CN102258213B - Complex enzyme bacterium preparation used for tobacco thread production and preparation method thereof - Google Patents
Complex enzyme bacterium preparation used for tobacco thread production and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a complex enzyme bacterium preparation used for tobacco thread production. The complex enzyme bacterium preparation contains a liquid A and a liquid B, wherein the liquid A is preserved in CGMCC (China general microbiological culture collection center), is enrolled as a fermentation liquor clear solution after fermentation of a strain and is numbered as CGMCC No.4121, and the liquid B is preserved in CGMCC, is enrolled as a fermentation liquor clear solution after fermentation of a strain and is numbered as CGMCC No.4120. By applying the preparation provided by the invention, indexes of cigarette such as colour and lustre, fragrance quality, fragrance quantity, strength, impurity gas, irritation, aftertaste, flammability, grey and the like are obviously improved.
Description
Technical field
The present invention relates to a kind of pipe tobacco production composite enzyme bacterium preparation and preparation method thereof.
Background technology
At present, in the production of cigarettes process, can add the not mature assorted gas that produces when additive is covered pipe tobacco and lighted, and change the sensation that the smog that produces when lighting can produce the smoker.But these additive prices are higher, and majority is synthetic.
Summary of the invention
The technical problem that the present invention mainly solves provides a kind of ecology of adding, nontoxic, cheap composite enzyme bacterium preparation in the pipe tobacco production process of cigarette, and preparation method thereof.
Pipe tobacco production composite enzyme bacterium preparation provided by the invention, described composite enzyme bacterium preparation contains liquid A and liquid B;
Described liquid A registers on the books zymotic fluid behind the strain fermentation that is numbered CGMCC No.4121 through pretreated treatment fluid for being preserved in the zymotic fluid behind CGMCC (the China Committee for Culture Collection of Microorganisms common micro-organisms center) strain fermentation that is numbered CGMCC No.4121 of registering on the books or being preserved in CGMCC, and described treatment fluid contains mycelium;
Described liquid B be preserved in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4120 zymotic fluid or register on the books and be numbered CGMCCNo.4120 for being preserved in CGMCC, classification name: bacillus subtilis, classification name Latin title: the zymotic fluid behind the strain fermentation of Bacillus subtilis is through pretreated treatment fluid, and described treatment fluid contains mycelium.The invention has the beneficial effects as follows, use additive of the present invention after, indexs such as the color and luster of cigarette, fragrance matter, perfume quantity, strength, assorted gas, excitant, pleasant impression, flammability, grey significantly improve.
Wherein, the volume ratio of liquid A and liquid B is 1: 2 in the described enzyme bacteria preparation.
The present invention also provides a kind of preparation method of described composite enzyme bacterium preparation, and described preparation method may further comprise the steps:
A. go bail for be hidden in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4121 zymotic fluid or be preserved in the zymotic fluid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCC No.4121 and make liquid A through pretreated treatment fluid;
Go bail for be hidden in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4120 zymotic fluid or make liquid B for being preserved in the zymotic fluid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCC No.4120 through pretreated treatment fluid;
B. liquid A is mixed with liquid B or liquid A is mixed the back with liquid B and add acceptable auxiliary liquid.
The composite enzyme bacterium preparation that goes out according to method for preparing can be applied to tobacco leaf processing, and indexs such as the color and luster of the cigarette that the leaf tobacco production of processing, fragrance matter, perfume quantity, strength, assorted gas, excitant, pleasant impression, flammability, grey significantly improve.
Wherein, the described CGMCC of being preserved in registers on the books the bacterial classification that is numbered CGMCC No.4121 with to be preserved in the register on the books strain fermentation process that is numbered CGMCC No.4120 of CGMCC specific as follows:
A, actication of culture: from the slant preservation pipe, get 1 ring and register on the books the lawn of the bacterial classification that is numbered CGMCCNo.4121 to the 30mL fluid nutrient medium, under 37 ℃, the 200--250rpm shaking table cultivated 20--24 hour, made the cultured bottle A that shakes;
From the slant preservation pipe, get 1 ring and register on the books the lawn of the bacterial classification that is numbered CGMCC No.4120 to the 30mL fluid nutrient medium, under 37 ℃, the 200--250rpm shaking table cultivated 20--24 hour, made the cultured bottle B that shakes;
B, fermentation tank culture: the cultured bottle A that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180--200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid A
The cultured bottle B that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180--200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid B;
The solid medium preparation method on inclined-plane is as follows: the bean sprouts of 15% (ratio of weight and deionized water) and 3% (weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner, supply volume, add 5% (weight) glucose, 2% (weight) agar powder dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made solid medium after 30 minutes
It is fluid nutrient medium with the fermentation tank culture culture medium that bacterial strain shakes bottle, the preparation method is as follows: 15% (weight) bean sprouts and 3% (weight) tobacco leaf add deionized water and boiled 20 minutes, filter cleaner, supply volume, add 5% (weight) glucose dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made after 30 minutes.
Wherein, the preparation method of described liquid A is, zymotic fluid A is got upper strata liquid with 4000--5000rpm after centrifugal 5 minutes, and the preparation method of described liquid B is that zymotic fluid B is got upper strata liquid with 4000--5000rpm after centrifugal 5 minutes.
The specific embodiment
By describing technology contents of the present invention, structural feature in detail, realized purpose and effect, give explanation below in conjunction with embodiment is detailed.
The method for preparing liquid A: will be preserved in the zymotic fluid Separation of Solid and Liquid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCCNo.4121 and make liquid A, specific as follows:
Actication of culture: from the slant preservation pipe, get 1 environmental protection and be hidden in CGMCC and register on the books the lawn of the bacterial classification (hereinafter to be referred as bacterial classification A) that is numbered CGMCC No.4120 to the 30mL fluid nutrient medium, under 37 ℃, the 200-250rpm shaking table cultivated 24 hours, made the cultured bottle A that shakes;
Fermentation tank culture: the cultured bottle A that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180-200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid A;
Zymotic fluid A is got upper strata liquid with 4000-5000rpm after centrifugal 5 minutes, make liquid A; Centrifugally removing most of mycelium, but also have the small part thalline, is not complete Separation of Solid and Liquid, and thalline has play a part indispensable in improving the quality of tobacco shreds process.
Used culture medium preparation respectively by the following method in the said process:
The solid medium preparation method on described inclined-plane is as follows: 15% (weight) (is annotated: among the present invention, indicate the percentage with deionized water weight of being of (weight)) bean sprouts and 3% (weight) tobacco leaf add deionized water and boiled 20 minutes, filter cleaner, supply volume, add 5% (weight) glucose, 2% (weight) agar powder dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made solid medium after 30 minutes
It is fluid nutrient medium with the fermentation tank culture culture medium that described bacterial strain shakes bottle, the preparation method is as follows: 15% (weight, ratio with deionized water) bean sprouts and 3% (weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner is supplied volume, adds 5% (weight) glucose dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made after 30 minutes.
The method for preparing liquid B: will be preserved in the zymotic fluid Separation of Solid and Liquid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCCNo.4120 and make liquid B, specific as follows:
Actication of culture: from the slant preservation pipe, get 1 environmental protection and be hidden in CGMCC and register on the books the lawn of the bacterial classification (hereinafter to be referred as bacterial classification B) that is numbered CGMCC No.4120 to the 30mL fluid nutrient medium, under 37 ℃, the 200-250rpm shaking table cultivated 20--24 hour, made the cultured bottle B that shakes;
Fermentation tank culture: the cultured bottle B that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180-200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid B;
Zymotic fluid B is got upper strata liquid with 4000-5000rpm after centrifugal 5 minutes, make liquid B; Centrifugally removing most of mycelium, but also have the small part thalline, is not complete Separation of Solid and Liquid, and thalline has play a part indispensable in improving the quality of tobacco shreds process.
The solid medium preparation method on described inclined-plane is as follows: 15% (weight, ratio with deionized water) bean sprouts and 3% (weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner, supply volume, add 5% (weight) glucose, 2% (weight) agar powder dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made solid medium after 30 minutes
It is fluid nutrient medium with the fermentation tank culture culture medium that described bacterial strain shakes bottle, the preparation method is as follows: 15% (weight, ratio with deionized water) bean sprouts and 3% (weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner is supplied volume, adds 5% (weight) glucose dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made after 30 minutes.
Seen from the above description used solid medium and fluid nutrient medium can adopt used culture medium in the liquid A preparation process in the liquid B preparation process.
The liquid A that makes is mixed with 1: 2 volume ratio with liquid B, make the composite enzyme bacterium preparation, when the reinforced leaves moisting of cigarette manufacturing process, add the composite enzyme bacterium preparation, enzyme in the preparation and bacterium are coordinated the inherent chemical composition of pipe tobacco, improve jealous (this is the professional term that the cigarette product are inhaled), improve quality of tobacco shreds, enter tobacco shreds drying technology after about 90 minutes, this moment, temperature reached more than 200 ℃, the deactivation in short-term of enzyme and bacterium, and effect stops.
Bacterial classification A of the present invention and bacterial classification B are the viable bacteria that fresh and alive tobacco leaf surface separates, wherein bacterial classification A is tobacco leaf flavouring bacterium, the liquid A that is made by its fermentation is the flavouring bacteria culture fluid, and bacterial classification B is the nicotine degradation bacterium, and the liquid B that is made by its fermentation is the nicotine degradation bacteria culture fluid.
Below be to adopt composite enzyme bacterium preparation of the present invention and the pipe tobacco of usual manner (interpolation chemical addition agent) production and the contrast test of clear water control group (not using composite enzyme bacterium preparation and chemical addition agent):
The listed bracketed numerical value of the index of more than smokeing panel test is the highest scoring of this index, index scoring more near this numeric representation smoke panel test estimate more high.
With the situation of change of 40 degree purging-capture-GC/MS analysis aroma components, the result is:
(1) 3-hydroxyl-2-butanone increases considerably, and is about 20 times;
(2) a plurality of fragrance components such as solanone, damascenone, damascone, geranyl acetone, phenmethylol, benzyl carbinol, acetyl pyrrole obviously increase;
(3) acetic acid obviously increases, and the attitude of always volatilizing nicotine obviously reduces.
From chemical terms, fragrance matter should obviously be improved.
The above only is embodiments of the invention; be not so limit claim of the present invention; every equivalent structure or equivalent flow process conversion that utilizes description of the present invention to do; or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.
Claims (5)
1. a pipe tobacco production composite enzyme bacterium preparation is characterized in that, described composite enzyme bacterium preparation contains liquid A and liquid B;
Described liquid A be preserved in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4121 zymotic fluid or be preserved in CGMCC and register on the books zymotic fluid behind the strain fermentation that is numbered CGMCC No.4121 through pretreated treatment fluid, described treatment fluid contains mycelium;
Described liquid B be preserved in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4120 zymotic fluid or register on the books zymotic fluid behind the strain fermentation that is numbered CGMCC No.4120 through pretreated treatment fluid for being preserved in CGMCC, described treatment fluid contains mycelium.
2. composite enzyme bacterium preparation according to claim 1 is characterized in that, the volume ratio of liquid A and liquid B is 1: 2 in the described composite enzyme bacterium preparation.
3. the preparation method of composite enzyme bacterium preparation according to claim 1 and 2 is characterized in that, described preparation method may further comprise the steps:
A. go bail for be hidden in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4121 zymotic fluid or be preserved in the zymotic fluid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCC No.4121 and make liquid A through pretreated treatment fluid;
Go bail for be hidden in CGMCC register on the books behind the strain fermentation that is numbered CGMCC No.4120 zymotic fluid or be preserved in the zymotic fluid that CGMCC registers on the books behind the strain fermentation that is numbered CGMCC No.4120 and make liquid B through pretreated treatment fluid;
B. liquid A is mixed with liquid B or liquid A is mixed the back with liquid B and add acceptable auxiliary liquid.
4. preparation method according to claim 3 is characterized in that, the described CGMCC of being preserved in registers on the books the bacterial classification that is numbered CGMCC No.4121 with to be preserved in the register on the books strain fermentation process that is numbered CGMCC No.4120 of CGMCC specific as follows:
A, actication of culture: from the slant preservation pipe, get 1 ring and register on the books the lawn of the bacterial classification that is numbered CGMCC No.4121 to the 30mL fluid nutrient medium, under 37 ℃, the 200--250rpm shaking table cultivated 20--24 hour, made the cultured bottle A that shakes;
From the slant preservation pipe, get 1 ring and register on the books the lawn of the bacterial classification that is numbered CGMCC No.4120 to the 30mL fluid nutrient medium, under 37 ℃, the 200--250rpm shaking table cultivated 20--24 hour, made the cultured bottle B that shakes;
B, fermentation tank culture: the cultured bottle A that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180--200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid A;
The cultured bottle B that shakes is inoculated into fermentation tank expansion cultivation, and vaccination ways adopts the flame inoculation, and inoculum concentration is 1%, 37 ℃ of cultivation temperature, speed of agitator: 180--200rpm, throughput: 1: 1, tank pressure: 0.02MPa, incubation time: 20--24h makes zymotic fluid B;
The solid medium preparation method on inclined-plane is as follows: 15%(weight, ratio with deionized water) bean sprouts and 3%(weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner, supply volume, add 5%(weight) glucose, 2%(weight) agar powder dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made solid medium after 30 minutes
It is fluid nutrient medium with the fermentation tank culture culture medium that bacterial strain shakes bottle, the preparation method is as follows: 15%(weight, ratio with deionized water) bean sprouts and 3%(weight, ratio with deionized water) tobacco leaf adds deionized water and boiled 20 minutes, filter cleaner is supplied volume, adds 5%(weight) glucose dissolving mixing, adjust pH value to 7.0,121 ℃ of sterilizations made after 30 minutes.
5. preparation method according to claim 3, it is characterized in that the preparation method of described liquid A is that zymotic fluid A is got upper strata liquid with 4000--5000rpm after centrifugal 5 minutes, the preparation method of described liquid B is that zymotic fluid B is got upper strata liquid with 4000--5000rpm after centrifugal 5 minutes.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007072224A3 (en) * | 2006-09-13 | 2008-04-17 | Nara Inst Science & Technology | Increasing levels of nicotinic alkaloids |
| CN101177669A (en) * | 2007-05-29 | 2008-05-14 | 云南烟草科学研究院 | Crude enzyme liquid prepared by microorganism and uses thereof |
| CN101818123A (en) * | 2010-02-02 | 2010-09-01 | 红云红河烟草(集团)有限责任公司 | Acinetobacter and application thereof |
| CN101824391A (en) * | 2010-02-02 | 2010-09-08 | 红云红河烟草(集团)有限责任公司 | Bacillus pumilus and application thereof |
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| US9102948B2 (en) * | 2006-11-17 | 2015-08-11 | 22Nd Century Limited, Llc | Regulating alkaloids |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007072224A3 (en) * | 2006-09-13 | 2008-04-17 | Nara Inst Science & Technology | Increasing levels of nicotinic alkaloids |
| CN101177669A (en) * | 2007-05-29 | 2008-05-14 | 云南烟草科学研究院 | Crude enzyme liquid prepared by microorganism and uses thereof |
| CN101818123A (en) * | 2010-02-02 | 2010-09-01 | 红云红河烟草(集团)有限责任公司 | Acinetobacter and application thereof |
| CN101824391A (en) * | 2010-02-02 | 2010-09-08 | 红云红河烟草(集团)有限责任公司 | Bacillus pumilus and application thereof |
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Effective date of registration: 20200513 Address after: 350800 Minqing province Jinsha Town, Fuzhou County, platinum Industrial Park Patentee after: NONGBOSHI (FUJIAN) BIOTECHNOLOGY Co.,Ltd. Address before: 350002, Jinshan technology business incubator No. 7, Jinzhou North Road, Fujian, Fuzhou, No. 7 Patentee before: FUZHOU NBS BIOTECHNOLOGY Co.,Ltd. |