CN109288124B - Fermented carbonized walnut shell particles and preparation method and application thereof - Google Patents

Fermented carbonized walnut shell particles and preparation method and application thereof Download PDF

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CN109288124B
CN109288124B CN201811089521.3A CN201811089521A CN109288124B CN 109288124 B CN109288124 B CN 109288124B CN 201811089521 A CN201811089521 A CN 201811089521A CN 109288124 B CN109288124 B CN 109288124B
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fermented
carbonized
walnut
walnut shell
shell particles
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CN109288124A (en
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者为
高莉
李文均
董立民
郭青
张健
赵升逵
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China Tobacco Yunnan Industrial Co Ltd
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    • AHUMAN NECESSITIES
    • A24TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
    • A24DCIGARS; CIGARETTES; TOBACCO SMOKE FILTERS; MOUTHPIECES FOR CIGARS OR CIGARETTES; MANUFACTURE OF TOBACCO SMOKE FILTERS OR MOUTHPIECES
    • A24D3/00Tobacco smoke filters, e.g. filter-tips, filtering inserts; Filters specially adapted for simulated smoking devices; Mouthpieces for cigars or cigarettes
    • A24D3/06Use of materials for tobacco smoke filters
    • A24D3/14Use of materials for tobacco smoke filters of organic materials as additive
    • CCHEMISTRY; METALLURGY
    • C01INORGANIC CHEMISTRY
    • C01BNON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
    • C01B32/00Carbon; Compounds thereof
    • C01B32/05Preparation or purification of carbon not covered by groups C01B32/15, C01B32/20, C01B32/25, C01B32/30
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor

Abstract

The invention provides fermented carbonized walnut shell particles and a preparation method and application thereof, and the fermented carbonized walnut shell particles are prepared by the method comprising the following steps: (1) preparing a tobacco soil bacterium agent L4-6; (2) fermenting walnut shells by using a tobacco soil bacterium L4-6 to obtain fermented walnut shells; (3) carbonizing the fermented walnut shells to obtain fermented carbonized walnut shells; (4) drying the fermented carbonized walnut shell powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain the fermented carbonized walnut shell particles. The particles take walnut shells as raw materials, are green and environment-friendly, and change waste into valuable; after fermentation treatment and application to cigarettes, the special aroma coordinated with the smoke of the cigarettes can be generated, the cigarette aroma of the cigarettes is enriched, and the smoking taste of the cigarettes is improved; through carbonization treatment, the specific surface area of the walnut shell can be obviously increased, the adsorption effect of the particles on harmful substances in cigarette smoke is enhanced, and the plant particles are endowed with special tar fragrance.

Description

Fermented carbonized walnut shell particles and preparation method and application thereof
Technical Field
The invention belongs to the technical field of cigarette materials, and particularly relates to fermented carbonized walnut shell particles and a preparation method and application thereof.
Background
The low coking is the inevitable trend of the development of the tobacco industry, the smoking taste of the cigarettes is reduced due to the low coking, and the insufficient aroma gradually becomes the prominent problem of the 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, silica gel micropowder and the like have a remarkable effect on adsorbing harmful substances in smoke, a large amount of fragrance components are also 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.
The microbial fermentation 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, meet the requirements of consumers on the cigarette quality, and the microbial fermentation technology is widely applied to tobacco essences. In addition, the fermentation of the plant raw material with the odor by the microorganism can also endow the plant 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.
The walnut with the title of 'king of woody oil' has extremely high nutritive value and good medical health care function, and is listed as the head of the world 4 big dried fruits. Yunnan is the main production area of walnuts in China, the current yield is over 30 ten thousand tons, and if the kernel taking rate is calculated according to 55%, at least 13.5 ten thousand tons of walnut shells are produced every year. The concentrated walnut shells are discarded or incinerated, which causes great waste of resources. The walnut shell has a loose and porous structure on the surface and has good adsorption performance. Therefore, the comprehensive utilization of the walnut shells is enhanced, the waste of resources can be avoided, the solid waste can be effectively treated, and the waste can be changed into valuable. The walnut shells are carbonized after being fermented by microorganisms and are used for preparing the cigarette filter tip particles, so that the adsorption performance of the walnut shells on harmful substances in cigarette smoke can be improved, and the particles can be endowed with unique fragrance different from the original fragrance to enrich the cigarette fragrance.
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 improve the effects of the granular materials on the aspects of increasing the harmony of cigarette smoke, promoting the secretion of saliva and the like while reducing the irritation of the cigarette smoke become the technical problems to be further solved.
Disclosure of Invention
Aiming at the defects that the adsorption effect of the existing plant particles on harmful substances in smoke is not obvious, cigarettes cannot be directly endowed with fragrance, or the coordination with the fragrance of the cigarettes is not good and the like, the invention provides the fermented carbonized walnut shell 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 cigarettes can be enhanced, the irritation of the smoke of the cigarettes can be reduced, the cigarette perfuming effect can be enhanced, and the smoking quality of the cigarettes 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 walnut shell particles prepared by a process comprising the steps of:
(1) preparing a tobacco soil bacterium agent L4-6;
(2) fermenting walnut shells by using a tobacco soil bacterium L4-6 to obtain fermented walnut shells;
(3) carbonizing the fermented walnut shells to obtain fermented carbonized walnut shells;
(4) drying the fermented carbonized walnut shell powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain the fermented carbonized walnut shell particles.
The fermented carbonized walnut shell particles specifically comprise the following steps (1): 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 walnut shell particles specifically comprises: weighing walnut shells with the water content balanced to 12% -14% in advance, spraying 20mL of tobacco soil-borne bacterium L4-6 to every 100g of walnut shells, and fermenting the treated walnut shells in a constant-temperature and constant-humidity box with the temperature of 22 ℃ and the concentration of 60% for 24-72 h to obtain the fermented walnut shells.
The step (3) of the above-mentioned fermented carbonized walnut shell particles specifically comprises: putting the fermented walnut shells into a carbonization furnace, and carrying out closed heating carbonization treatment on the fermented walnut shells, wherein the specific operation steps are that the furnace temperature is gradually increased to 80 ℃, the temperature is quickly increased to 120-300 ℃ after the fermented walnut shells are dried, and the temperature is kept for 1-6 hours, so that the fermented carbonized walnut shells are obtained.
The step (4) of the fermentation carbonization walnut shell particles specifically comprises the following steps:
1) detecting the water content of the fermented carbonized walnut shells, controlling the water content to be 8-12%, and crushing the walnut shells into powder with the particle size of 100-160 meshes;
2) weighing 95-99 parts of the fermented carbonized walnut shell powder and 1-5 parts of glucose powder in the step 1) according to parts by weight, and uniformly mixing for later use;
3) weighing 1-5 parts of gelatin and 3-7 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 walnut shell particles.
The step 4) of the fermentation carbonization walnut shell 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 binder solution prepared in the step 3) of the claim 5 by adopting a top spraying mode under the conditions of the fluidizing pressure of 0.12-0.25 Bar, the airflow temperature of 60-80 ℃, the spraying pressure of 0.08-0.15 Bar and the spraying speed of 8-12 g/min, wherein the dosage of the binder solution is 20-40% of the weight of the dry mixed powder material;
(c) and (d) repeating step (a) after the binder is applied to obtain the fermented carbonized walnut shell particles.
The fermented carbonized walnut shell particles obtained in the step (c) have the water content of 10-12%, the particle size of 20-60 meshes and the shape of round-like particles with rough surfaces.
In a third aspect, the present invention relates to a cigarette filter rod comprising the fermented carbonized walnut shell particles produced according to the second aspect of the present invention.
According to the cigarette filter rod, 1-2 mg of fermented carbonized walnut shell particles are added in each millimeter of cigarette filter rod.
The invention has the following beneficial effects:
(1) according to the invention, the walnut shells are fermented by utilizing the tobacco soil bacterium L4-6, and after the walnut shells are applied to cigarette filter rods, special aroma coordinated with the aroma of cigarettes can be generated in the smoking process of the cigarettes, so that the cigarette aroma is enriched, and the smoking taste of the cigarettes is improved;
(2) according to the method, the fermented walnut shells are carbonized, walnut shell cells are dehydrated and shaped at high temperature in a short time, the adsorption specific surface area in the walnut shells is increased, the filtration efficiency of the fermented carbonized walnut shell particles on cigarette smoke is obviously improved, harmful substances in the cigarette smoke can be selectively adsorbed, and the fermented carbonized walnut shell particles are endowed with special burnt flavor;
(3) the walnut shells are used as raw materials, so that the method is green and environment-friendly, changes waste into valuable, and increases the utilization rate of plant resources;
(4) 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 16S rDNA partial sequence of the tobacco soil bacterium L4-6 is shown in the description of the attached drawing, the sequence is compared and analyzed with known sequences in GenBank database by BLAST, and the 16S rDNA gene sequence of related species is obtained from the database,a phylogenetic tree was constructed, see FIG. 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 16SrDNA 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. Fermenting walnut shell
Weighing 1200g of walnut shells with the water content balanced to 13% in advance, spraying 240mL of tobacco soil rhizoctonia L4-6 microbial inoculum, and fermenting the treated walnut shells in a constant-temperature constant-humidity box with the temperature of 22 ℃ and the concentration of 60% for 72 hours to obtain the fermented walnut shells.
3. Carbonization of fermented walnut shells
Placing the fermented walnut shells into a carbonization furnace, gradually heating the furnace to about 80 ℃, keeping the temperature for 40min, after the fermented walnut shells are dried, quickly heating to 160 ℃, keeping the temperature for 4h, turning off a heat source after the carbonization degree is inspected to be qualified, and taking out the fermented carbonized walnut shell samples for later use.
4. Preparation of fermented carbonized walnut shell particles
Taking fermented carbonized walnut shells, detecting the water content of the fermented carbonized walnut shells, complementing the water content to 10%, and crushing the fermented carbonized walnut shells into powder with the particle size of 100 meshes; mixing 960g of walnut shell fermentation carbonized powder and 40g of glucose powder, granulating in Midi Glatt fluidized bed, and drying at 60 deg.C and fluidized pressure of 0.18Bar for 10 min; then 5 parts of gelatin and 5 parts of sodium carboxymethyl starch are weighed according to the parts by weight, a 10% adhesive solution is prepared by distilled water, and 350g of the adhesive solution is applied by adopting a top spraying mode under the conditions that the fluidizing air pressure is 0.2Bar, the air flow temperature is 60 ℃, the spraying pressure is 0.15Bar and the spraying speed is 10 g/min; after the adhesive is applied, the mixture is dried for 5 minutes under the conditions that the fluidizing air pressure is 0.18Bar and the air flow temperature is 60 ℃, and the fermented carbonized walnut shell particles with 10 percent of moisture content, rough surface and 40-mesh particle size are prepared.
Example 3
1. The culture method of the tobacco soil bacterium L4-6 is the same as that of example 2.
2. Fermenting walnut shell
Weighing 600g of walnut shells with the water content balanced to 12% in advance, spraying 120mL of tobacco soil rhizoctonia L4-6 microbial inoculum, and fermenting the treated walnut shells in a constant temperature and humidity box with the temperature of 22 ℃ and the concentration of 60% for 72 hours.
3. Carbonization of fermented walnut shells
Placing the fermented walnut shell into a carbonization furnace, gradually heating the furnace to 80 ℃, keeping the temperature for 60min, after the walnut shell fermentation sample is dried, rapidly heating to 180 ℃, keeping the temperature for 2.5h, turning off a heat source after the carbonization degree is inspected to be qualified, and taking out the fermented carbonized walnut shell sample for later use.
4. Preparation of fermented carbonized walnut shell particles
Taking fermented carbonized walnut shells, detecting the water content of the fermented carbonized walnut shells, complementing the water content to 8%, and crushing the fermented carbonized walnut shells into powder with the particle size of 120 meshes; uniformly mixing 480g of prepared walnut shell fermentation carbonized powder and 20g of glucose powder, and placing the mixture in a Midi Glatt fluidized bed for granulation, wherein the fluidized air pressure of the fluidized bed is 0.15Bar, and the airflow temperature is 60 ℃ for drying for 15 min; then 5 parts of gelatin and 7 parts of sodium carboxymethyl starch are weighed according to the parts by weight, a 12% adhesive solution is prepared by distilled water, and 200g of the adhesive solution is applied by adopting a top spraying mode under the conditions that the fluidizing air pressure is 0.2Bar, the air flow temperature is 80 ℃, the spraying pressure is 0.15Bar and the spraying speed is 12 g/min; after the adhesive is applied, drying for 5 minutes under the conditions that the fluidizing air pressure is 0.18Bar and the air flow temperature is 60 ℃, and preparing the fermented carbonized walnut shell particles with 12 percent of moisture content, rough surface and 20 meshes of particle size.
Example 4
In order to examine the uniqueness of the tobacco soil bacterium L4-6 for fermenting walnut shells, example 4 was different from example 3 in that walnut shells were fermented with the closely related strain Dyella soli JS12-10 (purchased from korean agricultural microbial cultures collection (KACC)) of the tobacco soil bacterium L4-6 instead of the tobacco soil bacterium L4-6, and the rest of the procedure was the same as example 3, to obtain fermented carbonized walnut shell particles having a water content of 12%, a rough surface, and a particle size of 20 mesh, which were obtained by fermentation with the Dyella soli JS 12-10.
Example 5
The fermented carbonized walnut shell particles prepared in the example 2 are applied to a cigarette composite filter stick, 2mg of the fermented carbonized walnut shell 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 added with the fermented carbonized walnut shell particles has obviously reduced smoke irritation, mellow and full aroma and increased richness of tobacco aroma; the detection of harmful components in the smoke shows that: in the smoke of the experimental cigarette, the tar is reduced by 9.3%, the nicotine is reduced by 14.6%, and the phenol is reduced by 21.6%; the detection of harmful components in the smoke shows that: the tar is reduced by 9.3%, the nicotine is reduced by 14.6%, and the phenol is reduced by 21.6%, which shows that the fermented carbonized walnut shell particles can obviously reduce the contents of tar, nicotine and phenol in the cigarette smoke.
Example 6
The fermented carbonized walnut shell particles prepared in the example 3 are applied to a cigarette composite filter stick, 1.5mg of the fermented carbonized walnut shell particles are added to each millimeter of the cigarette filter stick to prepare an experimental cigarette, and the cigarette without the fermented carbonized walnut shell 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 of the added fermented carbonized walnut shell particles is reduced, the special acid aroma and burnt aroma are added while the characteristic aroma of the original walnut shell is maintained, and the aroma richness is increased; the detection of harmful components in the smoke shows that: in the smoke of experimental cigarettes, the tar is reduced by 7.9%, the nicotine is reduced by 12.6%, and the phenol is reduced by 19.3%, which shows that the fermented carbonized walnut shell particles can obviously reduce the contents of tar, nicotine and phenol in the smoke of cigarettes.
Example 7
The fermented carbonized walnut shell particles prepared by further processing the dylla soli JS12-10 fermented walnut shells in the example 4 are applied to the cigarette composite filter stick, and 1.5mg of the fermented carbonized walnut shell particles are added to each millimeter of the cigarette 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 walnut shells with Dyella soli JS12-10 has no obvious reduction of smoke irritation, insufficient aftertaste improvement and single and thin aroma texture. The detection of harmful components in the smoke shows that: in the smoke of the experimental cigarette, the tar is reduced by 5.1%, the nicotine is reduced by 8.4%, and the phenol is reduced by 15.1%, compared with the data in the example 6, the reduction rate of the harmful components in the experimental cigarette is not as good as that in the control group of cigarettes. Thus, although the strain is a related strain, the strain Dyella soliJS12-10 can not achieve the effect similar to the tobacco soil bacterium L4-6 when being used for walnut shell 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:
Figure BDA0001803994340000101
sequence listing
<110> tobacco industry Limited liability company in Yunnan
The person is Gao Li Wen Li
<120> fermented carbonized walnut shell particles and preparation method and application thereof
<130> RIB180322
<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 walnut shell particle, characterized in that it is prepared by a process comprising the steps of:
(1) preparing a tobacco soil bacterium agent L4-6;
(2) fermenting walnut shells by using a tobacco soil bacterium L4-6 to obtain fermented walnut shells;
(3) carbonizing the fermented walnut shells to obtain fermented carbonized walnut shells;
(4) drying the fermented carbonized walnut shell powder on a fluidized bed, spraying an adhesive, bonding the powder to form particles, and continuously drying to obtain the fermented carbonized walnut shell particles;
the microbiological classification of the tobacco soil bacterium L4-6 is named as tobacco soil bacteriumDyellatabacisoliL4-6, which has been preserved in the general microbiological center of China Committee for culture Collection of microorganisms at 19.6.2018, with the preservation number of CGMCC 1.16273 and the address of the institute of microbiology, China academy of sciences, North West Lu No. 1 institute of North Chen, Ind.
The step (2) specifically comprises the following steps: weighing walnut shells with the water content balanced to 12% -14% in advance, spraying 20mL of tobacco soil-borne bacterium L4-6 to every 100g of walnut shells, and fermenting the treated walnut shells in a constant-temperature and constant-humidity box with the temperature of 22 ℃ and the concentration of 60% for 24-72 h to obtain fermented walnut shells;
the step (3) specifically comprises the following steps: putting the fermented walnut shells into a carbonization furnace, and carrying out closed heating carbonization treatment on the fermented walnut shells, wherein the specific operation steps are that the furnace temperature is gradually increased to 80 ℃, the temperature is quickly increased to 120-300 ℃ after the fermented walnut shells are dried, and the temperature is kept for 1-6 hours, so that the fermented carbonized walnut shells are obtained.
2. The fermented carbonized walnut shell particles of claim 1, wherein 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 walnut shell particles of claim 1, wherein step (4) specifically comprises the steps of:
1) detecting the water content of the fermented carbonized walnut shells, controlling the water content to be 8-12%, and crushing the walnut shells into powder with the particle size of 100-160 meshes;
2) weighing 95-99 parts of the fermented carbonized walnut shell powder and 1-5 parts of glucose powder in the step 1) according to parts by weight, and uniformly mixing for later use;
3) weighing 1-5 parts of gelatin and 3-7 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 walnut shell particles.
4. The fermented carbonized walnut shell particles of claim 3, wherein 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 binder solution prepared in the step 3) of the claim 3 by adopting a top spraying mode under the conditions of the fluidizing pressure of 0.12-0.25 Bar, the airflow temperature of 60-80 ℃, the spraying pressure of 0.08-0.15 Bar and the spraying speed of 8-12 g/min, wherein the dosage of the binder solution is 20-40% of the weight of the dry material of the mixed powder;
(c) and (d) repeating step (a) after the binder is applied to obtain the fermented carbonized walnut shell particles.
5. The fermented carbonized walnut shell particles of claim 4, wherein the fermented carbonized walnut shell particles obtained in step (c) have a moisture content of 10 to 12%, a particle size of 20 to 60 mesh, and a shape of quasi-round particles with rough surfaces.
6. A cigarette filter rod comprising the fermented carbonized walnut shell particles of any one of claims 1 to 5.
7. The cigarette filter rod of claim 6, wherein 1-2 mg of the fermented carbonized walnut shell particles are added per millimeter of the cigarette filter rod.
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CN105923631A (en) * 2016-07-13 2016-09-07 袁春华 Method for preparing activated carbon adsorbent on basis of nutshells

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CN105923631A (en) * 2016-07-13 2016-09-07 袁春华 Method for preparing activated carbon adsorbent on basis of nutshells

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