CN103756946B - A kind of bacterium enzyme combined preparation containing bacillus subtilis strain xp and the application in acceleration tobacco sheet in starch degradation thereof - Google Patents
A kind of bacterium enzyme combined preparation containing bacillus subtilis strain xp and the application in acceleration tobacco sheet in starch degradation thereof Download PDFInfo
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Abstract
The invention belongs to microbial technology field, be specifically related to a strain and produce diastatic bacillus subtilis strain xp, does is its Classification And Nomenclature <i>Bacillus? subtilis</i>, does is this bacterial strain at the preserving number of China typical culture collection center: CCTCC? No:M2011452.The invention also discloses a kind of utilize described bacterial strain xp to make bacterium enzyme combined preparation and accelerating the application in tobacco sheet in starch degradation.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of bacterium enzyme combined preparation containing bacillus subtilis strain xp and accelerating the application in tobacco sheet in starch degradation.
Background technology
Tobacco sheet is also known as regeneration tobacco leaf or homogeneous tobacco leaf, and it is the product of comprehensive utilization during tobacco industry is produced and retrofit, adds the composition such as sizing agent and other additives primarily of offal, fragment, offal or discarded tobacco leaf.In fact all tobacco materials may be used to the manufacturing of tobacco sheet, by offal discarded in whole production of cigarettes process, stalk label, fragment and part discarded tobacco leaf, short stalk through different working methods, make proterties close to the tobacco sheet being even better than natural tobacco, and it is low that the tobacco sheet made has cost, filling properties well also can reduce the advantages such as the coal-tar middle oil content of flue gas.According to the difference of production and processing technology, mainly contain paper process, thick slurry processes and roll-in method three productions technique.Wherein paper process refers to tobacco material first with water extraction, and enter paper machine after slurry made by insoluble substance or interpolation natural fiber and form thin slice sheet base, water-soluble extract is coated with in the lump and sheet base with additive after concentrated, is finished product after drying.During processing and manufacturing, can extract from water-soluble substances or remove some tobacco component, or add physical chemistry and the combustionproperty that some new compositions improve thin slice.The main component of thin slice or tobacco leaf and offal, if so these starting material are not through to ferment completely and degrade, innerly contain starch tobacco being burnt and sucked to side effect, greatly reduce the quality of tobacco sheet.
Tobacco fermentation is also called tobacco mellowing, its essence is under certain temperature and humidity condition, impelling the process of tobacco leaf physicochemical property generation profound change, is a kind of preliminary working method that cigarette industry is improved the quality of products, and is also a link very important in Cigarette processing.Without the former cigarette of fermentative processing and new cigarette, not only color and luster can not be satisfactory, and often have raw blue or green gas and region and to mix gas, and inhale taste coarse, pungency is large, and especially low-quality tobacco also has the shortcomings such as bitter, peppery, puckery.Process can overcome the bad quality factor of new cigarette and low-quality tobacco to some extent by fermentation, reduce or eliminate new cigarette with cyan, leaf look is made to turn dark, reduce blue foreign smell and soil to mix gas, fragrance appeared, smoke alcohol and, reduce pungency and peculiar smell, make suction taste and suitable, increase the elasticity of tobacco leaf, strengthen the incendivity of tobacco leaf and reduce its wettability power.Alcoholized tobacco or fermentation process are generally divided into artificial ageing and natural alcoholization.Artificial ageing's speed is fast, but alcoholization weak effect; Natural alcoholization is deposited in warehouse by tobacco leaf by certain requirement, and the alcoholization carried out under certain humidity (sometimes also needing control temperature) condition, the alcoholization time is 1-3.Although natural alcoholization effect is better, also come with some shortcomings, as: 1) because tobacco leaf is rich in nutritive substance and long-term and outside atmosphere close contact, tobacco leaf is easy to infested.2) in spring, summer, autumn and winter Four seasons change process, because the change of temperature, humidity greatly, tobacco leaf is easy to generation and goes mouldy.3) because tobacco leaf contacts with excess air for a long time, tobacco leaf color is easy to browning, deepens, so the natural alcoholization time can not be long, and generally can not more than 3 years.
The eighties in 19th century, little Shi Liejinge proposed microbial process hypothesis, what think tobacco fermentation primary action is microorganism, and subsequent process is only and carries out under organic catalyst effect.Because microbe species is various, reproduction speed is fast, metabolic capacity is strong and the kind of microbial enzyme is many, so can the various types of biochemical reaction of high-level efficiency catalysis.In the fermentation initial stage, number of bacteria rolls up, and when cigarette stack temperature raises, when oxygen lacks, number of bacteria reduces gradually.Further research also finds, under normal fermentation condition, bacterium increases, and fungi reduces, if fermentation is improper, the too high or anoxic of epidemic disaster, causes anaerobion quantity to increase, and fermentation afterwards tobacco leaf will produce peculiar smell.Not ferment or the flue-cured tobacco blade face of artificial fermentation exists a large amount of microflora, in Tobacco Fermentation Process the kind of microorganism and quantity totally on a declining curve, illustrate that the microorganism of tobacco leaf surface plays an important role in tobacco fermentation, thus affect fragrance and the quality of tobacco leaf.
No matter be spontaneous fermentation or artificial fermentation, in whole Tobacco Fermentation Process, the effect of microorganism be can not ignore.Microorganism to the raising of tobacco fermentation quality mainly through microorganism from growth metabolism process using the organic composition in tobacco leaf as matrix, be alcohols, ester class and other aromatoising substance carbohydrate breakdown, the fragrance matter of tobacco leaf is made to be able to abundant performance, be the small-molecule substances such as amino acid protein degradation, be water-soluble sugar starch degradation, pectin, xylogen, Mierocrystalline cellulose, phenols etc. are degraded to low molecular weight substance.On this basis, then through tobacco mailland reaction, tobacco leaf color and luster, suction taste and fragrance is made to be fully improved.
Tobacco fermentation zymin is specifically designed to the tobacco additive agent improving flue gas quality and characteristic, after tobacco leaf formulation reaches sufficient flue gas concentration and intensity, use this product can improve flue gas aerosol character, flue gas grain is attenuated mutually, thus reach flue gas exquisiteness, mellow and full effect.Increasing along with commercial enzyme preparation kind, quality product is also uneven, and have inferior zymin to occur time on market, therefore the vitality test of zymin just serves very important effect.The method of wherein conventional mensuration amylase activity has four classes, and one is the consumption measuring substrate starch, has viscosimetry, nephelometry and starch-Iodine colorimetry etc.; Two make a living sugared method, measure the growing amount of product glucose; Three is chromogen Substrate hydrolysis method, and four is enzyme coupling methods.It is easy and simple to handle rapid, practical that wherein starch-Iodine colorimetry surveys amylase activity.Its principle be starch through α-amylase catalytic hydrolysis, generation product is glucose, maltose and dextrin, and under the condition that substrate starch concentration is known and excessive, the starch adding the catalyzed hydrolysis of iodine liquid and end after reaction is combined into blue complex.The depth of its blueness is more proportional with the blank tube without enzymatic reaction, thus calculates amount of starch, calculates amylase activity.
Tobacco, after starter process, needs to carry out starch and determining the protein quantity, tentatively to determine whether fermented liquid creates effect to the alcoholization of tobacco.Conventional starch content measuring method has Iodine colorimetry, Fehling reagent, phynol method and perchloric acid extraction method.Wherein Iodine colorimetry adopts directly to boil tobacco powder and then measures its filtrate, and Measures compare is simple, but to destroy the effect that tobacco leaf cells wall makes starch expose undesirable due to simple heating, thus the value measured is smaller and error is larger.Fehling reagent, phynol method are the content of indirect measurement Starch Production product reductibility carbohydrate, due to consuming time, consumption reagent and error will not take greatly.Perchloric acid extraction method can be extracted starch in a large number with it and selectedly do prefered method.
Therefore, study and improve starch degradation technology in new tobacco sheet, cigarette quality is improved to cigarette enterprise, saves production cost, increasing economic efficiency has the extensively meaning of reality.
Summary of the invention
The object of the invention is to provide a kind of bacterium enzyme combined preparation of being mixed by subtilis and amylase, cellulase and saccharifying enzyme and is accelerating the application in tobacco sheet in starch degradation.
For achieving the above object, the present invention adopts following technical scheme:
One bacillus subtilis xp, its Classification And Nomenclature is
bacillussubtilis.xp, this bacterial strain at the preserving number of China typical culture collection center is: CCTCCNo:M2011452, preservation date: on December 9th, 2011, depositary institution address: No. 16, Luo Jia Shan road, Wuchang District, Wuhan City, Hubei Province (Wuhan University).
Utilize the bacterium enzyme combined preparation that above-mentioned subtilis xp makes, it is obtained by following step:
1) slant culture: suspended by subtilis xp LB liquid nutrient medium, then uses transfering loop picking bacterium liquid in 20-40 DEG C of slant culture 2-14h;
2) seed culture: step 1) is cultivated the bacterium obtained, be aseptically cultured to OD in LB liquid culture based on 30-38 DEG C of shaking table with transfering loop picking list colony inoculation
600value is 1.0-5.0, obtains seed liquor;
3) after by seed liquor and amylase, cellulase and saccharifying enzyme, 1:1:1:1 mixes by volume and get final product.
Concrete, the enzyme concentration alive of described amylase, cellulase and saccharifying enzyme is respectively 6-9U/ml, 700-900U/ml and 50-150U/ml.
The described bacterium enzyme combined preparation utilizing subtilis xp to make is accelerating the application in tobacco sheet in starch degradation.
Described bacterium enzyme combined preparation is accelerating the application in tobacco sheet in starch degradation, is specially: by this bacterium enzyme combined preparation temperature 28-32 DEG C, be evenly sprayed on tobacco sheet surface under humidity 55-65% condition or apply in the water-soluble vat liquor of paper process thin slice.
Bacterial strain uses therefor xp of the present invention is accredited as the diastatic subtilis of high yield through 16srDNA.The amylase selected, cellulase and saccharifying enzyme are common commercially available prod.The effect of bacterium and enzyme is common by the preliminary Partial digestion of tobacco cell wall material, enzyme is fully contacted with material in tobacco cell, reach the object starchy material affecting tobacco sucking quality being converted into sugar, alcohol and other small-molecule substances, improve tobacco sucking quality with short period of time, high-level efficiency and low cost.
Compared to the prior art, beneficial effect of the present invention: through the tobacco of bacterium enzyme combined preparation process, score of smokeing panel test total score adds 1.17 points, and thin slice grade is significantly improved.Starch degradation level in tobacco sheet is made in 30 days to reach the 1-2 of ageing originally effect.
Accompanying drawing explanation
Fig. 1 is the phylogenetic tree built based on microorganism strains xp16SrRNA gene order;
Fig. 2 is that bacterium enzyme combined preparation of the present invention sprays incubation time to the impact of starch content in tobacco sheet.
Embodiment
(1) subtilis (
bacillussubtilis) screening of bacterial strain xp:
Because the starch in tobacco 70% is amylopectin, the application take soluble Amylopectin as substrate, is made into agar plate.30 DEG C, use the vega soil aqueous solution to carry out coated plate cultivation under the condition of 24hr, plate culture is after the subzero treatment 24-72hr of 4 DEG C, and the substratum of periphery of bacterial colonies darkens, transparent, in the white background of starch agar medium, can directly be separated the bacterial strain be not bacterial contamination.The diastatic vigor that different strains produces is measured according to the relation of diastatic vigor and transparent circle diameter.Cultivate at the enterprising row filter of the agar plate of pH value 4.0-6.0, acid resistance bacterial strain xp can be obtained.
(2) subtilis (
bacillussubtilis) qualification of bacterial strain xp:
Screening the bacterial strain xp obtained is gram-positive microorganism, and this bacterium is bar shape, and long have gemma, has pod membrane outward, be about 1.2 μm, wide about 0.4 μm.Entrust China typical culture collection center to carry out physio-biochemical characteristics qualification to it, the results are shown in Table 1 and table 2, its 16SrRNA gene order comparison result is in table 3, and the phylogenetic tree data of structure are shown in accompanying drawing 1.
Table 3 is based on the comparison result of microorganism strains xp16SrRNA gene order
According to the phylogenetic tree of above-mentioned physio-biochemical characteristics, 16SrRNA gene order comparison result and structure, identify this bacterial strain
Xp belong to bacillus amyloliquefaciens (
bacillusamyloliquefaciens), bacterial strain xp at the preserving number of China typical culture collection center is: CCTCCNo:M2011452.
(3) application of bacterium enzyme combined preparation starchy compounds in degrading tobacco thin slice
If no special instructions, LB liquid culture based formulas used is following embodiment: contain in every 1000ml deionized water: yeast extract paste 5g, peptone 10g, NaCl10g, adjust pH to be 7.0 by NaOH solution.Slant culture selects LB solid medium, fills a prescription to be: contain in every 1000ml deionized water: agar powder 15g, yeast extract paste 5g, peptone 10g, NaCl10g, adjust pH to be 7.0 by NaOH solution.Substratum is before use in 121 DEG C of sterilizing 15min.
Amylase used, cellulase and saccharifying enzyme are purchased from Shanghai Kai Yang Bioisystech Co., Ltd, and its enzyme concentration alive is respectively 4000U/g, 400U/mg and 100U/mg.
embodiment 1
Prepare microbe-enzymatic preparation of the present invention and measure enzyme activity method
1, microbial preparation is prepared
1) slant culture: suspended by bacterial strain xp 0.5mlLB liquid nutrient medium, then uses transfering loop picking bacterium liquid in 37 DEG C of slant culture 14h;
2) seed culture: by step 1) cultivate obtain bacterium, aseptically with transfering loop picking list colony inoculation in the 300ml triangular flask that 100mlLB liquid nutrient medium is housed, be cultured to OD in 37 DEG C of shaking tables
600value is about 10h, rotating speed 200rpm for 5.0(), obtain seed liquor.Collect seed liquor, load in 250ml bacterium liquid receiving flask, centrifugal at 4 DEG C, centrifugal condition used is 8000rpm, 20min.Get supernatant liquor and carry out enzyme activity determination.Enzyme activity determination method is starch-Iodine colorimetry.
The enzyme of starch-Iodine colorimetry is lived definition: 1ml enzyme liquid in 60 DEG C, under the condition of pH7.0, the grams of the Zulkovsky starch that liquefies at 60min, counts 1 enzyme activity unit (mg/ml).Calculation formula is (60/T × A × B × N) ÷ D; Wherein 60 reaction times of living in definition for enzyme; T is reaction times (min); A is the milliliter number of Zulkovsky starch; B is Zulkovsky starch concentration; N is enzyme liquid extension rate; Enzyme liquid measure (ml) used when D is for measuring.
The enzyme activity of above-mentioned supernatant liquor is: 153.25mg/ml.
3) zymin is prepared
The appropriate pH of amylase used, cellulase, saccharifying enzyme is respectively 5.0-8.0,4.0-5.5 and 4.0-4.5.The vat liquor pH of test offal and offal is about 5, substantially in the appropriate pH of three kinds of enzyme liquid, the suitable fermentation after tobacco sheet surface sprays of these three kinds of enzymes is described.
Be mixed with the amylase solution of different concns, cellulase solution and Glucoamylase Solution respectively with deionized water, carry out enzyme activity determination and determine to act on the best use of concentration of standard substance under different enzyme concn condition.
Amylase activity measuring method is starch-Iodine colorimetry; Activity Determination of Cellulase is filter paper enzyme activity (FPA) assay method, and saccharifying enzyme (or glucoamylase) vigour-testing method is spectrophotometer method.
Amylase activity measures: 1. get 0.2g amylase and be dissolved in 50ml water, constant volume is to 100ml;
2. get the starch solution that above-mentioned enzyme liquid 0.2ml adds 1ml0.4g/ml, 40 DEG C of accurate response 10min, mensuration diastatic activity is 107.41mg/ml.Define the work of diastatic enzyme and under 660nm wavelength, change 0.001OD for per minute reaction system
660value is 1 enzyme activity unit 1U;
3. the degradation rate to starch when the amylase solution measuring 2-40U/ml acts on the substrate starch solution of 1ml0.4mg/ml, determines the best use of concentration to starch standard substance.Its measurement result is in table 4.
Enzyme in cellulase activity amylograph is lived and is defined: under appointment reaction conditions, per minute catalyzing cellulose hydrolysis becomes the enzyme amount of 1 μm of ol glucose.Its calculation formula is: enzyme activity (U/mL)=glucose/(60 × Ew), wherein: 60 is soaking time (enzyme-to-substrate action time, min); Ew is the volume (mL) of crude enzyme liquid.
Cellulase activity Force meansurement: the cellulase solution of compound concentration 0-800U/ml; Get 0.1M acetate buffer solution (pH4.6) 1ml in test tube, add 1ml cellulase solution, be preheating to 50 DEG C, add Xinhua's filter paper bar (1 × 6cm), 50 DEG C of insulation 1h, take out, boiling water bath deactivation 5min, be cooled to room temperature, 3mlDNS(dilutes 3 times) colour developing, survey OD
520nmlight absorption value.Its measurement result is in table 5.
Enzyme in saccharifying enzymic activity assay method is lived and is defined: under appointment reaction conditions, the milligram number of 1ml liquid enzymes 1h decomposing soluble starch, counts 1 enzyme activity unit.
Saccharifying enzymic activity is tested: the Glucoamylase Solution of preparation 0-200U/ml; Getting 1ml Glucoamylase Solution joins in 1ml substrate starch solution (0.4mg/ml), and 40 DEG C of reaction 30min, add 1ml potassiumiodide-iodine solution, measure OD
600nmlight absorption value.Its measurement result is in table 6.
Can be found out by the result of above-mentioned table 1 to 3, diastatic the best use of amount is 8U/ml, and the best use of amount of cellulase is 800U/ml, and the best use of amount of saccharifying enzyme is 100U/ml.
embodiment 2
Containing a bacterium enzyme combined preparation of bacillus subtilis strain xp, it is obtained by following step:
1) slant culture: suspended by bacterial strain xp LB liquid nutrient medium, then uses transfering loop picking bacterium liquid in 37 DEG C of slant culture 14h;
2) seed culture: step 1) is cultivated the bacterium obtained, be aseptically cultured to OD in LB liquid culture based on 37 DEG C of shaking tables with transfering loop picking list colony inoculation
600value is about 10h for 5.0(), obtain seed liquor;
3) after by seed liquor and amylase, cellulase and saccharifying enzyme, 1:1:1:1 mixes by volume and get final product.The enzyme concentration alive of described amylase, cellulase and saccharifying enzyme is respectively 8U/ml, 800U/ml and 100U/ml.
Above-mentioned for 100ml bacterium enzyme combined preparation is evenly sprayed on 500g tobacco sheet surface under temperature 30 DEG C, humidity 60% condition, in different time sampling, detects the content of starch in tobacco sheet.In tobacco sheet, the mensuration of starch content adopts perchloric acid-Iodine colorimetry to carry out.Control group is placed naturally, does not deal with.Test tobacco sheet used and be selected from the thin board thin slice of Xuchang, Henan in 2012 gold.
Test-results: bacterium enzyme combined preparation sprays the affect data of incubation time on tobacco sheet starch content and sees Fig. 2, can find out in figure spray cultivation in 30 days bacterium enzyme combined preparation to the degraded situation of starch, OD
575nmthe absorption peak of place's starch reduces in time gradually.Spray the tobacco sheet of cultivation after 30 days and press 20%(mass percent) addition, join and suck mouthfeel data in table 7 after standard cigarettes.As can be seen from Table 7, control group: total score 40.96; Test group: total score 42.14.To smoke panel test conclusion: control group: assorted gas is comparatively large, aroma quality, amount are general; Test group: slightly assorted gas, pleasant impression is better, and sugariness increases, and concentration, strength increase to some extent, and the soft and fine degree of flue gas is better, and aroma quality, amount increase; Tobacco is made to improve a grade.
Table 7 tobacco is smoked panel test scoring results
| Aroma quality | Perfume quantity | Concentration | Soft and fine degree | Pleasant impression | Assorted gas | Stimulation degree | Total score | |
| Control group | 5.71 | 5.88 | 5.79 | 6.21 | 5.67 | 5.46 | 6.25 | 40.96 |
| Test group | 5.89 | 5.95 | 6.10 | 6.15 | 5.92 | 5.90 | 6.23 | 42.14 |
embodiment 3
Containing a bacterium enzyme combined preparation of bacillus subtilis strain xp, it is obtained by following step:
1) slant culture: suspended by bacterial strain xp LB liquid nutrient medium, then uses transfering loop picking bacterium liquid in 37 DEG C of slant culture 14h;
2) seed culture: step 1) is cultivated the bacterium obtained, be aseptically cultured to OD in LB liquid culture based on 37 DEG C of shaking tables with transfering loop picking list colony inoculation
600value is about 10h for 5.0(), obtain seed liquor;
3) after by seed liquor and amylase, cellulase and saccharifying enzyme, 1:1:1:1 mixes by volume and get final product.The enzyme concentration alive of described amylase, cellulase and saccharifying enzyme is respectively 8U/ml, 800U/ml and 100U/ml.
Under the condition of temperature 30 DEG C, above-mentioned for 100ml bacterium enzyme combined preparation is put in the water-soluble vat liquor (pH value nature, about 6.0) of 10L paper process thin slice, in different time sampling, detect the content of starch in vat liquor.In tobacco vat liquor, the mensuration of starch content adopts perchloric acid-Iodine colorimetry to carry out.Control group does not deal with.The vat liquor of test group and control group carries out respectively concentrating and coats the sheet base of tobacco sheet, is made into standard 500g tobacco sheet.
Test-results: the starch content in tobacco sheet reduces in time gradually.In addition, bacterium enzyme combined preparation carries out concentrating to vat liquor and coats the sheet base of tobacco sheet, being made into standard tobacco thin slice after applying 4h.Tobacco sheet 20% is added in standard cigarettes and sucks mouthfeel data in table 8.As can be seen from Table 8, control group: total score 40.96; Test group: total score 42.15.To smoke panel test conclusion: control group: assorted gas is comparatively large, aroma quality, amount are general; Test group: slightly assorted gas, pleasant impression is better, and sugariness increases, and concentration, strength increase to some extent, and the soft and fine degree of flue gas is better, and aroma quality, amount increase; Tobacco is made to improve a grade.
Table 8 tobacco is smoked panel test scoring results
| Aroma quality | Perfume quantity | Concentration | Soft and fine degree | Pleasant impression | Assorted gas | Stimulation degree | Total score | |
| Control group | 5.71 | 5.88 | 5.79 | 6.21 | 5.67 | 5.46 | 6.25 | 40.96 |
| Test group | 5.86 | 5.97 | 6.07 | 6.16 | 5.94 | 5.88 | 6.27 | 42.15 |
Claims (2)
1. one kind is being accelerated the application in tobacco sheet in starch degradation containing the bacterium enzyme combined preparation of bacillus subtilis strain xp, it is characterized in that, this bacterium enzyme combined preparation temperature 28-32 DEG C, be evenly sprayed on tobacco sheet surface under humidity 55-65% condition or put in the water-soluble vat liquor of paper process thin slice; Described bacterium enzyme combined preparation is obtained by following step:
1) slant culture: suspended by bacillus subtilis strain xp LB liquid nutrient medium, then uses transfering loop picking bacterium liquid in 20-40 DEG C of slant culture 2-14h; Described bacillus subtilis strain xp Classification And Nomenclature is
bacillussubtilis, this bacterial strain at the preserving number of China typical culture collection center is: CCTCCNo:M2011452;
2) seed culture: step 1) is cultivated the bacterium obtained, be aseptically cultured to OD in LB liquid culture based on 30-38 DEG C of shaking table with transfering loop picking list colony inoculation
600value is 1.0-5.0, obtains seed liquor;
3) after by seed liquor and amylase, cellulase and saccharifying enzyme, 1:1:1:1 mixes by volume and get final product.
2. accelerating the application in tobacco sheet in starch degradation containing the bacterium enzyme combined preparation of bacillus subtilis strain xp as claimed in claim 1, it is characterized in that, the enzyme concentration alive of described amylase, cellulase and saccharifying enzyme is respectively 6-9U/ml, 700-900U/ml and 50-150U/ml.
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| CN106676035B (en) * | 2016-12-05 | 2020-10-30 | 河南中烟工业有限责任公司 | Application of bacillus subtilis in degrading pectin in tobacco products |
| CN106690395B (en) * | 2017-01-12 | 2018-05-22 | 河南中烟工业有限责任公司 | A kind of bacterium enzyme mix preparation of the SMXP-58 containing bacillus subtilis strain |
| CN107488614B (en) * | 2017-09-12 | 2020-06-23 | 河南中烟工业有限责任公司 | XC-3 strain for degrading starch 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 |
| CN109090224A (en) * | 2018-07-09 | 2018-12-28 | 湖州吴兴道场城乡建设发展有限公司 | A kind of microorganism fruit antistaling agent |
| CN109370937A (en) * | 2018-10-29 | 2019-02-22 | 河南中烟工业有限责任公司 | A kind of reconstituted tobacoo quality adjustment complex micro organism fungicide |
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