CN106723311B - Application of the bacillus subtilis in degrading tobacco product in terms of cellulose - Google Patents
Application of the bacillus subtilis in degrading tobacco product in terms of cellulose Download PDFInfo
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- CN106723311B CN106723311B CN201611102085.XA CN201611102085A CN106723311B CN 106723311 B CN106723311 B CN 106723311B CN 201611102085 A CN201611102085 A CN 201611102085A CN 106723311 B CN106723311 B CN 106723311B
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- bacillus subtilis
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/20—Biochemical treatment
Abstract
The invention belongs to technology field of tobacco preparation, specifically application of a kind of bacillus subtilis xp bacterial strains in degrading tobacco product in terms of cellulose.The bacillus subtilis is bacillus subtilis xp bacterial strains, which is now preserved in China typical culture collection center, and deposit number is CCTCC No：M 2011452；The bacterial strain is for reducing content of cellulose in tobacco product；In application, bacillus subtilis xp bacterial strains are prepared into microbial inoculum, it is uniformly sprayed on tobacco product surface by the mass ratio of tobacco product 1.2%, then 30 ~ 35 DEG C, under 55 ~ 65% damp conditions, 40 ~ 50d of storage alcoholization.The selection that the present invention passes through specific bacillus subtilis strain, it was demonstrated that it can effectively reduce content of cellulose in tobacco product, promote tobacco grade, ensuring to refine effect simultaneously and then accelerating tobacco mellowing process.
Description
Technical field
The invention belongs to technology field of tobacco preparation, and specifically a kind of bacillus subtilis xp bacterial strains are in degrading tobacco product
Application in terms of cellulose.
Background technology
Bacillus subtilis is one kind of Bacillus, is widely distributed in soil and the organic matter of corruption, easily withered
It is bred in grass leaching juice.Its bacterium colony rough surface is opaque, dirty white or yellowish, available protein, a variety of sugar and starch, point
It solves tryptophan and forms indoles, be widely used in genetics research.In addition, bacillus subtilis thalline itself can synthesize alphalise starch
The enzymes such as enzyme, neutral proteinase, lipase, cellulase have good application in terms of the complicated macromolecular compound of degradation
Foreground, therefore bacillus subtilis is increasingly becoming the hot spot for being concerned and studying in recent years.
Tobacco fermentation is also known as tobacco mellowing, and its essence is under certain temperature and damp condition, promote tobacco leaf physics and chemistry special
Property occur profound change process, i.e., will be unfavorable for the macromolecular substances component of quality of tobacco（Including protein, starch, pectin
Deng）Degradation and Transformation is at the small molecule component for being conducive to be promoted quality of tobacco（Reduced sugar, amino acid etc.）, and then reach and overcome new cigarette
Bad quality, improvement and the purpose for improving inherent quality of tobacco leaf and presentation quality.
Tobacco mellowing be cigarette industry improve product quality a kind of preliminary working method and Cigarette processing in it is particularly important
A link.Alcoholized tobacco or fermentation process are generally divided into two kinds of artificial ageing and natural alcoholization.Artificial ageing's speed is fast,
Period is short, but it is poor to refine effect；Natural alcoholization is to be deposited in tobacco leaf in reserve storage by certain requirement, under field conditions (factors) into
Capable alcoholization, time are generally 1 ~ 3 year.Although natural alcoholization effect is preferable, there is also some shortcomings, such as：1）Alcoholization process
In, tobacco leaf is easy to grow insect pest；2）Due to the variation of temperature, humidity, tobacco leaf, which easy tos produce, to go mouldy；3）The alcoholization period is longer, easily
Cause the occupancy of inventory's tobacco leaf overstock with substantial contribution.
Cellulose, hemicellulose and lignin are the main matters for constituting plant cell wall, account for the 50% of all polysaccharide of plant
More than.Some researches show that total cell wall substance content is even accounted for 20 ~ 30% or so in offal and reconstituted tobacoo in tobacco leaf
1/3 or more of gross mass.Cellulose has very directly the flammability of tobacco leaf and the flavor and taste of the Filling power of pipe tobacco and cigarette
Influence, it will usually a kind of excitement of spine and a kind of bad smell of similar burning paper are generated during burn and suck, from
And side effect is generated to cigarette quality.
Since cellulosic material structure is relatively stable, thus durings tobacco natural alcoholization, cigarette storage etc., content
Variation is little, and degradation rate is extremely slow.For this defect, inoculating microbe preparation or enzyme preparation are developed and used, is accelerated
The macromolecular substances such as the cellulose in degradation tobacco leaf shorten the tobacco mellowing period to accelerate tobacco mellowing process, look forward to cigarette
Industry improves cigarette quality, saves production cost, increases economic efficiency with the meaning of reality extensively.
Invention content
By the selection of specific bacillus subtilis xp bacterial strains, present invention aims at provide bacillus subtilis xp bacterium
New opplication of the strain in degrading tobacco product in terms of cellulose, to obtain the skill of content of cellulose in relatively stable reduction tobacco
Art effect, and then accelerate the alcoholization process of tobacco leaf, shorten the alcoholization period.
Details are as follows for the technical solution that the application is taken.
Application of the bacillus subtilis in degrading tobacco product in terms of cellulose, the bacillus subtilis are withered grass bud
Spore bacillus xp bacterial strains, Classification And Nomenclature areBacillus subtilisXp, the bacterial strain are now preserved in Chinese Typical Representative culture guarantor
Tibetan center, preservation address are：Wuhan City, Hubei Province Wuchang District Luo Jia Shan road 16（Wuhan University）, the deposit date is：2011
On December 9, in, deposit number are CCTCC No：M 2011452.
Application of the bacillus subtilis in degrading tobacco product in terms of cellulose, is used in degrading tobacco product
When cellulose, it is prepared into microbial inoculum first, then with the mass ratio of tobacco product 1 ~ 2%（It is preferred that 1.2%）Uniformly it is sprayed on tobacco
Product surface, finally under 30 ~ 35 DEG C, 55 ~ 65% damp conditions, 40 ~ 50d of storage alcoholization；
The microbial inoculum, the OD for specially using LB liquid medium to prepare600=1.0 ~ 5.0 suspending nutrient solution；
The tobacco product, concretely fresh picking or the different cultivars after toasted, drying, fermentation, different production
The complete stool tobacco on ground or portion of tissue organ, still further, specific for example, Flue-cured.
In general, the present invention is by the selection of specific bacillus subtilis strain, after being expanded and being prepared into microbial inoculum,
When applied to tobacco product, content of cellulose in tobacco product can be effectively reduced, improves its effect of smokeing panel test, promotes tobacco grade,
Ensure to refine effect simultaneously and then accelerates tobacco mellowing process.Due to the present invention can in a short time, at lower cost and compared with
High efficiency preferably improves the sucking quality of tobacco, in addition its operating process is relatively simple, so that the application is prepared in cigarette
Field shows preferable application value and application prospect.
Description of the drawings
Fig. 1 is that bacterium solution preparation of the present invention sprays influence of the after effect time to cellulose degradation rate in tobacco mellowing.
Specific implementation mode
Explanation is further explained to the application with reference to embodiment, before introducing specific embodiment, with regard to following implementations
Part Experiment background involved in example is briefly discussed below.
Experiment material：
For trying tobacco leaf：The table mountain of Henan in 2015 C3F tobacco leaves, are provided by China Tobacco Henan Industrial Co., Ltd；
Bacillus subtilis xp bacterial strains employed in following embodiments, Classification And Nomenclature areBacillus subtilis.
Xp, the bacterial strain are now preserved in China typical culture collection center, and deposit number is CCTCC No：M 2011452；Due to this
Bacterial strain is that applicant voluntarily screens acquisitions, and has had in other patents disclosed, thus belongs to the microorganism that can disclose acquisition
Resource, the application belong to the new application patent application of the bacterial strain；
Experiment reagent：
LB liquid medium is formulated conventionally and sterilizes（121 DEG C of 15 min of heat preservation）, specific formula is：Every 1000 mL
Contain in deionized water：5 g of yeast extract, peptone 10 g of 10 g, NaCL, with NaOH solution tune pH=7.0；
Major experimental equipment：
UV-2550 ultraviolet specrophotometers are Shimadzu Products.
Embodiment
It is appropriate for cellulose in degrading tobacco product, to need bacillus subtilis xp bacterial strains first to carry out the bacterial strain
Amplification cultivation, being prepared into can just be applied containing a certain number of active microbial inoculums, and the specific preparation method of microbial inoculum is as described below.
（1）Actication of culture utilizes oese picking slant medium（It is formulated same LB liquid medium, only in addition addition
15g/L agar powders）Upper strain, is placed in LB liquid medium the culture that suspends, and 30 DEG C of culture 12h carry out actication of culture；
（2）Strain expands, by step（1）Bacterium solution is forwarded in LB liquid mediums after middle actication of culture, 35 DEG C of shaking table trainings
It supports to OD600=3.5 or so, it can be used as degrading tobacco product microbial inoculum after suspending uniformly.
When above-mentioned microbial inoculum is specifically used for cellulose in degrading tobacco product, the mass ratio by tobacco product 1.2% is uniform
It is sprayed on tobacco product surface, then 40 ~ 50d of storage alcoholization or so under 30 DEG C, 55 ~ 65% damp conditions.
For the specific degraded cellulose effect of determination, specifically with sheet（3cm×3cm）Tobacco leaf（Remove the tobacco leaf of offal）As
Tobacco product（Every group of 200g）Alcoholization processing has been carried out, while blank control group is set（CK）（By the mass ratio of tobacco 1.2%
Spray distilled water）, after storing 45d after spraying, cellulose degradation rate in tobacco leaf is measured and carries out sensory quality assessment.
Cellulose degradation rate measures
Cellulose degradation rate uses flow analysis measuring method, is as follows：
（1）Accurate tobacco leaf powder after weighing 0.25g, the crushing of 40 mesh, is placed in 100 mL conical flasks, 25 mL, 5% matter is added
The acetum of score is measured, 30 min of shaking table oscillation extraction extracts the water-soluble sugar contained by sample itself；
（2）It is filtered with G3 funnels, filter residue distilled water flushing 3 times, every time 50 mL；
（3）Filter residue is transferred in 100 mL conical flasks, citric acid/lemon that 60 mL contain 0.25 g fiber rope enzymes is added
Lemon acid trisodium buffer solution (pH=4.7) is then placed in 37 DEG C of constant-temperature tables and hydrolyzes 24 h；
（4）Sample is cooled to room temperature after hydrolysis, filters, filtrate is settled to 100 mL with distilled water, utilizes stream
Glucose in dynamic analysis-e/or determining sample liquid is multiplied by conversion coefficient 0.9 to measure glucose content and (hydrolyzes equation by cellulose
Formula is calculated) up to content of cellulose；
（5）It after detection obtains each sample content of cellulose, is compared with initial control group tobacco leaf fiber cellulose content, conversion is corresponding
Cellulose degradation rate.
Measurement result is as shown in Figure 1.
It will be seen from figure 1 that as processing time increases, the content of cellulose of tobacco sample has no bright in blank control group
Aobvious variation；And after microbial inoculum is handled, in processing group tobacco sample content of cellulose occur significant change, and with processing when
Between extend, cellulose degradation rate persistently increases, and when storage alcoholization processing 40 days, cellulose degradation rate was up to peak in tobacco leaf
（11.5%）, tend towards stability later；
Tobacco product sensory quality assessment
Within the different storages alcoholization period, appropriate tobacco leaf chopping is taken, routinely prepares cigarette sample, constant temperature and humidity balance
Afterwards, by professional judging panel to the aroma quality of cigarette（A）, perfume quantity（B）, concentration（C）, soft and fine degree（D）, pleasant impression（E）, miscellaneous gas（F）, thorn
Swash property（G）Etc. indexs evaluated, each index press 9 points system give a mark, sensory evaluating smoking's total score（T）=（A+B）× 2.3 + C
× 1.5 + D + E + F + G。
Specific evaluation result is as shown in the table：
CK is the control tobacco leaf for spraying equivalent distilled water and storing 0 day in table（It should be noted that smokeing panel test after storing 45 days
As a result roughly the same with this data, so omitting）.
Upper table data analysis can be seen that compared with sample original state, after microbial inoculum is handled, when refining with storage
Between extend, the aesthetic quality of tobacco product can obtain the improvement of duration.After microbial inoculum is handled, storage is smoked panel test when refining 40 days
Total score reaches peak, and total score increases 1.06 points compared to control, in conjunction with above-mentioned tobacco leaf fiber element degradation rate result, it may be determined that bacterium
The optimization process time of liquid formulation is 40 days, can make tobacco leaf fiber element degradation 11.5% at this time, total score of smokeing panel test is improved compared to control
1.06 points, tobacco leaf is made to be integrally improved a grade, has dramatically speeded up the alcoholization process of tobacco leaf, thus the present invention is improving tobacco leaf
Overall quality has a good application prospect in terms of shortening the alcoholization period.
Claims (3)
1. application of the bacillus subtilis in degrading tobacco product in terms of cellulose, which is characterized in that the bacillus subtilis
Bacterium is bacillus subtilis xp bacterial strains, and Classification And Nomenclature isBacillus subtilisXp, the bacterial strain are now preserved in Chinese allusion quotation
Type culture collection, deposit number are CCTCC No：M 2011452；The bacterial strain is for reducing cellulose in tobacco product
Content；The tobacco product is fresh picking or toasted, dries, the complete stool cigarette of the different cultivars after fermentation, different sources
Grass or in which portion of tissue organ.
2. application of the bacillus subtilis as described in claim 1 in degrading tobacco product in terms of cellulose, which is characterized in that
Bacillus subtilis xp bacterial strains are prepared into microbial inoculum, tobacco product table is uniformly sprayed on by the mass ratio of tobacco product 1.2%
Face, then 30 ~ 35 DEG C, under 55 ~ 65% damp conditions, 40 ~ 50d of storage alcoholization；
The microbial inoculum, the OD for specially using LB liquid medium to prepare600=1.0 ~ 5.0 suspending nutrient solution.
3. application of the bacillus subtilis as claimed in claim 1 or 2 in degrading tobacco product in terms of cellulose, feature exist
In the tobacco product is Flue-cured.
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107488613B (en) * | 2017-09-12 | 2020-09-29 | 河南中烟工业有限责任公司 | XC-19-1 strain for degrading cellulose in tobacco leaves and application thereof |
| CN108081429A (en) * | 2017-11-16 | 2018-05-29 | 湖州南浔双杨木业有限公司 | A kind of ultralow water absorption rate bamboo and woods fiber decorative panel |
| CN108115805A (en) * | 2017-11-16 | 2018-06-05 | 湖州南浔双杨木业有限公司 | A kind of high-strength bamboo wood-fibred decorative panel |
| CN109602070B (en) * | 2018-10-29 | 2021-11-19 | 河南中烟工业有限责任公司 | Application of SMXP-58 strain in reducing content of cellulose in tobacco stems |
| CN109527640B (en) * | 2018-10-29 | 2021-11-19 | 河南中烟工业有限责任公司 | Application of SMXP-58 strain in degrading cellulose in tobacco leaves |
| CN110693075B (en) * | 2019-10-12 | 2021-10-08 | 中国科学院青岛生物能源与过程研究所 | Four-stage fermentation method of cigar core tobacco leaves |
| CN110876485A (en) * | 2019-11-21 | 2020-03-13 | 于曦华 | Agilawood cigarette and processing method thereof |
| CN110876486A (en) * | 2019-11-21 | 2020-03-13 | 于曦华 | Rosemary cigarette and processing method thereof |
| CN112043002A (en) * | 2020-09-07 | 2020-12-08 | 湖北中烟工业有限责任公司 | Application of Bacillus belgii in degrading cellulose in tobacco flavor raw material |
| CN112971191A (en) * | 2021-02-05 | 2021-06-18 | 河南中烟工业有限责任公司 | Tobacco redrying method capable of preserving fragrance and removing impurities |
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| JPS54160800A (en) * | 1978-06-09 | 1979-12-19 | Japan Tobacco Inc | Method for improving flavor and taste of tobacco |
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| CN102986391A (en) * | 2011-09-08 | 2013-03-27 | 郑州大学 | Method for improving content and aromas of tobacco secondary metabolite products |
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