C3F, X2F; Guizhou Bijie B2F, C2F, X3F; Yunnan Yuxi B3F, C3F, X2F; Yunnan Kunming B2F, C3F, X2F; Yunnan Dali B2F, C3F, X2F, including tobacco seed varieties such as Honghua Dajinyuan, Cuibi No. 1, and Qinyan 96. In addition, the inventors also used burley tobacco, aromatic tobacco, cigar tobacco, sun-cured tobacco, and cloves in their experiments. Unless otherwise specified below, this invention is also applicable to the above-mentioned raw materials.

[0062] II. Enzyme Preparations

[0063] 1. Pectinase: Activity 60000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., the main components are protopectinase, polygalacturonase, pectin lyase and pectin esterase.

[0064] 2. Hemicellulase: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., the main component is mannanase, its function is to degrade hemicellulose. The inventors found that after it acts on the tobacco cell wall, the products are mainly mannooligosaccharides and a small amount of mannose, etc.

[0065] 3. Amylase: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., contains multiple bioactive components such as cellulase, β-glucanase and xylanase. The inventors have discovered that it can degrade non-starch polysaccharides in tobacco, rapidly disintegrate the intertwined structure of fiber, protein and starch, promote the separation of each component, and improve the content of residual starch in fiber.

[0066] 4. Cellulase: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., Xiasheng Cellulase (CEL-01 type), mainly contains endonuclease (EG), and contains a small amount of exonuclease and a small amount of β-1,4 glucosidase.

[0067] 5. Flavor protease: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd. The inventors discovered that adding a suitable flavor protease to tobacco hydrolyzes flavor precursors and other existing enzymes in the tobacco, or the aforementioned added enzymes, thereby releasing flavor substances and enhancing and improving the flavor of the food. It can also control the bitterness of peptides. The principle is that an endopeptide breaks the peptide bonds within the polypeptide, forming short-chain peptides, some of which contain hydrophobic amino acids, thus becoming bitter peptides. An exopeptide is then used to cleave the polypeptide chain one amino acid at a time, releasing the bitter peptides completely into amino acids.

[0068] 6. Neutral protease: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd. Neutral proteases belong to the class of hydrolases. Based on the location of their hydrolytic sites on the peptide chain, they can be further subdivided into aminopeptidases, carboxypeptidases, and endopeptidases. Aminopeptidases and carboxypeptidases hydrolyze peptide bonds closest to the N-terminus or C-terminus of the protein substrate, while endopeptidases act on the interior of the protein, far from its N-terminus and C-terminus. These enzymes work together to hydrolyze the peptide bonds in the protein molecule, breaking down the protein into oligopeptides and amino acids. The inventors discovered that the neutral protease used primarily contains endopeptidase activity, with small amounts of aminopeptidase and carboxypeptidase activity. This can be used to hydrolyze residual enzymes in tobacco leaves after enzyme treatment, avoiding the generation of burnt feather odor during heating.

[0069] III. Other Major Materials, Reagents and Instruments

[0070] Experimental water: Grade I water (resistivity > 18.2 MΩ·cm) as specified in GB / T 6682-2008.

[0071] Instruments: ME204 electronic balance (sensitivity 0.0001 g, Mettler Toledo, USA); A33 continuous flow analyzer (BRAN LUEBBE, Germany); Fibertec 2010 fiber analyzer (FOSS, Denmark); Helios NanoLab 600i dual-beam focused ion beam scanning electron microscope (FEI, USA); Spirit 120kV transmission electron microscope (FEI, USA); ICS-3000 multi-functional ion chromatograph (Dionex, USA); 6890N-5975C gas chromatograph-mass spectrometer (Agilent Technologies, USA); TGA / DSC 3+ thermogravimetric analyzer; IST 16 thermal analysis system (Mettler-Toledo, Switzerland); Milli-Q Reference ultrapure water system (Millipore, USA); KBF 720 constant temperature and humidity chamber (Binder, Germany).

[0072] Blank Example A: Enzymatic hydrolysis without the addition of exogenous biological enzymes

[0073] According to traditional processes, the raw tobacco leaves are rehydrated, cut into shreds to obtain tobacco shreds, and then subjected to low-loss permeability treatment and harm reduction treatment, with each group consisting of 50 g.

[0074] Take 50 g of high-purity water (without active enzyme composition) and place it in a laboratory sprayer, then spray it evenly onto the tobacco product. Seal the sprayed tobacco product and place it in a constant temperature and humidity chamber (temperature 55℃, relative humidity 25%) for 4 h. Then place the reacted tobacco in an oven at 80℃ for 20 min. Finally, dry the final tobacco in an oven at 50℃ for 20 min, and use it as a tobacco sample for later use.

[0075] Examples B1-B4: Pectinase hydrolysis

[0076] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0077] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0078] In Examples B1-B4, pectinase was uniformly sprayed onto the tobacco shreds according to the pectinase-to-tobacco mass ratio specified in Table B-1. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0079] Table B-1: Mass ratio of pectinase and tobacco in each example

[0080]

[0081] Examples B5-B6: Pectinase hydrolysis (at different storage temperatures and humidity levels)

[0082] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0083] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0084] Pectinase was evenly sprayed onto the tobacco shreds at a ratio of 1.0% by weight of pectinase to tobacco, corresponding to examples B5-B7. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber for enzymatic hydrolysis for 4 hours according to the conditions set in Table B-2. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, drying time 20min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, drying time 20min) and used as tobacco shred samples for later use.

[0085] Table B-2: Setting conditions of the constant temperature and humidity chamber in each embodiment

[0086]

[0087] Examples B7-B12: Pectinase hydrolysis (different hydrolysis reaction times)

[0088] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0089] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0090] Pectinase was evenly sprayed onto the tobacco shreds at a ratio of 1.0% by weight of pectinase to tobacco, corresponding to examples B7-B12. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%), and the enzymatic hydrolysis reaction time was set according to Table B-3. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). For example B10, the spraying and enzymatic hydrolysis process was repeated three times to ensure enzyme activity; for example B11, the spraying and enzymatic hydrolysis process was repeated four times to ensure enzyme activity. The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) as tobacco shred samples for later use.

[0091] Table B-3: Enzymatic hydrolysis reaction time for each embodiment

[0092]

[0093] Examples B13-B15: (Different methods for terminating enzymatic hydrolysis reactions)

[0094] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0095] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0096] Pectinase was evenly sprayed onto the tobacco shreds at a ratio of 1.0% by weight of pectinase to tobacco, corresponding to examples B13-B15. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction, with the oven parameters set according to Table B-4. The finally enzymatically hydrolyzed tobacco shreds were then dried in the oven (set temperature 50℃, drying time 20 min) as tobacco shred samples for later use.

[0097] Table B-4: Conditions for terminating the enzymatic hydrolysis reaction in each example

[0098]

[0099] Examples C1-C4: Enzymatic hydrolysis of hemicellulase

[0100] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0101] Dilute the hemicellulase with ultrapure water to prepare a 200 U / g hemicellulase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0102] Hemicellulase and tobacco were uniformly sprayed onto the tobacco shreds according to the proportions in Table C-1, corresponding to examples C1-C4 of tobacco shreds, respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0103] Table C-1: Mass ratio of hemicellulase to tobacco in each example

[0104]

[0105] Examples D1-D4: Amylase hydrolysis

[0106] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0107] Dilute the amylase with ultrapure water to prepare a 200 U / g amylase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0108] Amylase and tobacco were sprayed evenly onto the tobacco shreds according to the proportions in Table D-1, corresponding to examples D1-D4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0109] Table D-1: Mass ratio of amylase and tobacco in each example

[0110]

[0111] Examples E1-E4: Cellulase hydrolysis

[0112] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0113] Dilute the cellulase with ultrapure water to prepare a 200 U / g cellulase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0114] Cellulase and tobacco were sprayed evenly onto the tobacco shreds according to the proportions in Table E-1, corresponding to examples E1-E4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0115] Table E-1: Mass ratio of cellulase and tobacco in each example

[0116]

[0117] Examples F1-F4: Enzymatic hydrolysis with flavor proteases

[0118] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0119] The flavor protease was diluted with ultrapure water to prepare a 200 U / g flavor protease solution. The solution was prepared and used immediately. The flavor protease solution was placed in a laboratory sprayer.

[0120] Flavor protease and tobacco were evenly sprayed onto the tobacco shreds according to the ratios shown in Table F-1, corresponding to examples F1-F4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0121] Table F-1: Mass ratio of flavor protease and tobacco in each example

[0122]

[0123] Examples F5-F8: Enzymatic hydrolysis with neutral protease

[0124] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0125] Dilute the neutral protease with ultrapure water to prepare a 200 U / g neutral protease solution. Prepare and use immediately. Place the neutral protease solution into a laboratory sprayer.

[0126] Neutral protease and tobacco were evenly sprayed onto the tobacco shreds according to the ratios shown in Table F-2, corresponding to examples F1-F4 respectively. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) as tobacco samples for later use.

[0127] Table F-2: Mass ratio of neutral protease and tobacco in each example

[0128]

[0129] Examples G1-G6: Enzymatic hydrolysis of enzyme compositions

[0130] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0131] Prepare the enzyme composition according to the following weight ratio, dilute the enzyme composition with ultrapure water to prepare an enzyme composition solution of 200 U / g, and use it immediately. Put the enzyme composition solution into a laboratory sprayer.

[0132] The enzyme composition formulations are shown in Table G-1, corresponding to examples G1-G6 of the tobacco shreds, respectively. The enzyme composition was uniformly sprayed onto the tobacco shreds at a ratio of 1.0% by mass of tobacco. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0133] Table G-1: Composition Ratio

[0134]

[0135] Examples H1-H5: Different nebulization doses

[0136] Add 10%, 15%, 17.5%, 20%, and 25% of the tobacco weight of the tobacco to the tobacco obtained in Example B5, respectively, and then use them as tobacco samples for Examples H1-H5.

[0137] Examples H6-H9: Different atomizing agent formulations

[0138] Add 15% by weight of atomizing agent, consisting of glycerol and propylene glycol, to the tobacco shreds obtained in Example B5. The formulation of the atomizing agent is shown in Table H-1. Then, use these as tobacco shreds samples for Examples H6-H9.

[0139] Table H-1: Proportions of atomizing agents in each embodiment

[0140]

[0141] To further demonstrate the beneficial effects of the present invention, the tobacco samples obtained from blank example A and each of the examples B to H were tested using the following test examples.

[0142] Experimental Example 1: Routine Chemical Composition Detection

[0143] The routine chemical composition of Comparative Example A and Samples B1-B4, C1-C4, and D1-D4 was determined using industry-standard methods, including the following assays:

[0144] YC / T 159―2019 Determination of water-soluble sugars in tobacco and tobacco products by continuous flow method;

[0145] YC / T 160-2002 Determination of total alkaloids in tobacco and tobacco products by continuous flow method;

[0146] YC / T 161-2002 Determination of total nitrogen in tobacco and tobacco products by continuous flow method;

[0147] YC / T 162-2011 Determination of chlorine in tobacco and tobacco products by continuous flow method;

[0148] YC / T 217-2007 Determination of potassium in tobacco and tobacco products by continuous flow method.

[0149] In this invention, the experimental results of each case were repeated three times, and the data are expressed as mean ± standard deviation. SPSS software (SPSS 13.0, SPSS Inc., USA) was used to perform statistical analysis on the differences between data groups, and one-way analysis of variance (ANOVA) was used to perform the least significant difference (LSD) test on the data. p <0.05 indicates a statistically significant difference. The data is represented by *. The results are shown in Tables 1-3.

[0150] Table 1: Effects of pectinase treatment on conventional chemical components of tobacco leaves

[0151]

[0152] Table 1 shows that pectinase treatment mainly produces sugar compounds. The higher the content of sugar compounds, the better the aroma of heated cigarettes and the smoother and more delicate the smoke.

[0153] Table 2: Effects of hemicellulase treatment on conventional chemical components of tobacco leaves

[0154]

[0155] Table 2 shows that hemicellulase treatment mainly produces carbohydrate compounds.

[0156] Table 3: Effects of amylase treatment on conventional chemical components of tobacco leaves

[0157]

[0158] Table 3 shows that amylase treatment mainly produces carbohydrate compounds and affects chlorides.

[0159] Experimental Example 2: Detection of Major Components of the Cell Wall

[0160] The main chemical components of the cell wall in Comparative Example A and Examples B1-B4, C1-C4, and D1-D4 were determined using industry-standard methods:

[0161] YC / T 347-2010 Determination of neutral detergent fibers, acid detergent fibers and acid-washed lignin in tobacco and tobacco products - Detergent method;

[0162] YC / T 346-2010 Determination of pectin in tobacco and tobacco products by ion chromatography.

[0163] Figure 4 The results shown are the pectin content test results of cell walls after pectinase treatment in Examples B1-B4. Pectinase decomposes pectin with a significant dose-response relationship. When the amount of pectinase added is 1.5%, a statistically significant difference is observed.

[0164] Figure 5 The results show the hemicellulose content of the cell wall after hemicellulase treatment. The hemicellulase decomposes hemicellulose with a significant dose-response relationship. When the amount of hemicellulase added is 1.0%, a statistically significant difference is observed.

[0165] Figure 6 The results show the starch content of the cell wall after amylase treatment. The amylase decomposes starch with a significant dose-response relationship. When the amount of amylase added was 1.0%, a statistically significant difference was observed.

[0166] Test Example 3: Maximum Atomizing Agent Load Test

[0167] Based on the products of Comparative Examples A and Examples B1-B19, C1-C4, D1-D4, E1-E4, F1-F8, and G1-G6, a certain mass of tobacco samples were weighed and placed in a constant temperature and humidity chamber, where they were equilibrated for 4 hours at 25°C and 20% humidity. Using a tobacco filling value measuring instrument, 20g of the equilibrated tobacco was accurately weighed, and 1% (by weight) of an atomizing agent (selected as 100% glycerol) was evenly sprayed onto the tobacco. Simultaneously, tests were conducted for Examples A and B1-B4 using 100% propylene glycol as the atomizing agent. Alternatively, 100% propylene glycol or a combination of two or more polyols can be used as the aerosol for the same experiment.

[0168] like Figure 7As shown, tobacco shreds 1 with 1% atomizing agent (relative to the initial tobacco mass) were placed in a compression container 2. The compression container 2 was a cylindrical barrel with a diameter of (60.0±0.1) mm and a height of not less than 100 mm. A force-applying probe 3 with a diameter of (55.0±0.1) mm and capable of generating a uniform pressure of (29.4±0.5) N on the sample was applied to the surface of the tobacco shreds. The pressure application speed of the force-applying probe was (19.5±0.5) mm / s, and the pressure application time was (30.0±0.5) s. After the pressure was applied, the tobacco shreds were poured out onto a flat piece of white paper and gently shaken. If the tobacco shreds could be dispersed well without obvious clumping, then 1% atomizing agent (relative to the initial tobacco mass) was applied again, and all other conditions remained unchanged, and the above experiment was repeated. The experiment was terminated when the tobacco shreds were poured onto a flat sheet of white paper and gently shaken. If the tobacco shreds could not disperse well and showed obvious clumping, it indicated that the atomizing agent had not been extracted. The amount of atomizing agent used at this point was recorded as B%. The maximum load of tobacco substances on the atomizing agent under these conditions was (B-1)%. The test results are shown in Tables 4-6.

[0169] Table 4: Maximum loading after different enzyme treatments

[0170]

[0171] Table 5: Maximum glycerol loading after treatment with different enzyme compositions

[0172]

[0173] Table 6: Maximum glycerol loading after different total enzymatic hydrolysis times

[0174]

[0175] Based on scanning electron microscopy experiments and maximum nebulizer loading experiments, it can be deduced that the maximum loading is determined by the cell wall rupture rate, intercellular space, and the viscosity of the nebulizer. Table 7 shows the results of intercellular space, cell wall rupture rate, and maximum nebulizer loading tests for Examples A and B1-B4.

[0176] Table 7: Relationship between intercellular space, cell wall rupture rate and maximum atomizing agent load

[0177]

[0178] Experimental Example 4: Thermogravimetric Analysis of Tobacco Shreds

[0179] This invention involves thermogravimetric analysis of tobacco leaves treated with 0-2.0% enzyme to investigate the effect of different cell wall rupture degrees on the heat release performance of tobacco leaves.

[0180] Approximately 5 mg of tobacco sample treated with enzyme solution was weighed, and 18% of atomizing agent (100% glycerol) was added. The sample was then placed in an alumina crucible and subjected to thermogravimetric analysis (TGA). Since the heating temperature of the cigarette core segment did not exceed 400℃, the heating program was selected to increase the temperature from 30℃ to 400℃ at a rate of 20℃ / min; gas flow rate: 40 mL / min; ambient gas: air. Each parallel sample was repeated three times.

[0181] The results of tobacco leaves treated with different concentrations of pectinase in Examples B1-B4 are as follows: Figure 8A and Figure 8B As shown in the figure, when the temperature is between 30 and 125°C, the weight loss ratio of tobacco leaves treated with different concentrations of pectinase is relatively small, ranging from 5% to 10%, and the corresponding weight loss rate of the DTG curve is also low. It is noteworthy that the temperature at which the maximum weight loss rate of tobacco leaves treated with pectinase occurs within this temperature range is 25°C lower than that of blank example A, suggesting that cell wall rupture within this temperature range promotes the evaporation and premature release of water and some small molecule chemical substances. When the temperature is between 125 and 250°C, the TG curve gradually becomes the stage of maximum weight loss of tobacco leaves with increasing pectinase concentration, with a weight loss ratio of 25% to 35%. The corresponding DTG curve shows a larger weight loss rate, with the maximum weight loss rate occurring at 210°C. When the pectinase treatment concentration exceeds 50 U / g, this stage becomes the range of maximum weight loss rate, and the weight loss ratio increases with increasing pectinase concentration. The inventors believe that this stage may be the release range of the main small molecule aroma substances. When the temperature was between 250 and 350℃, the TG curve showed that the weight loss percentage of the tobacco leaves was between the first two temperature ranges, ranging from 15% to 25%. The corresponding DTG curve showed the maximum weight loss rate at 310℃, and the weight loss percentage decreased with increasing pectinase concentration. The inventors believe that the weight loss at this stage may be due to the decomposition of large, non-volatile macromolecules in the tobacco. The enzymatic hydrolysis of pectin reduces the substrate in this stage, leading to a slight decrease in the weight loss percentage. With increasing enzyme addition, the proportion of weight loss due to heat in the tobacco-type smoking medium continuously increased. Compared to blank example A, the total weight loss of the experimental groups with enzyme additions of 0.5%, 1%, 1.5%, and 2% increased by 4.09%, 5.03%, 7.08%, and 10.06%, respectively, and the weight loss range was mainly between 170 and 220℃.

[0182] Taking 1% of different enzyme treatments as an example, thermogravimetric analysis was performed on Examples B2, C2, D2, F2, and F6. The results are as follows: Figure 9A and Figure 9B As shown, the TG / DTG results further confirm the sensory evaluation results. Pectinase, hemicellulase, and amylase can significantly increase the proportion of thermal weight loss, suggesting that they disrupt the cell wall structure and increase the release of aroma substances. Figures 10A-12B This further validates the conclusion.

[0183] Thermogravimetric analysis was performed on tobacco samples treated with 0-2% hemicellulase in Examples C1-C4. Figure 10A and Figure 10B As shown.

[0184] Thermogravimetric analysis was performed on tobacco samples treated with 0-2% concentration amylase in Examples D1-D4. Figure 11A and Figure 11B As shown.

[0185] Thermogravimetric analysis was performed on tobacco samples treated with the enzyme compositions in Examples G1-G6 as follows: Figure 12A and Figure 12B As shown.

[0186] Experimental Example 5: Thermogravimetric-Gas Mechanism Analysis of Tobacco Shreds

[0187] Approximately 5 mg of enzyme-treated tobacco sample was weighed and placed in a crucible. The thermogravimetric analysis (TGA) temperature program was as follows: temperature increased from 30°C to 400°C at a rate of 100°C / min; gas flow rate was 40 mL / min; and the ambient gas was air. An IST 16 coupled system was used to connect the TGA analyzer and the gas chromatography-mass spectrometer (GC-MS), employing a single-loop injection mode. The valve opening time was set according to the experimental conditions; sampling time: 10 s; loop volume: 120 μL.

[0188] The gas chromatography conditions are as follows:

[0189] The injection mode was external direct injection; the column was a DB-5MS capillary column (30 m × 0.25 mm, 0.25 μm) (Agilent Technologies, USA); the carrier gas was helium; the carrier gas flow rate was 1.0 mL / min; the injection port temperature was 260℃; the injection volume was 10 μL; and the split ratio was 10:1. The temperature program was: 60℃ for 1 min, then increased to 280℃ at a rate of 15℃ / min, and held at 280℃ for 15 min; the mass spectrometry ionization mode was EI; the ion source temperature was 230℃; the ionization energy was 70 eV; the quadrupole temperature was 150℃; the scan range was 30–450 amu; and the solvent delay was 2 min.

[0190] Thermogravimetric analysis (TGA) and gas chromatography-mass spectrometry (GC-MS) were performed on tobacco samples treated with different concentrations of pectinase in Examples B1-B4. Since there were significant differences in the thermal release of pectinase from tobacco leaves treated with different concentrations at 210°C, and the heating temperature range of most heated cigarette core segments included 210°C, the quantitative loop valve was set to open at 660 seconds, corresponding to a reaction temperature of 210°C. Gas samples released during TGA were collected for GC-MS analysis. Some of the detection results are shown below. Figure 13As shown, 18 tobacco-related compounds were identified in the releases from tobacco leaves, mainly including nicotine, furfural, furfuryl alcohol, solanone, cyclopentanone, 5-hydroxymethylfurfural, and neophytadiene. Among them, 5 compounds showed a significant dose-response effect with the amount of pectinase used. As the concentration of pectinase treatment increased, the peak area per unit weight of furfural, furfuryl alcohol, 5-hydroxymethylfurfural, and solanone increased to varying degrees, with the first three showing the most significant increase; the peak area per unit weight of neophytadiene did not change much. The results indicate that using pectinase to degrade pectin in tobacco leaves promotes the generation and release of browning reaction products, thereby further enriching the aroma of heated cigarettes. Studies have shown that pectinase can effectively degrade pectin in cell walls, increasing neutral aroma substances. Nicotine and some high-molecular-weight aroma substances are concentratedly released within the heating temperature range of 120℃ to 250℃. Therefore, this invention finds that treating tobacco leaves with pectinase can promote the thermal release of aroma substances in tobacco raw materials. Other active enzyme compositions, such as hemicellulase, amylase, or other enzyme compositions, can achieve similar effects to pectinase.

[0191] Thermogravimetric-gas chromatography-mass spectrometry (TGA) was used to further analyze the tobacco samples treated under different storage conditions in Examples B2, B5, and B6. Figure 14 As shown, when treated with biological enzymes, the cell walls of tobacco leaves are destroyed, making it easier for the cells to exchange substances with the external environment when heated. If the temperature is too high, the aroma substances in tobacco will be significantly lost. Using representative aroma substances megastigmatrienone, neophytadiene, and furfuryl alcohol as research subjects, it was demonstrated that among the storage conditions of the three examples, the storage environment of 29°C +70% in Example B5 was better.

[0192] Experimental Example 6: Scanning Electron Microscopy Experiment of Tobacco Shreds

[0193] This experimental case was tested at the Protein Science Research Platform of the Institute of Biophysics, Chinese Academy of Sciences.

[0194] The tobacco samples were placed in phosphate buffer (0.1 MPB, pH 7.2-7.4) of 2.5% glutaraldehyde + 2% paraformaldehyde at 4°C for 3 days. They were then washed four times on ice with 0.1 M PB for 10 min each time. The samples were then fixed at 4°C for 2 h with 1% osmium tetroxide solution (0.1 MPB). They were then washed three times on ice with redistilled water for 10 min each time. Gradient dehydration was performed using ethanol solution according to the conditions in Table 8, and gradient osmosis was performed according to the conditions in Table 9.

[0195] Table 8: Gradient elution conditions

[0196]

[0197] Table 9: Gradient Permeability Conditions

[0198]

[0199] The tobacco samples, after being permeated with the enzyme solution, were placed in a mold filled with resin (+0.7% DMAE) and polymerized at 45°C for 12 h and 70°C for 24 h, respectively. The polymerized tobacco samples were then cut into 70 nm thick tobacco sheets using a Leica UC6 slicer, and the tobacco sheet samples were backscattered electron images performed using a scanning electron microscope.

[0200] Results of cell wall rupture rate of tobacco leaf cells after treatment with different concentrations of pectinase in Examples B1-B4 ( Figure 15 The results showed that pectinase disrupts cell walls in a significant dose-response relationship. When the amount of pectinase added was greater than 1%, the cell wall rupture rate of the experimental group was significantly different from that of the blank sample A, indicating that under this concentration, pectinase can significantly disrupt the cell wall structure of tobacco leaves.

[0201] Meanwhile, the scanning electron microscopy results of the tobacco samples from Examples B1-B4, obtained using a Spirit 120kV transmission electron microscope, showed... Figure 1 ,in Figure 1 In this context, A is a blank example A; Figure 1 In this context, B refers to Example B1 (treated with 0.5% pectinase). Figure 1 C in the example is Example B2 (treated with 1.0% pectinase); Figure 1 D in the example is Example B3 (treated with 1.5% pectinase); Figure 1 E in the example is Example B4 (treated with 2.0% pectinase).

[0202] As can be seen from the figure, blank example A ( Figure 1 In A), the tobacco cells showed intact overall structure, with complete organelles and clear boundaries. However, with increasing pectinase concentration, the structures of the cells within the cells were significantly altered. Figure 1 In samples C, D, and E, cell wall rupture, organelle dissolution, and cell wall swelling were observed. The decomposition and rupture of the cell wall may be the mechanism by which heated cigarette tobacco-type smoke-generating media effectively adsorbs atomizing agents and releases flavor substances upon heating.

[0203] like Figure 2 and Figure 3 As shown, where Figure 2 In this context, A is a blank example A; Figure 2 B in the example is Example C1 (treated with 0.5% hemicellulase); Figure 2 C in this context refers to Example C2 (treated with 1.0% hemicellulase). Figure 2 D in the example is Example C3 (treated with 1.5% hemicellulase); Figure 2E in the example is Example C4 (treated with 2.0% hemicellulase);

[0204] in Figure 3 In this context, A is a blank example A; Figure 3 B in the example is Example C1 (treated with 0.5% hemicellulase); Figure 3 C in this context refers to Example C2 (treated with 1.0% hemicellulase). Figure 3 D in the example is Example C3 (treated with 1.5% hemicellulase); Figure 3 E in the example is Example C4 (treated with 2.0% hemicellulase);

[0205] Transmission electron microscopy (TEM) results of 10 μm and 2 μm intercellular gaps in tobacco samples treated with C1-C4 hemicellulase, compared with blank example A (…). Figure 2 Group A) and low concentration treatment group ( Figure 2 In B), the tobacco cells are arranged in an orderly and compact manner with clear cell wall boundaries. However, as the concentration of the gradient treatment increases, in Figure 2 In cell C, disordered cell arrangement, widened intercellular spaces, cell wall swelling, and further widening of intercellular spaces leading to cell detachment were observed. With increasing hemicellulase concentration, no obvious cell wall rupture was observed, the overall cell morphology changed from round to irregular, and organelles gradually dissolved. This indicates that although hemicellulase treatment did not cause cell wall rupture, it effectively penetrated the cell and destroyed organelles by disrupting the cell wall's microstructure, thus achieving the mechanism of effective adsorption and release of atomizing agents and flavoring substances upon heating.

[0206] Amylase or enzyme combinations can also achieve similar effects in adsorbing atomizing agents and effectively releasing flavoring substances upon heating.

[0207] Experiment Example 7: Sensory Evaluation Experiment

[0208] The tobacco samples from Examples H1-H5 were made into cigarettes, and the cigarette tasters used F5202-JS3 heating devices to heat and smoke them. Referring to GB5606.4-2005 Cigarettes Part 4 Sensory Technical Requirements, YC / T 564-2018 Sensory Evaluation Method for Chinese Cigarettes Based on Consumer Experience, and YC / T 497-2014 Sensory Evaluation Method for Chinese Cigarette Style, and in combination with the characteristics of heated cigarette smoke, key evaluation indicators were formulated, sensory evaluation was conducted, and the evaluation scores were recorded according to Table 10.

[0209] Table 10: Sensory Evaluation Record Form for Tobacco-Based Heated Cigarettes

[0210]

[0211] The sensory evaluation results of Examples H1-H5 are shown in Table 11:

[0212] Table 11: Sensory Evaluation Record Table for Examples H1-H5

[0213]

[0214] Based on the sensory evaluation in Table 11, adding 15% (by weight of tobacco) of atomizing agent to the tobacco sample to produce cigarettes is preferred. This amount of atomizing agent ensures sufficient smoke production while maintaining a relatively balanced distribution of other indicators, thus achieving a satisfactory smoke production effect. Furthermore, in actual production, thermogravimetric analysis of the cigarettes produced in Examples H1-H5 was conducted as shown in Experiment 4. The inventors found that as the amount of atomizing agent added increases, the weight loss of the tobacco-type smoke-producing medium gradually increases. When the addition amount is 25%, the weight loss becomes significantly greater, but this leads to severe tobacco adhesion, affecting large-scale production. Therefore, considering the sensory evaluation indicators in Table 12, a 15% atomizing agent addition amount is preferred.

[0215] Cigarettes were made by adding 15% (by weight) of atomizing agent to the tobacco samples from Examples B13-B15. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 12.

[0216] Table 12: Sensory Evaluation Record Table for Examples B13-B15

[0217]

[0218] As can be seen from the sensory evaluation record tables of Examples B13-B15, among the drying conditions of low temperature long drying (90℃, 10min), medium temperature short drying (170℃, 1min-3min) and high temperature fast drying (235℃, 3-5s), high temperature fast drying is more suitable for tobacco-type smoking medium. High temperature fast drying can quickly inactivate biological enzymes while avoiding the volatilization of aroma substances.

[0219] Cigarettes were made by adding 15% (by weight) of atomizing agent to the tobacco samples from Examples B13-B15. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 13.

[0220] Table 13: Sensory evaluation of different dosages of pectinase preparations

[0221]

[0222] Sensory evaluations after adding different doses of enzyme preparation showed that the overall sensory quality first increased and then decreased with increasing dose, with 1% or 1.5% of enzyme preparation being preferred.

[0223] Cigarettes were made by adding 15% (by weight) of atomizing agent to tobacco samples from different tobacco storage times. These samples were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 14.

[0224] Table 14: Sensory evaluation of different leaf storage times

[0225]

[0226] Sensory evaluations of different leaf storage times show that the optimal storage time is 4 hours, with a storage temperature of 29℃ and a humidity of 70%.

[0227] Cigarettes were made by adding 15% (by weight) of atomizing agent to tobacco samples from different enzyme preparation examples. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 15.

[0228] Table 15: Sensory evaluation of different enzyme preparations

[0229]

[0230] Based on the sensory evaluation results of different enzyme preparations in Table 15, different types of enzyme preparations were screened, and pectinase, hemicellulase and amylase were selected as the preferred enzyme preparation formulations for treating tobacco leaves.

[0231] Glycerin and propylene glycol, two of the most common atomizing agents, were used. Glycerin provides a better smoky sensation, while propylene glycol provides sweetness and improves smoke characteristics. Cigarettes were made by adding 15% of the tobacco weight of different atomizing agents to tobacco samples. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 16.

[0232] Table 16: Sensory evaluation of different atomizing agent formulations

[0233]

[0234] Based on the sensory evaluation in Table 16, and by adjusting the ratio of the two atomizing agents, the preferred atomizing agent formulation is 4:6.

[0235] To further demonstrate the application of this invention, the following uses single-origin tobacco from nine core production areas, configured into light-aroma, medium-aroma, and strong-aroma modules with distinct style characteristics.

[0236] Strongly aromatic modules: Chenzhou Guiyang C2F, Yongzhou Lanshan C2F, Sanmenxia Mianchi C3F, with a material ratio of 1:1:1;

[0237] Intermediate fragrance modules: Guizhou Zunyi C2F, Bijie Dafang C3F, and Guizhou Bijie C2F, with a material ratio of 1:1:1.

[0238] Lightly scented modules: Yunnan Yuxi C3F, Yunnan Kunming C3F, Yunnan Dali C3F, with a material ratio of 1:1:1.

[0239] The tobacco raw materials of the light aroma module, intermediate aroma module and strong aroma module were respectively made into tobacco shreds samples by the method of Example G2. 15% of the tobacco shred weight of atomizing agent was added to the tobacco shreds samples to make cigarettes. The atomizing agent was composed of glycerin and propylene glycol in a ratio of 4:6. The cigarettes were subjected to sensory evaluation tests, and the results are shown in Table 17.

[0240] Table 17: Sensory evaluation of different fragrance modules

[0241]

[0242] Based on the sensory evaluation results in Table 17, all three aroma types have a good taste. Among them, the strong aroma type has the best overall sensory quality. The tobacco raw materials have significant differences in style characteristics between shredded heated cigarettes and sheet heated cigarettes, and have a similar style performance to traditional cigarettes.

[0243] Sanmenxia Mianchi B2F was used as the upper tobacco leaf raw material, Sanmenxia Mianchi C3F as the middle tobacco leaf raw material, and Sanmenxia Mianchi X2F as the lower tobacco leaf raw material. Tobacco samples were prepared according to the method of Example G2. Cigarettes were made by adding 15% of the tobacco weight of atomizing agent to the tobacco samples. The atomizing agent was composed of glycerol and propylene glycol in a ratio of 4:6. Sensory evaluation tests were conducted on the prepared cigarettes, and the results are shown in Table 18.

[0244] Table 18: Sensory evaluation of different parts of tobacco leaves

[0245]

[0246] Sensory quality evaluation was conducted using tobacco leaves from different parts of the same production area. The results showed that the upper tobacco was superior to the middle and lower tobacco. This result is different from that of traditional cigarettes and can provide support for the comprehensive utilization of upper tobacco.

[0247] Test Example 8: Whole Filament Rate Test

[0248] The whole-shred rate and broken-shred rate of tobacco shreds in some of the above embodiments were determined according to "YC / T 178-2003 Method for Determination of Whole-shred Rate and Broken-shred Rate of Tobacco Shreds". For other forms of plant materials such as tobacco, tobacco sheets, and cloves, the whole-shred rate can also be measured after cutting them into shreds in a conventional manner. The whole-shred rate of each embodiment is shown in Table 19.

[0249] Table 19: Whole tobacco yield after different treatment methods

[0250]

[0251] Test Example 9: Production Line Test

[0252] refer to Figure 17 The process steps of this invention from natural plant raw materials to the formation of aerosol matrix can be combined in various ways. The essential steps include: pretreatment step, low-damage permeable treatment step, aerosol permeation step, harm reduction treatment step and morphological change step. However, the order of these steps can be adjusted, and some steps can be carried out in separate steps or in combination with other steps and carried out simultaneously.

[0253] In a specific embodiment, such as Example I1, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing, shredding) → low-damage permeable treatment step (enzymatic hydrolysis) → first harm reduction treatment step (high temperature rapid drying of smoke sheets) → second aerosol permeation step → formation of aerosol generation matrix.

[0254] In one specific embodiment, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing) → low-damage permeability treatment step (enzymatic hydrolysis) → first aerosol permeation step (conducted simultaneously with enzymatic hydrolysis) → first harm reduction treatment step (high-temperature rapid drying of smoke sheets) → second morphological change step (shredding) → second harm reduction treatment step (adding protease) → second aerosol permeation step (simultaneously with adding protease) → formation of aerosol generation matrix.

[0255] In one specific embodiment, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing) → low-damage permeability treatment step (enzymatic hydrolysis) → first aerosol permeation step (conducted simultaneously with enzymatic hydrolysis) → first harm reduction treatment step (high-temperature rapid drying of smoked sheets) → second morphological change step (shredding) → second harm reduction treatment step (addition of protease) → formation of aerosol generation matrix. Specifically, as follows:

[0256] like Figure 18 As shown, the tobacco shreds and formulation of this invention can be produced using a conventional cigarette production line. The production line includes: a tobacco sheet processing section 100, a tobacco shred processing section 200, a blending and flavoring device 250, a cigarette rolling section 300, and a production line control system. The production line control system includes one or more processors (CPUs), input / output devices or interfaces, wired or wireless network interfaces, and memory. The input devices include human-machine interface devices to receive control information from operators, including cigarette type and formula. The memory stores computer programs and / or computer instructions, which, when executed by the processor, control the production line.

[0257] The tobacco leaf processing section 100 includes an unpacking device, a metering device, a slicing device, a loosening and rehydration device, a tobacco leaf pre-mixing device, a screening device, a feeding device, and a leaf storage device. After the tobacco leaves have been threshed, re-dried, and aged, they are sent to the reuse production line in boxes and need to go through the following processes: unpacking and metering, slicing, loosening and rehydration, tobacco leaf pre-mixing, screening and feeding, leaf mixing and storage. First, according to the leaf group formula requirements of the product, the tobacco leaves are prepared for feeding. The unpacking device 110 is used to unpack the tobacco leaves through the unpacking process. After removing the outer packaging, the tobacco sheets are cut into several pieces of a specified thickness using a slicing device 130. Then, they are loosened and rehydrated using a loosening and rehydration device 150 and a tobacco sheet pre-mixing device to improve the processing resistance of the tobacco sheets and to initially and evenly mix the tobacco sheets. Then, a screening device is used to remove fragments, and tobacco slurry is sprayed onto the tobacco sheets using a feeding device 170. Finally, the selected tobacco sheets are evenly mixed and stored in a leaf storage device 180 for use in the next process stage.

[0258] The feeding device 170 includes a liquid source containing an enzyme combination (with the same proportions as in Example G2) and an atomizing agent composed of glycerol, propylene glycol, and water as the solvent. The feeding device 170 has a nozzle connected to the liquid source, thereby allowing the liquid to be selectively sprayed onto the tobacco sheet according to instructions issued by the production line control system. Adding the aerosol before the enzymatic hydrolysis reaction allows the aerosol to quickly fill the interior of the tobacco cell walls after enzymatic hydrolysis, ensuring full absorption of the aerosol by the tobacco and preventing clumping or failure to disperse during subsequent shredding by the shredder 210. A single enzymatic hydrolysis reaction is preferred on the production line, but multiple reactions are also possible. This means the aerosol can be sprayed before the first or last reaction, or evenly before each reaction. The minimal aerosol release on the tobacco surface prevents contamination of the production line, particularly hindering the cigarette rolling process 300. Furthermore, the absorption of glycerol and propylene glycol within the cell walls reduces their contact with airborne moisture, lowering apparent water absorption and thus extending shelf life.

[0259] The leaf storage device 180 can be a leaf storage cabinet. The tobacco leaves sprayed with liquid will be sent into the leaf storage cabinet for enzymatic degradation reaction. The temperature of the leaf storage cabinet is 28-33℃ and the relative humidity is 67-75%. The leaf storage time can be selected as needed.

[0260] Because the enzymatically hydrolyzed tobacco sheets of this invention have high processability, they can be directly processed into heated tobacco shreds through the tobacco processing section 200 of a traditional tobacco production line, using processes such as shredding. The tobacco processing section 200 includes a shredding device, a drying device, and a blending and flavoring device. First, the shredding device shreds the tobacco sheets 21 to form heated tobacco shreds of a certain width. Then, the drying device dries the tobacco shreds. This step, in addition to drying the heated tobacco shreds to remove some of the moisture (to achieve a specified moisture content, for example, 11.5%~14.0%), thereby improving the filling capacity and sensory quality of the shreds, also includes a high-temperature rapid drying stage (temperature 200-250℃, preferably 235℃; time 1-10s, preferably 3-5s) to inactivate residual enzyme combinations. Finally, the blending and flavoring device further adds flavorings and additives suitable for heating cigarettes to the tobacco, including proteases used to degrade biological enzymes to remove impurities, and then sends it to the cigarette rolling section 300.

[0261] The cigarette rolling section 300 includes a cigarette rolling device and a feeding device. The cigarette rolling device rolls heated cigarette tobacco into heated cigarette tobacco segments, and the feeding device feeds heated cigarette tobacco segments and heated cigarette filter segments into heated cigarettes 61.

[0262] This invention utilizes cell wall-degrading enzymes to quantitatively and controllably modify traditional cigarette tobacco, enabling it to absorb more aerosols and release more aroma compounds during heating while maintaining its original processability (especially the whole tobacco yield). This allows traditional cigarette production lines to be reused with heated tobacco with only minor modifications. By selecting specific enzyme compositions, the enzymatic degradation reaction can be carried out in the existing leaf storage environment without the need for additional high-temperature reaction equipment.

[0263] The terminology and expressions used herein are for descriptive purposes only, and the invention should not be limited to these terms and expressions. The use of these terms and expressions does not imply the exclusion of any illustrative and descriptive equivalents (or parts thereof), and it should be recognized that various modifications that may exist should also be included within the scope of the claims. Other modifications, variations, and substitutions may also exist. Accordingly, the claims should be considered to cover all such equivalents.

[0264] Similarly, it should be noted that although the present invention has been described with reference to the specific embodiments described above, the present invention is not limited to the detailed methods described above, that is, it does not mean that the present invention must rely on the detailed methods described above to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A natural plant aerosol generating matrix for electric heating smoke appliances, characterized in that, The natural plant aerosol generating matrix for electric heating smoke appliances includes: natural plant raw materials that have undergone quantified cell wall disruption treatment and exogenous aerosol generating agents loaded within the natural plant raw materials; The quantitative cell wall disruption treatment includes 1 to 5 enzymatic cell disruption steps, each of which includes: Step A: Spray the active cell wall breaking enzyme solution evenly onto the natural plant material; Step B: Seal the sprayed natural plant material and carry out enzymatic hydrolysis and cell wall breaking reaction under constant temperature and humidity conditions; The active cell-wall-breaking enzyme includes any one or a combination of at least two of pectinase, hemicellulase, cellulase or amylase. After step B is completed, step C is performed: the apparent loading and / or micro loading of the natural plant material are tested. When the apparent loading and / or micro loading of the aerosol generating agent of the natural plant material is greater than the target loading, the next enzymatic hydrolysis and cell wall breaking step will not be performed. Based on the difference between the apparent loading and / or microscopic loading of the aerosol generating agent of the natural plant raw material and the target loading, determine the enzymatic hydrolysis reaction temperature, relative humidity, and / or reaction time in the next enzymatic hydrolysis reaction step. The apparent loading test includes weighing the maximum loading of the exogenous aerosol generating agent; the microscopic loading test includes observing the cell wall and statistically analyzing the cell wall disruption rate. After each enzymatic hydrolysis and cell wall breaking reaction is completed, step E is performed: the whole fiber rate of the natural plant material is detected. If the whole fiber rate of the natural plant material is greater than the whole fiber rate warning value, the next enzymatic hydrolysis and cell wall breaking step will not be performed.

2. The natural plant aerosol generating matrix according to claim 1, characterized in that, In step A, the weight ratio of the active cell wall-breaking enzyme to the natural plant raw material is 0.5-3.0%.

3. The natural plant aerosol generating matrix according to claim 1, characterized in that, In step A, the active cell-wall-breaking enzyme is composed of pectinase, hemicellulase and amylase; Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, cellulase, and amylase; Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, hemicellulase, cellulase, and amylase, in the following weight ratio: Pectinase: 15-25%, Total hemicellulase and cellulase: 32-48%, Amylase: 32-48%.

4. The natural plant aerosol generating matrix according to claim 1, characterized in that, In step B, the conditions for each enzymatic hydrolysis and cell wall breaking reaction are independent: temperature of 25-60℃, relative humidity of 15-80%, and enzymatic hydrolysis and cell wall breaking reaction time of 0.5-24.0 h.

5. The natural plant aerosol generating matrix according to claim 1, characterized in that, After each enzymatic hydrolysis and cell wall breaking reaction is completed, step D is performed: the natural plant raw material is subjected to a harm reduction treatment.

6. The natural plant aerosol generating matrix according to claim 1, characterized in that, The exogenous aerosol generator is loaded into the natural plant material that has undergone quantitative cell wall disruption treatment in the following manner: The exogenous aerosol generating agent is sprayed onto the natural plant raw material that has undergone quantified cell wall disruption treatment, wherein: the weight of the exogenous aerosol generating agent is less than the weight of the natural plant raw material × maximum loading rate × whole fiber rate; Once the exogenous aerosol generating agent has completely penetrated into the natural plant raw material that has undergone quantified cell wall disruption treatment, the natural plant aerosol generating matrix is ​​prepared.

7. The natural plant aerosol generating matrix according to claim 1, characterized in that, The warning value for the whole yarn rate is above 60%.

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