CN101084800A - Tobacco biological quality-enhancer, preparation method and application thereof - Google Patents
Tobacco biological quality-enhancer, preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a biological Increasing the quality agent of tobacco and its preparation method and application. the Increasing the quality agent comprises (by weight parts) Bacillus megaterium ZXB1 (CGMCC 1.941) 180-200, Bacillus subtilis subsp. Subtilis (CGMCC 1.354) 180-200, diastase 5-15, amylase 50-200, and protease 450-550. The invention has the advantages of improved product fermentation quality, shortened aging period by 10-12 months, improved chemical ingredients in tobacco, improved tobacco flavor, reduced cost by 3.5 yuan for each box, and improved tobacco grade by one third.
Description
Technical field
The present invention relates to a kind of biological tobacco and increase the matter agent, simultaneously, also relate to preparation method and application thereof that a kind of this biological tobacco increases the matter agent.
Background technology
China is maximum in the world leaf tobacco production and cigarette consumer nation, 1,500 ten thousand mu of the whole nation year plantation tobacco leaf areas, purchase tobacco leaf 4,000 ten thousand loads, year cigarette consumption figure 3,500 ten thousand casees.Realize 9,000 hundred million yuans of taxes and profits China's tobacco business year, occupy position first of the national every profession and trade.But because China's flue-cured tobacco raw material is second-rate, particularly tabacco fragrance is of poor quality, and the sleep cycle long (2-3), occupied fund is many, the storage burden is heavy, is seriously restricting the development of China's tobacco business production and the further raising of industry benefit.Simultaneously, along with China joined WTO, the trend of global economic integration, the internationalization of the domestic market is obvious day by day, and this will bring serious impact to China's tobacco business.Therefore, the key that improves China's tobacco business productivity effect is to improve the tobacco fermentation quality, shortens the old one-tenth time, improves the aroma quality of tobacco leaf.
Tobacco fermentation is the key link in the tobacco processing course.Improve the tobacco fermentation technology, promote the fragrant jealous quality of tobacco leaf, become the important means that cigarette industry improves technology.Facts have proved,, can not directly enter the cigarette process process through just baking and redrying tobacco leaf afterwards, must could improve its use value through natural ageing or artificial fermentation after a while, reach and alleviate assorted gas, improve jealously, reduce excitant, improve the purpose of perfume quantity.Many studies show that, tobacco leaf is improved in ageing and fermentation back quality, and its main cause is that extremely complicated variation has taken place in tobacco tissue inside.
Koller (1858) studies the fermentation mechanism of tobacco the earliest, points out that the cigar fermentation is similar to alcohol fermentation in some aspects.Suchsland thought to microbiotic research in the cigar in 1889, and tobacco fermentation is caused that by microorganism Reid etc. (1937) find that also micro organism quantity increases severely in the cigar sweat, and are proportionate with some enzyme content.The result of study of Johnson (1934) thinks that then microorganism has just increased enzyme amount in the tobacco leaf to the effect of tobacco fermentation.Bodnar and Borta (1933) studies show that to flue-cured tobacco spontaneous fermentation the peroxidase of tobacco leaf, catalase, amylase activity obviously descend in the sweat, and peptidase activity significantly increases.But the research of Dixon etc. (1936) proves that then behind the flue-cured tobacco ageing, micro organism quantity reduces to some extent, and the effect of enzyme is minimum in the tobacco leaf, and the flue-cured tobacco fermentation is a pure chemistry process to a great extent.Frankenberg (1950) has proposed sleep and is the hypothesis of finishing on the basis of summing up forefathers' research under self enzyme, microorganism and autoxidation three acting in conjunction.Think that these three kinds of effects all may exist, can promote total chemical reaction simultaneously, but which kind of mechanism accounts for leading role, then depend on tobacco, type fermentation pre-treatment and fermentation condition.
The viewpoint that pure chemistry changes thinks that blue or green tobacco leaf is a highly active catalyst system and catalyzing, but after apoptosis finished, moisture content in leaves lacked, and color is by the blue-stain Huang, and most of cell is all dead, and the enzyme majority loses activity.People such as Arsenjian discover, under Turkish tobaccos natures ageing condition, between the oxidation enzyme-to-substrate without any effect.
Promote the mechanism of tobacco fermentation to cause some research workers' extensive interest about microorganism, and obtained some breakthrough progress.People such as Reid (1933) find that the activity of microorganism is more active in research cigar sweat.In the fermentation incipient stage, bacterial number rolls up, and when buttress temperature rise amount of oxygen reduced, bacterial number reduced gradually, and after turning over buttress, temperature reduces, and amount of oxygen increases, and bacterial number increases again.Temperature raises again, and oxygen exhausts again gradually.
The micro organism quantity of tobacco leaf surface and kind and fermentation condition have confidential relation.Tamyo (1953) studies show that Spain's tobacco fermentation temperature mainly is a bacterium under 30-45 ℃, fermentation temperature is on the 55-65 ℃ of next bacterium that a small amount of is arranged.Reid etc. (1944) are if report microbial host micrococcus and the bacillus in the flue-cured tobacco under the normal fermentation condition, and mould fades away.If under humidity, high temperature and anoxia condition, then anaerobic bacteria occupies advantage.Similarly, under suitable fermentation condition, cigar mainly has been the microorganism of the gas and the good gas of holding concurrently, and under anoxia condition, mainly is the gemma clostridium of anaerobism.
About the fragrance of microorganism in the tobacco leaf and tobacco leaf and the relation of quality, (1967) such as Reid etc. (1944) and English studies show that, under inappropriate conditions such as humidity, high temperature or anaerobic, ferment, if the microbial host mould in the tobacco leaf, anaerobism gemma clostridium then will cause tobacco leaf to rot and fragrance loss.
In early days about microbe inoculation in tobacco leaf to improve tabacco fragrance and jealous report, the result often can not be satisfactory.Koller (1958) has carried out adding microorganism the earliest to improve cigar fragrance and jealous trial.Add saccharomycete in cigar, can quicken fermentation greatly and carry out, it is more thorough to ferment, but also can promote the growth of bacterium and mould simultaneously.Pounds etc. 1972 are the different microorganism of inoculation in the tobacco leaf of storing, and wherein most microorganisms can be caused rotting and offending smell of tobacco leaf.
Then received good result to research work afterwards.Tamyo etc. (1953) report, inoculation Micrococcus autantiacus can improve the fragrance of tobacco leaf in the tobacco leaf.English etc. (1967) are difference or combined inoculation Bacillus subtilis and B. clrculans in the Pennsylvania tobacco leaf, produces a kind of fragrance of happiness rapidly.Reid etc. prove that also bacterium is more suitable for the fermentation of tobacco leaf than fungi.Studies show that further along with microbial growth, total carbon in the tobacco leaf and reduced sugar reduce, the pH value raises, and nicotine and total nitrogen change.Koiwai etc. (1971) studies show that, inoculation is from tobacco leaf isolated bacterial bacterial strain in the Nambu cigar, although obvious variation does not take place total nitrogen content, significant variation has taken place in the content of free amino acid, and free amino acid helps improving the fragrance of tobacco leaf.Some trimethyls in the report Turkish tobaccos such as Mikami cyclic group compound can obtain in carotenoid by microbial action.
Domestic Mechanism Study to tobacco fermentation is later, hydrogen cyanide, ethanol and high-temperature process tobacco leaf such as nineteen fifty-seven Zhang Jingxiang, activity with destruction enzyme that tobacco leaf was contained or inhibition tobacco leaf endoenzyme, carrying out the artificial fermentation then, the result shows, except that oxygen index obviously reduces, and tobacco leaf exterior quality and inherent quality and the no significant difference that normally ferments, in the proof sweat in the tobacco leaf effect of enzyme little, and oxidation is comparatively obvious; (1980) such as Qingdao light industry research institutes once killed the microorganism of tobacco leaf surface with oxirane, the discovery fermentation still can be carried out, and this result has denied the effect of microorganism in the tobacco fermentation process.(1990) such as Liu Baiheng (1980) and Yu Yongmao have also drawn identical conclusion with ozone and UV treatment tobacco leaf.The research of Ares etc. (1987) finds that also in the sweat, the nicotine content and the oxygen index of tobacco leaf are proportionate.
Other results of study then find to have the effect of microorganism and enzyme in the flue-cured tobacco sweat.
In recent years, along with deepening continuously of biotechnology research, microbial fermentation technology has obtained in fields such as food, medicine, health products using widely, and this also provides prerequisite and means for this technology in the application study aspect the tobacco fermentation.Yu Yongmao etc. (1988) utilize microbial enzyme to quicken the fermentation of low-quality tobacco, have obtained certain effect; Someone finds that peroxidase not only has activity during the fermentation, even in the cigarette products afterwards, peroxidase activity still continues to keep.Have enzymatic reaction to participate in ageing, fermentation and process, the possibility that this effect of the tobacco leaf of the system of more particularly drying in the air exists is bigger.
Xie He etc. (1990) report, the major microorganisms monoid in China flue-cured tobacco NC82 ageing tobacco leaf is a bacterium, accounts for more than 85% of sum.
In a word, the research of relevant tobacco fermentation mechanism, forefathers' work all is to lay particular emphasis on some aspects, and the variation of tobacco leaf each side in the sweat is also lacked deeply and systematic research, therefore, tobacco fermentation mechanism is difficult to make comprehensive explanation.
In recent years, along with deepening continuously of biotechnology research, microbial fermentation technology has obtained in fields such as food, medicine, health products using widely, and this also provides prerequisite and means for this technology in the application study aspect the tobacco fermentation.Microbial technique does not also obtain utilizing on the tobacco fermentation engineering, still adopts the method for traditional artificial fermentation or natural ageing to handle tobacco leaf on the cigarette industry but up to now.Yet because the artificial fermentation increases the matter weak effect, the natural ageing cycle is oversize again, and capital turnover is slow, is perplexing the raising of China's cigarette industry quality and benefits for many years always.
Tobacco leaf through a series of biochemical reactions, makes its main chemical composition that violent variation take place in modulated process, has formed different style, the visibly different former cigarette of interior quality.But the flue-cured tobacco that newly modulates, suncured tabacco or air-curing of tobacco leaves remain the defective in various degree on quality, as blue foreign smell is heavy, excitant is big, flue gas is coarse, cigarette fragrance is covered by assorted gas, especially that low-grade tobacco leaf also has is bitter, peppery, puckery, etc. shortcoming, such tobacco leaf should not directly be used for processing tobacco product, must be through suitable ageing, fermentation process.In ageing, sweat, a series of variations take place in the inherent chemical composition of tobacco leaf, and blue foreign smell and excitant are significantly reduced, and cigarette fragrance strengthens, and it is pure and mild to inhale flavor, and tobacco leaf color more evenly and is suitably deepened.Therefore, ageing, fermentation are the important steps that cigarette industry improves the quality.
Summary of the invention
The object of the present invention is to provide a kind of biological tobacco to increase the matter agent,, shorten digestion time, improve the chemical composition of tobacco leaf, improve the aroma quality of tobacco leaf to significantly improve the fermented quality of tobacco leaf.
Simultaneously, the present invention also aims to provide a kind of this biological tobacco to increase the preparation method of matter agent.
In addition, the present invention also aims to provide a kind of this biological tobacco to increase the application of matter agent.
To achieve these goals, technical program of the present invention lies in having adopted a kind of biological tobacco to increase the matter agent, comprise the composition of following parts by weight: bacillus megaterium ZXB
1(Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941,180-200 part; Bacillus subtilis ZXB
2(Bacillus subtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354,180-200 part; Carbohydrase 5-15 part; Amylase 50-200 part; Protease 450-550 part.
Described carbohydrase is an aspergillus niger saccharification enzyme.
Described amylase is bacterial.
Described protease can be the bacillus subtilis neutral proteinase.
Simultaneously, technical program of the present invention also lies in adopting a kind of biological tobacco to increase the preparation method of matter agent, may further comprise the steps:
(1) preparation of microbial inoculum;
(2) biological tobacco increases the preparation of matter agent:
Getting parts by weight is the bacillus megaterium ZXB of 180-200 part
1(Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941, the bacillus subtilis ZXB of 180-200 part
2(Bacillus subtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354 is through the bacterium powder of solid culture preparation; The protease of the carbohydrase of 5-15 part, amylase 50-200 part and 450-550 part mixes.
The preparation of described microbial inoculum may further comprise the steps:
1. bacterial classification: ZXB
1, bacillus megaterium (Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941; ZXB
2, bacillus subtilis (Bacillussubtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354;
2. the inclined-plane is cultivated: inoculation one ring lawn on fresh inclined-plane, with cultivation under 30 ℃ of conditions 2 days;
3. seed bottle enlarges cultivation: culture medium prescription is, wheat bran 70%, soya-bean cake powder 28.5%, urea 1.5%, the material: water=1: 1.0, pH are 9.0, by the prescription with raw material weigh up mix thoroughly after, add suitable quantity of water (material: water=1: 1.0). continue to turn, reach and hold agglomeratingly, the degree of touching of promptly loosing is 9.0 with 2%NaOH solution adjustment pH value again, in seed bottle, loading amount to shoulder slightly by down a bit, wipe the compost that bottleneck and body adhere to, then tampon beyond the Great Wall, the top is covered two-layer paperboard grade (stock) cotton rope and is tied, at 1.1-1.5kg/cm
2Pressure is sterilization 1.5h down;
Treat compost cooling back inoculation, in cultured slant strains, pour the 7ml sterilized water into, make bacteria suspension.The bacteria suspension that makes is poured into respectively in 3 seed bottle culture mediums, shaken up, place under 28 ℃ of conditions and cultivated 3 days, on the visible bottle wall lawn is arranged, taking off tampon has pungent little stink, and it is qualified that other living contaminants of observation nothing is;
4. tray is cultivated: the material that will prepare is sub-packed in the polybag, and every bag of about 1000g uses the cotton rope tying, notices that Zha Detai is not tight, otherwise unfavorable steam circulation sterilization is sent in the high-pressure sterilizing pot, at 1.4-1.5kg/cm after material installs immediately
2Pressure is sterilization 1.5h down, insert cultured bacterial classification in the seed bottle by 2% inoculum concentration when waiting to expect temperature drop to 40 ℃, fully mix thoroughly, evenly shakeout then, thickness 2-3cm, last lid layer calico, in order to preserving moisture, culturing room's temperature is controlled at 28 ℃-30 ℃, when the material temperature is higher than 35 ℃, to in time stir compost, and adjust the upper-lower position of tray.After waiting to cultivate 3 days, the material temperature descends gradually, and microscopy overwhelming majority thalline has formed gemma, can stop cultivating;
5. dry, pulverize: cultured solid fungicide is put into drying box, under 50 ℃ of conditions, dries, dry by the fire to water content less than 8%, pulverize with pulverizer then, smashing fineness requires to reach more than 95% by the bacterium powder of 80 mesh sieves;
6. quality inspection: the quality requirement of qualified bacterium powder is that total bacteria count should be greater than 8,000,000,000/g, and moisture must be by 80 mesh sieves less than 8%, 95% bacterium powder, and coliform count must be less than 30/100g, and salmonella and pseudomonas aeruginosa must not detect.
In addition, technical program of the present invention also lies in adopting a kind of biological tobacco to increase the application of matter agent, during use, earlier the 1000g preparation has been soaked 30min with the 5000ml clear water, stirred, made the abundant stripping of bacterium and enzyme while soaking; Use two-layer filtered through gauze afterwards, remove residue, filtrate is used for spraying and handles fermenting tobacco leaf.When using, choose the leaves moisting operation on primary processing line, utilize compressed air that filtrate is sprayed onto on the blade in the leaves moisting tube, store leaf 4h afterwards, following operation is same as normal technology for making tobacco threds.
Biological tobacco of the present invention increases matter agent tobacco leaf biologically active and blade face advantage flavouring bacterium in to flue-cured tobacco phylloplane microflora, fermentation or ageing process to be separated on the basis of evaluation, utilize 3 kinds of dominant strains to be mixed and made into fumet with relevant enzyme with high bioactivity, and in flue-cured tobacco artificial fermentation and natural ageing process, carried out Preliminary Applications, obtained good effect, this provides theoretical foundation and technical foundation for the application of biotechnology in tobacco fermentation technology.The present invention is technology and means such as applied microbiology, zymetology and molecular biology, fermenting mechanism to flue-cured tobacco is furtherd investigate, as guidance, from the flue-cured tobacco phylloplane microflora, filter out advantage flavouring bacterium, separating on the basis of identifying, further preferred flavouring bacterial strain, optimize the typing fermentation and increase the matter agent prescription, and carried out the laboratory applications experiment and verified with industry, obtained good effect, this provides technical support for biotechnology applying on tobacco fermentation technology and primary processing line.Because exist a large amount of microorganisms in the tobacco leaf, these microorganisms play an important role in the tobacco fermentation process, bacillus megaterium ZXB of the present invention
1(Bacillusmegaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941 and bacillus subtilis ZXB
2(Bacillus subtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354 helps improving fragrance in the tobacco leaf and jealous, thereby improves the quality of tobacco leaf.Biological tobacco of the present invention increases the matter agent and is used for the ageing of flue-cured tobacco nature, fermentation period can be foreshortened to 9 months, has shortened more than 1 year than conventional fermentation time; The tobacco microbial fermentation increases the matter agent and is used for online prescription blade processing, can obviously improve cigarette quality at short notice.
Biological tobacco of the present invention increase the matter agent in cigarette industry enterprises such as Xinzheng tobacco group, Xuchang Cigarette General Factory through repeatedly pilot scale, all obtained and significantly increased matter flavouring effect, the present invention can significantly improve the fermented quality of tobacco leaf, shorten digestion time, improve the chemical composition of tobacco leaf, improve the aroma quality of tobacco leaf.Can shorten sleep time 10-12 month, if calculated in 1 year by shortening digestion time, per tobacco leaf storage cost of can saving for ten thousand casees is ten thousand yuan of 10,000 casees * 40 kilogram/case * 26 yuan/kilogram (on average) * 4.5%=26.7.Average single case cigarette cost can reduce by 3.5 yuan, improves 1/3 grade of tobacco leaf service rating, and single case cigarette charging cost can be saved 3 yuan.
In addition, the raising of quality of tobacco can strengthen China's tobacco leaf and cigarette competitiveness in the international market, when reducing the tobacco leaf import, can also expand export and earn foreign exchange, and will produce remarkable economic efficiency and social benefit.
Biological tobacco increases matter agent heatproof experimental result
Process tobacco microbial fermentation increases the tobacco leaf after the matter agent is handled, and by heating 80 ℃ of 10min, 140 ℃ of 10min behind 140 ℃ of 20min, measure the viable count in the tobacco leaf, enzyme activity residual rate, the results are shown in Table 1.
Table 1 tobacco microbial fermentation increases matter agent heatproof experimental result
| Treatment conditions | Viable bacteria residual rate (%) | AMS residual rate (%) | Carbohydrase residual rate (%) | Protease residual rate (%) |
| 140 ℃ of 20min of 140 ℃ of 10min of (ck) 80 ℃ of 10min are untreated | 100 100 95 91 | 100 94 71 67 | 100 89 68 55 | 100 85 52 44 |
As can be seen from Table 1, after the heat treatment through three kinds of different conditions, in the tobacco leaf residual rate of viable count all more than 90%, this may with increase that the flavouring bacterium is to exist with the gemma form in the matter agent, gemma has stronger resistance to elevated temperatures relevant.The AMS residual rate is respectively 94%, 71% and 67% after the heat treatment; The carbohydrase residual rate is respectively 89%, 68% and 55%; The residual rate of protease is respectively 85%, 52% and 44%.The residual rate of AMS is higher, and carbohydrase takes second place, and the protease residual rate is minimum, and this is because the AMS heat endurance is stronger, and carbohydrase, and protease heat stability is relatively poor to be caused.It is little that the leaves moisting condition increases matter agent influence to tobacco fermentation, and the hot conditions in baking silk stage will exert an influence to the activity that increases the matter agent.
Biological tobacco increases the effect of matter agent
Biological tobacco increases the influence of matter agent to tobacco components
Utilize and separate the tobacco fermentation flavouring that superior microorganism flavouring bacterium that obtains and the relevant enzyme with high bioactivity are mixed with 4 kinds of formulations, and adopt 1% and 2% two kind of concentration the flue-cured tobacco blade face is sprayed, carry out spontaneous fermentation and artificial fermentation then respectively, the result shows, the prescription of fumet is formed different, and is also different to the action effect of tobacco fermentation.No matter being spontaneous fermentation or artificial fermentation, compared with the control, handling 4. various chemical composition change maximums in the tobacco leaf, secondly is to handle 2., is to handle 3. once more, handles 1. various chemical composition change amplitude minimums (table 2 and table 3) in the tobacco leaf.The variation tendency of chemical composition is in full accord with the variation tendency of not handling the direct fermentation tobacco leaf chemical composition in the fumet processing tobacco leaf, but the amplitude of variation of each composition obviously strengthens, illustrate and use fumet to help promoting that material transforms in the leaf during the fermentation, accelerate the tobacco fermentation process.
Biological tobacco increases the influence of matter agent to the tobacco leaf flue gas characteristic
The tobacco leaf characteristic smoking result that uses biology to increase the matter agent in spontaneous fermentation and the artificial fermentation's process is shown, the component characteristic difference of fumet, it increases matter flavouring effect and has evident difference.Compared with the control, the tobacco leaf that sprays fumet by fermentation after, fragrance matter is improved, perfume quantity increases, assorted gas, excitant alleviate, pleasant impression is pure comfortable, and total jealous quality of body note improves, and the flavouring effect between the different disposal shows as: handle 4.＞handle 2.＞handle 3.＞handle in 1. (table 4 and table 5).
Table 2 biological tobacco increases the influence of matter agent to the spontaneous fermentation tobacco leaf chemical composition
| Digestion time (moon) | Handle | Reduced sugar (%) | Total nitrogen (%) | K2O(％) | Reduced sugar/cigarette | Total nitrogen/nicotine |
| Alkali | ||||||
| 2 4 | Processing 1. ck 2. processes ck and processes 3. ck and process 4. ck and process 1. ck and process 2. ck and process 3. that ck processes 4. ck | 16.25 17.63 15.51 17.53 15.18 16.40 15.73 18.16 16.10 17.55 15.34 17.10 15.00 16.10 15.50 17.84 | 1.13 1.14 1.18 1.19 1.27 1.28 1.10 1.24 1.10 1.13 1.15 1.16 1.20 1.23 1.05 1.15 | 1.62 1.65 10.18 1.88 2.06 2.65 1.46 1.47 1.58 1.61 1.80 1.84 2.02 2.14 1.32 1.41 | 10.03 10.68 8.25 14.73 7.37 6.19 10.77 12.35 10.13 10.90 8.52 9.42 7.43 7.52 11.74 12.65 | 0.70 0.69 0.63 0.63 0.62 0.48 1.10 0.84 0.70 0.70 0.64 0.63 0.59 0.57 1.05 0.82 |
Table 3 biological tobacco increases the influence of matter agent to artificial fermentation's tobacco leaf chemical composition
| Handle | Reduced sugar (%) | Total nitrogen (%) | Nicotine (%) | K 2O (％) | Cl (％) | Reduced sugar/nicotine | Total nitrogen/nicotine | K 2O/Cl |
| 1. former cigarette is processed CK and is 1. processed 2. CK and 2. process 3. CK and 3. process 4. CK 4. | 15.76 15.25 15.88 14.27 14.40 14.02 41.10 12.69 14.39 | 1.73 1.40 1.64 1.66 1.71 1.52 1.67 1.63 1.70 | 2.57 2.13 2.38 2.28 2.32 2.39 2.42 2.43 2.57 | 1.49 1.24 1.21 1.24 1.29 1.27 1.47 1.48 1.37 | 0.42 0.30 0.33 0.25 0.30 0.21 0.22 0.27 0.42 | 6.13 7.15 6.67 6.26 6.21 5.87 5.83 5.22 5.60 | 0.67 0.66 0.69 0.73 0.74 0.64 0.69 0.69 0.66 | 2.83 4.13 3.67 4.96 4.30 6.05 6.68 5.48 3.26 |
Table 4 biological tobacco increases the influence of matter agent to spontaneous fermentation tobacco leaf flue gas characteristic
| Digestion time (moon) | Handle | Fragrance matter (15) | Perfume quantity (25) | Assorted gas (15) | Pleasant impression (15) | Concentration (9) | Excitant (8) | Strength (5) | Flammability (5) | Ash content (3) | General comment (100) |
| Handle 1. | 11.5 | 16.2 | 11.0 | 12.0 | 6.5 | 6.4 | 4.0 | 3.5 | 2.5 | 73.6 |
| 2 4 | 2. 1. CK process CK and 2. process 3. CK and 3. process 4. CK and 4. process 1. CK and 1. process 2. CK and 2. process 3. CK and 3. process 4. CK 4. | 10.5 12.0 11.0 11.5 11.0 13.0 11.5 12.0 10.8 13.0 11.5 12.5 11.3 13.5 12.0 | 15.8 16.0 14.7 16.5 13.9 16.7 15.2 17.0 16.5 18.0 16.3 17.5 15.5 19.5 16.7 | 10.6 11.5 11.0 11.3 10.7 12.0 10.8 12.0 11.4 12.5 11.5 12.8 11.1 13.0 11.0 | 11.0 12.0 11.5 12.5 11.7 12.5 11.6 12.7 11.4 12.5 12.0 13.1 12.4 13.2 12.6 | 6.0 6.7 6.0 6.2 5.7 6.7 6.0 7.0 6.5 7.0 6.3 7.0 6.0 7.0 6.1 | 6.0 6.8 6.1 6.3 6.0 7.0 6.2 6.8 6.3 7.2 6.6 7.0 6.4 7.3 6.7 | 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 | 3.5 3.5 3.5 3.5 3.5 4.0 3.5 3.7 3.5 3.8 3.5 3.6 3.4 4.0 3.6 | 2.5 2.5 2.5 2.5 2.5 2.5 2.5 3.0 3.5 3.5 3.5 3.0 3.0 3.0 3.0 | 69.9 75.0 70.3 74.3 69.0 78.4 71.3 78.2 73.4 81.0 74.7 80.5 73.1 83.5 75.7 |
Table 5 biological tobacco increases the influence of matter agent to artificial fermentation's tobacco leaf flue gas characteristic
| Handle | Fragrance matter (15) | Perfume quantity (25) | Assorted gas (15) | Pleasant impression (15) | Concentration (9) | Excitant (8) | Strength (5) | Flammability (5) | Grey (3) | General comment (100) |
| 1. former cigarette is processed CK and is 1. processed 2. CK and process 2. 3. that 4. 3. CK process | 12.0 13.3 12.5 13.0 13.0 13.0 12.5 13.5 | 20.5 21.8 21.5 22.5 22.0 21.8 21.5 22.5 | 11.0 12.0 13.0 12.0 11.8 13.0 12.0 13.0 | 11.5 13.0 13.0 13.5 13.0 14.0 12.5 13.8 | 6.7 7.5 7.5 7.5 6.8 7.5 7.0 7.5 | 6.8 7.5 7.5 7.5 6.8 7.5 7.0 7.5 | 4.8 5.0 5.0 5.0 5.0 5.0 5.0 5.0 | 4.7 5.0 5.0 5.0 5.0 5.0 5.0 5.0 | 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 | 81.0 87.7 87.3 89.0 86.6 88.8 85.0 90.3 |
| CK④ | 12.5 | 21.5 | 12.5 | 12.5 | 7.0 | 7.0 | 5.0 | 5.0 | 3.0 | 85.5 |
Biological tobacco increases the influence of matter agent to amino acid in the cigarette and higher fatty acids content
(1) biological tobacco increases the matter agent influence of amino acid content in the cigarette be it is reported that amino acid all has material impact to the fragrant jealous quality of tobacco leaf and the style of sucking of cigarette in the tobacco leaf.Show that by measurement result (table 6) amino acid content in the cigarette after processing reduces to amino acid content in the cigarette, but the variation tendency difference of different types of its content of amino acid.Compare with the contrast cigarette, proline content shows as increase, winter propylhomoserin, threonine, serine, glutamic acid, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, lysine, the content of 13 seed amino acids such as arginine all has decline in various degree, and cystine, the benzo propylhomoserin, histidine, 4 seed amino acids such as tryptophan show as handle with contrast between not there are differences this result and show, it is closely related that the variation of amino acid content and microbial fermentation increase the decomposition and the transforming degree of matter agent acceleration tobacco leaf inherence composition in the cigarette.Show in the leaves moisting link and use flavouring bacterial classification and enzyme preparation mixed processing prescription blade, make the protein in the tobacco leaf under the effect of microorganism and protease, be degraded to amino acid gradually, afterwards through overbaking silk heating process, the amino acid of these new generations easily and the carbohydrate in the pipe tobacco non-enzyme browning reaction takes place, form complicated flavor matter, thus most of amino acid minimizing in various degree in the final strong-willed cigarette.Majority studies show that the reduction of amino acid content helps the cigarette color burn in the cigarette process, fragrant jealous improving.
Table 6 biological tobacco increases the influence of matter agent to amino acid content in the cigarette
| The amino acid kind | The microbial fermentation cigarette | The contrast cigarette | Amplitude of variation (%) |
| Aspartic acid (ASP) threonine (THR) serine (SER) glutamic acid (GLU) glycine (GLY) alanine (ALA) cystine (CYS) valine (VAL) methionine (MET) isoleucine (ILE) leucine (LEU) tyrosine (TYR) benzo propylhomoserin (PHE) lysine (LYS) | 1.31 0.26 0.24 1.34 0.42 0.46 0.17 0.48 0.15 0.28 0.46 0.24 0.41 0.31 | 1.37 0.27 0.27 1.42 0.44 0.53 0.17 0.51 0.18 0.37 0.52 0.26 0.41 0.34 | -4.38 -3.70 -11.11 -5.63 -4.55 -13.2 0 -5.88 -16.67 -24.32 -11.54 -7.69 0 -8.82 |
| Histidine (HIS) arginine (ARG) proline (PRO) tryptophan (TRY) summation | 0.15 0.16 0.54 0.18 7.56 | 0.15 0.20 0.52 0.18 8.11 | 0 -20.00 3.84 0 -6.78 |
(2) biological tobacco increases the influence of matter agent to the middle-and-high-ranking content of fatty acid of cigarette
Higher fatty acids in the tobacco is the important non-volatile organic acid of a class, and the height of its content is determining the Main Ingredients and Appearance of tobacco flavor, directly or indirectly to quality of tobacco and cigarette quality thereof exert an influence [
19,20].In the leaves moisting process, in the prescription blade, add microorganism flavouring bacterial classification and enzyme compound formulation, quickened the variation of the middle-and-high-ranking content of fatty acid of cigarette, some aliphatic acid changes very violently relatively, and some is small, from measurement result (table 7), myristic acid, linoleic acid, 3 kinds of aliphatic acid of arachidonic acid increase compared with the control, wherein myristic acid, arachidonic acid increases large percentage, and comparison is according to increasing by 12.4% and 10% respectively.Laurate, palmitic acid, stearic acid and oleic acid content compared with the control descend, and wherein laurate has descended 23.1%, and palmitic acid has descended 22.8%, and stearic acid is relative with the oleic acid fall less, reaches 15.4% and 11.1% respectively.This shows that the activity and the growth conditions of the growth and decline of various content of fatty acid and flavouring bacterial classification are closely related in the cigarette.
Table 7 biological tobacco increases the influence of matter agent to the middle-and-high-ranking content of fatty acid of cigarette
| Handle | Laurate (12: 0) | Myristic acid (14: 0) | Palmitic acid (16: 0) | Stearic acid (18: 0) | Oleic acid (18: 1) | Linoleic acid (18: 2) | Arachidonic acid (20: 4) |
| Microbial fermentation cigarette contrast cigarette amplitude of variation (%) | 14.50 18.84 -23.04 | 44.42 39.56 12.28 | 9.36 12.13 -22.84 | 2.04 2.41 -15.35 | 3.70 4.16 -11.06 | 7.7 7.31 6.29 | 17.12 15.57 9.96 |
Biological tobacco increases the influence of matter agent to flavor matter in the cigarette and chemical composition content
(1) biological tobacco increases the influence of matter agent to cigarette aroma component content
Flavor matter Determination on content result (table 8) in the cigarette is shown the tobacco leaf after microbial fermentation increases matter agent processing has 17 kinds of flavor matter component contents to be higher than contrast, has only 7 kinds of flavor matter component contents to be lower than contrast.Wherein in the cigarette after increasing the matter agent and handling flavor matter content amplification maximum be benzaldehyde (35.48%), phenylacetaldehyde (46.95%), benzyl carbinol (43.97%), solanone (30.23%), dorinone (29.76%), huge beans-4,6 (Z), 8 (E)-three ketenes-3 (27.12%) huge beans-4,6 (E), 8 (Z)-three ketenes-3 (19.86%), huge beans-4,6, (E), 8 (E)-three ketenes-3 (28.28%), eggplant any scholar's ketone (22.65%), (1S, 2E, 4R, 6E, 8R, 11S, 12E)-8,11-oxygen support-2,6,12-west cypress triolefin-4-alcohol etc.It is reported, the most important perfuming cigarette raw materials of above-mentioned these compositions, the increase of their content in cigarette is very favourable to improving jealous quality of the total body note of cigarette and style undoubtedly.
(2) biological tobacco increases the influence of matter agent to cigarette chemical composition and inherent quality
Result of study (table 9) shows, the content of water-soluble total reducing sugar and total alkaloid is compared the increase that presents in various degree with the contrast cigarette in the cigarette after microbiological treatment, increasing degree is respectively 14.88% and 2.69%, and total nitrogen content then shows as decline (2.92%) in the cigarette.From chemical ratio, the sugar of microbial fermentation cigarette/alkali ratio, nitrogen/alkali contrast more reasonable, the coordination of cigarette frequently, illustrate that tobacco fermentation increases the acid-base balance that the matter agent helps promoting flue gas, also make the suction quality more plentiful and mellow and full.Further smoking result (table 10) also illustrate, microbial fermentation increase the matter agent to the perfume quantity that increases cigarette, improve fragrance matter, reduce assorted gas and excitant, the distinguish the flavor of effect of quality of the whole fragrance that improves cigarette is fairly obvious.
Table 8 microbial fermentation increases the influence of matter agent to cigarette aroma component content
| Peak number | The flavor matter title | Molecular formula | Molecular weight | The microbial fermentation cigarette | The contrast cigarette | Amplitude of variation (%) |
| 170 203 279 308 343 375 477 799 842 856 892 | 2 furan carboxyaldehyde ethylbenzene benzaldehyde 2,3,5-trimethyl ethane 1-ethyl-cyclohexene phenylacetaldehyde benzyl carbinol nicotine solanone 1-(2,6,6-trimethyl-1, the 3-cyclohexadiene)-2-butene-1-ketone dorinone | C5H4O2 C8H10 C7H6O C9H2O C8H14 C8H8O C8H10O C10H14N2 C3H22O C13H18O C12H18O | 96 106 106 128 110 120 122 162 194 190 190 | 0.601 0.095 0.042 0.024 0.167 0.723 0.203 24.319 2.856 0.924 0.109 | 0.517 0.089 0.031 0.028 0.152 0.492 0.141 27.187 2.193 0.897 0.084 | 16.25 6.74 35.48 -8.57 9.87 46.95 43.97 -10.55 30.23 3.01 29.76 |
| 1061 1035 1118 1135 1027 1352 1381 1435 1564 1820 1846 1895 1916 | Huge beans-4,6 (Z), 8 (Z)-three ketenes-3 huge beans-4,6 (Z), 8 (E)-three ketenes-3 huge beans-4,6 (E), 8 (Z)-three ketenes-3 huge beans-4,6 (E), 8 (E)-three ketenes-3 eggplants any scholar's ketone ten tetra-carbonic isoprene solanone neophytadiene geranyl spiceleaf diene 3,7,11-triethyl group-1,3,6,10-encircles 14 carbon tetraene (1S, 2E, 4R, 6R, 7E, 11S)-2,7,12-west cypress triolefin-4,6,11-triol 10-isopropyl-3,17,13-trimethyl-2,6,11,3-14 carbon tetraene (1S, 2E, 4R, 6E, 8R, 11S, 12E)-8,11-oxygen support-2,6,12-west cypress triolefin-4-alcohol | C13H18O C13H18O C13H18O C13H18O C15H22O C14H28O2 C18H29O1 C20H38 C20H32 C20H32 C20H34O3 C20H32O C20H32O2 | 190 190 190 190 218 228 262 278 272 272 322 288 304 | 0.075 0.209 0.501 0.095 0.287 0.107 0.518 28.475 0.582 3.059 3.912 12.567 3.721 | 0.059 0.187 0.418 0.074 0.234 0.128 0.453 34.232 0.519 3.482 3.974 14.372 3.152 | 27.12 11.76 19.86 28.38 22.65 -16.41 14.35 -16.82 12.14 -12.15 -15.60 -12.56 18.05 |
Table 9 microbial fermentation increases the influence of matter agent to chemical composition content in the cigarette
| Handle | Water-soluble total reducing sugar | Total alkaloid | Total nitrogen | Sugar/alkali | Nitrogen/alkali |
| Microbial fermentation cigarette contrast cigarette amplitude of variation (%) | 9.88 8.60 14.88 | 2.67 2.60 2.69 | 2.66 2.74 -2.92 | 3.70 3.31 11.78 | 0.996 1.054 -5.5 |
Table 10 microbial fermentation increases the influence of matter agent to the cigarette inherent quality
| Handle | Gloss (6) | Fragrance (36) | Harmony (6) | Assorted gas (16) | Excitant (16) | Pleasant impression (20) | Add up to (100) |
| Microbial fermentation cigarette contrast cigarette amplitude of variation (%) | 4.0 4.0 0 | 32.38 30.25 5.30 | 4.88 4.81 1.46 | 13.75 13.38 2.77 | 14.00 13.50 3.70 | 15.75 15.12 4.17 | 84.80 81.60 3.92 |
During use, the accessible prescription tobacco leaf of 1000g preparation is 100kg.
The specific embodiment
(1) preparation of microbial inoculum:
May further comprise the steps:
1. bacterial classification: ZXB
1, bacillus megaterium (Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941; ZXB
2, bacillus subtilis (Bacillussubtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354;
2. the inclined-plane is cultivated: inoculation one ring lawn on fresh inclined-plane, with cultivation under 30 ℃ of conditions 2 days;
3. seed bottle enlarges cultivation: culture medium prescription is, wheat bran 70%, soya-bean cake powder 28.5%, urea 1.5%, the material: water=1: 1.0, pH are 9.0, by the prescription with raw material weigh up mix thoroughly after, add suitable quantity of water (material: water=1: 1.0). continue to turn, reach and hold agglomeratingly, the degree of touching of promptly loosing is 9.0 with 2%NaOH solution adjustment pH value again, in seed bottle, loading amount to shoulder slightly by down a bit, wipe the compost that bottleneck and body adhere to, then tampon beyond the Great Wall, the top is covered two-layer paperboard grade (stock) cotton rope and is tied, at 1.1-1.5kg/cm
2Pressure is sterilization 1.5h down;
Treat compost cooling back inoculation, in cultured slant strains, pour the 7ml sterilized water into, make bacteria suspension.The bacteria suspension that makes is poured into respectively in 3 seed bottle culture mediums, shaken up, place under 28 ℃ of conditions and cultivated 3 days, on the visible bottle wall lawn is arranged, taking off tampon has pungent little stink, and it is qualified that other living contaminants of observation nothing is;
4. tray is cultivated: the material that will prepare is sub-packed in the polybag, and every bag of about 1000g uses the cotton rope tying, notices that Zha Detai is not tight, otherwise unfavorable steam circulation sterilization is sent in the high-pressure sterilizing pot, at 1.4-1.5kg/cm after material installs immediately
2Pressure is sterilization 1.5h down, insert cultured bacterial classification in the seed bottle by 2% inoculum concentration when waiting to expect temperature drop to 40 ℃, fully mix thoroughly, evenly shakeout then, thickness 2-3cm, last lid layer calico, in order to preserving moisture, culturing room's temperature is controlled at 28 ℃-30 ℃, when the material temperature is higher than 35 ℃, to in time stir compost, and adjust the upper-lower position of tray.After waiting to cultivate 3 days, the material temperature descends gradually, and microscopy overwhelming majority thalline has formed gemma, can stop cultivating;
5. dry, pulverize: cultured solid fungicide is put into drying box, under 50 ℃ of conditions, dries, dry by the fire to water content less than 8%, pulverize with pulverizer then, smashing fineness requires to reach more than 95% by the bacterium powder of 80 mesh sieves;
6. quality inspection: the quality requirement of qualified bacterium powder is that total bacteria count should be greater than 8,000,000,000/g, and moisture must be by 80 mesh sieves less than 8%, 95% bacterium powder, and coliform count must be less than 30/100g, and salmonella and pseudomonas aeruginosa must not detect.
(2) biological tobacco increases the preparation of matter agent:
Getting parts by weight is the bacillus megaterium ZXB of 200 grams
1(Bacillus meggaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, the bacillus subtilis ZXB of deposit number 1.941,185 grams
2(Bacillus subtilis subsp.subtilis), China common micro-organisms culture presevation administrative center CGMCC, the commodity bacillus subtilis neutral proteinase of the commodity aspergillus niger saccharification enzyme of deposit number 1.354,15 grams, the commodity bacterial of 100 grams and 500 grams mixes.
It should be noted last that: above embodiment is the unrestricted technical scheme of the present invention in order to explanation only, although the present invention is had been described in detail with reference to the foregoing description, those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not breaking away from any modification or partial replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (7)
1, a kind of biological tobacco increases the matter agent, it is characterized in that: the composition that comprises following parts by weight: bacillus megaterium ZXB
1(Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941,180-200 part; Bacillus subtilis ZXB
2(Bacillus subtilissubsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354,180-200 part; Carbohydrase 5-15 part; Amylase 50-200 part; Protease 450-550 part.
2, biological tobacco according to claim 1 increases the matter agent, it is characterized in that: described carbohydrase is an aspergillus niger saccharification enzyme.
3, biological tobacco according to claim 1 increases the matter agent, it is characterized in that: described amylase is bacterial.
4, biological tobacco according to claim 1 increases the matter agent, it is characterized in that: described protease can be the bacillus subtilis neutral proteinase.
5, a kind of biological tobacco as claimed in claim 1 increases the preparation method of matter agent, it is characterized in that: may further comprise the steps:
(1) preparation of microbial inoculum;
(2) biological tobacco increases the preparation of matter agent:
Getting parts by weight is the bacillus megaterium ZXB of 180-200 part
1(Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941 is through the bacterium powder of solid culture preparation; The bacillus subtilis ZXB of 180-200 part
2(Bacillus subtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354 is through the bacterium powder of solid culture preparation; The protease of the carbohydrase of 5-15 part, amylase 50-200 part and 450-550 part mixes.
6, biological tobacco according to claim 5 increases the preparation method of matter agent, it is characterized in that: the preparation of described microbial inoculum may further comprise the steps:
1. bacterial classification: ZXB
1, bacillus megaterium (Bacillus megaterium), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.941; ZXB
2, bacillus subtilis (Bacillussubtilis subsp.subtilis), Chinese common micro-organisms culture presevation administrative center CGMCC, deposit number 1.354;
2. the inclined-plane is cultivated: inoculation one ring lawn on fresh inclined-plane, with cultivation under 30 ℃ of conditions 2 days;
3. seed bottle enlarges cultivation: culture medium prescription is, wheat bran 70%, soya-bean cake powder 28.5%, urea 1.5%, the material: water=1: 1.0, pH are 9.0, by the prescription with raw material weigh up mix thoroughly after, add suitable quantity of water (material: water=1: 1.0). continue to turn, reach and hold agglomeratingly, the degree of touching of promptly loosing is 9.0 with 2%NaOH solution adjustment pH value again, in seed bottle, loading amount to shoulder slightly by down a bit, wipe the compost that bottleneck and body adhere to, then tampon beyond the Great Wall, the top is covered two-layer paperboard grade (stock) cotton rope and is tied, at 1.1-1.5kg/cm
2Pressure is sterilization 1.5h down;
Treat compost cooling back inoculation, in cultured slant strains, pour the 7ml sterilized water into, make bacteria suspension.The bacteria suspension that makes is poured into respectively in 3 seed bottle culture mediums, shaken up, place under 28 ℃ of conditions and cultivated 3 days, on the visible bottle wall lawn is arranged, taking off tampon has pungent little stink, and it is qualified that other living contaminants of observation nothing is;
4. tray is cultivated: the material that will prepare is sub-packed in the polybag, and every bag of about 1000g uses the cotton rope tying, notices that Zha Detai is not tight, otherwise unfavorable steam circulation sterilization is sent in the high-pressure sterilizing pot, at 1.4-1.5kg/cm after material installs immediately
2Pressure is sterilization 1.5h down, insert cultured bacterial classification in the seed bottle by 2% inoculum concentration when waiting to expect temperature drop to 40 ℃, fully mix thoroughly, evenly shakeout then, thickness 2-3cm, last lid layer calico, in order to preserving moisture, culturing room's temperature is controlled at 28 ℃-30 ℃, when the material temperature is higher than 35 ℃, to in time stir compost, and adjust the upper-lower position of tray.After waiting to cultivate 3 days, the material temperature descends gradually, and microscopy overwhelming majority thalline has formed gemma, can stop cultivating;
5. dry, pulverize: cultured solid fungicide is put into drying box, under 50 ℃ of conditions, dries, dry by the fire to water content less than 8%, pulverize with pulverizer then, smashing fineness requires to reach more than 95% by the bacterium powder of 80 mesh sieves;
6. quality inspection: the quality requirement of qualified bacterium powder is that total bacteria count should be greater than 8,000,000,000/g, and moisture must be by 80 mesh sieves less than 8%, 95% bacterium powder, and coliform count must be less than 30/100g, and salmonella and pseudomonas aeruginosa must not detect.
7, a kind of biological tobacco as claimed in claim 1 increases the application of matter agent, it is characterized in that: earlier the 1000g preparation is soaked 30min with the 5000ml clear water, stir while soaking, make the abundant stripping of bacterium and enzyme; Use two-layer filtered through gauze afterwards, remove residue, filtrate is used for spraying and handles fermenting tobacco leaf.When using, choose the leaves moisting operation on primary processing line, utilize compressed air that filtrate is sprayed onto on the blade in the leaves moisting tube, store leaf 4h afterwards, following operation is same as normal technology for making tobacco threds.
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