A reconstituted tobacco leaf coating liquid and a method of preparing the same

Abstract

The application discloses a kind of reconstituted tobacco coating liquid and preparation method thereof, preparation method includes the following steps: S1 picks up the sub-pseudomonas ZY-03 single colony of preservation number CGMCC NO.24422 is inoculated to culture seed liquid, seed liquid removes supernatant, with sterile water redissolving, obtains sub-pseudomonas bacterial agent;S2 tobacco raw material is exploded;S3 after explosion tobacco raw material is mixed with water, and mechanically beaten into slurry;S4 sub-pseudomonas bacterial agent is added to slurry and fermented;S5 fermentation slurry is filtered ethanol extraction, and extract liquid is combined to obtain fermentation slurry residue extract liquid;S6 fermentation slurry residue extract liquid is mixed with fermentation filtrate, and reduced pressure concentration, it is reconstituted tobacco coating liquid.This application is prepared reconstituted tobacco coating liquid, and the amount of aroma and aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the amount of aroma quality increase, and the

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and in particular to a reconstituted tobacco coating solution and its preparation method. Background Technology

[0002] Reconstituted tobacco, also known as reconstituted tobacco leaf or tobacco sheet, is produced by recombining tobacco stems, dust, and some secondary tobacco leaves generated during tobacco or cigarette production using physical or chemical methods. It can be used as a substitute for cigarette raw materials. Reconstituted tobacco used in cigarettes can effectively reduce tar and CO content and improve the flavor quality. The leaf base of reconstituted tobacco is mostly composed of supporting substances such as cellulose and lignin, providing the framework for the reconstituted tobacco. The reconstituted tobacco coating liquid is a liquid substance obtained after a series of treatments of the water-soluble components extracted from the tobacco raw materials; it is an important factor in imparting aroma and improving the sensory quality of reconstituted tobacco.

[0003] The coating solution for reconstituted tobacco has the following defects: First, the concentrate contains many macromolecular substances, which not only affect the coating effect but also have a negative effect on the smoking quality of reconstituted tobacco. Second, the aroma substances are severely lost during the concentration process of the extract, resulting in poor aroma and insufficient aroma concentration in the reconstituted tobacco.

[0004] Currently, the main methods to improve the above-mentioned defects of reconstituted tobacco coating liquid include: 1. Low-temperature concentration, which concentrates tobacco extract under low-temperature conditions to reduce aroma loss, but this method is less effective and has a long processing time; 2. High-temperature Maillard reaction, which promotes the formation of Maillard products in the concentrate system under high-temperature conditions, producing new aroma substances, but the process conditions are harsh and the results are unstable; 3. Membrane retention, which can non-selectively retain all macromolecules, but at the same time, many macromolecules that are beneficial to aroma are also removed; 4. Biological methods, mainly including microbial fermentation and enzymatic methods, which can utilize macromolecules to convert into new aroma substances, but due to the high nicotine and high osmotic pressure of the concentrate, many microorganisms and enzymes are difficult to function. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a reconstituted tobacco coating liquid and its preparation method, which can efficiently degrade lignin and has better sensory evaluation effect.

[0006] The technical problem to be solved by the present invention is achieved through the following technical solution:

[0007] A method for preparing a reconstituted tobacco coating liquid includes the following steps:

[0008] S1: Select a single colony of Pseudomonas paraxanthii ZY-03 with preservation number CGMCC NO.24422 and inoculate it into liquid culture medium to culture into seed culture. Remove the supernatant from the seed culture, add sterile water to reconstitute it, and make the OD value 1.8-2.1 to obtain Pseudomonas paraxanthii inoculum.

[0009] S2: After wetting the tobacco raw material, use saturated steam as the medium to detonate the tobacco raw material. The working pressure is 1-2.0 MPa, the pressure retention time is 30-80 s, the steam detonation time is less than 0.01 seconds, and the cycle is 5-10 times.

[0010] S3: Mix the blasted tobacco raw material with water at a mass-to-volume ratio of 1:12-1:20, mechanically beat it into a slurry, and sterilize the slurry at 121℃ for 15 minutes.

[0011] S4: Add 2-4% of Pseudomonas paraxanthii inoculum to the slurry, ferment in a fermenter at 150-200 r / min and 30-37℃ for 48-96 h with stirring, and introduce sterile air at 0.2-0.8 V / V·min.

[0012] S5: Filter the fermented slurry. The slurry residue is loaded into the filter paper tube of the Soxhlet extractor. Ethanol is added to the flask according to the volume-mass ratio of ethanol to slurry residue of 5:1-10:1. Soxhlet extraction is carried out at 90℃ for 2-4 hours. The operation is repeated 2-4 times. The extracts are combined to obtain the fermented slurry residue extract.

[0013] S6: Mix the fermented pulp residue extract with the fermented filtrate, and concentrate under reduced pressure at 50-80℃ and 70-90mbar until the density is 1.2-1.25g / mL, which is the reconstituted tobacco coating solution.

[0014] Preferably, in the above technical solution, S1 specifically involves: inoculating a single colony of *Pseudomonas paraxanthizobium* ZY-03 with preservation number CGMCC NO.24422 into YPD liquid medium, and culturing it for 12-72 hours at 22-37℃ and a shaking speed of 100-250 r / min to obtain a seed culture; centrifuging the seed culture at 8000-12000 r / min and 4℃ for 10-20 minutes, removing the supernatant, and reconstituteing it with sterile water to make the OD value 1.8-2.1, thereby obtaining *Pseudomonas paraxanthizobium* inoculum.

[0015] Preferably, in the above technical solution, the YPD liquid culture medium is composed of 10 g / L yeast powder, 20 g / L peptone and 20 g / L glucose, and is prepared by autoclaving at 121°C for 15 min.

[0016] Preferably, in the above technical solution, in S2, the tobacco stem is detonated at a working pressure of 2 MPa, a pressure retention time of 80 s, a steam detonation time of less than 0.01 seconds, and the process is repeated 8 times.

[0017] Preferably, in the above technical solution, in S3, the tobacco raw material is one or more of tobacco stems, low-grade tobacco leaves, tobacco leaf fragments, tobacco shreds, and tobacco dust, and the tobacco raw material is mixed with water at a mass-volume ratio of 1:15.

[0018] Preferably, in the above technical solution, in S4, 3% of Pseudomonas paraxanthii inoculum is added, and fermentation is carried out in a fermenter at 200 r / min and 30°C for 48-96 hours with stirring, while 0.3 V / V·min of sterile air is introduced.

[0019] Preferably, in the above technical solution, in S5, the volume-to-mass ratio of ethanol to slurry residue is 8:1, and Soxhlet extraction is performed at 90℃ for 3 hours, repeated 4 times; in S6, the vacuum concentration conditions are: 70℃, 80mbar, and vacuum concentration is performed until the density is 1.24g / mL.

[0020] A method for preparing a reconstituted tobacco coating liquid.

[0021] Application of a reconstituted tobacco coating liquid in improving tobacco quality.

[0022] Preferably, in the above technical solution, the total alkali of the reconstituted tobacco coating liquid remains unchanged compared with the control, the total nitrogen decreases, and the total sugar and reducing sugar increase. The aroma quality and aroma quantity of the reconstituted tobacco with added reconstituted tobacco coating liquid are increased, the off-flavors and irritation are reduced, and the aftertaste in the mouth is improved.

[0023] The above-described technical solution of the present invention has the following beneficial effects:

[0024] This application utilizes steam flash explosion to disrupt the lignin, cellulose, and hemicellulose entanglement structure of tobacco raw materials. It then employs a Pseudomonas paraxanthizobium ZY-03 preparation for solid-liquid mixed fermentation of the tobacco raw material pulp to amplify the fermentation effect. After fermentation, the pulp residue and fermentation liquid are treated separately to maximize the extraction of tobacco aroma components. The resulting coating liquid exhibits reduced impurities, increased aroma quality and quantity, and reduced irritation. Detailed Implementation

[0025] Various exemplary embodiments of the present invention will now be described in detail. It should be noted that, unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps set forth in these embodiments do not limit the scope of the invention.

[0026] The tobacco materials used in this application are obtained as follows: tobacco leaves and other tobacco raw materials are obtained from a cigarette factory under Henan China Tobacco Industry Co., Ltd., and are provided by Henan China Tobacco Co., Ltd.

[0027] The *Pseudomonas parafulva* ZY-03 of this application is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCCNo.24422 on February 23, 2022.

[0028] This strain was isolated from the surface of tobacco leaf fragments. It can tolerate the complex environment of tobacco itself, efficiently degrade lignin macromolecules, and has a certain aroma-enhancing effect.

[0029] Example 1

[0030] A method for preparing a reconstituted tobacco coating liquid includes the following steps:

[0031] (1) Select a single colony of Pseudomonas paraxanthizobium ZY-03 with preservation number CGMCC NO.24422 and inoculate it into YPD liquid medium. Incubate at 30℃ and shaker speed of 120r / min for 24h.

[0032] YPD medium: 10 g / L yeast extract, 20 g / L peptone, 20 g / L glucose, autoclaved at 121°C for 15 min.

[0033] The seed culture was centrifuged at 10,000 r / min and 4℃ for 15 min, the supernatant was removed, and sterile water was added to reconstitute it to make the OD value 1.8, thus obtaining the Pseudomonas paraxanthii inoculum.

[0034] (2) After wetting the tobacco leaf fragments, use saturated steam as the medium to detonate them. The working pressure is 1 MPa, the pressure retention time is 60 s, the steam detonation time is less than 0.01 seconds, and the cycle is repeated 6 times.

[0035] (3) Mix the blasted tobacco raw materials with water at a mass-to-volume ratio of 1:12, mechanically beat them into a slurry, and sterilize the slurry at 121℃ for 15 minutes.

[0036] (4) Add 2% Pseudomonas paraxanthizobium to the slurry, stir and ferment in the fermenter at 180 r / min and 30 ℃, and introduce sterile air at 0.2 V / V·min.

[0037] (5) Filter the fermented slurry. The slurry residue is placed in the filter paper tube of a Soxhlet extractor. Ethanol is added to the flask according to the volume-to-mass ratio of ethanol to slurry residue of 5:1. Soxhlet extraction is carried out at 90°C for 4 hours, and the operation is repeated 4 times. The volume of organic solvent is in mL, and the mass of fermented slurry is in g. The extracts are combined to obtain the fermented slurry residue extract.

[0038] (6) The fermented pulp residue extract and the fermented filtrate are mixed and concentrated under reduced pressure at 60°C and 70mbar to a density of 1.23g / ml, which is the reconstituted tobacco coating liquid.

[0039] Example 2

[0040] A method for preparing a reconstituted tobacco coating liquid includes the following steps:

[0041] (1) Select a single colony of Pseudomonas paraxanthizobium ZY-03 with preservation number CGMCC NO.24422 and inoculate it into YPD liquid medium. Incubate at 30℃ and shaker speed of 120r / min for 24h.

[0042] YPD medium: 10 g / L yeast extract, 20 g / L peptone, 20 g / L glucose, autoclaved at 121°C for 15 min.

[0043] The seed culture was centrifuged at 10,000 r / min and 4℃ for 15 min, the supernatant was removed, and sterile water was added to reconstitute it to make the OD value 2.1, thus obtaining the Pseudomonas paraxanthii inoculum.

[0044] (2) After wetting the tobacco leaf fragments, use saturated steam as the medium to detonate them. The working pressure is 1.5 MPa, the pressure retention time is 30 s, the steam detonation time is less than 0.01 seconds, and the cycle is repeated 10 times.

[0045] (3) Mix the blasted tobacco raw materials with water at a mass-volume ratio of 1:20, mechanically beat them into a slurry, and sterilize the slurry at 121℃ for 15 minutes.

[0046] (4) Add 2% Pseudomonas paraxanthii to the slurry, stir and ferment in the fermenter at 180 r / min and 30 ℃, and introduce sterile air at 0.8 V / V·min.

[0047] (5) Filter the fermented slurry. The slurry residue is placed in the filter paper tube of a Soxhlet extractor. Ethanol is added to the flask according to a volume-to-mass ratio of ethanol to slurry residue of 10:1. Soxhlet extraction is performed at 90°C for 2 hours, and this process is repeated twice. The volume of the organic solvent is in mL, and the mass of the fermented slurry is in g. The extracts are combined to obtain the fermented slurry residue extract.

[0048] (6) The fermented pulp residue extract and the fermented filtrate are mixed and concentrated under reduced pressure at 50°C and 90mbar to a density of 1.2g / ml, which is the reconstituted tobacco coating solution.

[0049] Example 3

[0050] (1) Select a single colony of Pseudomonas paraxanthizobium ZY-03 with preservation number CGMCC NO.24422 and inoculate it into YPD liquid medium. Incubate at 30℃ and shaker speed of 120r / min for 24h.

[0051] Prepare YPD medium: 10 g / L yeast extract, 20 g / L peptone, 20 g / L glucose, autoclave at 121°C for 15 min.

[0052] The seed culture was centrifuged at 8000 r / min and 4℃ for 20 min, the supernatant was removed, and sterile water was added to reconstitute it to make the OD value 2.0, thus obtaining the Pseudomonas paraxanthii inoculum.

[0053] (2) After wetting the tobacco stems, use saturated steam as a medium to explode the tobacco stems. The working pressure is 2.0 MPa, the pressure retention time is 80 s, the steam explosion time is less than 0.01 seconds, and the cycle is repeated 8 times.

[0054] (3) Mix the blasted tobacco raw material with water at a ratio of 1:15, mechanically grind it into a slurry, and sterilize the slurry at 121℃ for 15 minutes.

[0055] (4) Add 3% Pseudomonas paraxanthizobium to the slurry, stir and ferment in the fermenter at 200 r / min and 30℃, and introduce sterile air at 0.3 V / V·min.

[0056] (5) Filter the fermented slurry. The slurry residue is placed in the filter paper tube of a Soxhlet extractor. Ethanol is added to the flask according to the volume-to-mass ratio of ethanol to slurry residue of 8:1. Soxhlet extraction is carried out at 90°C for 3 hours, and the operation is repeated 4 times. The volume of organic solvent is in mL, and the mass of fermented slurry is in g. The extracts are combined to obtain the fermented slurry residue extract.

[0057] (6) Mix the fermented pulp residue extract with the fermented filtrate at 70°C and 80mbar, and concentrate under reduced pressure to a density of 1.24g / mL, which is the reconstituted tobacco coating solution.

[0058] The reconstituted tobacco coating solutions prepared in the above three examples and the unfermented reconstituted tobacco coating solutions were analyzed by conventional chemical index analysis using flow analysis. It can be seen that the total alkali of the fermented reconstituted tobacco coating solution remains unchanged, the total nitrogen decreases, and the total sugar and reducing sugar increase (the results are shown in Table 1).

[0059] Table 1. Conventional Chemical Properties of Coating Solution

[0060]

[0061] The results showed that solid-liquid mixed fermentation of tobacco raw material pulp using the Pseudomonas paraxanthizobium ZY-03 preparation promoted the degradation of lignin in the tobacco raw material.

[0062] Smoking effect evaluation test: The reconstituted tobacco coating liquid prepared in the above three examples was coated on both sides of the thin sheet base at a coating rate of 30%. After drying, it was cut into shreds and rolled into cigarettes. The sheet base with added unfermented reconstituted tobacco coating liquid was used as a blank control for smoking evaluation. The specific smoking results are shown in Table 2.

[0063] Table 2 Sensory evaluation results

[0064]

[0065] The evaluation results showed that the aroma quality and aroma quantity of the reconstituted tobacco leaves with added fermentation coating liquid were increased, while the off-flavors and irritation were significantly reduced, and the aftertaste in the mouth was also improved, resulting in a good overall effect.

[0066] Although the present invention has been disclosed above with reference to embodiments, it is not intended to limit the present invention. Any person skilled in the art can make various different choices and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention is defined by the claims and their equivalents.

Claims

1. A method for preparing a reconstituted tobacco coating liquid, characterized in that, Includes the following steps: S1: Select a single colony of *Pseudomonas paraxanthii* ZY-03 with preservation number CGMCC NO. 24422 and inoculate it into YPD liquid medium. Incubate at 22-37℃ and 100-250 r / min for 12-72 h to obtain seed culture. Centrifuge the seed culture at 8000-12000 r / min and 4℃ for 10-20 min, remove the supernatant, and reconstitute with sterile water to make the OD value 1.8-2.1 to obtain *Pseudomonas paraxanthii* inoculum. The YPD liquid medium is composed of 10 g / L yeast extract, 20 g / L peptone, and 20 g / L glucose, and is prepared by autoclaving at 121℃ for 15 min. S2: After wetting the tobacco raw material, use saturated steam as the medium to detonate the tobacco raw material. The working pressure is 1-2.0 MPa, the pressure retention time is 30-80 s, the steam detonation time is less than 0.01 seconds, and the cycle is 5-10 times. S3: Mix the blasted tobacco raw material with water at a mass-to-volume ratio of 1:12-1:20, mechanically beat it into a slurry, and sterilize the slurry at 121℃ for 15 minutes. S4: Add 2-4% of Pseudomonas paraxanthii inoculum to the slurry, ferment in a fermenter at 150-200 r / min and 30-37℃ for 48-96 h with stirring, and introduce sterile air at 0.2-0.8 V / V·min. S5: Filter the fermented slurry. The slurry residue is loaded into the filter paper tube of the Soxhlet extractor. Ethanol is added to the flask according to the volume-mass ratio of ethanol to slurry residue of 5:1-10:

1. Soxhlet extraction is carried out at 90℃ for 2-4 hours. The operation is repeated 2-4 times. The extracts are combined to obtain the fermented slurry residue extract. S6: Mix the fermented pulp residue extract with the fermented filtrate, and concentrate under reduced pressure at 50-80℃ and 70-90mbar until the density is 1.2-1.25 g / mL, which is the reconstituted tobacco coating solution.

2. The method for preparing the reconstituted tobacco coating liquid according to claim 1, characterized in that, In S2, the tobacco stems are detonated at a working pressure of 2 MPa, a pressure retention time of 80 s, and a steam detonation time of less than 0.01 seconds, for a total of 8 cycles.

3. The method for preparing the reconstituted tobacco coating liquid according to claim 1, characterized in that, In S3, the tobacco raw material is one or more of tobacco stems, low-grade tobacco leaves, tobacco leaf fragments, tobacco shreds, and tobacco dust, and the tobacco raw material is mixed with water at a mass-volume ratio of 1:

15.

4. The method for preparing the reconstituted tobacco coating liquid according to claim 1, characterized in that, In S4, add 3% of Pseudomonas paraxanthii inoculum, ferment in a fermenter at 200 r / min and 30℃ for 48-96 h with stirring, and introduce sterile air at 0.3 V / V·min.

5. The method for preparing the reconstituted tobacco coating liquid according to claim 1, characterized in that, In S5, the volume-to-mass ratio of ethanol to slurry residue was 8:1, and Soxhlet extraction was performed at 90℃ for 3 hours, repeated 4 times. In S6, the vacuum concentration conditions were 70℃ and 80mbar, and the concentration was carried out under reduced pressure until the density was 1.24 g / mL.

6. A reconstituted tobacco coating solution prepared by the method for preparing the reconstituted tobacco coating solution according to any one of claims 1-5.

7. The application of the reconstituted tobacco coating solution prepared by the method according to any one of claims 1-5 or the reconstituted tobacco coating solution according to claim 6 in improving tobacco quality.

8. The application according to claim 7, characterized in that, Compared with the control, the total alkali of the reconstituted tobacco coating solution remained unchanged, the total nitrogen decreased, and the total sugar and reducing sugar increased. The aroma quality and aroma quantity of the reconstituted tobacco with added reconstituted tobacco coating solution were increased, the off-flavors and irritation were reduced, and the aftertaste in the mouth was improved.

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