CN106957868A - A kind of immobilized yeast prepares method of the cigarette with tuberose extract - Google Patents
A kind of immobilized yeast prepares method of the cigarette with tuberose extract Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
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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
- A24B3/00—Preparing tobacco in the factory
- A24B3/12—Steaming, curing, or flavouring tobacco
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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Abstract
The present invention provides a kind of immobilized yeast and prepares method of the cigarette with tuberose extract.The preparation method is concretely comprised the following steps：(1) take tuberose to add water using Microwave Extraction repeatedly, merge extract solution, obtain tuberose extract A；(2) wet thallus is obtained after being inoculated into broth bouillon culture centrifugation using saccharomycete, in the mixed solution that wet thallus is added to polyvinyl alcohol and sodium alginate, and with containing CaCl2Boric acid saturated solution and saturation Na2SO4Fixing lactic acid bacteria bead is crosslinked to obtain in solution；(3) by tuberose extract A and fixing lactic acid bacteria bead mixing after fermentation, clarified broth is filtered to obtain, successively after freeze-drying, alcohol precipitation, filtering, concentration, molecular distillation, it is tuberose extract to take light component.The present invention has abundant cigarette perfume, and fragrant and sweet rhythm and the fragrance of a flower rhythm is more obvious, and cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.
Description
Technical field
The present invention relates to cigarette additive field, and in particular to prepares cigarette tuberose extract using immobilized yeast
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 tuberose extract.
Tuberose also known as cordate telosma, fragrant below the moon, Amaryllidaceae tuberose platymiscium is distributed in the areas such as Mexico, South America.With
Tuberose is the cigarette additive commonly used in cigarette for the spices tuberose essential oil and tuberose absolute of raw material, is had in cigarette
Abundant cigarette perfume, the increase effect such as the fragrance of a flower and fragrant and sweet note.
Saccharomycete is some unicellular fungis, is sac fungus, the common name of basidiomycetes Deng Ji sections unicellular fungi, typically refers to
The fungi of energy fermenting carbohydrate, available for production is brewageed, the bacterium can produce the fragrance thing such as ethyl acetate, benzyl carbinol during the fermentation
Matter, these aroma substances can assign the fragrant and sweet fragrance of cigarette feature, be a kind of high-quality aroma former for producing tobacco aromaticss.
The content of the invention
Method of the cigarette with tuberose extract is prepared it is an object of the invention to provide a kind of immobilized yeast, with reference to micro- life
The technologies such as thing immobilization fermentation, freeze-drying and molecular distillation, develop a kind of Novel tobacco tuberose extract.
The technical scheme that the present invention is provided：A kind of immobilized yeast prepares method of the cigarette with tuberose extract,
It is characterized in that specifically including following steps：
(1) tuberose is taken, the water of 3~5 times of its weight is added, Microwave Extraction 2~3 times, are carried every time at 50~65 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 late perfume (or spice)
Beautiful extract A；
(2) saccharomycete 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 saccharomycete：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 saccharomycete wet thallus prepared in liquid, the step of the 1~3% of total solution volume is 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%, 1h being crosslinked, then in saturation Na2SO43-5h is crosslinked in solution, PVA-SA is obtained and fixes
Change saccharomycete bead, then using aseptic water washing, dry for standby；
(4) the tuberose extract A extracted in step (1) and the immobilized yeast bead of preparation in step (3) are pressed
According to mass ratio 5~10:1 mixing, is 20~25 DEG C in temperature, and speed of agitator is 3~5d of fermentation under conditions of 1~5rpm, then
It is placed in 7~14d of preservation in 4 DEG C of refrigerators；
(5) step (4) fermentation after fermentation liquid is filtered, obtains clarified broth, zymotic fluid is freeze-dried, be freeze-dried
Feed liquid thickness be 1~2 centimetre, vacuum is 10Pa, and drying time is 6~12h；
(6) zymotic fluid after being freeze-dried in step (5) is taken, 80~95% alcoholic solutions of 2~4 times of its quality are added,
Precipitation 24~48 hours, 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
1.25~1.30, propane diols is then added in concentrate, tuberose extract B is obtained, the addition of the propane diols is total matter
The 20% of amount；
(7) by the tuberose extract B in step (6), in 0.1~5Pa of pressure, feeding temperature is 50 DEG C~65 DEG C, is entered
Material speed is 10~30ml/min, and scraper plate rotating speed is 50~90rpm, carries out molecular distillation, and it is tuberose extract to take light component.
The saccharomycete used in the present invention is saccharomyces cerevisiae (Saccharomyces cerevisiae) HHL-1, in 2009
It is preserved in Wuhan University's China typical culture collection center on November 17, in, deposit number is CCTCC NO:M 209272.
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 immobilized yeast fermentation tuberose extract, obtains a kind of product using tuberose as raw material
Good cigarette tuberose extract of fine quality, the extract has abundant cigarette perfume in cigarette, increases fragrant and sweet rhythm and fragrance of a flower rhythm, increase
The cigarette flavor property sent out thoroughly, oral cavity pleasant impression is relatively comfortable.Compared with existing tuberose concrete and tuberose extract, the tuberose is extracted
Thing fragrance in cigarette more enriches, and fresh and sweet note and the fragrance of a flower rhythm is more obvious, and the property sent out is more preferable thoroughly for cigarette flavor；With suspended yeast
Bacterium fermentation is compared, and esters fragrance content of material adds 14.85%~19.64% in tuberose extract prepared by this method,
Benzyl carbinol content is added in 24.35%~28.52%, cigarette effect assessment, and fragrance more enriches, and fragrant and sweet rhythm and the fragrance of a flower rhythm is more
Plus substantially, cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.This method instead of the traditional extractive technique of tuberose extract,
It has updated biofermentation preparation technology so that it applicability can increase in cigarette, to preparation of the cigarette with tuberose extract
There is provided a new way.
Embodiment
With reference to embodiment to further detailed description of the present invention.
PVA-SA (polyvinyl alcohol-sodium alginate) immobilized yeast bead in following examples of the present invention be according to
Lower method is obtained using polyvinyl alcohol (PVA) and sodium alginate (SA) to being fixed of saccharomycete：
(1) saccharomycete 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；It is conventional that agar culture is supported 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 saccharomycete：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 saccharomycete wet thallus prepared in liquid, the step of the 1~3% of total solution volume is 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 is then added to saturation Na by crosslinking 1h formation beads2SO4Solution
Middle crosslinking 3-5h, obtains PVA-SA immobilized yeast beads, then using aseptic water washing, dry for standby；It is described to contain 1-3%
CaCl2Boric acid saturated solution amount and saturation Na2SO4The amount of solution has no particular/special requirement, contains 1-3%CaCl2Boric acid satisfy
Only need to that mixed liquor can be made to produce precipitation with solution, meet the immobilization requirement of lactic acid bacteria, Na2SO4Solution is only needed to
Bead is completely soaked and just may be used；The boric acid saturated solution, saturation Na2SO4Solution is saturated aqueous solution.
The saccharomycete used in the present invention is saccharomyces cerevisiae (Saccharomyces cerevisiae) HHL-1, in 2009
It is preserved in Wuhan University's China typical culture collection center on November 17, in, deposit number is CCTCC NO:M 209272.
A kind of embodiment 1, immobilized yeast prepares method of the cigarette with tuberose extract, comprises the following steps：
(1) tuberose is taken, the water of 3 times of its weight is added, Microwave Extraction 2 times at 50 DEG C, each extraction time is 2 small
When, room temperature is cooled to after having extracted, and extract solution is filtrated to get, merge multiple extract solution, obtain tuberose extract A；Twice
The condition of extraction is identical；
(2) the tuberose extract A extracted in step (1) and the immobilized yeast bead of preparation in step (3) are pressed
According to mass ratio 5:1 mixing, is 20 DEG C in temperature, speed of agitator is the 3d that fermented under conditions of 1rpm, is subsequently placed in 4 DEG C of refrigerators and protects
Deposit 7d；
(3) step (2) fermentation after fermentation liquid is filtered, obtains clarified broth, zymotic fluid is freeze-dried, be freeze-dried
Feed liquid thickness be 1~2 centimetre, vacuum is 10Pa, and drying time is 6h；
(4) zymotic fluid after being freeze-dried in step (3) is taken, 80% alcoholic solution of 2 times of its quality is added, precipitation 24 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 1.25, then dense
Propane diols is added in contracting thing, tuberose extract B is obtained, the addition of the propane diols is the 20% of gross mass；
(5) by the tuberose extract B in step (4), in pressure 1Pa, feeding temperature is 50 DEG C, and charging rate is
10ml/min, scraper plate rotating speed is 50rpm, carries out molecular distillation, and it is tuberose extract to take light component.
The tuberose extract prepared in embodiment 1 has abundant cigarette perfume in cigarette, and fragrance more enriches, fragrant and sweet rhythm and
Fragrance of a flower rhythm is more obvious, and cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.
A kind of embodiment 2, immobilized yeast prepares method of the cigarette with tuberose extract, comprises the following steps：
(1) tuberose is taken, the water of 4 times of its weight is added, Microwave Extraction 2 times at 55 DEG C, each extraction time is 2.5
Hour, room temperature is cooled to after having extracted, and extract solution is filtrated to get, merge multiple extract solution, obtain tuberose extract A；Two
The condition of secondary extraction is identical；
(2) the tuberose extract A extracted in step (1) and the immobilized yeast bead of preparation in step (3) are pressed
According to mass ratio 8:1 mixing, is 22 DEG C in temperature, speed of agitator is the 4d that fermented under conditions of 3rpm, is subsequently placed in 4 DEG C of refrigerators and protects
Deposit 10d；
(3) step (2) fermentation after fermentation liquid is filtered, obtains clarified broth, zymotic fluid is freeze-dried, be freeze-dried
Feed liquid thickness be 1~2 centimetre, vacuum is 10Pa, and drying time is 8h；
(4) zymotic fluid after being freeze-dried in step (3) is taken, 90% alcoholic solution of 3 times of its quality is added, precipitation 24 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 1.25, then dense
Propane diols is added in contracting thing, tuberose extract B is obtained, the addition of the propane diols is the 20% of gross mass；
(5) by the tuberose extract B in step (4), in pressure 3Pa, feeding temperature is 60 DEG C, and charging rate is
20ml/min, scraper plate rotating speed is 70rpm, carries out molecular distillation, and it is tuberose extract to take light component.
The tuberose extract prepared in embodiment 2 has abundant cigarette perfume in cigarette, and fragrant and sweet rhythm and the fragrance of a flower rhythm is brighter
Aobvious, cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.
A kind of embodiment 3, immobilized yeast prepares method of the cigarette with tuberose extract, comprises the following steps：
(1) tuberose is taken, the water of 5 times of its weight is added, Microwave Extraction 3 times at 65 DEG C, each extraction time is 3 small
When, room temperature is cooled to after having extracted, and extract solution is filtrated to get, merge multiple extract solution, obtain tuberose extract A；Three times
The condition of extraction is identical；
(2) the tuberose extract A extracted in step (1) and the immobilized yeast bead of preparation in step (3) are pressed
According to mass ratio 10:1 mixing, is 25 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 14d；
(3) step (2) fermentation after fermentation liquid is filtered, obtains clarified broth, zymotic fluid is freeze-dried, be freeze-dried
Feed liquid thickness be 1~2 centimetre, vacuum is 10Pa, and drying time is 9h；
(4) zymotic fluid after being freeze-dried in step (3) is taken, 95% alcoholic solution of 4 times of its quality is added, precipitation 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, then dense
Propane diols is added in contracting thing, tuberose extract B is obtained, the addition of the propane diols is the 20% of gross mass；
(5) by the tuberose extract B in step (4), in pressure 5Pa, feeding temperature is 65 DEG C, and charging rate is 30
Ml/min, scraper plate rotating speed is 90rpm, carries out molecular distillation, and it is tuberose extract to take light component.
The tuberose extract prepared in embodiment 3 has abundant cigarette perfume in cigarette, and fragrant and sweet rhythm and the fragrance of a flower rhythm is brighter
Aobvious, cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.
Contrast experiment, the tuberose extract that applicant prepares the present invention and evening prepared by existing traditional extraction process are fragrant
Beautiful extract, the tuberose extract prepared that fermented using suspended yeast bacterium are added in pipe tobacco according to normal perfuming mode, are made
Into cigarette, and smoked panel test, its effect of smokeing panel test is as follows：
The sensory evaluation effect that tuberose 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 tuberose 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 suspended yeast bacterium is fermented, ester in tuberose extract prepared by this method
Class Aromatic Matter Contents add 14.85%~19.64%, and benzyl carbinol content adds 24.35%~28.52%, cigarette effect
During fruit is evaluated, fragrance more enriches, and fragrant and sweet rhythm and the fragrance of a flower rhythm is more obvious, and cigarette perfume is sent out more thoroughly, and oral cavity pleasant impression is more comfortable.
Claims (2)
1. a kind of immobilized yeast prepares method of the cigarette with tuberose extract, it is characterised in that specifically include following steps：
(1) tuberose is taken, the water of 3~5 times of its weight is added, Microwave Extraction 2~3 times at 50~65 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, the multiple extract solution of merging obtains tuberose and carried
Take thing A；
(2) saccharomycete 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 saccharomycete：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 saccharomycete wet thallus prepared in the step of adding the 1~3% 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 immobilized yeast beads, then using aseptic water washing, dry for standby；
(4) by the middle immobilized yeast bead prepared of the tuberose extract A extracted in step (1) and step (3) according to matter
Measure ratio 5~10:1 mixing, is 20~25 DEG C in temperature, and speed of agitator is 3~5d of fermentation under conditions of 1~5rpm, is subsequently placed in 4
7~14d is preserved in DEG C refrigerator；
(5) step (4) fermentation after fermentation liquid is filtered, obtains clarified broth, zymotic fluid is freeze-dried, the material of freeze-drying
Liquid thickness is 1~2 centimetre, and vacuum is 10Pa, and drying time is 6~12h；
(6) zymotic fluid after being freeze-dried in step (5) is taken, 80~95% alcoholic solutions of 2~4 times of its quality are added, precipitated
24~48 hours, it was 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 1.25
~1.30, propane diols is then added in concentrate, tuberose extract B is obtained, the addition of the propane diols is gross mass
20%；
(7) by the tuberose extract B in step (6), in 0.1~5Pa of pressure, feeding temperature is 50 DEG C~65 DEG C, charging speed
Spend for 10~30ml/min, scraper plate rotating speed is 50~90rpm, carries out molecular distillation, it is tuberose extract to take light component.
2. a kind of immobilized yeast according to claim 1 prepares method of the cigarette with tuberose extract, its feature exists
In:Saccharomycete in the step (2) is saccharomyces cerevisiae (Saccharomyces cere visiae) HHL-1, and deposit number is
CCTCC NO:M 209272。
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2016
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| Publication number | Priority date | Publication date | Assignee | Title |
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Application publication date: 20170718 |