CN103504461A - Biological coloring and flavoring method for tobacco products - Google Patents
Biological coloring and flavoring method for tobacco products Download PDFInfo
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- CN103504461A CN103504461A CN201210207933.9A CN201210207933A CN103504461A CN 103504461 A CN103504461 A CN 103504461A CN 201210207933 A CN201210207933 A CN 201210207933A CN 103504461 A CN103504461 A CN 103504461A
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Abstract
The invention provides a biological coloring and flavoring method for tobacco products. In the tobacco processing procedure, tobacco or tobacco extract liquid is used as a main raw material, the main raw material undergoes microbial fermentation to biosynthesize natural pigment, and the natural pigment is added back to the tobacco products to obtain colored and flavored tobacco products. The method includes the steps of tobacco extract liquid culture medium preparation, fermentation seed liquid preparation, natural pigment synthesis, pigment extraction and back adding of the pigment to the tobacco products. The method is suitable for coloring and flavoring of the tobacco products in the technology of flavoring of tobacco sheets, cut stems, flue-cured tobacco laminas and cigarettes. According to the method, tobacco waste is converted into the natural pigment in a biological mode, the natural pigment is used as a coloring adjunct to be used in the tobacco products and replaces chemical pigment, color and flavor are enhanced at the same time, and the two purposes are achieved; on-line digestion and utilization of the tobacco waste and liquid waste can be achieved, pollutant discharge is reduced, and good social benefits, environment benefits and economic benefits are generated.
Description
Technical field
The present invention relates to tobacco processing and Comprehensive Utilization of Tobacco Waste and utilize field, particularly the hyperchromic flavouring method of the biology of a tobacco articles.
Background technology
The appearance color of tobacco product is one of importance of cigarette quality, golden yellow, orange tone is the appearance characteristics of High Quality Tobacco, therefore in tobacco processing course, keep and improve tobacco appearance color, outstanding golden yellow, orange tone is the quality standard that processing technology must be paid close attention to.In reconstituted tobacoo, stem, tobacco leaf, pipe tobacco process, conventionally adopt reinforced mode to add food coloring, to improve the appearance luster of tobacco product.But the food coloring adding in tobacco processing course is at present chemical synthesis pigment often, because molecular structure, the character of chemical synthesis pigment are different from natural colouring matter, some or even harmful or potential hazard, even the chemical synthesis pigment identical with natural pigment member structure, also subject to from the impurity in building-up process, the especially pollution of harmful substance used in synthetic reaction, therefore say from security standpoint, natural products need to be provided.But so far, be suitable for the orange colour natural colouring matter product category of tobacco seldom, it is mainly carotenoids, in existing product, the natural carotenoid product of low-purity is added with a large amount of additives conventionally, tobacco aroma is had to harmful effect, and highly purified use cost is too high, so that existing market is applicable to the natural colouring matter product of tobacco product is few.
Already knew, natural colouring matter especially carotenoid is important fragrance component and the fragrance precursor naturally existing in tobacco, and the appearance luster of tobacco product and flavouring essence quality are had to important function.Therefore, adopt natural colouring matter especially the pigment such as carotenoid as tobacco product toner, both can improve appearance color quality, can increase again fragrance, there is good application prospect.
Adopting microorganism fermenting organism synthesis of natural pigment, is one of important sources of natural colouring matter, has been widely used at present food and feed industry.The raw materials for production of microorganism fermenting organism synthesis of natural pigment are mainly the agricultural resources such as carbohydrate, starchiness class grain at present.In tobacco processing course, conventionally there is a large amount of tobacco wastes to produce, comprise waste tobacco, offal, offal waste liquid etc., for example, in tobacco sheets by paper making method processing technology, conventionally the even whole stem extracting solutions of part are discarded, and as discharging after wastewater treatment; In cut rolled stem processing procedure, also exist the part phenomenon that even all stem extracting solution is discarded, cause the wasting of resources and environmental pollution.These discarded objects are rich in protein, carbohydrate, cellulose, organic acid, pectic substance etc., are the living resources of a preciousness, are not yet rationally utilized.
So far there is not yet utilize the tobacco waste that produces in tobacco processing course, waste liquid as primary raw material fermenting and producing natural colouring matter the report for the hyperchromic flavouring of tobacco product.
Summary of the invention
The object of the present invention is to provide the hyperchromic flavouring method of biology of a tobacco articles, another object of the present invention is to provide the circulation utilization method of waste material in a kind of tobacco processing course, waste liquid resource.
The present invention adopts following technical scheme for achieving the above object: the hyperchromic flavouring method of biology of a tobacco articles, in tobacco product process, take tobacco or tobacco leaching liquor is primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in tobacco product, the tobacco product that obtains hyperchromic flavouring, comprises the following steps:
(1) prepare tobacco extract culture medium;
(2) prepare fermentation seed liquid;
(3) the tobacco extract culture medium fermentation synthesis of natural pigment that utilizes step (1) to prepare;
(4) the zymotic fluid of preparing from step (3), extract natural colouring matter;
(5) natural colouring matter of being prepared by step (4) returns and is added in tobacco product.
Above-mentioned tobacco product process comprises the existing processing technology that respectively grows tobacco, as reconstituted tobacoo processing technology, cut rolled stem processing procedure, tobacco threshing redrying technique, cigarette process etc.
In tobacco sheets by paper making method processing technology, the present invention be take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in reconstituted tobacoo, the reconstituted tobacoo product that obtains hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal leachate that tobacco sheets by paper making method process Raw extraction process produces, add adequate nutrition source, pH is controlled at 4.0～6.5, obtains offal leachate culture medium, standby after sterilizing;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in reconstituted tobacoo
The natural colouring matter that step (4) is obtained is delivered to feed liquid dip coating apparatus or the feeding device of tobacco sheets by paper making method production line, evenly adds in reconstituted tobacoo sheet base, obtains hyperchromic flavouring reconstituted tobacoo.
In cut rolled stem processing procedure, the present invention be take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in stem, obtains the stem product of hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal leaching liquor that stem process Raw extraction process produces, add adequate nutrition source, pH is controlled at 4.0～6.5, obtains offal leachate culture medium, standby after sterilizing;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in stem
The natural colouring matter that step (4) is obtained is delivered to feed liquid dip coating apparatus or the feeding device of stem production line, evenly adds in stem, obtains hyperchromic flavouring stem product.
In tobacco threshing redrying technique, the present invention be take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in tobacco leaf, obtains the sheet smoke product of hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal producing in tobacco threshing redrying technical process, add the water of 5～50 times of weight, in 30～100 ℃ of lixiviate 0.2～24h, make the abundant stripping of DDGS; Then Separation of Solid and Liquid, obtains offal leaching liquor and solid phase stalk slag; Offal leaching liquor is added to adequate nutrition source, and pH is controlled at 4.0～6.5, obtains offal leachate culture medium, standby after sterilizing; Solid phase stalk slag separately uses it for anything else;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in tobacco leaf
The natural colouring matter that step (4) is obtained is delivered to the tobacco feeding device of tobacco threshing redrying production line, evenly adds in tobacco leaf, obtains hyperchromic flavouring sheet smoke product.
The natural colouring matter product of the above-mentioned preparation of the present invention also can be used in cigarette primary processing technique tobacco leaf or tobacco perfuming is reinforced, obtains hyperchromic flavouring cigarette product.
The maximum absorption wavelength of the biosynthetic natural colouring matter of the present invention is 380～520nm, color is mainly yellow to red tone, as lemon yellow, orange, reddish yellow, redness etc., can be used for all kinds tobacco product, mainly be applicable to the tobacco products such as reconstituted tobacoo, stem, sheet cigarette, pipe tobacco, cigarette, strengthen the golden yellow and orange tone of product.The biosynthetic natural colouring matter kind of the present invention preferred class carrotene, monascorubin and yeast red pigment, because the color of these pigments and High Quality Tobacco coordinates very much, and carotenoid has good flavouring effect; Described carotenoid comprises one or more pigments in beta carotene, alpha-carotene, lutein, lycopene, zeaxanthin, astaxanthin, canthaxanthin, β-kryptoxanthin; Described monascorubin comprises one or more pigments in erythema Monascin, red colouring agent for food, also used as a Chinese medicine rubine element, Monascin, ankaflavin, erythema red colouring agent for food, also used as a Chinese medicine amine, red colouring agent for food, also used as a Chinese medicine rubine amine.
The inventive method can adopt the bacterial classification of the known above-mentioned natural colouring matter of various products as fermented bacterium, and wherein a kind of the or many bacterial classification in preferred monascus ruber (Monascus), rhodotorula (Rhodotorula), rhodothece rubra (Rhodotorula rubra), rhodotorula glutinis (Rhodotorula glutinis), phaffia rhodozyma (Phaffia rhodozyma), trispore Bruce mould (Blakeslea trispora), phycomyces blakesleeanus (Phycomyces blakesleeanus) and secondary coccus (Paracoccus carotinifaciens) is used in conjunction with.
For improving the adaptive capacity of bacterial strain to tobacco extract, can adopt offal extract culture medium to carry out domestication and improvement to the microorganism fungus kind of selected product natural colouring matter, method is: get offal extract culture medium and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, under 20～40 ℃ of conditions on shaking table shaken cultivation 20～100h, then be transferred in identical culture medium and cultivate, method is transferred 3～5 times continuously like this, to after the nutrient solution dilution of acquisition, be applied on previously prepared offal leaching liquor solid medium, tobacco offal leaching liquor solid medium is by offal leaching liquor fluid nutrient medium, to add 1.9% agar to make, then under 20～40 ℃ of conditions, cultivate 20～100h, after growing bacterium colony, picking macrocolony is rule on same solid plate culture medium, select single bacterium colony and be transferred to solid slant medium, cultivation obtains naturalized strain, save backup, bacterial strain after domestication produces high degree of adaptability to offal extract culture medium, and fermenting property significantly improves.
The inventive method can be utilized various tobacco waste resources, wherein preferably in tobacco sheets by paper making method process or the stem extracting solution producing in stem process, or the waste tobacco stalk producing in tobacco threshing redrying process; In addition, cigarette stalk is processed through hydrolysis and saccharification, also can be used as the fermentation raw material of natural colouring matter of the present invention.
The advantage of the inventive method is:
1, the natural colouring matter that employing the inventive method is produced especially carotenoid, as tobacco product toner, both can improve appearance color quality, can increase fragrance again, killed two birds with one stone;
2, by microorganism biological synthesis, by tobacco waste bio-transformation, be natural colouring matter, and be used for tobacco product quality-improving, can realize the efficient circulation utilization of tobacco biomass waste material, reduce environmental pollution, can produce good social benefit and environmental benefit;
3, in the present invention, fermentation raw material used is mainly tobacco waste, the waste liquid that tobacco machining production line produces, raw material sources are concentrated, steady quality, pigment product directly or indirectly returns to tobacco product, can realize the online digestibility and utilization of tobacco waste, waste liquid, compare with other agricultural, grain, industrial waste, can reduce production costs, enhance productivity and economic benefit; Can reduce the interference of non-tobacco component to pigment product or tobacco product quality simultaneously;
4, the nutrition of tobacco waste, waste liquid and physico chemical factor are applicable to growth and the pigment biosynthesis of pigment formation bacterium very much, and pigment productive rate is higher than conventional agriculture, grain, industrial waste.
The specific embodiment
Below will by embodiment, the invention will be further described, these descriptions are not to be further limited content of the present invention; What the technical characterictic of content of the present invention was done is equal to replacement, or improves accordingly, within still belonging to protection scope of the present invention.
Embodiment mono-:
In tobacco sheets by paper making method process, take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in reconstituted tobacoo, the reconstituted tobacoo product that obtains hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Directly get the one-level offal leaching liquor that tobacco sheets by paper making method process Raw extraction process produces, described one-level offal leaching liquor is generally at 60 ℃, with the flooding offal 5～30min of 3～10 times of weight, obtain, usually used as waste liquid, deliver to sewage disposal system and process; In the present embodiment, leaching liquor is added by weight to 1% sucrose, 0.8% NaH
2pO
4, natural pH, makes offal extract culture medium, standby after 110 ℃ of sterilizing 20min;
(2) prepare fermentation seed liquid
The domestication of slant strains: get above-mentioned offal extract culture medium 50mL and be placed in triangular flask, rhodothece rubra (the Rhodotorula rubra sp.) slant strains of carotenoid is produced in access, under 28 ℃ of conditions, cultivate 48h, then be transferred in identical culture medium and cultivate, method is transferred 3 times continuously like this, to after the nutrient solution dilution of acquisition, be applied on previously prepared offal leaching liquor solid medium, offal leaching liquor solid medium is by offal extract culture medium, to add 1.9% agar to make, then under 28 ℃ of conditions, cultivate 48h, picking macrocolony is rule on same solid plate culture medium, select single bacterium colony and be transferred to solid slant medium, cultivation obtains naturalized strain, save backup,
Fermentation seed is cultivated: get above-mentioned tobacco waste fluid nutrient medium 100mL and be placed in triangular flask, access, through rhodothece rubra (the Rhodotorula rubra sp.) slant strains of the product carotenoid of domestication, is cultivated 30h at 28 ℃, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) the synthetic carotenoid of fermentation
Get above-mentioned offal extract culture medium in fermentation tank, the fermentation seed liquid that cut-in quality mark is 5%, in 28 ℃ of fermentation 96h of temperature, obtains fermentative carotenoid liquid;
(4) extract carotenoid
Above-mentioned fermentative carotenoid liquid is concentrated, self-dissolving broken wall, sterilizing, make the thick product of paste carotenoid, standby;
(5) carotenoid is returned and is added in reconstituted tobacoo
The thick product of above-mentioned paste carotenoid is delivered to feed liquid dip coating apparatus or the feeding device of tobacco sheets by paper making method production line, after ratio in 1%～10% and normal leaching liquor or coating fluid premix, evenly add in reconstituted tobacoo sheet base, drying, obtains reconstituted tobacoo product; Through outward appearance and the evaluation of smokeing panel test, the orange colour color and luster of this reconstituted tobacoo product obviously strengthens, and xylon gas alleviates, and perfume quantity increases, and fresh and sweet note is outstanding.
Embodiment bis-:
In cut rolled stem processing procedure, the present invention be take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in stem, obtains the stem product of hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal leaching liquor that stem process Raw extraction process produces, described offal leaching liquor is generally at 40～80 ℃, obtains, usually used as liquid waste processing with the flooding offal 5～60min of 3～10 times of weight; In the present embodiment, offal leaching liquor is added by weight to 0.5% sucrose, 0.5% NaH
2pO
4, natural pH, makes offal leachate culture medium, standby after sterilizing;
(2) prepare fermentation seed liquid
The domestication of slant strains: get above-mentioned offal extract culture medium 50mL and be placed in triangular flask, phaffia rhodozyma (the Phaffia rhodozyma sp.) slant strains of carotenoid is produced in access, under 22 ℃ of conditions, cultivate 72h, then be transferred in identical culture medium and cultivate, method is transferred 3 times continuously like this, to after the nutrient solution dilution of acquisition, be applied on previously prepared offal leaching liquor solid medium, offal leaching liquor solid medium is by offal extract culture medium, to add 1.9% agar to make, then under 22 ℃ of conditions, cultivate 72h, picking macrocolony is rule on same solid plate culture medium, select single bacterium colony and be transferred to solid slant medium, cultivation obtains naturalized strain, save backup,
Get above-mentioned tobacco waste fluid nutrient medium 100mL and be placed in triangular flask, access, through phaffia rhodozyma (the Phaffia rhodozyma sp.) slant strains of the product carotenoid of domestication, is cultivated 80h at 22 ℃, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) the synthetic carotenoid of fermentation
Get above-mentioned offal extract culture medium in fermentation tank, the fermentation seed liquid that cut-in quality mark is 10%, in 22 ℃ of fermentation 120h of temperature, obtains fermentative carotenoid liquid;
(4) extract carotenoid
Above-mentioned fermentative carotenoid liquid is concentrated, broken wall, sterilizing, make the thick product of paste carotenoid, standby;
(5) carotenoid is returned and is added in stem
The thick product of above-mentioned paste carotenoid is delivered to feed liquid dip coating apparatus or the feeding device of stem production line, after the ratio in 1%～10% and other feed liquid premix, evenly adds in stem, drying, obtains stem product; Through outward appearance and the evaluation of smokeing panel test, the orange colour color and luster of this stem product obviously strengthens, and assorted gas alleviates, and perfume quantity increases, and fresh and sweet note is outstanding.
Embodiment tri-:
In tobacco threshing redrying technique, take offal leaching liquor as primary raw material, by the synthetic monascorubin of microorganism fermenting organism, then the monascorubin making is returned and is added in sheet cigarette, obtain the sheet smoke product of hyperchromic flavouring, specifically comprise following operating procedure:
(1) prepare offal extract culture medium
Get the offal producing in tobacco threshing redrying technical process, add the water of 10 times of weight, in 30 ℃ of lixiviate 12h, make the abundant stripping of DDGS; Then Separation of Solid and Liquid, obtains offal leaching liquor and solid phase stalk slag; Leaching liquor adds by weight 1% soluble starch, 0.3% NaH
2pO
4, make offal extract culture medium, standby after 100 ℃ of sterilizing 30min; Solid phase stalk slag is for the fermenting and producing peat composed of rotten mosses;
(2) prepare fermentation seed liquid
Fermentation seed is cultivated: get above-mentioned offal extract culture medium 100mL and be placed in triangular flask, access, through monascus ruber (Monascus sp.) slant strains of the product monascorubin of domestication, is cultivated 48h at 32 ℃, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate; Strain domestication method is with reference to embodiment mono-;
(3) the synthetic monascorubin of fermentation
Get above-mentioned offal extract culture medium in fermentation tank, the fermentation seed liquid that cut-in quality mark is 15%, in 32 ℃ of fermentation 96h of temperature, obtains monascorubin zymotic fluid;
(4) extract monascorubin
Above-mentioned monascorubin zymotic fluid is concentrated into paste, adds the ethanol of 10 times of volumes 95%, 50 ℃ are stirred extraction 1～2h, and filtering thalline residue, reclaims ethanol, makes paste or powdery monascorubin product, standby;
(5) monascorubin is returned and is added in tobacco leaf
The above-mentioned monascorubin making is delivered to the tobacco feeding device of tobacco threshing redrying production line, after suitably diluting with 50%～90% ethanolic solution, ratio in 0.1%～5% is evenly added in tobacco leaf, through outward appearance and the evaluation of smokeing panel test, the appearance luster of the sheet smoke product after processing improves 1～2 grade, perfume quantity increases, and fresh and sweet note is outstanding.
Embodiment tetra-:
In tobacco processing course, the offal that the beating and double roasting operation of take produces is primary raw material, synthetic by microorganism fermenting organism; Carotenoid, then adds the carotenoid making in the pipe tobacco of silk producing procedures to, obtains the cigarette product of hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal producing in tobacco threshing redrying technical process, add the water of 10 times of weight, in 60 ℃ of lixiviate 1h, make the abundant stripping of DDGS; Then Separation of Solid and Liquid, obtains offal leaching liquor and solid phase stalk slag; Leaching liquor adds by weight 0.5% soluble starch, 0.3% NaH
2pO
4, make offal extract culture medium, standby after 110 ℃ of sterilizing 30min; Solid phase stalk slag is for the fermenting and producing peat composed of rotten mosses;
(2) prepare fermentation seed liquid
The domestication of slant strains: get above-mentioned offal extract culture medium 50mL and be placed in triangular flask, the positive bacterium slant strains of Blakeslea trispora (Blakeslea trispora zust sp.) of carotenoid is produced in access, under 28 ℃ of conditions, cultivate 36h, then be transferred in identical culture medium and cultivate, method is transferred 3 times continuously like this, to after the nutrient solution dilution of acquisition, be applied on previously prepared offal leaching liquor solid medium, offal leaching liquor solid medium is by offal waste material fluid nutrient medium, to add 1.9% agar to make, then under 28 ℃ of conditions, cultivate 72h, picking macrocolony is rule on same solid plate culture medium, select single bacterium colony and be transferred to solid slant medium, cultivation obtains the positive bacteria strain of Blakeslea trispora of domestication, save backup, adopt same method domestication to obtain the negative bacteria strain of Blakeslea trispora, save backup,
Fermentation seed is cultivated: get above-mentioned offal extract culture medium 100mL and be placed in triangular flask, access respectively the above-mentioned slant strains through domestication, positive bacterium is cultivated 20h at 28 ℃, obtains positive bacterium shake-flask seed liquid; Negative bacterium is cultivated 35h at 28 ℃, must bear bacterium shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) the synthetic carotenoid of fermentation
Get above-mentioned tobacco waste Liquid Culture based in fermentation tank, the above-mentioned positive bacterium shake-flask seed liquid that cut-in quality mark is 3% and 7% above-mentioned negative bacterium shake-flask seed liquid, in 28 ℃ of fermentation 120h of temperature, obtain zymotic fluid;
(4) extract carotenoid
By above-mentioned zymotic fluid centrifugation, collect thalline, thalline, through vacuum drying, broken wall, then, with acetone or ethyl acetate extraction, reclaims solvent, obtains salmon pink paste or powdery natural colouring matter, standby;
(5) carotenoid is returned and is added in pipe tobacco
The above-mentioned carotenoid making is delivered in the flavoring and casing device of cigarette primary processing operation, in evenly adding to and cut filler in 0.001%～0.1% ratio after evenly mixing with other feed liquid, obtains the cigarette product of hyperchromic flavouring; Through outward appearance and the evaluation of smokeing panel test, the appearance luster of the cigarette product after processing improves 1～2 grade, and fragrance matter, perfume quantity improve, and assorted gas excitant alleviates, and the fragrance of a flower, fresh and sweet note are given prominence to.
Claims (10)
1. the hyperchromic flavouring method of the biology of a tobacco articles, it is characterized in that: in tobacco product process, take tobacco or tobacco leaching liquor is primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in tobacco product, the tobacco product that obtains hyperchromic flavouring, comprises the following steps:
(1) prepare tobacco extract culture medium;
(2) prepare fermentation seed liquid;
(3) the tobacco extract culture medium fermentation synthesis of natural pigment that utilizes step (1) to prepare;
(4) the zymotic fluid of preparing from step (3), extract natural colouring matter;
(5) natural colouring matter of being prepared by step (4) returns and is added in tobacco product.
2. the hyperchromic flavouring method of the biology of a tobacco articles according to claim 1, it is characterized in that: in tobacco sheets by paper making method process, take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in reconstituted tobacoo, the reconstituted tobacoo product that obtains hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal leaching liquor that tobacco sheets by paper making method process Raw extraction process produces, add adequate nutrition source, pH is controlled at 4.0～6.5, obtains offal leachate culture medium, standby after sterilizing;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in reconstituted tobacoo
The natural colouring matter that step (4) is obtained is delivered to feed liquid dip coating apparatus or the feeding device of tobacco sheets by paper making method production line, evenly adds in reconstituted tobacoo sheet base, obtains hyperchromic flavouring reconstituted tobacoo product.
3. the hyperchromic flavouring method of the biology of a tobacco articles according to claim 1, it is characterized in that: in stem process, take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in stem, the stem product that obtains hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal leaching liquor that stem process Raw extraction process produces, add adequate nutrition source, pH is controlled at 4.0～6.5, obtains offal leachate culture medium, standby after sterilizing;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in stem
The natural colouring matter that step (4) is obtained is delivered to feed liquid dip coating apparatus or the feeding device of stem production line, evenly adds in stem, obtains hyperchromic flavouring stem product.
4. the hyperchromic flavouring method of the biology of a tobacco articles according to claim 1, it is characterized in that: in tobacco threshing redrying process, take offal leaching liquor as primary raw material, by microorganism fermenting organism synthesis of natural pigment, then the natural colouring matter making is returned and is added in tobacco leaf, the sheet smoke product that obtains hyperchromic flavouring, specifically comprises following operating procedure:
(1) prepare offal extract culture medium
Get the offal producing in tobacco threshing redrying technical process, add the water of 5～50 times of weight, in 30～100 ℃ of lixiviate 0.2～24h, make the abundant stripping of DDGS; Then Separation of Solid and Liquid, obtains offal leaching liquor and solid phase stalk slag; Offal leaching liquor is added to adequate nutrition source, and pH is controlled at 4.0～6.5, obtains offal extract culture medium, standby after sterilizing; Solid phase stalk slag separately uses it for anything else;
(2) prepare fermentation seed liquid
Get the described offal extract culture medium of step (1) and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, at 20～40 ℃, cultivates 20～100h, obtains shake-flask seed liquid; According to production scale, can further in seeding tank, carry out the amplification of fermentation seed and cultivate;
(3) fermentation synthesis of natural pigment
Get the described offal extract culture medium of step (1) in fermentation tank, the fermentation seed liquid of being prepared by described step (2) that cut-in quality mark is 5%～20%, in 20～40 ℃ of fermentation 24～200h of temperature, obtains natural colouring matter zymotic fluid;
(4) extract natural colouring matter
The natural colouring matter zymotic fluid that step (3) is obtained concentrates, sterilizing, makes the thick product of natural colouring matter, standby; Or the natural colouring matter zymotic fluid that step (3) is obtained obtains thalline through precipitation, filtration or centrifugation, thalline is made medicinal extract through broken wall, or thalline is dry, organic solvent extraction, makes high-purity natural pigment product, standby;
(5) natural colouring matter is returned and is added in tobacco leaf
The natural colouring matter that step (4) is obtained is delivered to the tobacco feeding device of tobacco threshing redrying production line, evenly adds in tobacco leaf, obtains hyperchromic flavouring sheet smoke product.
5. according to the hyperchromic flavouring method of biology of the tobacco articles described in claim 1,2,3,4 any one, it is characterized in that: the natural colouring matter product of preparation can be used in cigarette primary processing technique tobacco leaf or tobacco perfuming is reinforced, obtains hyperchromic flavouring cigarette product.
6. according to the hyperchromic flavouring method of biology of the tobacco articles described in claim 1,2,3,4,5 any one, it is characterized in that: the maximum absorption wavelength of described biosynthetic natural colouring matter is 380～520nm, pigment appearance color is yellow to red.
7. according to the hyperchromic flavouring method of biology of the tobacco articles described in claim 1,2,3,4,5,6 any one, it is characterized in that: described biosynthetic natural colouring matter comprises one or more pigments in carotenoid, monascorubin and yeast red pigment.
8. the hyperchromic flavouring method of the biology of a tobacco articles according to claim 7, is characterized in that: described biosynthetic carotenoid comprises one or more pigments in beta carotene, alpha-carotene, lutein, lycopene, zeaxanthin, astaxanthin, canthaxanthin, β-kryptoxanthin; Described biosynthetic monascorubin comprises one or more pigments in erythema Monascin, red colouring agent for food, also used as a Chinese medicine rubine element, Monascin, ankaflavin, erythema red colouring agent for food, also used as a Chinese medicine amine, red colouring agent for food, also used as a Chinese medicine rubine amine.
9. according to claim 1, 2, 3, 4, 5, the hyperchromic flavouring method of biology of the tobacco articles described in 6 any one, it is characterized in that: the microorganism fungus kind of described product natural colouring matter is selected from monascus ruber (Monascus), rhodotorula (Rhodotorula), rhodothece rubra (Rhodotorula rubra), rhodotorula glutinis (Rhodotorula glutinis), phaffia rhodozyma (Phaffia rhodozyma), trispore Bruce mould (Blakeslea trispora), phycomyces blakesleeanus (Phycomyces blakesleeanus) and secondary coccus (Paracoccuscaro tinifaciens).
10. the hyperchromic flavouring method of the biology of a tobacco articles according to claim 9, it is characterized in that: adopt offal extract culture medium to carry out domestication and improvement to the microorganism fungus kind of selected product natural colouring matter, method is: get offal extract culture medium and be placed in triangular flask, the slant strains of natural colouring matter is produced in access, under 20～40 ℃ of conditions on shaking table shaken cultivation 20～100h, then be transferred in identical culture medium and cultivate, method is transferred 3～5 times continuously like this, to after the nutrient solution dilution of acquisition, be applied on previously prepared offal leaching liquor solid medium, offal leaching liquor solid medium is by offal extract culture medium, to add 1.9% agar to make, then under 20～40 ℃ of conditions, cultivate 20～100h, after growing bacterium colony, picking macrocolony is rule on same solid plate culture medium, select single bacterium colony and be transferred to solid slant medium, cultivation obtains naturalized strain, save backup, bacterial strain after domestication produces high degree of adaptability to offal extract culture medium, and fermenting property significantly improves.
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