CN102119784B - Spice extract prepared from waste tobacco leaves and preparation method and application thereof - Google Patents
Spice extract prepared from waste tobacco leaves and preparation method and application thereof Download PDFInfo
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- 241000208125 Nicotiana Species 0.000 title claims abstract description 89
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 89
- 239000002699 waste material Substances 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 235000013599 spices Nutrition 0.000 title abstract 3
- 238000000855 fermentation Methods 0.000 claims abstract description 31
- 230000004151 fermentation Effects 0.000 claims abstract description 31
- 235000019504 cigarettes Nutrition 0.000 claims abstract description 24
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 claims abstract description 18
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229960002715 nicotine Drugs 0.000 claims abstract description 18
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 13
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 12
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 12
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 claims abstract description 12
- CHWNEIVBYREQRF-UHFFFAOYSA-N 4-Ethyl-2-methoxyphenol Chemical compound CCC1=CC=C(O)C(OC)=C1 CHWNEIVBYREQRF-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000002994 raw material Substances 0.000 claims abstract description 4
- 241000588624 Acinetobacter calcoaceticus Species 0.000 claims description 11
- 239000002253 acid Substances 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 2
- 238000012360 testing method Methods 0.000 abstract description 11
- 230000000694 effects Effects 0.000 abstract description 9
- 239000002304 perfume Substances 0.000 abstract description 6
- IMKHDCBNRDRUEB-UHFFFAOYSA-N Dihydroactinidiolide Natural products C1CCC(C)(C)C2=CC(=O)OC21C IMKHDCBNRDRUEB-UHFFFAOYSA-N 0.000 abstract description 4
- YKVWPZJHENXDAJ-VOTSOKGWSA-N Megastigmatrienone Chemical compound CC1=CC(=O)CC(C)(C)C1\C=C\C=C YKVWPZJHENXDAJ-VOTSOKGWSA-N 0.000 abstract description 4
- NIDGCIPAMWNKOA-WOJBJXKFSA-N Neophytadiene Natural products [C@H](CCC[C@@H](CCCC(C)C)C)(CCCC(C=C)=C)C NIDGCIPAMWNKOA-WOJBJXKFSA-N 0.000 abstract description 4
- POIARNZEYGURDG-FNORWQNLSA-N beta-damascenone Chemical compound C\C=C\C(=O)C1=C(C)C=CCC1(C)C POIARNZEYGURDG-FNORWQNLSA-N 0.000 abstract description 4
- POIARNZEYGURDG-UHFFFAOYSA-N beta-damascenone Natural products CC=CC(=O)C1=C(C)C=CCC1(C)C POIARNZEYGURDG-UHFFFAOYSA-N 0.000 abstract description 4
- IMKHDCBNRDRUEB-LLVKDONJSA-N dihydroactinidiolide Chemical compound C1CCC(C)(C)C2=CC(=O)O[C@@]21C IMKHDCBNRDRUEB-LLVKDONJSA-N 0.000 abstract description 4
- NIDGCIPAMWNKOA-UHFFFAOYSA-N neophytadiene Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(=C)C=C NIDGCIPAMWNKOA-UHFFFAOYSA-N 0.000 abstract description 4
- PQDRXUSSKFWCFA-CFNZNRNTSA-N solanone Chemical compound CC(=O)CC[C@@H](C(C)C)\C=C\C(C)=C PQDRXUSSKFWCFA-CFNZNRNTSA-N 0.000 abstract description 4
- PQDRXUSSKFWCFA-UHFFFAOYSA-N solanone Natural products CC(=O)CCC(C(C)C)C=CC(C)=C PQDRXUSSKFWCFA-UHFFFAOYSA-N 0.000 abstract description 4
- 229930007850 β-damascenone Natural products 0.000 abstract description 4
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 abstract description 3
- 229940114124 ferulic acid Drugs 0.000 abstract description 3
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 abstract description 3
- 235000001785 ferulic acid Nutrition 0.000 abstract description 3
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 abstract description 3
- 241000589291 Acinetobacter Species 0.000 abstract description 2
- 230000000391 smoking effect Effects 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 239000000779 smoke Substances 0.000 abstract 2
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 abstract 2
- 235000012141 vanillin Nutrition 0.000 abstract 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 1
- 125000003118 aryl group Chemical group 0.000 abstract 1
- 150000002009 diols Chemical class 0.000 abstract 1
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 abstract 1
- 239000003205 fragrance Substances 0.000 abstract 1
- 238000009499 grossing Methods 0.000 abstract 1
- 230000007794 irritation Effects 0.000 abstract 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 abstract 1
- 238000007670 refining Methods 0.000 abstract 1
- 229920003266 Leaf® Polymers 0.000 description 38
- 241000894006 Bacteria Species 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 14
- 238000012545 processing Methods 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- 239000007921 spray Substances 0.000 description 10
- 230000015556 catabolic process Effects 0.000 description 9
- 238000006731 degradation reaction Methods 0.000 description 9
- 239000000796 flavoring agent Substances 0.000 description 9
- 244000005700 microbiome Species 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 239000002068 microbial inoculum Substances 0.000 description 7
- 101001065065 Aspergillus awamori Feruloyl esterase A Proteins 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 235000019634 flavors Nutrition 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 4
- 239000003546 flue gas Substances 0.000 description 4
- 235000013355 food flavoring agent Nutrition 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- LHGVFZTZFXWLCP-UHFFFAOYSA-N guaiacol Chemical compound COC1=CC=CC=C1O LHGVFZTZFXWLCP-UHFFFAOYSA-N 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 238000004064 recycling Methods 0.000 description 4
- 241001585562 Callitris columellaris Species 0.000 description 3
- 108090000371 Esterases Proteins 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 150000002083 enediols Chemical class 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 229920005610 lignin Polymers 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000012797 qualification Methods 0.000 description 3
- MYKUKUCHPMASKF-VIFPVBQESA-N (S)-nornicotine Chemical compound C1CCN[C@@H]1C1=CC=CN=C1 MYKUKUCHPMASKF-VIFPVBQESA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- MYKUKUCHPMASKF-UHFFFAOYSA-N Nornicotine Natural products C1CCNC1C1=CC=CN=C1 MYKUKUCHPMASKF-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- -1 ferulic acid ester Chemical class 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
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- 239000001963 growth medium Substances 0.000 description 2
- 229960001867 guaiacol Drugs 0.000 description 2
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- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
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Landscapes
- Manufacture Of Tobacco Products (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Disinfection, Sterilisation Or Deodorisation Of Air (AREA)
Abstract
The invention discloses a spice extract prepared from waste tobacco leaves, and a preparation method and application thereof. Waste tobacco leaves are used as raw materials, nicotine in the waste tobacco leaves is reduced through sequential fermentation of acinetobacter, bacillus pumilus and bacillus subtilis, ferulic acid in the tobacco leaves is converted to generate 4-ethylguaiacol, vanillin and other aroma components, and after the fermentation is finished, the aroma components in the waste tobacco leaves are extracted to prepare the spice extract. The aromatic substances with high content in the extract mainly include benzyl alcohol, solanone, vanillin, BETA-damascenone, dihydroactinidiolide, megastigmatrienone, cembretriene diol and neophytadiene. The result of the perfuming and smoke panel test shows that the perfume extract has the effects of obviously improving and adjusting the cigarette fragrance, refining and smoothing the alcohol and smoke, reducing the irritation, covering the offensive odor and improving the cigarette smoking taste.
Description
Technical field
The invention belongs to the technical field of cigarette additive, more particularly, the present invention relates to a kind of concrete flavouring for tobacco that utilizes the waste tobacco leaf preparation.Simultaneously, the invention still further relates to the preparation method of described concrete and the application in tobacco industry.
Background technology
Tobacco is one of mainstay of China's finance.State Tobacco Monopoly Bureau explicitly points out the 7th of Eleventh Five-Year Plan Chinese cigarette development in science and technology outline, " capture the key technology that affects tobacco aroma quality, perfume quantity ", dwindles and external quality of tobacco gap, competes power to improve the domestic and international market.
In tobacco planting and process, produced the tobacco wastes such as a large amount of discarded tobacco leafs, offal and offal, account for 10%～25% of tobacco leaf total output.At present, considerably less to the comprehensive Utilization Ways of these discarded objects, comprehensive utilization degree is very low, and the overwhelming majority is dropped as refuse, and environment has been caused very large pollution.Yet these discarded objects but are potential valuable resources.Identified now 300 multiple compounds from tobacco leaf, comprised the materials such as alkaloid, protein, amino acid, glucoside, carbohydrate, organic acid, wherein protein, Salanesol, nicotine, amino acid, organic acid, sugar etc. all are of great value materials.Therefore, in today that shortage of resources and problem of environmental pollution become increasingly conspicuous, adopt modern microbial fermentation technology and modern biochemical technology, carry out the comprehensive intensive processing of discarded tobacco leaf resource and the research and development of series of products thereof, significant.
In addition, the upper tobacco leaf that domestic cigarette enterprise is stored at present is more, generally is difficult to morely for tobacco leaf formulation because its nicotine content is higher, not only takies a large amount of storage spaces, and causes a large amount of wastes.This problem has caused the attention of the parties concerned, and the utilization rate that how to improve these tobacco leaves has become the realistic problem that each cigarette enterprise faces.In a word, at present lower to the recycling degree of waste tobacco leaf, how effectively to utilize these tobacco leaf resources, turn waste into wealth, become tobacco enterprise and save cost, the key of raising scientific and technological capability for innovation.
Have a large amount of aroma components in the waste tobacco leaf, simultaneously the forulic acid in the plant cell wall of tobacco leaf with the form of ester exist between lignin and the lignin, form between lignin and the cellulose crosslinked.Feruloyl esterase can be hydrolyzed the ester bond in the ferulic acid ester class, makes forulic acid free out.Forulic acid is a kind of most important precursor substance that generates vanillic aldehyde.Therefore can produce the microorganism of feruloyl esterase for decomposing the waste materials generation forulic acids such as waste tobacco leaf and offal by screening, and then generate the natural flavor material such as guaiacol, vanillic aldehyde, thereby reduce production costs the raising quality of tobacco, reduce the application of essence and flavoring agent product in cigarette in non-tobacco source, realize the recycling of waste tobacco leaf.
Summary of the invention
The inventor's early-stage Study discloses, screening obtains a collection of bacterium with higher nornicotine ability from tobacco leaf surface and tobacco cultivation soil, effect that can be by these bacteriums is the high concentration nicotine degradation in the waste tobacco leaf or the concentration of nicotine is reduced, thereby provides condition for other microorganisms (such as aroma formers) fermentation.Forulic acid is a kind of important precursor that generates vanillic aldehyde, it extensively is present in the plant cell wall with the form of ferulic acid ester, can utilize the microorganism that produces feruloyl esterase that forulic acid is dissociated out from the tobacco leaf cell membrane, and further generated the natural flavor material such as guaiacol and vanillic aldehyde by aroma formers, be used for perfuming cigarette.Thereby reduce production costs the raising quality of tobacco, realize the recycling of waste tobacco leaf.
Based on above-mentioned cognition, the object of the present invention is to provide a kind of novel concrete flavouring for tobacco.
Another object of the present invention provides the preparation method of described concrete flavouring for tobacco.
Further aim of the present invention provides the application of described concrete flavouring for tobacco in production of cigarettes.
Purpose of the present invention is achieved by the following technical programs.
* except as otherwise noted, the percentage that adopts among the present invention is mass percent.
A. the invention provides a kind of concrete flavouring for tobacco, described medicinal extract is as raw material with waste tobacco leaf, ferment successively by acinetobacter calcoaceticus, bacillus pumilus and bacillus subtilis, reduce the nicotine in the waste tobacco leaf, the forulic acid that transforms in the tobacco leaf generates the aroma components such as 4-ethyl guaiacol and vanillic aldehyde, after the fermentation ends, extract the aroma component in the waste tobacco leaf, the preparation concrete.
Microorganism involved in the present invention is as follows:
1. acinetobacter calcoaceticus (
AcinetobacterSp.) ND12: from the tobacco cultivation soil of Kunming, Yunnan Province, separated the bacterium ND12 that a strain has degraded nicotine ability, it has been carried out morphology, Physiology and biochemistry character and 16S rRNA analyzes, qualification result shows that this bacterium belongs to acinetobacter, this bacterial strain called after
AcinetobacterSp. ND12.Should be kept at China Committee for Culture Collection of Microorganisms common micro-organisms center by bacterial strain; Address: China. No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date: on November 5th, 2009; Preserving number is CGMCC No. 3410.Acinetobacter calcoaceticus ND12 has stronger nicotine degradation ability.
2. bacillus pumilus (
Bacillus pumilus) Van35: this bacterium obtains from the separation of flue-cured tobacco blade face, and qualification result shows that this bacterium belongs to bacillus pumilus, this bacterial strain called after
Bacillus pumilusThis bacterial strain has been kept at China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date: on November 5th, 2009; Preserving number is CGMCC No. 3411.Bacillus pumilus Van35 can produce feruloyl esterase.
3. bacillus subtilis (
Bacillus subtilis) Van3: this bacterium obtains from the separation of flue-cured tobacco blade face, and qualification result shows that this bacterium belongs to bacillus subtilis, this bacterial strain called after
Bacillus subtilisThis bacterial strain has been kept at China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date: on 08 16th, 2010; Preserving number is CGMCC No. 4091.Bacillus subtilis Van 3 can transform forulic acid and form the aroma components such as 4-ethyl guaiacol and vanillic aldehyde.
B. the invention provides a kind of preparation method of described concrete, the method adopts following steps:
(1) at first, acinetobacter calcoaceticus ND12 is enlarged fermented and cultured, inoculation waste tobacco leaf fermentation 10 days;
(2) will produce ester bacteria bacillus pumilus Van35 and enlarge fermented and cultured, be inoculated in the waste tobacco leaf after the acinetobacter calcoaceticus ND12 fermentation process fermentation 10 days;
(3) the aroma formers bacillus subtilis is enlarged fermented and cultured, be inoculated into the waste tobacco leaf fermentation 20 days after the bacillus pumilus Van35 fermentation process;
(4) after the fermentation ends, extract the aroma component in the sample, be required concrete.
C. the invention provides concrete as the application of tobacco aromatics using in production of cigarettes.
The concrete that the present invention obtains contains multiple volatile ingredient, and the flavor matter that content is higher mainly contains phenmethylol, solanone, vanillic aldehyde, BETA-damascenone, dihydroactinidiolide, Megastigmatrienone, western cypress three enediols and neophytadiene.The perfuming experimental result of smokeing panel test shows, this concrete has obvious promoting or transferring cigarette perfume, fine and smooth pure and mild flue gas, reduce excitant, cover assorted gas and improve the effect of cigarette odor-absorbing.
The perfuming experimental result of smokeing panel test shows, this concrete has obvious promoting or transferring cigarette perfume, fine and smooth pure and mild flue gas, reduce excitant, cover assorted gas and improve the effect of cigarette odor-absorbing.
Compared with prior art, the present invention has following beneficial effect:
1. adopt distinctive biotechnology, solved a difficult problem that perplexs for a long time production of cigarettes, realized the recycling of waste tobacco leaf.Both effectively avoided the wasting of resources.Reduce again the pollution to environment, simultaneously, can reduce the use amount of the essence and flavoring agent in non-tobacco source, save cost, achieve many things at one stroke.
2. the timely processing of waste tobacco leaf also makes tobacco enterprise saving storage room and time, improves the utilization rate in place, is conducive to the benign development of tobacco enterprise.
3. simple, the operability of production technology, standardization and controllability are strong, are convenient to realize large-scale industrial production, and can turn waste into wealth to have good development prospect.
The specific embodiment
Example only in order to explain the present invention, is not intended to limit the present invention.
Embodiment 1
1. culture medium and condition of culture
The used bacterial strain acinetobacter calcoaceticus ND12 philosophy of experiment is inoculated in the LB fluid nutrient medium of 50 mL, cultivates 12 h for 37 ℃, obtain liquid spawn.Liquid spawn is transferred in the LB fluid nutrient medium of 1 L 37 ℃, 150 r/min shaken cultivation, 12 h by 5% inoculum concentration.Nutrient solution, with the vibration of 20 mL sterilized waters evenly, dilutes 10 times during use and is microbial inoculum at last with sterilized water washing precipitation, centrifugal with centrifugal 10 min of 3500 rpm/min.
The waste tobacco leaf fermentation process
1) nicotine degradation bacterial fermentation
Per 200 g discarded tobacco leafs spray nicotine degradation bacterium (acinetobacter calcoaceticus ND12) microbial inoculum 40 mL(control samples and spray isopyknic sterilized water).Tobacco sample after processing is put into 22 ℃, and fermentation 10 d are used for next step experiment in 60% the climatic chamber.
2) producing the esterase bacterial fermentation processes
The sample of processing through the nicotine degradation bacterial fermentation, spray product feruloyl esterase bacterium (bacillus pumilus Van35) microbial inoculum 40 mL(control samples and spray isopyknic sterilized water).Tobacco sample after processing is put into 22 ℃, and fermentation 10 d are used for next step experiment in 60% the climatic chamber.
3) aroma formers fermentation process
Sample through nicotine degradation bacterium and the processing of product esterase bacterial fermentation sprays aroma formers (bacillus subtilis) microbial inoculum 40 mL(control samples and sprays isopyknic sterilized water).Tobacco sample after processing is put into 22 ℃, and fermentation 20 d in 60% the climatic chamber are used for medicinal extract and extract.
The extraction of tobacco extract
1) will be ground into through the discarded tobacco leaf that microbial bacterial agent mixed culture fermentation is processed powder below 2 mm with pulverizer, and then use the water of 6 ~ 8 times of amounts to finish extraction with offal mix and blend 1 ~ 2 h at ambient temperature.
2) (centrifugal condition is water extract: rotating speed 1500 ~ 3000 rmin through centrifugal filtration
-1, times 15 ~ 20 min) after, with the dilute sulfuric acid of 0.5 M the pH of filtrate is adjusted into 3, finish acidization (times 90 min).
3) under reduced pressure, carry out distillation and concentration through the filtrate of acidifying.
4) concentrate of distillation and concentration is tobacco extract.
Product feature and physicochemical property
This product is rufous stream shape lotion, does not clarify.The physicochemical property of product see the following form (table 1)
The physicochemical property of table 1. biochemical additive
5. perfuming cigarette experiment
The concrete that utilization prepares is processed tobacco leaf leaf group, feeds in raw material in the process by the throwing of adding to of different gradients (5 g/ casees, 10 g/ casees and 30 g/ casees), and with the tobacco leaf that do not add medicinal extract in contrast.
The test of smokeing panel test
After the pipe tobacco of processed group and control group put into climatic chamber inner equilibrium 24 h, make cigarette, sample and the equal random number of contrast.The group of smokeing panel test that is comprised of the expert that smokes panel test carries out the sensory evaluating smoking.
The concrete aroma component detects and the effect evaluation
The aroma component testing result shows: concrete has nearly 70 kinds of volatile ingredients, and the flavor matter that content is higher mainly contains phenmethylol, solanone, vanillic aldehyde, BETA-damascenone, dihydroactinidiolide, Megastigmatrienone, western cypress three enediols and neophytadiene.The perfuming experimental result of smokeing panel test shows, this concrete has obvious promoting or transferring cigarette perfume, fine and smooth pure and mild flue gas, reduce excitant, cover assorted gas and improve the effect of cigarette odor-absorbing.Simultaneously, the application of this concrete has reduced the consumption of essence and flavoring agent in cigarette composition in non-tobacco source, realizes the reasonable utilization of waste tobacco leaf.
Embodiment 2
1. culture medium and condition of culture:With embodiment 1.
The waste tobacco leaf fermentation process
1) nicotine degradation bacterial fermentation
Per 200 g discarded tobacco leafs spray nicotine degradation bacterium (acinetobacter calcoaceticus ND12) microbial inoculum 40 mL(control samples and spray isopyknic sterilized water).Tobacco sample after processing is put into 22 ℃, and fermentation 10 d are used for next step experiment in 60% the climatic chamber.
2) producing esterase bacterium and aroma formers mixed culture fermentation processes
The sample of processing through the nicotine degradation bacterial fermentation, spray each 20 mL(control sample of product feruloyl esterase bacterium (bacillus pumilus Van35) microbial inoculum and aroma formers (bacillus subtilis) microbial inoculum and spray isopyknic sterilized water).Tobacco sample after processing is put into 22 ℃, and fermentation 20 d in 60% the climatic chamber are used for medicinal extract and extract.
The extraction of tobacco extract:Method is with embodiment 1.
The concrete aroma component detects and the effect evaluation
The aroma component testing result shows: concrete has nearly 70 kinds of volatile ingredients, and the flavor matter that content is higher mainly contains phenmethylol, solanone, vanillic aldehyde, BETA-damascenone, dihydroactinidiolide, Megastigmatrienone, western cypress three enediols and neophytadiene.The perfuming experimental result of smokeing panel test shows, this concrete has obvious promoting or transferring cigarette perfume, fine and smooth pure and mild flue gas, reduce excitant, cover assorted gas and improve the effect of cigarette odor-absorbing.Simultaneously, the application of this concrete has reduced the consumption of essence and flavoring agent in cigarette composition in non-tobacco source, realizes the reasonable utilization of waste tobacco leaf.
Because the physiological metabolism activity of different microorganisms is inconsistent, different nornicotine microbial fermentation is processed the nicotine content that makes in the waste tobacco leaf and is obtained in various degree reduction, and the height of the nicotine content in the tobacco leaf has determined to produce energy for growth and the metabolic activity of ester bacterium and aroma formers, causes the final concrete that obtains to have different flavouring essence qualities.
Claims (3)
1. concrete flavouring for tobacco, it is characterized in that, this medicinal extract is to be prepared by following method: with waste tobacco leaf as raw material, ferment with acinetobacter calcoaceticus, bacillus pumilus and bacillus subtilis successively, reduce the nicotine in the waste tobacco leaf, transform 4-ethyl guaiacol and vanillic aldehyde aroma component that the forulic acid in the tobacco leaf generates, after the fermentation ends, extract the aroma component in the waste tobacco leaf, be required concrete flavouring for tobacco; The preserving number of wherein said acinetobacter calcoaceticus is CGMCC No.3410; The preserving number of described bacillus pumilus is CGMCC No.3411; The preserving number of described bacillus subtilis is CGMCC No.4091.
2. the preparation method of a concrete flavouring for tobacco claimed in claim 1, the method adopts following step:
(1) described acinetobacter calcoaceticus is enlarged fermented and cultured, inoculation waste tobacco leaf fermentation 10 days;
(2) described bacillus pumilus is enlarged fermented and cultured, be inoculated in the gains of above-mentioned steps 1, fermented 10 days;
(3) the aroma formers bacillus subtilis is enlarged fermented and cultured, be inoculated in the gains of above-mentioned steps 2, fermented 20 days;
(4) after the fermentation ends, extract the aroma component in the sample, be required concrete.
3. concrete flavouring for tobacco claimed in claim 1 is used for perfuming cigarette.
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