A process for the preparation of a tobacco flavouring agent, products and uses thereof

Abstract

The present application relates to a kind of tobacco flavor preparation method and its product and application, comprising the following steps: tobacco, sorghum and water are mixed to be sterilized, obtain tobacco-sorghum liquid medium, enzyme preparation and bacteria liquid are added to tobacco-sorghum liquid medium and are cultured fermentation, the bacteria liquid is yeast bacteria liquid and / or lactobacillus bacteria liquid, obtain the tobacco flavor.The present application uses sorghum and tobacco as common fermentation substrate, gives the double flavor of final flavoring liquor and tobacco flavor, using enzyme treatment combined with liquid fermentation method, solve the problem that the flavor brought by traditional stacking solid fermentation often needs to be passed through long time and flavor is insufficient etc., process is simple, condition is mild, green and environmental protection, cost is low, significantly improve the flavor quality of low quality cigar, improve the economic value of low quality cigar, with wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of natural flavoring and fragrance development technology for tobacco, specifically relating to a method for preparing tobacco flavoring agents, their products, and applications. Background Technology

[0002] In the cigar industry, the quality of tobacco leaves is considered fundamental to producing the complex and rich aromas and flavors of premium cigars. Traditionally, substandard tobacco leaves are disposed of by companies inefficiently, such as through simple scrapping or discarding, which not only wastes company resources but can also pollute the environment. With the increasing demand for diversified cigar products and the urgent need to improve the economic efficiency of low-quality tobacco leaves, the development of novel cigar flavoring agents has become extremely important. As an important auxiliary material in cigar processing, cigar flavoring agents can impart richer aromas and layers to cigars, thereby satisfying consumers' pursuit of unique tastes. Extracting aroma components from low-quality tobacco leaves using specific processes and using them as flavoring agents can not only enhance their economic value but also provide new possibilities for cigar product innovation.

[0003] Fermentation is an excellent method for improving the quality of tobacco leaves, but traditional solid-state fermentation often results in flavors that require a long fermentation time and are insufficient. Brewing-type tobacco flavoring agents are a new type of flavoring agent that combines the brewing aroma of baijiu (Chinese liquor) with the natural aroma of tobacco. Sorghum is often used as a primary raw material in the fermentation of Chinese baijiu and can be used together with tobacco as a fermentation substrate. However, traditional methods for producing tobacco-sorghum flavoring agents mainly employ dry fermentation or mechanical mixing, which suffers from uneven distribution of flavor compounds and insufficient flavor.

[0004] Therefore, developing a tobacco flavoring agent that is simple to produce, operates under mild conditions, is environmentally friendly, has low cost, and can significantly improve the flavor quality of low-quality cigars has broad market prospects and benefits. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for preparing tobacco flavoring agents, as well as the products and applications thereof.

[0006] To achieve this objective, the present invention employs the following technical solution:

[0007] In a first aspect, the present invention provides a method for preparing a tobacco flavoring agent, the method comprising the following steps:

[0008] (1) Mix tobacco shreds, sorghum and water and sterilize to obtain tobacco-sorghum liquid culture medium;

[0009] (2) The enzyme preparation and bacterial solution are then added to a tobacco-sorghum liquid culture medium for fermentation. The bacterial solution is a yeast solution and / or a lactobacillus solution to obtain the tobacco flavoring agent.

[0010] This invention uses sorghum and tobacco as co-fermentation substrates, imparting a dual flavor profile of brewed aroma and natural tobacco aroma to the final flavoring agent, achieving an innovative taste experience with significant aroma enhancement and strong smokeability. This invention employs synergistic liquid fermentation with yeast and lactobacillus, providing abundant alcohols and acids to improve cigar flavor, increasing the yield and uniformity of flavor compounds, and solving the problems of traditional solid-state fermentation, which often requires a long fermentation time and results in insufficient flavor. This invention also creatively combines liquid fermentation with enzyme treatment, which accelerates the degradation of macromolecules in the substrate, such as proteins, starches, and pectins, greatly improving fermentation efficiency. The preparation method of this invention is based on microbial fermentation technology, featuring a simple process, mild conditions, secondary utilization of fermentation sediment, environmental friendliness, and low cost. It significantly improves the flavor quality of low-quality cigars, increases their economic value, and has broad application prospects.

[0011] Preferably, after the culture and fermentation in step (2), the product is further freeze-dried to obtain a freeze-dried tobacco flavoring agent.

[0012] Preferably, after the culture and fermentation in step (2), the mixture is further centrifuged to obtain the supernatant, which is then used to obtain a liquid form of tobacco flavoring agent.

[0013] Preferably, after the culture and fermentation in step (2), the mixture is centrifuged and the supernatant is added to a thickener to obtain a tobacco flavor extract.

[0014] The precipitate obtained after centrifugation can also be added to a tobacco-sorghum liquid culture medium for secondary fermentation, allowing the fermented precipitate to be reused, which can achieve the effects of being green and environmentally friendly and reducing costs.

[0015] Preferably, the centrifugation speed is independently 3000-8000 rpm, and the centrifugation time is independently 5-10 min.

[0016] The centrifugation speed can be selected from 3000rpm, 3500rpm, 4000rpm, 4500rpm, 5000rpm, 5500rpm, 6000rpm, 6500rpm, 7000rpm, 7500rpm, 8000rpm, etc., and the centrifugation time can be selected from 5min, 5.5min, 6min, 7min, 8min, 9min, 10min, etc. Other specific values ​​within the above range can also be selected, and will not be elaborated here.

[0017] Preferably, the thickener is low-ester pectin and / or gellan gum.

[0018] The low-ester pectin refers to pectin with an esterification degree of less than 50%.

[0019] Preferably, the concentration of tobacco shreds in the tobacco-sorghum liquid culture medium in step (1) is 0.01-0.2 g / mL, and the concentration of sorghum is 0.01-0.2 g / mL.

[0020] The concentration of the tobacco shreds can be selected from 0.01g / mL, 0.02g / mL, 0.03g / mL, 0.04g / mL, 0.05g / mL, 0.07g / mL, 0.1g / mL, 0.15g / mL, 0.2g / mL, etc., and the concentration of the sorghum can be selected from 0.01g / mL, 0.02g / mL, 0.03g / mL, 0.04g / mL, 0.05g / mL, 0.07g / mL, 0.1g / mL, 0.15g / mL, 0.2g / mL, etc. Other specific values ​​within the above ranges can be selected, and will not be elaborated here.

[0021] Preferably, the enzyme preparation in step (2) includes any one or a combination of at least two of amylase, flavor enzyme or saccharifying enzyme.

[0022] This invention creatively discovers that using enzyme preparations with a specific composition can effectively promote the production of flavor substances, thereby improving the flavor of cigars. This is because the substrate after enzyme treatment contains many sugars that can support the growth and reproduction of microorganisms, which is conducive to rapid fermentation and the production of more flavor substances. At the same time, the enzyme preparation with this specific composition does not have an adverse effect on the ash formed during cigar combustion.

[0023] Preferably, the enzyme preparation comprises a combination of amylase, flavor enzyme, and saccharifying enzyme.

[0024] Preferably, the enzyme activity in the enzyme preparation is 100-1000 U / mL, such as 100 U / mL, 200 U / mL, 300 U / mL, 400 U / mL, 500 U / mL, 600 U / mL, 700 U / mL, 800 U / mL, 900 U / mL, 1000 U / mL, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0025] Preferably, the amylase activity in the enzyme preparation is not less than 40%, such as 40%, 45%, 50%, 55%, 60%, 65%, 70%, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0026] Preferably, the yeast culture in step (2) is obtained by culturing yeast strains at 25-35℃ (e.g., 25℃, 26℃, 28℃, 30℃, 32℃, 35℃, etc.) and 150-250r / min (e.g., 150r / min, 160r / min, 180r / min, 200r / min, 220r / min, 250r / min, etc.) with constant temperature shaking for 24-48h (e.g., 24h, 26h, 28h, 30h, 35h, 40h, 48h, etc.). Other specific point values ​​within the above range can be selected, and will not be elaborated here.

[0027] Preferably, the Lactobacillus culture solution is obtained by culturing Lactobacillus strains at 35-40℃ (e.g., 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc.) and at 150-250 r / min (e.g., 150 r / min, 160 r / min, 180 r / min, 200 r / min, 220 r / min, 250 r / min, etc.) with constant temperature shaking for 12-24h (e.g., 12h, 14h, 16h, 18h, 20h, 22h, 24h, etc.). Other specific point values ​​within the above range can be selected, and will not be elaborated here.

[0028] Preferably, the inoculation amount of yeast culture in step (3) is 0.5%-1.5%, such as 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.3%, 1.5%, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0029] Preferably, the inoculation amount of the Lactobacillus bacterial solution is 0.25%-0.75%, such as 0.25%, 0.3%, 0.35%, 4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0030] Preferably, the fermentation conditions described in step (2) are constant temperature oscillation at 25-35℃ (e.g., 25℃, 26℃, 28℃, 30℃, 32℃, 35℃, etc.) and 150-250r / min (e.g., 150r / min, 160r / min, 180r / min, 200r / min, 220r / min, 250r / min, etc.) for 1-10 days (e.g., 1 day, 2 days, 3 days, 5 days, 7 days, 10 days, etc.). Other specific values ​​within the above range can be selected, and will not be elaborated here.

[0031] In a second aspect, the present invention provides a tobacco flavoring agent prepared according to the preparation method described in the first aspect.

[0032] Thirdly, the present invention provides an application of the tobacco flavoring agent according to the second aspect in tobacco production.

[0033] Preferably, the tobacco is a cigar.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] This invention uses sorghum and tobacco as co-fermentation substrates, imparting a dual flavor profile of brewed aroma and natural tobacco aroma to the final flavoring agent, achieving an innovative taste experience with significant aroma enhancement and strong smokeability. This invention employs synergistic liquid fermentation with yeast and lactobacillus, providing abundant alcohols and acids to improve cigar flavor, increasing the yield and uniformity of flavor compounds, and solving the problems of traditional solid-state fermentation, which often requires a long fermentation time and results in insufficient flavor. This invention also creatively combines liquid fermentation with enzyme treatment, which accelerates the degradation of macromolecules in the substrate, such as proteins, starches, and pectins, greatly improving fermentation efficiency. The preparation method of this invention is based on microbial fermentation technology, featuring a simple process, mild conditions, secondary utilization of fermentation sediment, environmental friendliness, and low cost. It significantly improves the flavor quality of low-quality cigars, increases their economic value, and has broad application prospects. Attached Figure Description

[0036] Figure 1 This is a process flow diagram of the preparation method of tobacco flavoring agents. Detailed Implementation

[0037] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0038] Unless otherwise specified, the reagents and consumables used in the following embodiments were purchased from conventional reagent manufacturers in the art; unless otherwise specified, the experimental methods and techniques used were conventional methods and techniques in the art.

[0039] The yeast involved in the following examples is Pichia manshurica JNC004.001, classified as Pichia manshurica JNC004.001. It is deposited at the China Center for Type Culture Collection (CCTCC) with accession number M 2024637, on April 7, 2024, at Wuhan University, Wuhan, China.

[0040] The lactobacillus involved in the following examples is Lactobacillus plantarum, type JNC003.001, classified and named Lactobacillus plantarum JNC003.001. It is deposited at the China Center for Type Culture Collection, with accession number CCTCC NO: M 2024447, deposited on March 11, 2024, and deposited at Wuhan University, Wuhan, China.

[0041] The amylase was purchased from Shanghai Yuanye Biotechnology Co., Ltd., model S10002; the flavor enzyme was purchased from Shanghai Yuanye Biotechnology Co., Ltd., model S10153; the saccharifying enzyme was purchased from Shanghai Yuanye Biotechnology Co., Ltd., model S10017; and the acidic protease was purchased from Shanghai Yuanye Biotechnology Co., Ltd., model S10012.

[0042] Example 1

[0043] This embodiment provides a tobacco flavor extract, the process flow diagram of which is shown below. Figure 1 As shown, the preparation method is as follows:

[0044] (1) Pichia pastoris JNC004.001 was inoculated into YPD liquid medium (10 g / L peptone, 5 g / L yeast extract, 20 g / L glucose, 0.1 g / L chloramphenicol, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 30℃ and 200 r / min for 48 h to obtain yeast culture solution;

[0045] (2) Lactobacillus plantarum JNC003.001 was inoculated into MAS liquid medium (10 g / L peptone, 10 g / L beef extract, 5 g / L yeast extract, 20 g / L glucose, 2 g / L sodium citrate, 2 g / L anhydrous sodium acetate, 0.1 g / L magnesium sulfate, 0.05 g / L manganese sulfate, 2 g / L dipotassium hydrogen phosphate, 1 mL / L Tween 80, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 37℃ and 200 r / min for 24 h to obtain Lactobacillus bacterial culture;

[0046] (3) Prepare an amylase solution with an enzyme activity of 500 U / mL and a flavor enzyme solution with an enzyme activity of 500 U / mL.

[0047] (4) Mix tobacco shreds, sorghum powder and distilled water to make the concentration of tobacco shreds reach 0.05 g / mL and the concentration of sorghum powder reach 0.05 g / mL. Sterilize at 121℃ for 20 minutes to obtain tobacco-sorghum liquid culture medium. Mix the amylase solution and flavor enzyme solution in step (3) in a 1:1 ratio to obtain an enzyme solution. Then add the enzyme solution according to the volume ratio of enzyme solution to liquid culture medium of 1:10.

[0048] (5) Inoculate 1% by volume of yeast culture and 0.5% by volume of lactobacillus culture in liquid culture medium and culture in a shaker at 200 r / min for 7 days at 30℃.

[0049] (6) Centrifuge the liquid culture medium after fermentation in step (5) at 4℃ and 5000rpm for 7min, take the supernatant and add 6.5% low-ester pectin, stir in a 60℃ water bath to obtain tobacco flavor extract.

[0050] Example 2

[0051] This embodiment provides a tobacco flavor extract, the process flow diagram of which is shown below. Figure 1 As shown, its preparation method is as follows:

[0052] (1) Pichia pastoris JNC004.001 was inoculated into YPD liquid medium (10 g / L peptone, 5 g / L yeast extract, 20 g / L glucose, 0.1 g / L chloramphenicol, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 25℃ and 240 r / min for 60 h to obtain yeast culture solution;

[0053] (2) Lactobacillus plantarum JNC003.001 was inoculated into MAS liquid medium (10 g / L peptone, 10 g / L beef extract, 5 g / L yeast extract, 20 g / L glucose, 2 g / L sodium citrate, 2 g / L anhydrous sodium acetate, 0.1 g / L magnesium sulfate, 0.05 g / L manganese sulfate, 2 g / L dipotassium hydrogen phosphate, 1 mL / L Tween 80, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 35℃ and 240 r / min for 30 h to obtain Lactobacillus bacterial culture;

[0054] (3) Prepare an amylase solution with an enzyme activity of 100 U / mL and a flavor enzyme solution with an enzyme activity of 100 U / mL.

[0055] (4) Mix tobacco shreds, sorghum powder and distilled water to make the concentration of tobacco shreds reach 0.01 g / mL and the concentration of sorghum powder reach 0.01 g / mL. Sterilize at 121℃ for 20 minutes to obtain tobacco-sorghum liquid culture medium. Mix the amylase solution and flavor enzyme solution in step (3) in a ratio of 1.25:1 to obtain enzyme solution. Then add enzyme solution according to the volume ratio of enzyme solution to liquid culture medium of 1:10.

[0056] (5) Inoculate 0.5% by volume of yeast culture and 0.25% by volume of lactobacillus culture in liquid culture medium and culture in a shaker at 230 r / min for 10 days at 25℃.

[0057] (6) Centrifuge the liquid culture medium after fermentation in step (5) at 4°C and 3000 rpm for 10 min, take the supernatant and add 6.5% low-ester pectin, stir in a 60°C water bath to obtain tobacco flavor extract.

[0058] Example 3

[0059] This embodiment provides a tobacco flavor extract, the process flow diagram of which is shown below. Figure 1 As shown, its preparation method is as follows:

[0060] (1) Pichia pastoris JNC004.001 was inoculated into YPD liquid medium (10 g / L peptone, 5 g / L yeast extract, 20 g / L glucose, 0.1 g / L chloramphenicol, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 35℃ and 160 r / min for 24 h to obtain yeast culture solution;

[0061] (2) Lactobacillus plantarum JNC003.001 was inoculated into MAS liquid medium (10 g / L peptone, 10 g / L beef extract, 5 g / L yeast extract, 20 g / L glucose, 2 g / L sodium citrate, 2 g / L anhydrous sodium acetate, 0.1 g / L magnesium sulfate, 0.05 g / L manganese sulfate, 2 g / L dipotassium hydrogen phosphate, 1 mL / L Tween 80, autoclaved at 121℃ for 20 minutes) and cultured in a shaker at 40℃ and 160 r / min for 12 h to obtain Lactobacillus bacterial culture;

[0062] (3) Prepare an amylase solution with an enzyme activity of 1000 U / mL and a flavor enzyme solution with an enzyme activity of 1000 U / mL.

[0063] (4) Mix tobacco shreds, sorghum powder and distilled water to make the concentration of tobacco shreds reach 0.15 g / mL and the concentration of sorghum powder reach 0.15 g / mL. Sterilize at 121℃ for 20 minutes to obtain tobacco-sorghum liquid culture medium. Mix the amylase solution and flavor enzyme solution in step (3) in a ratio of 1.5:1 to obtain enzyme solution. Then add enzyme solution according to the volume ratio of enzyme solution to liquid culture medium of 1:10.

[0064] (5) Inoculate 1.5% by volume of yeast culture and 0.75% by volume of lactobacillus culture in liquid culture medium and culture in a shaker at 160 r / min for 4 days at 35℃.

[0065] (6) Centrifuge the liquid culture medium after fermentation in step (5) at 4°C and 8000rpm for 5 minutes, take the supernatant and add 1.5% gellan gum, stir in a 60°C water bath to obtain tobacco flavor extract.

[0066] Example 4

[0067] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that in step (3), only an amylase solution with an enzyme activity of 500 U / mL is prepared, and in step (4), an amylase solution is added according to a volume ratio of amylase solution to liquid culture medium of 1:10, while other conditions remain unchanged.

[0068] Example 5

[0069] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that in step (3), only a flavor enzyme solution with an enzyme activity of 500 U / mL is prepared, and in step (4), the flavor enzyme solution is added at a volume ratio of 1:10 of flavor enzyme solution to liquid culture medium, while other conditions remain unchanged.

[0070] Example 6

[0071] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that in step (3), an amylase solution with an enzyme activity of 500 U / mL, a flavor enzyme solution with an enzyme activity of 500 U / mL and a saccharifying enzyme solution with an enzyme activity of 500 U / mL are prepared. In step (4), the amylase solution, flavor enzyme solution and saccharifying enzyme solution from step (3) are mixed in a ratio of 1:0.5:0.5 to obtain an enzyme solution. Then, the enzyme solution is added according to the volume ratio of enzyme solution to liquid culture medium of 1:10. All other conditions remain unchanged.

[0072] Example 7

[0073] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that the flavor enzyme is replaced with an acidic protease of equal activity, while all other conditions remain unchanged.

[0074] Example 8

[0075] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that in step (5), only 1.5% by volume of yeast culture is inoculated in the liquid culture medium, while other conditions remain unchanged.

[0076] Example 9

[0077] This embodiment provides a tobacco flavor extract, which differs from Example 1 only in that in step (5), only 1.5% by volume of Lactobacillus bacterial solution is inoculated in the liquid culture medium, while other conditions remain unchanged.

[0078] Comparative Example 1

[0079] This comparative example provides a tobacco flavor extract, which differs from Example 1 only in that no enzyme preparation is added. That is, step (4) is replaced by mixing tobacco shreds, sorghum powder and distilled water to make the concentration of tobacco shreds reach 0.05 g / mL and the concentration of sorghum powder reach 0.05 g / mL. The mixture is then autoclaved at 121°C for 20 minutes to obtain a tobacco-sorghum liquid culture medium. All other conditions remain unchanged.

[0080] Application Example 1

[0081] This application example provides a cigar, the preparation method of which is as follows:

[0082] The tobacco flavor extract prepared in Example 1 was applied to the low-quality cigar at a mass ratio of 1:5, and the cigar was dried in an oven at 60°C for 2 hours to equilibrate the moisture content. Then it was rolled into a commercial cigar.

[0083] Application Example 2-9

[0084] This application example provides eight types of cigars, which differ from Application Example 1 only in that the tobacco flavor extract prepared in Example 1 is replaced in equal amounts with the tobacco flavor extract prepared in Examples 2-9, while all other conditions remain unchanged.

[0085] Comparative Application Example 1

[0086] This comparative application example provides a cigar that differs from Application Example 1 only in that the tobacco flavor extract prepared in Example 1 is replaced in equal amounts with the tobacco flavor extract prepared in Comparative Example 1, while all other conditions remain unchanged.

[0087] Test Example 1

[0088] Cigar sensory evaluation:

[0089] The cigars produced in corresponding use cases 1-9 and comparative application example 1 were subjected to sensory evaluation according to YC / T 415-2011 "Sensory Evaluation Method for Tobacco Products". The specific standards are shown in Table 1, and the results are shown in Table 2. The control group is the low-quality cigar described in application example 1 that has not been treated.

[0090] Table 1

[0091]

[0092] Table 2

[0093] Color aroma Mixed gases Irritating Aftertaste grayscale Total Score control group 6.00±0.00 21.00±0.00 7.00±0.00 10.00±0.00 13.00±0.00 4.00±0.00 61.00±0.00 Application Example 1 8.20±0.45 31.00±0.71 10.00±0.71 12.60±0.55 20.60±0.55 4.60±0.55 87.00±2.00 Application Example 2 8.03±0.02 28.36±0.81 9.56±0.25 13.22±0.45 19.56±0.24 4.44±0.02 83.17±0.65 Application Example 3 7.95±0.33 28.96±0.25 9.75±0.15 12.05±0.35 19.00±0.02 4.50±0.02 82.21±0.55 Application Example 4 6.00±0.71 27.00±0.71 8.60±0.55 9.20±0.55 15.41±0.55 4.41±0.55 70.62±1.95 Application Example 5 8.20±0.45 31.20±0.84 9.40±0.89 12.20±0.55 18.40±0.89 4.40±0.89 83.80±1.87 Application Example 6 8.85±0.02 32.59±0.24 11.25±0.12 12.86±0.25 21.25±0.15 5.22±0.02 91.99±0.24 Application Example 7 6.86±0.02 22.25±0.05 8.78±0.22 9.14±0.18 18.85±0.15 4.12±0.12 70.00±0.65 Application Example 8 7.56±0.05 25.68±0.06 10.82±0.24 11.02±0.24 21.24±0.02 4.56±0.22 80.88±0.34 Application Example 9 7.24±0.02 24.56±0.24 10.02±0.16 11.12±0.14 20.56±0.02 4.25±0.05 77.75±0.25 Comparative Application Example 1 4.40±0.89 19.00±0.71 4.80±0.45 8.20±0.00 12.00±0.71 4.00±0.71 52.40±0.55

[0094] The data in the table show that the quality of the untreated low-quality cigars in the control group is in the Grade II rating range; there is no significant difference in ash size among the groups, indicating that enzyme treatment does not have an adverse effect on the ash formation during cigar combustion.

[0095] Comparing the control group, Example 1, and Examples 4-7 reveals that amylase-treated cigars exhibit improved aroma, off-flavors, irritation, and aftertaste. Flavor enzyme-treated cigars perform well across all indicators, particularly in aroma and off-flavors. The combined use of amylase and flavor enzymes achieves a synergistic effect, bringing the cigars to or near Grade I standards in aroma, off-flavors, irritation, and aftertaste. The combined use of amylase, flavor enzyme, and saccharifying enzyme further enhances the synergistic effect, improving the overall quality of the cigars. When the flavor enzyme in Example 1 is replaced with acidic protease, the overall quality of the cigars declines, further demonstrating that the present invention achieves a synergistic effect in improving cigar quality through a specific combination of enzyme preparations.

[0096] Comparing the control group, Example 1, and Examples 8-9, it can be found that yeast and lactobacillus fermentation have a synergistic effect on improving cigar quality.

[0097] Comparing the control group and the comparative application example 1, it can be found that the cigars produced by omitting the enzyme treatment step and using tobacco flavor extract have lower quality in terms of color, aroma and off-flavors compared to low-quality cigars. The aftertaste and ash are slightly better, but the overall quality is not as good as that of low-quality cigars.

[0098] Test Example 2

[0099] Detection of flavor compounds in tobacco flavor extract:

[0100] Accurately weigh 8.0 g of the supernatant obtained in step (6) of Examples 1, 4-7, and Comparative Example 1 into headspace vials, add 1.0 g of NaCl and 1 μL of 118.2 mg / L 2-methyl-3-heptanone as internal standards, and seal quickly. Then expose the solid-phase microextraction fiber head above the headspace vial. Place the headspace vial in a heating mantle at 70°C and extract the odor components for 30 min using a simultaneous distillation extraction device. Subsequently, desorb at the gas chromatography (GC) inlet for 7 min. The GC column was a DB-WAX capillary column (30 mm × 0.25 mm × 0.25 μm), the carrier gas (He) flow rate was 0.8 mL / min, and the inlet (split) temperature was 250°C. The column temperature was 40°C for 3 min, then increased to 90°C at a rate of 5°C / min, and then increased to 250°C at a rate of 10°C / min. Mass spectrometry conditions: Electron impact (EI) mode, ion energy 70 eV, detector voltage 350 V, scan range 33–450 m / z, scan frequency 3.00 scans / second. Qualitative and quantitative analyses were performed using the NIST08 mass library and an Agilent ChemWorkstation, and the results are shown in Table 3.

[0101] Table 3

[0102]

[0103]

[0104]

[0105] The data in the table show that enzyme treatment can effectively promote the production of flavor compounds, resulting in a significant increase in the content of flavor compounds such as esters, alcohols, aldehydes, ketones, and organic acids, while the concentration of nicotine decreases significantly.

[0106] Comparing Examples 1, 4-5, and Comparative Example 1, it was found that the total content of aldehydes and ketones in the fermentation broth treated with a mixture of amylase and flavor enzymes was lower than that in the untreated broth or in the broth treated with flavor enzymes or amylase alone. This may help reduce unpleasant odors and increase the richness and balance of the flavor. Regarding organic acids, the fermentation broth treated with the mixture of amylase and flavor enzymes showed increased levels of acids such as glacial acetic acid, resulting in a more delicate, balanced, and rich aroma profile. These acids may also play a positive role in the overall flavor's body and richness.

[0107] Comparing Examples 1 and 6, it can be found that the total content of aldehydes and ketones in the fermentation broth treated with a mixture of amylase, flavor enzyme, and saccharifying enzyme is lower, while the content of organic acids is further increased.

[0108] Comparing Examples 1 and 7, it can be found that the combination of amylase and flavor enzyme is superior to the flavor effect of fermentation treated with a mixture of amylase and acid protease.

[0109] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0110] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0111] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for preparing a tobacco flavoring agent for cigars, characterized in that, The preparation method includes the following steps: (1) Mix tobacco, sorghum and water and sterilize to obtain tobacco-sorghum liquid culture medium; (2) The enzyme preparation and bacterial solution are then added to the tobacco-sorghum liquid culture medium for fermentation. The bacterial solution is yeast solution and lactobacillus solution to obtain the tobacco flavoring agent. The enzyme preparation in step (2) comprises a combination of amylase, flavor enzyme, and saccharifying enzyme; The enzyme activity in the enzyme preparation described in step (2) is 100-1000 U / mL; The enzyme activity of amylase in the enzyme preparation shall not be less than 40%; The yeast culture in step (2) is obtained by culturing yeast strains at 25-37℃ and 150-250 r / min for 24-60 h with constant temperature shaking. The Lactobacillus bacterial solution was obtained by culturing Lactobacillus strains at 30-40℃ and 150-250 r / min for 12-30 h with constant temperature shaking. The inoculation amount of yeast culture in step (2) is 0.5%-1.5%; The inoculation amount of the Lactobacillus bacterial solution is 0.25%-0.75%; The fermentation conditions described in step (2) are constant temperature shaking at 25-35℃ and 150-250 r / min for 1-10 days.

2. The preparation method according to claim 1, characterized in that, After the culture and fermentation in step (2), the product is freeze-dried to obtain a freeze-dried tobacco flavoring agent.

3. The preparation method according to claim 1, characterized in that, After the culture and fermentation described in step (2), the mixture is centrifuged and the supernatant is taken to obtain a liquid tobacco flavoring agent.

4. The preparation method according to claim 1, characterized in that, After the culture and fermentation described in step (2), the mixture is centrifuged and the supernatant is added to a thickener to obtain tobacco flavor extract.

5. The preparation method according to claim 3, characterized in that, The centrifugation speed is independently 3000-8000 rpm, and the centrifugation time is independently 5-10 min.

6. The preparation method according to claim 4, characterized in that, The thickener is low-ester pectin and / or gellan gum.

7. The preparation method according to claim 1, characterized in that, In step (1), the concentration of tobacco shreds in the tobacco-sorghum liquid culture medium is 0.01-0.2 g / mL, and the concentration of sorghum is 0.01-0.2 g / mL.

8. A tobacco flavoring agent prepared by the preparation method according to any one of claims 1-7.

9. An application of the tobacco flavoring agent according to claim 8 in tobacco production, characterized in that, The tobacco in question is a cigar.

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