CN109315829B - Fermented carbonized tobacco leaf particles and preparation method and application thereof - Google Patents
Fermented carbonized tobacco leaf particles and preparation method and application thereof Download PDFInfo
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- A24D—CIGARS; CIGARETTES; TOBACCO SMOKE FILTERS; MOUTHPIECES FOR CIGARS OR CIGARETTES; MANUFACTURE OF TOBACCO SMOKE FILTERS OR MOUTHPIECES
- A24D3/00—Tobacco smoke filters, e.g. filter-tips, filtering inserts; Filters specially adapted for simulated smoking devices; Mouthpieces for cigars or cigarettes
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Abstract
The invention provides fermented carbonized tobacco leaf particles and a preparation method and application thereof, and the fermented carbonized tobacco leaf particles are prepared by the method comprising the following steps: (1) preparing a tobacco soil bacterium agent L4-6; (2) fermenting tobacco leaves by using a tobacco soil bacterium L4-6 microbial inoculum to obtain fermented tobacco leaves; (3) carbonizing the fermented tobacco leaves to obtain fermented carbonized tobacco leaves; (4) drying the fermented carbonized tobacco powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain the fermented carbonized tobacco particles. The granule is fermented, so that irritation of smoke is reduced, richness of cigarette fragrance is increased, and sweet feeling of smoke is increased; through carbonization treatment, the specific surface area of the interior of the tobacco leaves can be obviously increased, the adsorption effect of the particles on harmful substances in cigarette smoke is enhanced, and meanwhile, the plant particles are endowed with special tar fragrance.
Description
Technical Field
The invention belongs to the technical field of cigarette materials, and particularly relates to fermented carbonized tobacco particles and a preparation method and application thereof.
Background
Cigarette aroma is the core content of cigarette quality, but with the development of low-coking cigarettes, the smoking taste of cigarettes is reduced, and insufficient aroma and the like become the prominent problems of tobacco development. How to overcome the key technology of influencing the aroma quality and the aroma quantity of the tobacco leaves becomes the core for improving the competitiveness of cigarette production. The addition of tobacco flavors and fragrances into tobacco shreds is a conventional and necessary technical means for supplementing fragrance, but the space for improving the quality of cigarettes is relatively limited, so that the development trend of finding related novel materials for improving and improving the fragrance of cigarettes and reducing the irritation of cigarette smoke is new from the non-combustion stage of cigarettes. Wherein, some particle materials with porosity and special aroma are added in the cigarette filter stick to improve or promote the quality of cigarette products, which is a novel and effective research direction. However, the advantages of the existing granular material additive cannot be fully exerted due to some problems in the preparation method and the selection and combination of raw materials. For example, although particles made of adsorptive materials such as activated carbon and silica gel micropowder have a remarkable advantage in adsorbing harmful substances in smoke, a large amount of fragrance components can be adsorbed, so that the smoking quality of cigarettes is remarkably reduced. The plant granular material obtained by the granulation method can introduce fragrant substances which are coordinated with the fragrance of the cigarettes into the smoke of the cigarettes, but the holes of the plant material are embedded due to the addition of the adhesive, so that the adsorption sites of harmful substances are reduced, and the tar and harm reducing effects are reduced. In addition, for the selection and treatment of raw materials, the coordination between various plant materials disclosed in the prior art and the fragrance of cigarettes needs to be improved, and the fragrance of tobacco is not rich enough, so that the continuous search for plant raw materials which are coordinated with the original fragrance of tobacco is also a direction which needs to be continuously researched.
The microbial fermentation and enzyme preparation treatment can increase the content of various aroma substances in the traditional extracted spice, improve the aroma richness, further produce cigarettes with rich aroma and high quality, and meet the requirements of consumers on the cigarette quality. The method utilizes microorganisms to ferment the plant raw materials with intrinsic odor and can also endow the plants with unique fragrance different from the original odor.
The plants are carbonized, the plants can be dehydrated in a short time through high temperature, the internal gaps of the plants are reserved, the adsorption of harmful substances of the plants to smoke is increased, and meanwhile, the partial fragrance characteristics of the plant raw materials are reserved, and the special burnt fragrance is given to the plant raw materials. At present, no research related to the preparation of the cigarette filter tip particles by combining the microbial fermentation technology with the carbonization technology is available.
Aiming at the defects, how to select proper plant raw materials and modify the plant raw materials to make up the defects caused by the prior art, and the technical problem to be further solved is to improve the effects of the granular materials on increasing the harmony of cigarette smoke, promoting the secretion of saliva or body fluid and the like while reducing the irritation of the cigarette smoke.
Disclosure of Invention
Aiming at the defects that the adsorption effect of the existing plant particles on harmful substances in smoke is not obvious, the cigarette cannot be directly endowed with fragrance, or the coordination with the fragrance of the cigarette is not good and the like, the invention provides the fermented carbonized tobacco particles and the preparation method and the application thereof, and the plant raw materials are modified by utilizing the fermentation and carbonization process of the tobacco soil bacterium L4-6, so that the harm and tar reducing effect of the cigarette can be enhanced, the irritation of the smoke of the cigarette can be reduced, the cigarette perfuming effect can be enhanced, and the smoking quality of the cigarette can be improved.
Unless otherwise stated, all percentages used in the present invention are mass percentages.
The invention relates to a tobacco soil bacteria L4-6, which is separated from tobacco soil samples in Yunnan Kunming tobacco planting base, the morphological, physiological and biochemical properties and 16S rDNA analysis are carried out on the tobacco soil bacteria L4-6, the identification result shows that the tobacco soil bacteria L4-6 belongs to the tobacco soil bacteria, the tobacco soil bacteria Dyellatabacilia L4-6 is classified and named, the tobacco soil bacteria L4-6 is preserved in China general microbiological culture Collection center (CGMCC for short) in 6 and 19 days in 2018, the preservation number is CGMCC 1.16273, the address: the institute of microbiology, national academy of sciences, west road No. 1, north Chen, Chaozhou, Chaoyang.
The main morphological characteristics and physiological and biochemical characteristics of the tobacco soil bacterium L4-6 strain obtained by separation are as follows: the growth is good on most culture media, and on the R2A agar culture medium, the colony has neat edges and a convex middle part and is yellow. The gelatin is liquefied, the oxidase, the catalase, the Tween 20, the Tween 40, the Tween 60 and the Tween 80 are hydrolyzed positively, the starch is hydrolyzed, the cellulose is hydrolyzed, the casein is hydrolyzed negatively, and hydrogen sulfide and melanin are not generated. Maltose, trehalose, cellobiose, lactose, mannose, glycerol, aspartic acid, glutamic acid, and histidine can be used. The polar lipid component of cell membrane mainly comprises phosphatidyl glycerol, diphosphatidyl glycerol and phosphatidyl ethanolamine, and the respiratory quinone of cell is ubiquinone-8 (Q-8).
The nucleotide sequence of the 16S rDNA gene of the tobacco soil bacterium L4-6 obtained by separation is shown in a sequence table, the sequence is submitted to an international nucleotide sequence database (GenBank), and the sequence retrieval number is as follows: MF 370623.
A second aspect of the invention relates to a fermented carbonized tobacco leaf pellet prepared by a process comprising the steps of:
(1) preparing a tobacco soil bacterium agent L4-6;
(2) fermenting tobacco leaves by using a tobacco soil bacterium L4-6 microbial inoculum to obtain fermented tobacco leaves;
(3) carbonizing the fermented tobacco leaves to obtain fermented carbonized tobacco leaves;
(4) drying the fermented carbonized tobacco powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain the fermented carbonized tobacco particles.
The step (1) of the above-mentioned fermented carbonized tobacco leaf particles specifically includes: inoculating the tobacco soil bacterium L4-6 liquid strain into a fermentation culture medium according to the inoculation amount of 10%, and performing shake-flask culture at 28 ℃ for 3-7 days to obtain a culture solution; and (3) performing centrifugal separation on every 1000mL of culture solution, washing the precipitate with sterile water, shaking the precipitate uniformly with 20mL of sterile water, and diluting by 10 times to obtain the microbial inoculum.
The step (2) of the above-mentioned fermented carbonized tobacco leaf particles specifically includes: weighing tobacco leaves with the water content balanced to 11% -14% in advance, spraying 20mL of tobacco soil bacterium agent L4-6 per 100g of tobacco leaves, and fermenting the treated tobacco leaves in a constant temperature and humidity box with the temperature of 22 ℃ and the concentration of 60% for 24-72 h to obtain the fermented tobacco leaves.
The step (3) of the above-mentioned fermented carbonized tobacco leaf particles specifically includes: putting the fermented tobacco leaves into a carbonization furnace, and carrying out closed heating carbonization treatment on the tobacco leaves, wherein the specific operation steps comprise gradually raising the temperature of the furnace to 80 ℃, quickly raising the temperature to 120-300 ℃ after the fermented tobacco leaves are dried, and keeping the temperature for 1-6 hours to obtain the fermented carbonized tobacco leaves.
The step (4) of the above-mentioned fermented carbonized tobacco leaf particles specifically comprises the following steps:
1) detecting the water content of the fermented carbonized tobacco leaves, controlling the water content to be 8-12%, and crushing the tobacco leaves into powder with the particle size of 100-160 meshes;
2) weighing 90-99 parts of tobacco powder and 1-10 parts of cane sugar or mannitol powder according to parts by weight, and uniformly mixing for later use;
3) weighing 5-10 parts of gelatin, 0-5 parts of polyvinylpyrrolidone and 0-2 parts of sodium carboxymethyl starch according to parts by weight, and preparing a 5-12% adhesive solution by using distilled water;
4) and weighing 300-1000 g of the mixed powder in the step 2), placing the mixed powder in a fluidized bed for granulation, and spraying an adhesive solution in the granulation process to obtain the fermented carbonized tobacco particles.
The step 4) of the above-mentioned fermented carbonized tobacco leaf particles specifically comprises the following steps:
(a) drying the mixed powder for 5-15 min under the conditions that the fluidizing air pressure is 0.12-0.2 Bar and the air flow temperature is 50-60 ℃;
(b) applying the adhesive solution prepared in the step 3) by adopting a top spraying mode under the conditions that the fluidizing pressure is 0.12-0.25 Bar, the airflow temperature is 60-80 ℃, the spraying pressure is 0.08-0.15 Bar and the spraying speed is 8-12 g/min, wherein the using amount of the adhesive solution is 20-50% of the weight of the fermented carbonized tobacco powder dry material;
(c) and (c) repeating the step (a) after the adhesive is applied, so as to obtain the fermented carbonized tobacco particles.
The fermented carbonized tobacco leaf particles obtained in the step (c) have a moisture content of 8-10%, a particle size of 20-60 meshes, and are round-like particles with rough surfaces.
In a third aspect, the present invention relates to a cigarette filter rod comprising particles of fermented carbonized tobacco leaf prepared according to the second aspect of the present invention.
According to the cigarette filter rod, 1-2 mg of fermented carbonized tobacco particles are added into each millimeter of cigarette filter rod.
The invention has the following beneficial effects:
(1) according to the invention, tobacco leaves are fermented by utilizing the tobacco soil bacterium L4-6, and after the tobacco leaves are applied to cigarette filter rods, the irritation of smoke can be reduced, the richness of cigarette fragrance is increased, and the effect of increasing the sweet feeling of smoke is achieved;
(2) according to the invention, the fermented tobacco leaves are carbonized, and the tobacco leaves cells are dehydrated and shaped at high temperature in a short time, so that the specific adsorption surface area in the tobacco leaves is increased, the filtering efficiency of the fermented carbonized tobacco leaves particles on cigarette smoke is obviously improved, harmful substances in the cigarette smoke can be selectively adsorbed, and the fermented carbonized tobacco leaves particles are endowed with special burnt flavor;
(3) the invention does not adjust the production process of plant particles, realizes the dual effects of improving aroma enhancement and reducing coke only by modifying the raw materials, has easily obtained raw materials, low cost, convenient realization of industrial production and good commercial application value.
Drawings
FIG. 1 is an electron micrograph of the tobacco soil bacterium L4-6 in R2A medium;
FIG. 2 shows a phylogenetic tree constructed by the tobacco soil bacterium L4-6 and some related strains according to the 16S rDNA gene sequence.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
1. Separation, culture and identification of tobacco soil bacterium L4-6
1.1 isolation of tobacco soil-borne bacterium L4-6
Taking a tobacco soil sample from a Yunnan Kunming tobacco planting base, sealing the tobacco soil sample with a plastic bag, and storing the tobacco soil sample at 4 ℃ for later use. Accurately weighing 10g of soil sample, adding 90mL of sterile water, shaking the mixture for 30min at 30 ℃ and 180rpm by a shaking table; diluting the concentrate to 10 with sterile water-1、10-2、10-3、10-4、10-5And (2) respectively taking 0.2mL of bacterial liquid with 5 concentrations, coating the bacterial liquid with each concentration on an LB plate culture medium, performing 3 parallel tests on the bacterial liquid with each concentration, culturing for 48h at 30 ℃, selecting different single bacterial colonies from the plate of each soil sample, purifying on the LB plate culture medium, adding glycerol according to 20% of the volume of the bacterial liquid, and storing in a refrigerator at-80 ℃ for later use.
The separated microorganisms are respectively inoculated in LB liquid culture medium, shaking culture is carried out on a shaking table at the temperature of 30 ℃ and the rpm of 160, and the microorganisms are cultured until the concentration of the bacteria liquid is OD which is 2.0 and are used as seed liquid for standby. Inoculating 1% of seed liquid of each strain to be tested in a screening culture medium, shaking and culturing for 3-5 days at 30 ℃ and 160rpm in a shaking table, observing the appearance of fermentation liquor and the change condition of aroma every day, and screening to obtain a strain of microorganism capable of producing aroma, wherein the strain is numbered L4-6.
1.2 identification of tobacco soil bacterium L4-6
Morphological, physiological and biochemical properties and 16S rDNA analysis are carried out on the separated and purified strain L4-6, and the identification result shows that the strain belongs to the tobacco soil worn bacteria, and the microbiological classification is named as the tobacco soil worn bacteria Dyellatabacilia L4-6.
An electron micrograph of the tobacco soil bacterium L4-6 on the R2A medium is shown in figure 1.
The tobacco soil bacterium L4-6 grows well on most culture media, and on the R2A agar culture medium, the colony is neat in edge, convex in the middle and yellow. The gelatin is liquefied, the oxidase, the catalase, the Tween 20, the Tween 40, the Tween 60 and the Tween 80 are hydrolyzed positively, the starch is hydrolyzed, the cellulose is hydrolyzed, the casein is hydrolyzed negatively, and hydrogen sulfide and melanin are not generated. Maltose, trehalose, cellobiose, lactose, mannose, glycerol, aspartic acid, glutamic acid, and histidine can be used. The polar lipid component of cell membrane mainly comprises phosphatidyl glycerol, diphosphatidyl glycerol and phosphatidyl ethanolamine, and the respiratory quinone of cell is ubiquinone-8 (Q-8).
The physiological and biochemical characteristics of the tobacco soil bacterium L4-6 are shown in Table 1:
TABLE 1 physiological and biochemical characteristics of T.nicotianae L4-6
| Experiment of | Growth reaction | Experiment of | Growth reaction |
| Liquefaction of gelatin | +++++ | Tween 20,40,60, and 80 | +++++ |
| Starch hydrolysis | -- | Growth on cellulose | -- |
| Hydrolysis of cellulose | -- | Alkaline phosphatase | ++++ |
| Casein hydrolysis | -- | Valine arylamine enzyme | ++++ |
Note: "+" indicates a positive result, and "-" indicates a negative result
The carbon and nitrogen utilization of the tobacco soil bacterium L4-6 is shown in Table 2:
TABLE 2 utilization of carbon and nitrogen sources by tobacco soil bacterium L4-6
| Carbon source utilization | Results | Carbon or nitrogen source utilization | Results |
| Dextrin | + | N-acetyl-D-glucosamine | + |
| Maltose | + | Mannitol | + |
| Trehalose | + | Fructose | + |
| Cellobiose | + | Galactose | + |
| Sucrose | - | Rhamnose | - |
| Cotton seed candy | - | Alanine | - |
| Melibiose | + | Histidine | + |
Note: "+" indicates a positive result, and "-" indicates a negative result
The partial sequence of 16S rDNA of the tobacco soil bacterium L4-6 is shown in the description of the attached figure, the sequence is compared and analyzed with the known sequence in the GenBank database by BLAST, and the 16S rDNA gene sequence of the related species is obtained from the database, and a phylogenetic tree is constructed, which is shown in figure 2. Through comparative analysis, the tobacco soil bacterium L4-6 and the strain (Dyella soli JS 12-10) are foundT) The genetic relationship is recent, independent branches are formed on the phylogenetic tree, and the characteristics of comprehensive morphology, physiology and biochemistry, cytochemistry, phylogenetic analysis and the like are obvious in difference, so that the tobacco soil bacterium L4-6 is a new species and is named as Dyellatabacili.
The nucleotide sequence accession number of the 16S rDNA gene of the tobacco soil bacterium L4-6 in a GenBank database is MF370623, and the preservation number of the China general microbiological culture Collection center is CGMCC 1.16273. The phylogenetic tree constructed by the 16S rDNA gene sequence of the tobacco soil bacterium L4-6 and the related species is shown in figure 2.
Example 2
1. Culture of tobacco soil bacterium L4-6
(1) Slant culture in test tubes
The culture medium is a slant preservation culture medium which is an R2A agar culture medium, and the formula of the culture medium is as follows: 0.5g of glucose, 0.5g of yeast extract, 0.5g of peptone, 0.5g of acid hydrolyzed casein, 0.5g of soluble starch, 0.3g of sodium pyruvate, 0.3g of dipotassium phosphate, 0.05g of magnesium sulfate, 15g of agar, and distilled water with the volume of 1000mL and the pH value of 7.2. Sterilizing the culture medium at 121 ℃ for 25 minutes, placing the culture medium into a slope, inoculating a tobacco soil-borne bacterium L4-6 strain, and culturing at 28 ℃ for 1 week to obtain a test tube strain;
(2) seed culture
The seed culture medium is adopted, and the formula of the seed culture medium is as follows: 120g of dextrin, 40g of soybean meal, 2g of yeast extract, 0.5g of tryptophan, 5g of beta-alanine, 0.5g of magnesium sulfate, 0.2g of ammonium phosphate and distilled water to reach the volume of 1000mL and the pH value of 7.2. Sterilizing the culture medium at 121 ℃ for 25 minutes, picking part of mycelia from the test tube inclined plane in the step (1) to be inoculated into seed liquid, and performing shake-flask culture at 28 ℃ for 36 hours to obtain liquid strains;
(3) preparation of tobacco soil bacterium L4-6
Transferring the liquid strain prepared in the step (2) into a fermentation medium according to the inoculation amount of 10%, shaking the flask at 28 ℃ for 6 days, centrifuging the obtained culture solution at 3500r/min for 10min, washing the precipitate with sterile water, uniformly shaking the precipitate with 20mL of sterile water, and diluting by 10 times to obtain the tobacco soil bacterium L4-6.
The formula of the fermentation medium is as follows: 10g of soybean meal, 10g of glucose, 3g of peptone, 2.5g of sodium chloride, 2g of calcium carbonate and distilled water with constant volume of 1000mL, pH7.2, and sterilizing at 121 ℃ for 25 minutes to obtain the culture medium.
2. Fermentation of tobacco leaves
Weighing 1200g of tobacco leaves with the water content balanced to 13% in advance, spraying 240mL of tobacco soil bacterium L4-6, putting the treated tobacco leaves into a constant temperature and humidity box with the temperature of 22 ℃ and the concentration of 60%, and fermenting for 48 hours to obtain the fermented tobacco leaves.
3. Carbonization of fermented tobacco leaves
Putting the fermented tobacco leaves into a carbonization furnace, gradually heating the furnace to about 80 ℃, keeping the temperature for 30min, quickly heating the fermented tobacco leaves to 160 ℃ after the fermented tobacco leaves are dried, keeping the temperature for 2h, turning off a heat source after the carbonization degree is inspected to be qualified, and taking out the fermented carbonized tobacco leaves for later use.
4. Preparation of fermented carbonized tobacco particles
Taking fermented carbonized tobacco leaves, detecting the moisture content of the fermented carbonized tobacco leaves, complementing the moisture content to 10 percent, and crushing the fermented carbonized tobacco leaves into powder with the particle size of 120 meshes; mixing 950g of prepared tobacco leaf fermentation carbonization powder and 50g of sucrose powder uniformly, placing in a Midi Glatt fluidized bed for granulation, and drying for 10min under the conditions that the fluidized air pressure of the fluidized bed is 0.15Bar and the air flow temperature is 60 ℃; then 5 parts of gelatin, 2 parts of polyvinylpyrrolidone and 1 part of sodium carboxymethyl starch are weighed according to the parts by weight, distilled water is used for preparing an adhesive solution with the concentration of 8 percent, and 300g of the adhesive solution is applied by adopting a top spraying mode under the conditions that the fluidization air pressure is 0.2Bar, the air flow temperature is 80 ℃, the spraying pressure is 0.15Bar and the spraying speed is 10 g/min; after the adhesive is applied, drying for 5 minutes under the conditions that the fluidizing air pressure is 0.15Bar and the air flow temperature is 60 ℃ to obtain the fermented carbonized tobacco leaves with 8 percent of moisture content, rough surface and 60-mesh particle size.
Example 3
1. The culture method of the tobacco soil bacterium L4-6 is the same as that of example 2.
2. Fermentation of tobacco leaves
Weighing 600g of tobacco leaves with the water content balanced to 12% in advance, spraying 120mL of tobacco soil bacterium L4-6, and fermenting the treated tobacco leaves in a constant temperature and humidity box with the temperature of 22 ℃ and the concentration of 60% for 72 hours.
3. Carbonization of fermented tobacco leaves
Putting the fermented tobacco leaves into a carbonization furnace, gradually heating the furnace to 80 ℃, keeping the temperature for 30min, quickly heating to 130 ℃ after the tobacco leaf fermentation sample is dried, keeping the temperature for 3h, turning off a heat source after the carbonization degree is inspected to be qualified, and taking out the fermented carbonized tobacco leaf sample for later use.
4. Preparation of fermented carbonized tobacco particles
Taking fermented carbonized tobacco leaves, detecting the moisture content of the fermented carbonized tobacco leaves, complementing the moisture content to 8 percent, and crushing the fermented carbonized tobacco leaves into powder with the particle size of 140 meshes; uniformly mixing 470g of prepared tobacco leaf fermentation carbonized powder and 30g of mannitol powder, placing in a Midi Glatt fluidized bed for granulation, and drying for 5min under the conditions that the fluidized air pressure of the fluidized bed is 0.18Bar and the air flow temperature is 60 ℃; then 6 parts of gelatin, 2 parts of polyvinylpyrrolidone and 2 parts of sodium carboxymethyl starch are weighed according to the parts by weight, and distilled water is used for preparing a 10% adhesive solution; applying 200g of adhesive solution by adopting a top spraying mode under the conditions that the fluidizing air pressure is 0.2Bar, the air flow temperature is 70 ℃, the spraying pressure is 0.15Bar and the spraying speed is 12 g/min; after the adhesive is applied, drying for 10 minutes under the conditions that the fluidizing air pressure is 0.18Bar and the air flow temperature is 60 ℃, and preparing the fermented carbonized tobacco leaf particles with the moisture content of 10 percent, rough surface and 40 meshes of particle size.
Example 4
In order to examine the uniqueness of the T.nicotianae L4-6 for fermenting tobacco leaves, example 4 was different from example 3 in that the tobacco leaves were fermented using the closely related strain Dyella soli JS12-10 (purchased from the Korean agricultural microbial cultures Collection (KACC)) of the T.nicotianae L4-6 instead of the T.nicotianae L4-6, and the remaining operation steps were the same as example 3, to obtain fermented carbonized tobacco leaves having a moisture content of 10%, a rough surface, and a particle size of 40 meshes, which were fermented using the Dyella soli JS 12-10.
Example 5
The fermented carbonized tobacco particles prepared in the example 2 are applied to a cigarette composite filter stick, 1.5mg of the fermented carbonized tobacco particles are added to each millimeter of the cigarette filter stick to prepare an experimental cigarette, and the cigarette without the particles is used as a control group for sensory evaluation and detection of harmful components in smoke. Sensory evaluation results show that: compared with a control group, the cigarette smoke throat pungent irritation feeling of the cigarette added with the fermented carbonized tobacco particles is obviously reduced, the richness of the cigarette fragrance is increased, and the sweet feeling of the smoke is obvious; the detection of harmful components in the smoke shows that: the reduction rate of phenol in the smoke of the experimental cigarettes is 24.3%, which shows that the fermented carbonized tobacco particles can obviously reduce the content of phenol in the smoke of the cigarettes.
Example 6
The fermented carbonized tobacco particles prepared in the example 3 are applied to a cigarette composite filter stick, 1mg of the fermented carbonized tobacco particles are added to each millimeter of the cigarette filter stick to prepare an experimental cigarette, and the cigarette without the particles is used as a control group for sensory evaluation and detection of harmful components in smoke. Sensory evaluation results show that: compared with a control group, the cigarette smoke irritation and the bitter and spicy taste of the added fermented carbonized tobacco particles are obviously reduced, the natural tobacco fragrance is richer, and the smoke sweet feeling is obvious; the detection of harmful components in the smoke shows that: the reduction rate of phenol in the smoke of the experimental cigarettes is 22.5%, which shows that the fermented carbonized tobacco particles can obviously reduce the content of phenol in the smoke of the cigarettes.
Example 7
Applying the fermented carbonized tobacco particles prepared by further processing the tobacco leaves fermented by Dyella soli JS12-10 in the example 4 into a cigarette composite filter stick, and adding 1mg of the fermented carbonized tobacco particles into each millimeter of the cigarette composite filter stick to prepare an experimental cigarette; the experimental cigarettes in example 6 were used as a control group for sensory evaluation and detection of harmful components in smoke. Sensory evaluation results show that: compared with a control group, the experimental cigarette prepared by fermenting the tobacco leaves with Dyella soli JS12-10 has no obvious reduction in smoke irritation and bitter and hot taste, and insufficient sweet and moist feeling of the smoke. The detection of harmful components in the smoke shows that: the reduction rate of phenol in the smoke of the experimental cigarette is 16.9 percent, and the comparison with the data in the example 6 shows that the reduction rate of harmful components in the experimental cigarette is inferior to that of the control group of cigarettes. Thus, the strain Dyella soli JS12-10, although it is a related strain, could not achieve the effect similar to the tobacco soil bacterium L4-6 when it is used for tobacco leaf fermentation.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
A sequence table is attached:
the 16S rDNA partial sequence of the tobacco soil bacterium L4-6 is as follows:
sequence listing
<110> tobacco industry Limited liability company in Yunnan
Li Yuan dong Gao Li Yang Hongming He Liang jiao Jun Su Yong Xia Jian Jun
<120> fermented carbonized tobacco leaf particles and preparation method and application thereof
<130> RIB180325
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1508
<212> DNA
<213> tobacco soil bacterium (Dyellatabacisis L4-6)
<400> 1
agagtttgat cctggctcag attgaacgct ggcggcatgc ctaacacatg caagtcgaac 60
ggcagcacag cagtagcaat actgtgggtg gcgagtggcg gacgggtgag taatgcatcg 120
ggacctacct agacgtgggg gataacgtag ggaaacttac gctaataccg catacgtcct 180
acgggagaaa gcgggggatc ttcggacctc gcgcggttag acggaccgat gttcgattag 240
ctagttggta gggtaatggc ctaccaaggc gacgatcgat agctggtctg agaggatgat 300
cagccacact ggaactgaga cacggtccag actcctacgg gaggcagcag tggggaatat 360
tggacaatgg gcgcaagcct gatccagcaa tgccgcgtgt gtgaagaagg ccttcgggtt 420
gtaaagcact tttatcagga gcgaaatgcc attggctaat acccggtgga gctgacggta 480
cctgaggaat aagcaccggc taacttcgtg ccagcagccg cggtaatacg aagggtgcaa 540
gcgttaatcg gaattactgg gcgtaaagcg tgcgtaggcg gttcgttagg tccgtcgtga 600
aatccccggg ctcaacctgg gaatggcgat ggatactggc gagctagagt gtgatagagg 660
atggtggaat tcccggtgta gcggtgaaat gcgtagagat cgggaggaac atcagtggcg 720
aaggcggcca tctggatcaa cactgacgct gaggcacgaa agcgtgggga gcaaacagga 780
ttagataccc tggtagtcca cgcccccaaa cgatgcgaac tggatgttgg tctcaactcg 840
gagatcagtg tcgaaagcta acgcgttaag ttcgccgcct ggggagtacg gtcgcaagac 900
tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagtatgtgg tttaattcga 960
tgcaacgcga agaaccttac ctggccttga catgtctgga atcctgcaga gatgcgggag 1020
tgccttcggg aatcagaaca caggtgctgc atggctgtcg tcagctcgtg tcgtgagatg 1080
ttgggttaag tcccgcaacg agcgcaaccc ttgtccttag ttgccagcac gtaatggtgg 1140
gaactctaag gagactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaagtcatc 1200
atggccctta cggccagggc tacacacgta ctacaatggt cggtacagag ggttgcaata 1260
ccgcgaggtg gagccaatcc cagaaagccg atcccagtcc ggatcgtagt ctgcaactcg 1320
actacgtgaa gtcggaatcg ctagtaatcg cggatcagct atgccgcggt gaatacgttc 1380
ccgggccttg tacacaccgc ccgtcacacc atgggagtga gttgctccag aagccgttag 1440
tctaaccgca agggggacga cgaccacgga gtggttcatg actggggtga agtcgtaaca 1500
aggtaacc 1508
Claims (7)
1. A fermented carbonized tobacco leaf pellet characterized in that it is prepared by a process comprising the steps of:
(1) preparing a tobacco soil bacterium agent L4-6;
(2) fermenting tobacco leaves by using a tobacco soil bacterium L4-6 microbial inoculum to obtain fermented tobacco leaves;
(3) carbonizing the fermented tobacco leaves to obtain fermented carbonized tobacco leaves;
(4) drying the fermented carbonized tobacco powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain fermented carbonized tobacco particles;
the tobacco soil bacterium L4-6 microbiological classification is named as tobacco soil bacteriumDyellatabacisoliL4-6, which has been cultivated in Chinese microbial strains in 6 and 19 months in 2018The preservation management committee common microorganism center has the preservation number of CGMCC 1.16273, address: the institute of microbiology, national academy of sciences, west road No. 1, north Chen, Chaozhou, Chaoyang;
the step (2) specifically comprises the following steps: weighing tobacco leaves with the water content balanced to 11% -14% in advance, spraying 20mL of tobacco soil bacterium agent L4-6 per 100g of tobacco leaves, and fermenting the treated tobacco leaves for 24-72 h in a constant temperature and humidity box with the temperature of 22 ℃ and the concentration of 60% to obtain fermented tobacco leaves;
the step (3) specifically comprises the following steps: putting the fermented tobacco leaves into a carbonization furnace, and carrying out closed heating carbonization treatment on the tobacco leaves, wherein the specific operation steps comprise gradually raising the temperature of the furnace to 80 ℃, quickly raising the temperature to 120-300 ℃ after the fermented tobacco leaves are dried, and keeping the temperature for 1-6 hours to obtain the fermented carbonized tobacco leaves.
2. The fermented carbonized tobacco leaf particles according to claim 1, wherein the step (1) specifically comprises: inoculating the tobacco soil bacterium L4-6 liquid strain into a fermentation culture medium according to the inoculation amount of 10%, and performing shake-flask culture at 28 ℃ for 3-7 days to obtain a culture solution; and (4) centrifugally separating the culture solution, washing the precipitate with sterile water, uniformly shaking with sterile water, and diluting by 10 times to obtain the microbial inoculum.
3. The fermented carbonized tobacco leaf particles according to claim 1, wherein the step (4) specifically comprises the steps of:
1) detecting the water content of the fermented carbonized tobacco leaves, controlling the water content to be 8-12%, and crushing the tobacco leaves into powder with the particle size of 100-160 meshes;
2) weighing 90-99 parts of tobacco powder and 1-10 parts of cane sugar or mannitol powder according to parts by weight, and uniformly mixing for later use;
3) weighing 5-10 parts of gelatin, 0-5 parts of polyvinylpyrrolidone and 0-2 parts of sodium carboxymethyl starch according to parts by weight, and preparing a 5-12% adhesive solution by using distilled water;
4) and weighing 300-1000 g of the mixed powder in the step 2), placing the mixed powder in a fluidized bed for granulation, and spraying an adhesive solution in the granulation process to obtain the fermented carbonized tobacco particles.
4. The fermented carbonized tobacco leaf particles according to claim 3, wherein the step 4) specifically comprises the steps of:
(a) drying the mixed powder for 5-15 min under the conditions that the fluidizing air pressure is 0.12-0.2 Bar and the air flow temperature is 50-60 ℃;
(b) applying the adhesive solution prepared in the step 3) of the claim 3 by adopting a top spraying mode under the conditions that the fluidization air pressure is 0.12-0.25 Bar, the air flow temperature is 60-80 ℃, the spraying pressure is 0.08-0.15 Bar and the spraying speed is 8-12 g/min, wherein the dosage of the adhesive solution is 20-50% of the weight of the fermented carbonized tobacco powder dry material;
(c) and (c) repeating the step (a) after the adhesive is applied, so as to obtain the fermented carbonized tobacco particles.
5. The fermented carbonized tobacco leaf particles according to claim 4, wherein the moisture content of the fermented carbonized tobacco leaf particles obtained in step (c) is 8 to 10%, the particle size is 20 to 60 mesh, and the particles are in the shape of quasi-circular particles with rough surfaces.
6. A cigarette filter rod, characterized in that it comprises the fermented carbonized tobacco leaf particles according to any one of claims 1 to 5.
7. The cigarette filter rod of claim 6, wherein 1-2 mg of the fermented carbonized tobacco particles are added to each millimeter of the cigarette filter rod.
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