CN107034162A - A kind of bacillus BA 01 and its application in terms of tobacco - Google Patents
A kind of bacillus BA 01 and its application in terms of tobacco Download PDFInfo
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- CN107034162A CN107034162A CN201710343989.XA CN201710343989A CN107034162A CN 107034162 A CN107034162 A CN 107034162A CN 201710343989 A CN201710343989 A CN 201710343989A CN 107034162 A CN107034162 A CN 107034162A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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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
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Abstract
The invention discloses a kind of bacterial strain for increasing aroma of tobacco shreds and accelerating pipe tobacco ageing, the bacterial strain obtains from tobacco leaf surface separation screening and is named as bacillus BA 01, and the method for being prepared into biological agent and its application is disclosed, belong to microorganism and its applied technical field.The biological agent viable count is not less than 5.0 × 109CFU/g.The biological agent of the present invention can increase the simultaneously fragrance of fine and smooth pipe tobacco, reduction excitant, the miscellaneous gas of decrease, purification pleasant impression, pipe tobacco sucking quality is lifted.
Description
Technical field
The present invention relates to field of agricultural microbial technology and tobacco ageing technical field, specifically, the present invention relates to profit
With a kind of bacterial strain manually separated, and its application in pipe tobacco ageing process.
Background technology
Dry the tobacco bred that cigarette is earliest incoming China, specific cultivation step, natural conditions and the modulation of different regions
The difference of details determines that drying smoking set has more unique quality and local flavor.The modulation system of airing tobacco is the nature system of drying in the air,
Modulated environments more gently make it that former cigarette carries a variety of quality defects to some extent, and such as tobacco leaf life is blue or green and miscellaneous gas is heavier, stimulate
Property larger cause that flue gas is soft, aroma quality is dull and perfume quantity is not enough etc. and do not reach also the quality for being directly used in industrial processes will
Ask, it is therefore desirable to carry out fermentation process to improve these quality defects.
During the fermentation, in tobacco leaf starch and cell wall substance (protein, cellulose, pectin etc.) microorganism,
Under the collective effect of enzyme, gradually it is dissipated and decomposition and inversion, so as to reduce tobacco leaf cells wall material to the bad of quality of tobacco
Influence.Tobacco leaf surface is isolated beneficial bacterial strain and is prepared into biological agent or complex enzyme formulation etc. by domestic Many researchers to be sprayed to
The purpose for improving quality of tobacco is reached in tobacco leaf.Huang Jingwen etc. has found that bacillus can effectively improve quality of tobacco；Yang Peixiang etc.
Isolated bacillus subtilis is added on pipe tobacco and clearly enhances cigarette perfume, miscellaneous gas and excitant is reduced；Xu Guowang
It is aged Deng by adding complex enzyme formulation to tobacco leaf, can also effectively improves quality of tobacco.At present, biological agent is in tobacco leaf
The time of application in fermentation is not also long, and scope is not also wide, and is applied on flue-cured tobacco, and it is contemplated that searching out
It is a kind of to improve the dominant strain added as biological agent for drying tobacco quality.
The content of the invention
For the weak point of above-mentioned technology, drying cigarette can be improved the technical problem to be solved in the present invention is to provide one kind old
Change speed and lift the dominant strain of its sucking quality, i.e. a bacillus BA-01, and be prepared into biological agent and add
It is added to progress ageing application in pipe tobacco.Described biological agent can accelerate pipe tobacco ageing, increase aroma quality, reduce excitant, subtract
Weak miscellaneous gas, the sucking quality for significantly improving pipe tobacco.
The present invention provides the screening of Congjiang Seat color " black tiger " cured tobacco leaf surface has industrialization in terms of tobacco leaf flavouring
The Bacillus strain BA-01 of application prospect, it is isolated to be coated on meat soup solid medium after being enriched with through culture medium, and passes through
Zytase, pectase, cellulase, protease, amylase screening and culturing medium determine its enzymatic productivity size, as a result show this
Bacterial strain possesses the ability for producing a variety of enzymes, is a kind of potential airing tobacco flavouring upgrading bacterium, and the artificial aging to airing tobacco has
Facilitation.
Jiangxi characteristic " black tiger " cured tobacco leaf is the Guangchang flue-cured tobacco cultivars for originating from Jiangxi Province's Fuzhou Shi.
It is a kind of to improve the Bacillus strain BA-01 for drying tobacco quality, it is characterised in that Gram's staining is positive, bar
Shape, size is 2.0-2.5 μm of 0.5-0.6 μ ms, brood cell's intermediate green or end life, and peritrichous can be moved.
Above-mentioned bacillus BA-01 physio-biochemical characteristics, it is characterised in that NaCl tolerances 2%-7%；Hydrogen peroxide
Enzyme, citrate produce experiment, V.P experiments using experiment, Starch Hydrolysis experiment, casein hydrolysis experiment, indoles, are the positive；
Growth test is feminine gender under pH5.7.
Described bacillus BA-01 (Bacillus amyloliquefaciens), is preserved in Guangdong Province's microbial bacteria
Collection is planted, address is 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100, and the preservation time is on April 26th, 2017, is protected
It is GDMCC No.60171 to hide numbering.
The bacillus BA-01 of present invention application process, it is characterised in that main flow is fermented and cultured, collects bacterium
Body, prepares bacteria suspension, adds Trehalosc protection agent, and vacuum freeze drying is sprayed on drying pipe tobacco into pulvis, carries out artificial old
Change.
Bacillus BA-01 fermentation medium composition：Peptone 1%, dusty yeast 0.5%, sodium chloride 0.5%, grape
2%, pH7.2 of sugar.Optimal conditions of fermentation：42 DEG C, 200rpm, 24h.
The collection of Bacillus strain BA-01 fermentation thalli after fermented and cultured terminates, 7000 rpm-10000rpm from
Heart 7min-15min makes bacterial sediment, bacteria suspension is obtained after washing 1-5 thalline using sterile saline, to above-mentioned bacteria suspension
The middle trehalose that adds makes it be 0.5%-2% in the final mass concentration of bacteria suspension, is placed in -80 DEG C of refrigerators and freezes, makes
It is without working fluid, then is positioned in vacuum freeze drier, is freeze-dried 8-15h, and it is standby to be preserved in 4 DEG C of refrigerators, produces life
Thing preparation.
Bacillus pumilus BA-01 biological agent viable count reaches 109-1011CFU/g, takes 0.001g- when using
0.015g biological agents are dissolved in 1ml-10ml sterilized waters, and it is uniformly sprayed on to 10g-40g and dries pipe tobacco.Artificial aging is dried
The condition of tobacco leaf is 20 DEG C -50 DEG C for the treatment of temperature, relative humidity 40%-70%, 8-12 days processing time.
The identification of strain classification status of the present invention, reference《Common bacteria system identification handbook》Content.According to
Its morphological feature and physiological and biochemical property, and 16SrRNA gene order comparison results, through multinomial taxonomic identification, the present invention divides
From bacterial strain be one plant of bacillus BA-01 (Bacillus amyloliquefaciens).
The advantage of the invention is that：The bacillus BA-01 of offer is that a kind of excellent cigarette uses flavouring microorganism, can be significantly
The artificial aging effect for drying cigarette is improved, it is particularly more sufficient on fragrance, excitant is reduced, is also obtained on miscellaneous gas
Weaken, while shortening drying cigarette aging period, reduce the production cost for drying cigarette.
Brief description of the drawings
Fig. 1：Bacillus BA-01 Gram-stained microphoto (× 100).
Fig. 2：Transparent circles of the Bacillus strain BA-01 on production xylanase plate.
Fig. 3：Transparent circles of the Bacillus strain BA-01 on cellulase-producing flat board.
Fig. 4：Transparent circles of the Bacillus strain BA-01 on production pectase flat board.
Fig. 5：Transparent circles of the Bacillus strain BA-01 on production protease flat board.
Fig. 6：Transparent circles of the Bacillus strain BA-01 on production amylase flat board.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, by the following examples and experiment number
According to the present invention is described in more detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention,
It is not intended to limit the present invention.
Embodiment 1:Bacillus BA-01 separation and producing enzyme analysis.
10-40g Jiangxi characteristic " black tiger " cured tobacco leaf is shredded with sterilized scissors, be immersed in 50mL-200mL without
In bacterium water, in shaking 30min on 100rpm-300rpm shaking tables, the 0.1mL cured tobacco leaf suspensions are taken after concussion is uniform, are diluted
It is coated on LB flat boards (peptone 10g/L, dusty yeast 5g/L, sodium chloride 10g/L).The microbionation of separation is analyzed to producing enzyme and cultivated
Base.Solid medium including producing zytase：Xylan 1g, meat soup 1.8g, agar 2g, natural pH；Produce the solid of pectase
Culture medium：Pectin 0.4g, peptone 1g, NaCl 0.05g, agar 2g, pH7.2-7.4；Produce the solid medium of cellulase：Carboxylic
Sodium carboxymethylcellulose pyce 0.8g, meat soup 1.8g, agar 2g, natural pH；Produce the solid medium of protease：Peptone 1g, beef extract
1g, NaCl 0.5g, casein 1g, agar 2g, adjust pH7.2-7.4；Produce the solid medium of amylase：Peptone 1g, ox
Meat extract 0.3g, NaCl 0.5g, soluble starch 0.2g, agar 2g, adjust pH7.2-7.4.
Cultivate and transparent circle and bacterium colony on Congo red measure zytase, cellulase, pectase flat board are utilized after 10h-40h
Diameter ratio, iodine solution determines transparent circle and colony diameter ratio on starch flat board, directly observes transparent circle and bacterium colony on protease flat board
Diameter ratio (HC values).As shown in table 1, isolated bacterial strain has produces zytase, cellulase, pectase, starch simultaneously
The ability of enzyme and protease.
Table 1：Strain enzyme-producing situation analysis
Embodiment 2:Bacillus BA-01 form and the identification of physiological and biochemical property.
Reference《Common bacteria system identification handbook》Experimental method carry out, detect its Gram's staining situation, thalline is big
Small and form, there is atrichia and gemma, NaCl tolerances, catalase test, Starch Hydrolysis, cellulose hydrolysis, xylan
Hydrolysis, pectin enzyme hydrolysis, casein degraded, V-P experiments, citrate test, indoles is produced, growth test under pH5.7.
Test result indicates that：Gram's staining is positive, shaft-like, and size is 2.0-2.5 μm of 0.5-0.6 μ ms, Ya Baozhong
Between raw or end life, peritrichous can move.Its physiological and biochemical property is：NaCI tolerances 2%-7%, catalase, lemon
Hydrochlorate utilizes experiment, Starch Hydrolysis experiment, casein hydrolysis experiment, cellulose hydrolysis, xylan hydrolysis, pectin enzyme hydrolysis, indoles
Produce experiment, V.P experiments, be the positive.Growth test is feminine gender under pH5.7.
Embodiment 3:The 16SrDNA of bacillus amplification and sequencing.
Bacillus strain BA-01 is inoculated in broth bouillon, 100rpm-300rpm, 20 DEG C of -40 DEG C of concussion and cultivates
10-30 hours, thalline is collected by centrifugation.Genomic DNA is extracted using bacterial genomes extracts kit, is drawn with 16SrDNA is general
Thing forward primer 27F (5 `-AGAGTTTGATCCTGGCTCAG-3`) and reverse primer 1492R (5`-GGTTAC
CTTGTTACGACTT-3`), its 16SrDNA is expanded, and the product of amplification is handed in into the limited public affairs of the raw work bioengineering in sea
Department is sequenced.PC amplification conditions are:94℃4mi n；94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 90s, 30 circulations；72℃10min.
PCR primer length is the Bacillus amyloliquefaciens S QR9 (bacillus in 1484bp, with ncbi database
SQR9) similitude is 99%.
Embodiment 4:The fermentation of bacillus.
The Bacillus strain of the present invention is seeded on meat soup solid plate, 10-20h are activated in 20 DEG C -40 DEG C, then
The Bacillus strain BA-01 of activation is transferred in fermentation medium and carries out shake flask fermentation, optimal conditions of fermentation is：Temperature is
42 DEG C, shaking speed is 200rpm, and fermentation time is 24h.
Embodiment 5:Bacillus BA-01 biological agents add the artificial aging selected Xuanwei piece and dry pipe tobacco.
Bacillus strain BA-01 zymotic fluid is taken, is centrifuged, is taken precipitation to be prepared into biological agent, weigh 0.001g-
0.015g biological agent is dissolved in 1ml-10ml sterilized waters, sets distilled water to compare, Bacillus strain biological agent is sprayed
It is applied to the piece choosing of 10-40g Xuanweis to dry on pipe tobacco, pipe tobacco is through ultraviolet irradiation 10min-40min before sprinkling, in constant temperature and humidity culture
Temperature is set as 20 DEG C -50 DEG C, relative humidity 40%-70%, artificial aging 8-12 days in case.It is aged after terminating again through ultraviolet
10min-40min is irradiated, to stop the effect for adding microorganism, sample presentation carries out expert sensory and smoked panel test, and smoking result is shown in Table 2 and table
3, after drying pipe tobacco through the bacterial strain artificial aging that separates of the present invention as can be seen from the table, its tobacco leaf smoke panel test fragrance it is more sufficient,
Finer and smoother, excitant is reduced, and miscellaneous gas weakens, and pleasant impression is neter more comfortable.
Table 2：Bacillus BA-01 selects Xuanwei piece pipe tobacco artificial aging sensory evaluating smoking's result
Table 3：Bacillus BA-01 selects Xuanwei piece pipe tobacco artificial aging sensory evaluating smoking to describe
Embodiment 6:Bacillus BA-01 biological agents dry choosing to Chenzhou the artificial aging of pipe tobacco.
Bacillus strain BA-01 zymotic fluid is taken, is centrifuged, is taken precipitation to be prepared into biological agent, weigh 0.001g-
0.015g biological agent is dissolved in 1ml-10ml sterilized waters, sets distilled water to compare, Bacillus strain biological agent is sprayed
It is applied to 10-40g Chenzhou to dry on pipe tobacco choosing, pipe tobacco is through ultraviolet irradiation 10min-40min before sprinkling, in constant temperature and humidity culture
Temperature is set as 20 DEG C -50 DEG C, relative humidity 40-70%, artificial aging 8-12 days in case.Ageing is shone through ultraviolet again after terminating
10min-30min is penetrated, to stop the effect for adding microorganism, sample presentation carries out expert sensory and smoked panel test, and smoking result is shown in Table 4 and table 5,
After drying pipe tobacco through the bacterial strain artificial aging that separates of the present invention as can be seen from the table, its tobacco leaf fragrance of smokeing panel test is more sufficient, more
Exquisiteness, excitant is reduced, and miscellaneous gas weakens, and pleasant impression is neter more comfortable.
The bacillus BA-01 of the present invention of table 4 selects pipe tobacco artificial aging sensory evaluating smoking's result to Chenzhou handle
The bacillus BA-01 of the present invention of table 5 selects cigarette pipe tobacco artificial aging sensory evaluating smoking to describe Chenzhou handle
Tobacco leaf carry out common artificial aging for up to the several years, result of study shows the old of biological agent of the present invention
Change effect and be better than control group, can substantially shorten the aging period for drying cigarette.Being embodied in can increase and fine and smooth pipe tobacco
Fragrance, reduction excitant, the miscellaneous gas of decrease, purification pleasant impression；Meanwhile, bacillus BA-01 is detected by gas chromatography combined with mass spectrometry
Material change after being aged on added to two kinds of pipe tobaccos, it is found that fragrance matter is generated after ageing includes 5 hydroxymethyl furfural, hydroxyl
Base maltol, 5 methyl furfural, furfuryl alcohol, furfural, 2 (5H)-furanones, 4- cyclopentene -1,3- diketone are significantly improved compared with control group,
And the content of harmful substance nicotine, formic acid, acetic acid, propionic acid, phenol is reduced, it is effectively improved the sucking property of airing tobacco.
1 gagtttgatc ctggctcagg acgaacgctg gcggcgtgcc taatacatgc aagtcgagcg
61 gacagatggg agcttgctcc ctgatgttag cggcggacgg gtgagtaaca cgtgggtaac
121 ctgcctgtaa gactgggata actccgggaa accggggcta ataccggatg gttgtttgaa
181 ccgcatggtt cagacataaa aggtggcttc ggctaccact tacagatgga cccgcggcgc
241 attagctagt tggtgaggta acggctcacc aaggcaacga tgcgtagccg acctgagagg
301 gtgatcggcc acactgggac tgagacacgg cccagactcc tacgggaggc agcagtaggg
361 aatcttccgc aatggacgaa agtctgacgg agcaacgccg cgtgagtgat gaaggttttc
421 ggatcgtaaa gctctgttgt tagggaagaa caagtgccgt tcaaataggg cggcaccttg
481 acggtaccta accagaaagc cacggctaac tacgtgccag cagccgcggt aatacgtagg
541 tggcaagcgt tgtccggaat tattgggcgt aaagggctcg caggcggttt cttaagtctg
601 atgtgaaagc ccccggctca accggggagg gtcattggaa actggggaac ttgagtgcag
661 aagaggagag tggaatttcc acgtgtagcg gtgaaatgcg tagagatgtg gaggaacacc
721 agtggcgaag gcgactctct ggtctgtaac tgacgctgag gagcgaaagc gtggggagcg
781 aacaggatta gataccctgg tagtccacgc cgtaaacgat gagtgctaag tgttaggggg
841 tttccgcccc ttagtgctgc agctaacgca ttaagcactc cgcctgggga gtacggtcgc
901 aagactgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa
961 ttcgaagcaa cgcgaagaac cttaccaggt cttgacatcc tctgacaatc ctagagatag
1021 gacgtcccct tcgggggcag agtgacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg
1081 agatgttggg ttaagtcccg caacgagcgc aacccttgat cttagttgcc agcattcagt
1141 tgggcactct aaggtgactg ccggtgacaa accggaggaa ggtggggatg acgtcaaatc
1201 atcatgcccc ttatgacctg ggctacacac gtgctacaat ggacagaaca aagggcagcg
1261 aaaccgcgag gttaagccaa tcccacaaat ctgttctcag ttcggatcgc agtctgcaac
1321 tcgactgcgt gaagctggaa tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt
1381 tcccgggcct tgtacacacc gcccgtcaca ccacgagagt ttgtaacacc cgaagtcggt
1441 gaggtaacct ttatggagcc agccgccgaa ggtgacagat ttgg
Claims (3)
1. a bacillus BA-01 (Bacillus amyloliquefaciens), Guangdong is preserved on April 26th, 2017
Culture Collection is saved, deposit number is GDMCC No.60171.
2. applications of the bacillus BA-01 as claimed in claim 1 in pipe tobacco ageing.
3. applications of the bacillus BA-01 as claimed in claim 2 in pipe tobacco ageing, it is characterised in that：Application process is：
After Bacillus strain BA-01 fermented and cultureds terminate, 7000rpm-10000rpm, 7min-15min centrifugation thalline are used
Sterile saline obtains bacteria suspension after washing 1-5 thalline, and trehalose is added into above-mentioned bacteria suspension makes it in bacteria suspension
Final mass concentration is 0.5%-2%, is placed in -80 DEG C of refrigerators and freezes, makes it without working fluid, then to be positioned over vacuum cold
In lyophilizer, 8-15h is freeze-dried, it is standby to be preserved in 4 DEG C of refrigerators, produces biological agent；Bacillus pumilus BA-01 life
Thing preparation viable count reaches 109-1011CFU/g, takes 0.001g-0.015g biological agents to be dissolved in 1ml-10ml sterilized waters when using
In, it is uniformly sprayed on to 10g-40g and dries pipe tobacco.
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| CN201610717824 | 2016-08-25 | ||
| CN201610717824X | 2016-08-25 |
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| CN109097404A (en) * | 2018-08-23 | 2018-12-28 | 江西中烟工业有限责任公司 | A kind of preparation and its application of the dry composite fermentation extract of loquat |
| CN109554306A (en) * | 2018-11-14 | 2019-04-02 | 河南中烟工业有限责任公司 | A kind of compound preparation of the protease with protein degradation bacterium and its application in cigar |
| CN109907360A (en) * | 2019-01-30 | 2019-06-21 | 云南中烟工业有限责任公司 | A method of strengthening Tobacco Fermentation Process |
| CN109929778A (en) * | 2019-03-08 | 2019-06-25 | 湖北中烟工业有限责任公司 | A kind of efficient flavored type bacterial strain and its application in raising cigarette quality |
| CN111616403A (en) * | 2020-05-28 | 2020-09-04 | 江南大学 | Biological agent for accelerating tobacco mellowing and improving tobacco quality and application thereof |
| CN111718878A (en) * | 2020-07-10 | 2020-09-29 | 湖北中烟工业有限责任公司 | Bacillus gene engineering bacterium for high yield of cellulase and amylase and preparation method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097404A (en) * | 2018-08-23 | 2018-12-28 | 江西中烟工业有限责任公司 | A kind of preparation and its application of the dry composite fermentation extract of loquat |
| CN109554306A (en) * | 2018-11-14 | 2019-04-02 | 河南中烟工业有限责任公司 | A kind of compound preparation of the protease with protein degradation bacterium and its application in cigar |
| CN109907360A (en) * | 2019-01-30 | 2019-06-21 | 云南中烟工业有限责任公司 | A method of strengthening Tobacco Fermentation Process |
| CN109907360B (en) * | 2019-01-30 | 2022-02-11 | 云南中烟工业有限责任公司 | Method for strengthening tobacco leaf fermentation process |
| CN109929778A (en) * | 2019-03-08 | 2019-06-25 | 湖北中烟工业有限责任公司 | A kind of efficient flavored type bacterial strain and its application in raising cigarette quality |
| CN111616403A (en) * | 2020-05-28 | 2020-09-04 | 江南大学 | Biological agent for accelerating tobacco mellowing and improving tobacco quality and application thereof |
| CN111616403B (en) * | 2020-05-28 | 2021-11-02 | 江南大学 | Biological agent for accelerating tobacco mellowing and improving tobacco quality and application thereof |
| CN111718878A (en) * | 2020-07-10 | 2020-09-29 | 湖北中烟工业有限责任公司 | Bacillus gene engineering bacterium for high yield of cellulase and amylase and preparation method thereof |
| CN111718878B (en) * | 2020-07-10 | 2022-05-06 | 湖北中烟工业有限责任公司 | Bacillus gene engineering bacterium for high yield of cellulase and amylase and preparation method thereof |
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Application publication date: 20170811 |