CN107674885A - A kind of method that immobilized yeast prepares cigarette feltwort extract - Google Patents
A kind of method that immobilized yeast prepares cigarette feltwort extract Download PDFInfo
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
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Abstract
The present invention provides a kind of method that immobilized yeast prepares cigarette feltwort extract.The preparation method concretely comprises the following steps：(1) take feltwort to add water multiple using Microwave Extraction, merge extract solution, obtain feltwort extract A；(2) wet thallus is obtained after being inoculated into broth bouillon culture centrifugation using saccharomycete, wet thallus is added in the mixed solution of 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 feltwort extract A and fixing lactic acid bacteria bead according to mass ratio 5~10:1 mixing after fermentation, then filters to obtain clarified broth, successively after freeze-drying, alcohol precipitation, filtering, concentration, molecular distillation, it is feltwort extract to take light component.The present invention has abundant cigarette perfume, and fresh and sweet note and the fragrance of a flower rhythm is more obvious, increases the cigarette flavor property sent out thoroughly, reduces excitant.
Description
Technical field
The present invention relates to cigarette additive field, and in particular to prepares cigarette feltwort 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.Essence spice for cigarette is main at present 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 produce.
Immobilized microorganism technology, it 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, suspension microorganism fermentation process can be made up
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 very living
Jump, development is very fast, has application to many industrial departments such as food fermentation industry, environmental protection, and new energy development and biology pass
Sensor etc., and deep development is achieved, show powerful superiority.It is mainly concerned with the following aspects：Sent out in food
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 to go deep into and be widely applied in terms of wastewater treatment, have and more deepen
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 feltwort extract.
Feltwort, Scrophulariaceae Verbascum biennial herb plant, also known as hygrometric boea herb, big hair leaf, common anotis herb, speedwell,
Rattle stick etc., it is distributed widely in the ground such as the Northern Hemisphere, Xinjiang of China, Tibet, Yunnan, Sichuan, Zhejiang, Jiangsu.Like monarch according to Dong,
Spices feltwort essential oil using feltwort as raw material is the cigarette additive commonly used in cigarette, have in cigarette abundant cigarette perfume,
Increase delicate fragrance rhythm, reduce irritating effect.
Saccharomycete is some unicellular fungis, is the common name of sac fungus, 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
It is an object of the invention to provide a kind of method that immobilized yeast prepares cigarette feltwort extract, with reference to micro- life
The technologies such as thing immobilization fermentation, freeze-drying and molecular distillation, develop a kind of Novel tobacco feltwort extract.
Technical scheme provided by the invention is：A kind of immobilized yeast prepares the side of cigarette feltwort extract
Method, it is characterised in that specifically include following steps：
(1) feltwort is taken, adds the water of 3~8 times of its weight, Microwave Extraction 2~3 times at 50~65 DEG C, extraction every time
Time is 2~3 hours, and room temperature is cooled to after having extracted, and is filtrated to get extract solution, merges multiple extract solution, obtains feltwort
Extract A；
(2) saccharomycete is inoculated into broth bouillon, cultivated under conditions of being 50rpm in 37 DEG C of temperature, speed of agitator
48 hours, nutrient solution is taken to be 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
The mixing that liquid prepares the sodium alginate of polyvinyl alcohol and mass percent 1%~3% containing mass percent 6%~10% is molten
Liquid, then into mixed solution add total solution volume 1~3% the step of (2) in prepare saccharomycete wet thallus, be configured to
Mixed liquor, the mixed liquor prepared is added dropwise to CaCl by the syringe needle in 1~5mm apertures with peristaltic pump2Quality 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, be dried for standby；
(4) the feltwort extract A of extraction in step (1) is pressed with the immobilized yeast bead prepared in step (3)
According to mass ratio 5~10:1 mixing, it is 15~22 DEG C in temperature, 5~7d of fermentation under conditions of speed of agitator is 1~5rpm, then
It is placed in 7~14d of preservation in 4 DEG C of refrigerators；
(5) by step (4) fermentation after fermentation liquid filtering, clarified broth is obtained, zymotic fluid is freeze-dried, and it freezes dry
Dry feed liquid thickness is 1~2 centimetre, vacuum 10Pa, and drying time is 6~12h；
(6) zymotic fluid after being freeze-dried in step (5) is taken, adds 80~95% alcoholic solutions of 2~4 times of its quality,
Precipitation 24~48 hours, it is 0.1MPa to filter to take supernatant in pressure, and temperature is concentrated into relative density under conditions of being 45 DEG C and is
1.20~1.25, propane diols is then added in concentrate, obtains feltwort extract B, the addition of the propane diols is total matter
The 20% of amount；
(7) the feltwort extract B that will be prepared in step (6), in 1~10Pa, feeding temperature is 50 DEG C~60 DEG C, charging
Speed is 1~10ml/min, and scraper plate rotating speed is 100~150rpm, carries out molecular distillation, takes light component to be extracted for feltwort
Thing.
The saccharomycete used in the present invention is saccharomyces cerevisiae (Saccharomyces cerevisiae) HHL-1, in 2009
On November 17, in is preserved in Wuhan University's China typical culture collection center, and deposit number is CCTCC NO:M 209272.
The immobilized microorganism technology that the present invention uses has that fermentation process efficiency high, separation of solid and liquid are easy, mithridatism is strong
The advantages that, the deficiency of traditional biological technique can be made up.
The present invention, using immobilized yeast fermentation feltwort extract, obtains a kind of product using feltwort as raw material
Good cigarette feltwort extract of fine quality, the extract have abundant cigarette perfume in cigarette, increase fragrant and sweet and floral notes, increase
The property sent out, oral cavity pleasant impression are relatively comfortable thoroughly for cigarette flavor.Compared with existing feltwort medicinal extract and feltwort extract, feltwort extraction
Thing fragrance in cigarette is more abundant, 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 11.15%~16.89% in feltwort extract prepared by this method,
Benzyl carbinol content adds 19.58%~20.31%, and in cigarette effect assessment, fragrance is more abundant, and fresh and sweet fragrant and fragrance of a flower rhythm is more
Add substantially, oral cavity pleasant impression is more comfortable.This method instead of the traditional extractive technique of feltwort extract, have updated biofermentation
Preparation technology so that it applicability can increase in cigarette, and a new way is provided with the preparation of feltwort extract to cigarette
Footpath.
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 being 50rpm in 37 DEG C of temperature, speed of agitator
48 hours, nutrient solution is taken to be 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 specific formula is (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
The mixing that liquid prepares the sodium alginate of polyvinyl alcohol and mass percent 1%~3% containing mass percent 6%~10% is molten
Liquid, then into mixed solution add total solution volume 1~3% the step of (2) in prepare saccharomycete wet thallus, be configured to
Mixed liquor, the mixed liquor prepared is added dropwise to CaCl by the syringe needle in 1~5mm apertures with peristaltic pump2Quality percentage
Than in the boric acid saturated solution for 1~3%, crosslinking 1h forms bead, then bead is taken out and is then added to saturation Na2SO4Solution
Middle crosslinking 3-5h, PVA-SA immobilized yeast beads are obtained, then using aseptic water washing, are dried 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 it is required to make mixed liquor produce precipitation with solution, meets the immobilization requirement of lactic acid bacteria, Na2SO4Solution only needs
Bead is completely soaked；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
On November 17, in is preserved in Wuhan University's China typical culture collection center, and deposit number is CCTCC NO:M 209272.
Embodiment 1, a kind of method that immobilized yeast prepares cigarette feltwort extract, comprises the following steps：
(1) feltwort is taken, adds the water of 3 times of its weight, Microwave Extraction 2 times at 65 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 feltwort extract A, twice
The condition of extraction is identical；
(2) the PVA-SA immobilized yeasts bead for preparing feltwort extract A and the above method is according to mass ratio 5:1
Mixing, it is 22 DEG C in temperature, speed of agitator is the 6d that fermented under conditions of 5rpm, is subsequently placed in 4 DEG C of refrigerators and preserves 14d；
(3) by step (2) fermentation after fermentation liquid filtering, clarified broth is obtained, zymotic fluid is freeze-dried, feed liquid thickness
For 1~2 centimetre, vacuum 10Pa, drying time 6h；
(4) zymotic fluid after step (3) is freeze-dried, adds 80% alcoholic solution of 2 times of its quality, and precipitation 24 is small
When, it is 0.1MPa to filter to take supernatant in pressure, and it is 1.20 that temperature is concentrated into relative density under conditions of being 45 DEG C, then dense
The propane diols of gross mass 20% is added in contracting thing, is well mixed to obtain feltwort extract B；
(5) the feltwort extract B of step (4) is taken, sets the parameter of molecular distillation：Pressure 5Pa, feeding temperature 50
DEG C, charging rate 10ml/min, scraper plate rotating speed is 100rpm, carries out molecular distillation, it is feltwort flower extract to take light component.
The feltwort extract prepared in embodiment 1, which is added in cigarette, has abundant cigarette perfume, fresh and sweet note and the fragrance of a flower rhythm
It is more obvious, increase the cigarette flavor property sent out thoroughly, reduce excitant.
Embodiment 2, a kind of method that immobilized yeast prepares cigarette feltwort extract, comprises the following steps：
(1) feltwort is taken, adds the water of 6 times of its weight, Microwave Extraction 3 times at 60 DEG C, each extraction time is 2.5 small
When, room temperature is cooled to after having extracted, and extract solution is filtrated to get, merge multiple extract solution, obtain feltwort extract A, three times
The condition of extraction is identical；
(2) the PVA-SA immobilized yeasts bead for preparing feltwort extract A and the above method is according to mass ratio 7:1
Mixing, it is 20 DEG C in temperature, speed of agitator is the 7d that fermented under conditions of 4rpm, is subsequently placed in 4 DEG C of refrigerators and preserves 10d；
(3) by step (2) fermentation after fermentation liquid filtering, clarified broth is obtained, zymotic fluid is freeze-dried, feed liquid thickness
For 1~2 centimetre, vacuum 10Pa, drying time 10h；
(4) zymotic fluid after step (3) is freeze-dried, adds 90% alcoholic solution of 3 times of its quality, and precipitation 48 is small
When, it is 0.1MPa to filter to take supernatant in pressure, and it is 1.25 that temperature is concentrated into relative density under conditions of being 45 DEG C, then dense
The propane diols of gross mass 20% is added in contracting thing, is well mixed to obtain feltwort extract B；
(5) the feltwort extract B of step (4) is taken, sets the parameter of molecular distillation：Pressure 10Pa, feeding temperature 60
DEG C, charging rate 6ml/min, scraper plate rotating speed is 130rpm, carries out molecular distillation, it is feltwort flower extract to take light component.
The feltwort extract prepared in embodiment 2, which is added in cigarette, has abundant cigarette perfume, fresh and sweet note and the fragrance of a flower rhythm
It is more obvious, increase the cigarette flavor property sent out thoroughly, reduce excitant.
Embodiment 3, a kind of method that immobilized yeast prepares cigarette feltwort extract, comprises the following steps：
(1) feltwort is taken, adds the water of 8 times of its weight, Microwave Extraction 2 times at 50 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 feltwort extract A, twice
The condition of extraction is identical；
(2) the PVA-SA immobilized yeasts bead for preparing feltwort extract A and the above method is according to mass ratio 10:
1 mixing, it is 15 DEG C in temperature, speed of agitator is the 5d that fermented under conditions of 2rpm, is subsequently placed in 4 DEG C of refrigerators and preserves 7d；
(3) by step (2) fermentation after fermentation liquid filtering, clarified broth is obtained, zymotic fluid is freeze-dried, feed liquid thickness
For 1~2 centimetre, vacuum 10Pa, drying time 6h；
(4) zymotic fluid after step (3) is freeze-dried, adds 95% alcoholic solution of 4 times of its quality, and precipitation 48 is small
When, it is 0.1MPa to filter to take supernatant in pressure, and it is 1.25 that temperature is concentrated into relative density under conditions of being 45 DEG C, is then added
Propane diols obtains feltwort extract B；The addition of propane diols is the 20% of gross mass；
(5) the feltwort extract B of step (4) is taken, sets the parameter of molecular distillation：Pressure 2Pa, feeding temperature 55
DEG C, charging rate 2ml/min, scraper plate rotating speed is 150rpm, carries out molecular distillation, it is feltwort flower extract to take light component.
The feltwort extract prepared in embodiment 3, which is added in cigarette, has abundant cigarette perfume, fresh and sweet note and the fragrance of a flower rhythm
It is more obvious, increase the cigarette flavor property sent out thoroughly, reduce excitant.
Contrast experiment, the hair stamen that applicant prepares feltwort extract prepared by the present invention with existing traditional extraction process
Flower extract, the feltwort extract prepared of being 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 feltwort 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 feltwort 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 feltwort extract prepared by this method
Class Aromatic Matter Contents add 11.15%~16.89%, and benzyl carbinol content adds 19.58%~20.31%, cigarette effect
In fruit evaluation, fragrance is more abundant, and fresh and sweet fragrant and fragrance of a flower rhythm is more obvious, and oral cavity pleasant impression is more comfortable.
Claims (2)
1. a kind of method that immobilized yeast prepares cigarette feltwort extract, it is characterised in that specifically include following steps：
(1) feltwort is taken, adds the water of 3~8 times of its weight, Microwave Extraction 2~3 times at 50~65 DEG C, each extraction time
For 2~3 hours, room temperature is cooled to after having extracted, and is filtrated to get extract solution, merges multiple extract solution, obtains feltwort extraction
Thing A；
(2) saccharomycete is inoculated into broth bouillon, culture 48 is small under conditions of being 50rpm in 37 DEG C of temperature, speed of agitator
When, take nutrient solution to be 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 and sodium alginate aqueous solution
The mixed solution of the sodium alginate of polyvinyl alcohol and mass percent 1%~3% containing mass percent 6%~10% 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 mix
Liquid, the mixed liquor prepared is added dropwise to CaCl by the syringe needle in 1~5mm apertures with peristaltic pump2Mass percent be 1
In~3% boric acid saturated solution, crosslinking 1h forms bead, then takes out bead and adds saturation Na2SO4It is crosslinked in solution
3-5h, PVA-SA immobilized yeast beads are obtained, then using aseptic water washing, are dried for standby；
(4) by the immobilized yeast bead prepared in the feltwort extract A of extraction in step (1) and step (3) according to matter
Measure ratio 5~10:1 mixing, it is 15~22 DEG C in temperature, 5~7d of fermentation, is subsequently placed in 4 under conditions of speed of agitator is 1~5rpm
7~14d is preserved in DEG C refrigerator；
(5) by step (4) fermentation after fermentation liquid filtering, clarified broth is obtained, zymotic fluid is freeze-dried, what it was freeze-dried
Feed liquid thickness is 1~2 centimetre, vacuum 10Pa, and drying time is 6~12h；
(6) zymotic fluid after being freeze-dried in step (5) is taken, adds 80~95% alcoholic solutions of 2~4 times of its quality, precipitation
24~48 hours, it was 0.1MPa to filter to take supernatant in pressure, and it is 1.20 that temperature is concentrated into relative density under conditions of being 45 DEG C
~1.25, propane diols is then added in concentrate, obtains feltwort extract B, the addition of the propane diols is gross mass
20%；
(7) the feltwort extract B that will be prepared in step (6), in 1~10Pa of pressure, feeding temperature is 50 DEG C~60 DEG C, charging
Speed is 1~10ml/min, and scraper plate rotating speed is 100~150rpm, carries out molecular distillation, takes light component to be extracted for feltwort
Thing.
2. the method that a kind of immobilized yeast according to claim 1 prepares cigarette feltwort extract, its feature exist
Saccharomycete in the step (2) is saccharomyces cerevisiae (Saccharomyces cerevisiae) HHL-1, and deposit number is
CCTCC NO:M 209272。
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