CN106957838A - A kind of method that immobilization acetic acid bacteria prepares cigarette iris extract - Google Patents
A kind of method that immobilization acetic acid bacteria prepares cigarette iris extract Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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Abstract
The present invention provides a kind of method that immobilization acetic acid bacteria prepares cigarette iris extract.The preparation method is concretely comprised the following steps：(1) orrisroot water extraction at 95~100 DEG C is taken repeatedly, to merge extract solution, obtain iris extract A；(2) wet thallus is obtained after being inoculated into broth bouillon culture centrifugation using acetic acid bacteria, in the mixed solution that wet thallus is added to polyvinyl alcohol and sodium alginate, and with containing 1~3%CaCl2Boric acid saturated solution and saturation Na2SO4PVA SA immobilization acetic acid bacteria beads are crosslinked to obtain in solution；(3) by iris extract A and immobilization acetic acid bacteria bead according to mass ratio 5~10:1 mixing after fermentation, then carries out molecular distillation, it is cigarette iris extract to take light component after filtering, precipitation, concentration.The present invention, which is added in cigarette, has abundant cigarette perfume, assigns the fragrant note of iris fragrance and acid of cigarette feature, increases cigarette perfume richness, soft fine and smooth flue gas reduces excitant, increases the sweet sense of flue gas.
Description
Technical field
The present invention relates to cigarette additive field, and in particular to prepares cigarette iris extract using immobilization acetic acid bacteria
Preparation method.
Background technology
Flavoring and casing is the key link in cigarette process, is the determinant and raising cigarette flavor for improving product style
One of effective means of quality.Current essence spice for cigarette is main or passes through the extraction to natural animal-plant, chemical synthesis system
It is standby.Due to by natural plant resource is limited and the harsher limitation of chemical synthesis condition.With the development of biotechnology, in recent years
Come, prepare cigarette flavor using microbe fermentation method, an economy, efficient approach are provided for the production of cigarette flavor,
Develop the new cigarette flavor that some conventional methods can not be produced.
Immobilized microorganism technology, is to be fixed on special microorganism on the carrier of choosing card, makes its highly dense and protect
Bioactivity is held, the biotechnology that can quickly, largely breed under condition of compatibility can make up suspension microorganism fermentation process
The shortcomings of efficiency is low, separation of solid and liquid is difficult, mithridatism is poor.Therefore, in recent years, the research of immobilized microorganism technology is lived very much
Jump, development is very fast, has application to many industrial departments such as food fermentation industry, environmental protection, new energy development and biological biography
Sensor etc., and deep development is achieved, show powerful superiority.It is mainly concerned with the following aspects：In food hair
It is used to produce ethanol, beer, Yoghourt etc. in ferment industry；It is used for environmental monitoring and the processing of industrial wastewater in environmental protection；
It is used to produce hydrogen, methane, biochemical fuel cell, microorganism battery etc. in new energy development；Immobilized microorganism is alternatively arranged as molecule
Recognition component, for making various biology sensors.And it is continuous ripe, perfect with fixation of microbe technology, gradually to height
The direction such as effect low energy consumption, low cost, simplification, safety automation, pollution-free is developed, dirty in energy crisis, shortage of resources, environment
In the case that dye is on the rise, it thus will be promoted more deeply and to be widely applied in terms of wastewater treatment, with more deepening
Remote meaning.Especially in terms of wastewater treatment, have great application prospect.But preparing cigarette using immobilized microorganism technology
Report is but had no with iris extract.
Iris (scientific name：Iris tectorum Maxim.) also known as blue butterfly, The purple butterfly, flat bamboo etc., belong to Iridaceae kite
Tail belongs to herbaceos perennial, originates in central China and Japan, is mainly distributed on the Chinese middle and south.Iris is available for viewing and admiring, flower
Fragrance is simple and elegant, can modulate perfume, and its rhizome can do Chinese medicine, and whole year can adopt, with antiinflammation.Using iris as the iris of raw material
Medicinal extract, iris extract, orris absolute etc. are widely used in cigarette, can assign the iris fragrance of cigarette feature, increase cigarette
Fragrant richness, the effect such as soft fine and smooth flue gas.
Acetic acid bacteria is that a class can produce the common name of organic acetic bacterials such as a large amount of acetic acid, acetic acid using fermentable carbohydrate
Bacterium is widely used in the brewing industries such as vinegar, fruit vinegar, but the seldom report in the preparation of cigarette natural perfume material.Acetic acid bacteria exists
A large amount of organic acids are produced in fermentation process, these organic acids have the work of reduction excitant and soft fine and smooth flue gas in cigarette
With simultaneously, moreover it is possible to assign the sour fragrant breeze lattice of cigarette feature, therefore, acetic acid bacteria fermentation is in the quality side of lifting cigarette natural perfume material
Face has a good application prospect.
The content of the invention
It is an object of the invention to provide the method that a kind of immobilization acetic acid bacteria prepares cigarette iris extract, using fixation
Change acetic acid bacteria fermentation technique, develop a kind of new iris extract, the extract can be used in cigarette, can be assigned
The fragrant note of iris fragrance and acid of cigarette feature, increases cigarette perfume richness, and soft fine and smooth flue gas reduces excitant, increases flue gas
Sweet sense.
The present invention provides the method that a kind of immobilization acetic acid bacteria prepares cigarette iris extract, it is characterised in that specific bag
Include following steps：
(1) orrisroot is taken, the water of 5~10 times of its weight is added, refluxing extraction 2~3 times, are carried every time at 95~100 DEG C
It is 2~3 hours to take the time, and room temperature is cooled to after having extracted, and is filtrated to get extract solution, merges multiple extract solution, obtains iris
Extract A；
(2) acetic acid bacteria is inoculated into broth bouillon, cultivated under conditions of 37 DEG C of temperature, speed of agitator are 50rpm
48 hours, take nutrient solution in being centrifuged under 8000rpm, the wet thallus after centrifugation is standby；
(3) polyvinyl alcohol-Immobilization in Sodium Alginate of acetic acid bacteria：It is water-soluble with sodium alginate using polyvinyl alcohol water solution
Liquid prepares the polyvinyl alcohol containing mass percent 6%~10% and the mixing of the sodium alginate of mass percent 1%~3% is molten
The acetic acid bacteria wet thallus prepared in liquid, the step of the 2~5% of total solution volume are then added into mixed solution (2), is configured to
Mixed liquor, CaCl is added dropwise to peristaltic pump by the syringe needle in 1~5mm apertures by the mixed liquor prepared2Quality percentage
Than in the boric acid saturated solution for 1~3%, then bead is taken out and adds saturation Na by crosslinking 1h formation beads2SO4In solution
3-5h is crosslinked, PVA-SA immobilization acetic acid bacteria beads, finally using aseptic water washing, dry for standby is obtained；
(4) by the iris extract A prepared in step (1) and the immobilization acetic acid bacteria bead that is prepared in step (3) according to
Mass ratio 5~10:1 mixing, is 30~37 DEG C in temperature, and speed of agitator is 2~4d of fermentation under conditions of 1~5rpm, is then put
7~14d is preserved in 4 DEG C of refrigerators；
(5) by the zymotic fluid of step (4), 80~95% alcoholic solutions of 2~4 times of its quality are added, precipitation 24~48 is small
When, it is 0.1MPa to filter to take supernatant in pressure, and temperature is that to be concentrated into relative density under conditions of 45 DEG C be 1.30~1.35, is obtained
Iris extract B；
(6) by the iris extract B of step (5), propane diols is added, in 0.1~10Pa of pressure, feeding temperature is 60 DEG C~
75 DEG C, charging rate is 2~20ml/min, and scraper plate rotating speed takes light component to carry out molecular distillation under conditions of 100~130rpm
For iris extract；The percentage that the addition of wherein propane diols accounts for molecular distillation thing gross mass is 10%~30%.
The acetic acid bacteria used in the present invention is acetobacter aceti (Acetobacter aceti) PDH2.05, in 2016 8
It is preserved within 29th Wuhan University's China typical culture collection center the moon, preserving number is CCTCC NO：M 2016436.
The immobilized microorganism technology that the present invention is used has that fermentation process efficiency high, separation of solid and liquid be easy, mithridatism is strong
The advantages of, the deficiency of traditional biological technique can be made up.
The present invention, using immobilization acetic acid bacteria fermentation iris extract, obtains a kind of quality excellent using orrisroot as raw material
Good cigarette iris extract, the extract has the iris fragrance for assigning cigarette feature in cigarette, increases cigarette perfume richness,
Increase the sweet sense of flue gas, lower the effect of excitant and soft fine and smooth flue gas, while also assigning the sour fragrant breeze lattice of cigarette characteristic；With
The fermentation of suspension acetic acid bacteria is compared, and iris extract organic acid content prepared by this method adds 5.28%~9.89%, cigarette
In effect assessment, oral cavity pleasant impression is more comfortable, and fragrance more enriches.This method instead of the traditional extraction skill of iris extract
Art, have updated biofermentation preparation technology so that it being capable of applicability increase, the preparation to cigarette iris extract in cigarette
There is provided a new way.
Embodiment
With reference to embodiment to further detailed description of the present invention.
The acetic acid bacteria used in the present invention is acetobacter aceti (Acetobacter aceti) PDH2.05, in 2016 8
It is preserved within 29th Wuhan University's China typical culture collection center the moon, preserving number is CCTCC NO：M 2016436.
PVA-SA (polyvinyl alcohol-sodium alginate) immobilization acetic acid bacteria bead in following examples of the present invention be according to
Lower method is obtained using polyvinyl alcohol (PVA) and sodium alginate (SA) Dichlorodiphenyl Acetate being fixed of bacterium：
(1) acetic acid bacteria is inoculated into broth bouillon, cultivated under conditions of 37 DEG C of temperature, speed of agitator are 50rpm
48 hours, take nutrient solution in being centrifuged under 8000rpm, the wet thallus after centrifugation is standby；Broth bouillon is conventional in the above method
Broth medium, its Ju Ti Pei Fang be (every liter)：Peptone 10g, beef extract powder 3g, sodium chloride 5g, final pH 7.3 ±
0.1；
(2) polyvinyl alcohol-Immobilization in Sodium Alginate of acetic acid bacteria：It is water-soluble with sodium alginate using polyvinyl alcohol water solution
Liquid prepares the mixed solution of the polyvinyl alcohol of the 6%-10% containing mass percent and mass percent 1%-3% sodium alginate,
Then the acetic acid bacteria wet thallus prepared in the step of 2-5% of total solution volume is added into mixed solution (2), is configured to mixing
Liquid, CaCl is added dropwise to peristaltic pump by the syringe needle in 1-5mm apertures by the mixed liquor prepared2Mass percent be 1-
In 3% boric acid saturated solution, then bead is taken out and adds saturation Na by crosslinking 1h formation beads2SO43- is crosslinked in solution
5h, obtains PVA-SA immobilization acetic acid bacteria beads, finally using aseptic water washing, dry for standby；It is described to contain 1-3%CaCl2's
The amount and saturation Na of boric acid saturated solution2SO4The amount of solution has no particular/special requirement, contains 1-3%CaCl2Boric acid saturated solution
Only need to that mixed liquor can be made to produce precipitation, meet the immobilization requirement of acetic acid bacteria, Na2SO4Solution is only needed to bead
It is completely soaked and just may be used；The boric acid saturated solution and saturation Na2SO4Solution is saturated aqueous solution.
A kind of embodiment 1, immobilization acetic acid bacteria prepares the method for cigarette iris extract, comprises the following steps：
(1) orrisroot is shredded, the water of 5 times of its weight is then added into the orrisroot of chopping, being flowed back at 100 DEG C carries
Take 2 times, each extraction time is 3 hours, room temperature be cooled to after having extracted, and be filtrated to get extract solution, merge extract solution twice,
Iris extract A is obtained, the condition extracted twice is identical；
(2) the PVA-SA immobilization acetic acid bacteria beads for preparing the iris extract A prepared in step (1) with the above method
According to mass ratio 5:1 mixing, is 30 DEG C in temperature, speed of agitator is the 4d that fermented under conditions of 5rpm, is subsequently placed in 4 DEG C of refrigerators
Preserve 7d；
(3) step (2) fermentation after fermentation liquid is filtered, obtains clarified broth, 95% alcohol for adding 3 times of its quality is molten
Liquid, is precipitated 24 hours, and it is 0.1MPa to filter to take supernatant in pressure, and temperature is to be concentrated into relative density under conditions of 45 DEG C to be
1.3, obtain iris extract B；
(4) the iris extract B for preparing step (3), adds propane diols, carries out molecular distillation, and it is iris to take light component
Extract；The Parameter Conditions of molecular distillation are：0.1~10Pa of pressure, feeding temperature be 60 DEG C~75 DEG C, charging rate be 2~
20ml/min, scraper plate rotating speed is 100~130rpm；The percentage that the addition of propane diols accounts for molecular distillation thing gross mass is
20%.
The iris extract prepared in embodiment 1 has abundant cigarette perfume in cigarette, assigns the iris fragrance of cigarette feature
With sour fragrant note, increase cigarette perfume richness, soft fine and smooth flue gas reduces excitant, increases the sweet sense of flue gas.
A kind of embodiment 2, immobilization acetic acid bacteria prepares the method for cigarette iris extract, comprises the following steps：
(1) orrisroot is taken, the water of 8 times of its weight is added after being crushed, refluxing extraction 3 times at 96 DEG C are extracted every time
Time is 2 hours, and room temperature is cooled to after having extracted, and is filtrated to get extract solution, merges No. three extract solutions, obtains iris extract
A；The condition of three extractions is identical；
(2) iris extract A is pressed with the PVA-SA immobilization acetic acid bacteria beads that prepare in the fixation above method for preparing
According to mass ratio 8:1 mixing, is 35 DEG C in temperature, speed of agitator is the 3d that fermented under conditions of 4rpm, is subsequently placed in 4 DEG C of refrigerators and protects
Deposit 14d；
(3) step (3) fermentation after fermentation liquid is filtered, obtains clarified broth, 80% alcohol for adding 4 times of its quality is molten
Liquid, is precipitated 48 hours, and it is 0.1MPa to filter to take supernatant in pressure, and temperature is to be concentrated into relative density under conditions of 45 DEG C to be
1.3, obtain iris extract B；
(4) the iris extract B for preparing step (3), adds propane diols, carries out molecular distillation, and it is iris to take light component
Extract；The Parameter Conditions of molecular distillation are：0.1~10Pa of pressure, feeding temperature be 60 DEG C~75 DEG C, charging rate be 2~
20ml/min, scraper plate rotating speed is 100~130rpm；The percentage that the addition of propane diols accounts for molecular distillation thing gross mass is
40%.
The iris extract prepared in embodiment 2 has abundant cigarette perfume, and the iris fragrance and acid for assigning cigarette feature are fragrant
Rhythm, increases cigarette perfume richness, and soft fine and smooth flue gas reduces excitant, increases the sweet sense of flue gas.
A kind of embodiment 3, immobilization acetic acid bacteria prepares the method for cigarette iris extract, comprises the following steps：
(1) orrisroot is taken, the water of 10 times of its weight is added after being crushed, refluxing extraction 3 times at 95 DEG C are extracted every time
Time is 3 hours, and room temperature is cooled to after having extracted, and is filtrated to get extract solution, merges No. three extract solutions, obtains iris extract
A；Three times extraction conditions are identical；
(2) the PVA-SA immobilization acetic acid bacteria beads for preparing the iris extract A prepared in step (1) with the above method
According to mass ratio 10:1 mixing, is 33 DEG C in temperature, speed of agitator is the 5d that fermented under conditions of 5rpm, is subsequently placed in 4 DEG C of refrigerators
Preserve 9d；
(3) step (2) fermentation after fermentation liquid is filtered, obtains clarified broth, 95% alcohol for adding 2 times of its quality is molten
Liquid, is precipitated 48 hours, and it is 0.1MPa to filter to take supernatant in pressure, and temperature is to be concentrated into relative density under conditions of 45 DEG C to be
1.35, obtain iris extract B；
(4) the iris extract B for preparing step (3), adds propane diols, carries out molecular distillation, and it is iris to take light component
Extract；The Parameter Conditions of molecular distillation are：0.1~10Pa of pressure, feeding temperature be 60 DEG C~75 DEG C, charging rate be 2~
20ml/min, scraper plate rotating speed is 100~130rpm；The percentage that the addition of propane diols accounts for molecular distillation thing gross mass is
30%.
The iris extract prepared in embodiment 3 has abundant cigarette perfume, and the iris fragrance and acid for assigning cigarette feature are fragrant
Rhythm, increases cigarette perfume richness, and soft fine and smooth flue gas reduces excitant, increases the sweet sense of flue gas.
Contrast experiment, applicant carries the iris that iris extract prepared by the present invention is prepared with existing traditional extraction process
Take thing, the iris extract prepared of being fermented using suspension acetic acid bacteria to be added to according to normal perfuming mode in pipe tobacco, cigarette be made,
And smoked panel test, its effect of smokeing panel test is as follows：
The sensory evaluation effect that iris prepared by three kinds of different process is extracted in cigarette compares
It can be seen that to come from above-mentioned contrast experiment, the iris extract for preparing is added in cigarette effect more in the present invention
It is good, and found according to specific experiment analysis, compared with suspension acetic acid bacteria ferments, iris extract organic acid prepared by this method
Content is added in 5.28%~9.89%, cigarette effect assessment, and oral cavity pleasant impression is more comfortable, and fragrance more enriches.
Claims (2)
1. a kind of method that immobilization acetic acid bacteria prepares cigarette iris extract, it is characterised in that specifically include following steps：
(1) orrisroot is taken, the water of 5~10 times of its weight is added, refluxing extraction 2~3 times at 95~100 DEG C, when extracting every time
Between be 2~3 hours, be cooled to room temperature after having extracted, and be filtrated to get extract solution, merge multiple extract solution, obtain iris and extract
Thing A；
(2) acetic acid bacteria is inoculated into broth bouillon, culture 48 is small under conditions of 37 DEG C of temperature, speed of agitator are 50rpm
When, take nutrient solution in being centrifuged under 8000rpm, the wet thallus after centrifugation is standby；
(3) polyvinyl alcohol-Immobilization in Sodium Alginate of acetic acid bacteria：Matched somebody with somebody using polyvinyl alcohol water solution with sodium alginate aqueous solution
The mixed solution of the polyvinyl alcohol containing mass percent 6%~10% and the sodium alginate of mass percent 1%~3% is made, so
The acetic acid bacteria wet thallus prepared in the step of adding the 2~5% of total solution volume in backward mixed solution (2), is configured to mixing
Liquid, CaCl is added dropwise to peristaltic pump by the syringe needle in 1~5mm apertures by the mixed liquor prepared2Mass percent be 1
In~3% boric acid saturated solution, then bead is taken out and adds saturation Na by crosslinking 1h formation beads2SO4It is crosslinked in solution
3-5h, obtains PVA-SA immobilization acetic acid bacteria beads, finally using aseptic water washing, dry for standby；
(4) by the immobilization acetic acid bacteria bead prepared in the iris extract A and step (3) that are prepared in step (1) according to quality
Than 5~10:1 mixing, is 30~37 DEG C in temperature, and speed of agitator is 2~4d of fermentation under conditions of 1~5rpm, is subsequently placed in 4 DEG C
7~14d is preserved in refrigerator；
(5) zymotic fluid after refrigerating in step (4) is taken, 80~95% alcoholic solutions of 2~4 times of its quality are added, precipitation 24~
48 hours, it was 0.1MPa to filter to take supernatant in pressure, temperature be concentrated under conditions of 45 DEG C relative density be 1.30~
1.35, obtain iris extract B；
(6) the iris extract B of step (5) is taken, propane diols is added, in 0.1~10Pa of pressure, feeding temperature is 60 DEG C~75
DEG C, charging rate is 2~20ml/min, and scraper plate rotating speed is to carry out molecular distillation under conditions of 100~130rpm, takes the light component to be
Iris extract；The percentage that the addition of wherein propane diols accounts for molecular distillation thing gross mass is 10%~30%.
2. the method that a kind of immobilization acetic acid bacteria according to claim 1 prepares cigarette iris extract, it is characterised in that：
Acetic acid bacteria in the step (2) is acetobacter aceti (Acetobacter aceti) PDH2.05, and deposit number is CCTCC
NO：M 2016436.
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Cited By (1)
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| CN108165371A (en) * | 2018-01-10 | 2018-06-15 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications of iris extract |
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| CN102771864A (en) * | 2012-07-31 | 2012-11-14 | 烟台绿杰饮料股份有限公司 | Fermenting type apple vinegar beverage and preparation method thereof |
| CN104585868A (en) * | 2015-01-20 | 2015-05-06 | 川渝中烟工业有限责任公司 | Human body essential amino acid-rich electronic cigarette juice |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108165371A (en) * | 2018-01-10 | 2018-06-15 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications of iris extract |
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