CN102492729A - Leavening with violet perfume and production method and purpose thereof - Google Patents
Leavening with violet perfume and production method and purpose thereof Download PDFInfo
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- CN102492729A CN102492729A CN2011104001404A CN201110400140A CN102492729A CN 102492729 A CN102492729 A CN 102492729A CN 2011104001404 A CN2011104001404 A CN 2011104001404A CN 201110400140 A CN201110400140 A CN 201110400140A CN 102492729 A CN102492729 A CN 102492729A
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- ionol
- dihydro
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Abstract
The invention relates to a microbial leavening with violet perfume and a preparation method thereof. The method includes Paenibacillus amylolyticus BM-B-340 seed culture, shaking fermentation, leavening separation and other steps. The leavening containing 7,8-dihydro-Beta-ionol has the perfume of natural tobacco plant. Compared with the perfume of the conventional tobacco essence Beta-irisone, the perfume of the product is more elegant and unique and milder, so the product has a good market prospect and high application value.
Description
[technical field]
The present invention relates to microbial fermentation technology.More specifically; The present invention relates to a kind of utilization separates starch based genus bacillus (Paenibacillus amylolyticus) fermentative prodn and contains 7; The method of 8-dihydro-β-ionol fermented product; Also relate to according to what said method prepared and contain 7, the fermented product of 8-dihydro-β-ionol and as the purposes of tobacco aromatics using.
[background technology]
In plant flower, tealeaves, grape, rose and the tobacco material, ubiquity ionol and jononeionone fragrance verivates such as (the pentadiene class fall in C13).This analog derivative is generated through the peroxo-fission reaction by precursor substances such as carrotenoid such as β-Hu Luobusu, xenthophylls, zeaxanthin, astaxanthins usually.In the perfume industry production process, ionol and jononeionone are one of important synthetic spices, and the further reduction of jononeionone can obtain ionol.According to the variation of jononeionone position of double bond, mainly divide α and β two kinds of isomer, both all have cypress fragrance, have the fragrance of VT after the dilution.The fragrance of jononeionone is very graceful, soft, in makeup and food flavor(ing), uses very wide.In addition, alpha, beta-lonone also is widely used in tobacco, food and fodder additives, is very important industrial raw material.
Ionol and jononeionone are widely used in the cigarette industry.It is reported that the ionol Ester burns and sucks under the state at cigarette, and scission reaction can take place, therefore a kind of flavor matter---Megastigmatrienone is arranged in flue gas.Megastigmatrienone plays an important role to the fragrance of cigarette, has become to pass judgment on a good and bad important indicator of cigarette.This shows that the cracking of materials such as ionol is the important channel that tobacco flavor material forms, and such fragrance precursor substance directly is added in the tobacco as feed liquid, can significantly improve the tobacco aroma quality.
At present, ionol and jononeionone preparation method mainly are industrial synthesis methods, and this method industry synthesis yield is merely 25%, and synthesis material costs an arm and a leg and is difficult for buying.In addition; Plant extract method equipment cost is high; Extraction yield is extremely low; Consider the factor such as industrial formation condition, production cost, purchasing of raw materials price of ionol, the applicant attempts adopting microbe fermentation method to prepare a kind of material with VT fragrance first, for the essence industry provides a kind of novel tobacco aromatics using.
[summary of the invention]
[problem that invention will solve]
The purpose of this invention is to provide a kind of preparation method with fermented product of VT fragrance.
Another object of the present invention provides the fermented product with VT fragrance that adopts said working method to obtain.
Another object of the present invention provides said purposes with fermented product of VT fragrance as tobacco aromatics using
[technical scheme]
The present invention relates to a kind of working method with fermented product of VT fragrance.
Said working method may further comprise the steps:
A, seed culture
Use yeast extract, NaCl, glucose, peptone, liquid microelement and deionized water preparation to contain the seed culture medium of yeast extract 4-6g/L, NaCl 25-32g/L, glucose 0.1-0.4g/L, peptone 8-15g/L, liquid microelement 1.0mL/L; Described liquid microelement is to contain H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O0.03g/L, ZnCl
20.02g/L, CoCl
26H
2The aqueous solution of O 0.04g/L;
Be added to said seed culture medium in the culturing bottle; With mineral alkali the pH value of this seed culture medium is adjusted to weakly alkaline; Sterilization then; Insert again with said seed culture medium volumeter 0.1-5% and separate starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340, at room temperature cultivated then 80-120 hour, obtain seed liquor;
B, shake flask fermentation
Use yeast extract, NaCl, peptone, xenthophylls, above-mentioned liquid microelement and deionized water preparation to contain the shake flask fermentation substratum of yeast extract 5.0-5.5g/L, NaCl 25-32g/L, peptone 8.5-12.0g/L, xenthophylls 180-200mg/L, liquid microelement 1.0mL/L;
Be added to said shake flask fermentation substratum in the culturing bottle; With mineral alkali the pH value of this shake flask fermentation substratum is adjusted to weakly alkaline; Sterilization at room temperature inserts the seed culture medium that obtains in steps A more then, and inoculum size is in said shake flask fermentation culture volume 5-10%; In the 50-80rpm shaking table, cultivate 80-120h down, obtain fermented liquid at temperature 28-34 ℃;
C, separation
The fermented liquid that step B is obtained carries out spinning, in its supernatant, adds with 1: 1 organic solvent of said supernatant volumeter, at room temperature carries out oscillation extraction, separates the organic phase that obtains and volatilizes, and obtains having the fermented product of VT fragrance.
A preferred embodiment of the invention, described mineral alkali are NaOH, KOH or Ca (OH)
2
According to another kind of preferred implementation of the present invention, in steps A and B, said weakly alkaline pH value is 7.2-7.4.
According to another kind of preferred implementation of the present invention, in step C, said organic solvent is selected from normal hexane, chloroform, acetone, ether, tetracol phenixin and/or sherwood oil.
According to another kind of preferred implementation of the present invention, in step C, it is nitrogen flushing method, vacuum decompression method or distillation method that organic phase volatilizes.
The invention still further relates to the fermented product that adopts said method to obtain, it is characterized in that it contains 60.0-70.0mg/L 7,8-dihydro-β-ionol with VT fragrance.
A preferred embodiment of the invention has 7 in the fermented product of VT fragrance, and 8-dihydro-β-ionol is to adopt the following analysis method to measure:
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing said fermented product 1.0mg, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 μ L sample and carry out gas-matter coupling analysis according to condition same as described above;
These are measured the result and obtain steps A through computes) in the sample solution 7, the content of 8-dihydro-β-ionol:
7,8-dihydro-β-ionol content (mg/L)=7,8-dihydro-β-ionol quality (mg)/0.1 μ L
The invention still further relates to said purposes with fermented product of VT fragrance as tobacco aromatics using.
Below the present invention will be described in more detail.
The present invention relates to a kind of working method with fermented product of VT fragrance.
Said working method may further comprise the steps:
A, seed culture
Use yeast extract, NaCl, glucose, peptone, liquid microelement and deionized water preparation to contain the seed culture medium of yeast extract 4-6g/L, NaCl 25-32g/L, glucose 0.1-0.4g/L, peptone 8-15g/L, liquid microelement 1.0mL/L.
Be added to said seed culture medium in the culturing bottle; With mineral alkali the pH value of this seed culture medium is adjusted to weakly alkaline; Sterilization then; Insert again with said seed culture medium volumeter 0.1-5% and separate starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340, at room temperature cultivated then 80-120 hour, obtain seed liquor.
Purchase obtains starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340 that separates that the present invention uses from market, is fungus industry preservation center (IIBM-UNAM) preservation of Mexico Investigaciones Biomedicas institute.
The morphological feature of separating starch based genus bacillus BM-B-340 is:
This bacterial strain belongs to genus bacillus, from soil, separates obtaining, and cell is direct rod shape, 0.5～2.5 μ m * 1.2～10 μ m, and with paired or catenation, tool nose circle or square end.Cell dyeing presents Gram-positive.Aerobic or amphimicrobian has heat, pH and the various multifarious physiological properties of salt, tool fermentation or respiratory metabolism type.
Described liquid microelement is to contain H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2The aqueous solution of O 0.04g/L.
In the present invention, C
4H
4KNaO
64H
2O should be appreciated that it is Rochelle salt.
It is pointed out that when the content of a certain composition in the liquid microelement surpasses or is lower than above-mentioned content, can't cause cultivating failure, just might influence culture effect or efficient.The preparation of liquid microelement is non-existent hell and high water to those skilled in the art, does not give unnecessary details at this.
B, shake flask fermentation
Use yeast extract, NaCl, peptone, xenthophylls, above-mentioned liquid microelement and deionized water preparation to contain the shake flask fermentation substratum of yeast extract 5.0-5.5g/L, NaCl 25-32g/L, peptone 8.5-12.0g/L, xenthophylls 180-200mg/L, liquid microelement 1.0mL/L;
Be added to said shake flask fermentation substratum in the culturing bottle; With mineral alkali the pH value of this shake flask fermentation substratum is adjusted to weakly alkaline; Sterilization at room temperature inserts in steps A more then) seed culture medium that obtains, inoculum size is in said shake flask fermentation culture volume 5-10%; In the 50-80rpm shaking table, cultivate 80-120h down, obtain fermented liquid at temperature 28-34 ℃.
Steps A) and B), described sterilization is with various bacteriums (comprising genus bacillus) and fungi deactivation in the physical environment.
The shaking table that this step is used is a normally used shaking table in the present technique field, for example the glad stamen automatic equipment in Shanghai ltd sell with trade(brand)name HYG-II constant temperature speed governing shaking table.
Owing to contain photosensitive material in the fermenting culture, therefore for avoiding these materials to see photolysis, the culturing bottle of use is used aluminium foil and is carried out lucifuge.
C, separation
With step B) fermented liquid that obtains carries out spinning, in its supernatant, adds with 1: 1 organic solvent of said supernatant volumeter, at room temperature carries out oscillation extraction, separates the organic phase that obtains and volatilizes, and obtains having the fermented product of VT fragrance.
In aforementioned production method, the mineral alkali that is used to regulate the pH value is NaOH, KOH or Ca (OH)
2
Preferably, in steps A) and B) described in weakly alkaline pH value be 7.2-7.4.
According to optimal technical scheme of the present invention, the organic solvent in the step (3) is selected from normal hexane, chloroform, acetone, ether, tetracol phenixin and/or sherwood oil.
More preferably, organic solvent is selected from normal hexane, acetone, tetracol phenixin and/or sherwood oil.
Preferably, volatilizing of organic solvent is to be selected from nitrogen flushing method, vacuum decompression method or distillation method in step (3).
The method that the technician that described nitrogen flushing method is the present technique field knows promptly adopts the automatic concentrating instrument method such as Turbo Vap.
The method that the technician that described vacuum decompression method is the present technique field knows is promptly carried out the pressure reducing and steaming organic solvent under vacuum tightness 0.06-0.08MPa.
Described distillation method also is the method that the technician in present technique field knows, i.e. the heating in water bath distillating method.
Confirm that through multiple analytical technology the fermented product with VT fragrance that adopts working method of the present invention to obtain contains 7,8-dihydro-β-ionol, its content is 60.0-70.0mg/L.
In the present invention, the fermented product with VT fragrance all should be appreciated that it is a kind of 60.0-70.0mg/L of containing 7, the fermented product of 8-dihydro-β-ionol.Certainly, 7,8-dihydro-β-ionol content or the higher or lower fermented product with VT fragrance of the present invention of also all should thinking.
Described fermented product with VT fragrance also contains a spot of other VT verivate, and for example 2.5-2.7mg/L 7,8-dihydro-, 0.7-0.8mg/L alpha, beta-lonone.
Have 7 in the fermented product of VT fragrance, 8-dihydro-β-ionol is to adopt the following analysis method to measure.Said analytical procedure may further comprise the steps:
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing said fermented product 1.0mg, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 μ L sample and carry out gas-matter coupling analysis according to condition same as described above;
These are measured the result and obtain steps A through computes) in the sample solution 7, the content of 8-dihydro-β-ionol:
7,8-dihydro-β-ionol content (mg/L)=7,8-dihydro-β-ionol quality (mg)/0.1 μ L
The present invention also provides according to aforementioned production method and prepares fermented product with the VT fragrance purposes as tobacco aromatics using.
Fermented product with VT fragrance should be appreciated that it is can contain the fermented product with VT fragrance of the present invention in this tobacco product gross weight 0.01-0.10% at tobacco product as the purposes of tobacco aromatics using.
In the present invention, described tobacco product is cigarette, cigar, pipe tobacco or redried leaf tobacco.
Can the fermented product with VT fragrance of the present invention is sprayed at pipe tobacco or be coated on the tobacco sheet according to conventional perfuming process program; In climatic chamber balance 45-50 hour then; Play cigarette or chopping again and beat cigarette and process cigarette and prop up, carry out sensory evaluating smoking's evaluation at last.
Described sensory evaluating smoking's authentication method is that five cigarettes of magnificent precious essence and flavoring agent company are formed evaluation group with the perfumer by our company; Adopt the method for secretly commenting; Fragrance matter, amount, harmony, assorted gas, stimulation, strength and six index grade standards of pleasant impression according to our company sets up are carried out grade estimation, so that relatively more existing essence and the influence with fermented product of VT fragrance to the tobacco product quality of the present invention.
Fermented product with VT fragrance of the present invention can highly significant ground promotes the quality of tobacco product.
[beneficial effect]
The inventive method adopts microorganism biological to transform xenthophylls and produces novel tobacco aromatics using, contains 7, the fermented product with VT fragrance of 8-dihydro-β-ionol, and this fermented product product has the fragrance of natural baccy plant.Compare with present tobacco aromatics using alpha, beta-lonone, this product fragrance is more elegant, unique, soft.The inventive method will change the chemosynthesis situation that VT alcohol ketone series product rely on poor efficiency, fill up the blank of bio-transformation High-efficient Production, and market outlook and using value are good.
[embodiment]
To illustrate in greater detail the present invention through embodiment below.
Embodiment 1: preparation has the fermented product of VT fragrance
Preparation process is following:
A, seed culture
It is yeast extract 5.0g/L, NaCl 30.0g/L, glucose 0.2g/L, peptone 10.0g/L, liquid microelement 1.0mL/L that seed culture medium is formed; Wherein the liquid microelement composition is H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L, all the other are deionized water.
Be added to the said seed culture medium of 80mL in the culturing bottle, stir into the culture medium solution of homogeneous.With the NaOH aqueous solution pH value of this seed culture medium is adjusted to pH=7.2; Add to 100mL with deionized water; The cotton plug mouth, the 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes inserts with said seed culture medium volumeter 1% again and separates starch based genus bacillus Paenibacillus amylolyticus BM-B-340; At room temperature cultivated then 90 hours, and obtained seed liquor;
B, shake flask fermentation
Replace glucose as carbon source with xenthophylls, it is yeast extract 5.5g/L, NaCl 30.0g/L, peptone 10.0g/L, xenthophylls 190.0mg/L, liquid microelement 1.0mL/L that its fermention medium is formed; It is H that liquid microelement is formed
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O0.04g/L;
Be added to said shake flask fermentation substratum 300mL in the culturing bottle; With the NaOH aqueous solution pH value of this fermention medium is adjusted to pH=7.2; The 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes then at room temperature inserts the seed culture medium that obtains in steps A again, and inoculum size is in said shake flask fermentation culture volume 10%; In the 60rpm shaking table, cultivate 96h down for 29 ℃, obtain fermented liquid in temperature;
See photolysis for fear of xenthophylls (carrotenoid), culturing bottle is used the aluminium foil shading.
C, separation
With step B) fermented liquid that obtains carries out centrifugal (cf-5400g) and separates 20min, again in its supernatant adding with 1: 1 organic solvent (organic solvent is acetone, ether and sherwood oil mixture, and its ratio is by volume 1: 1: 1) of said supernatant volumeter; Extract 10min down for 20 ℃ in temperature; Separate then and contain 7, the organic phase of 8-dihydro-β-ionol, it volatilizes again; Obtain having the fermented product of VT fragrance, its fermented product carries out following analysis.
D, product analysis
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing the fermented product 1.0mg of this embodiment preparation, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 its sample of μ L again and carry out gas-matter coupling analysis according to condition same as described above;
These are measured the result and obtain steps A through computes) in the sample solution 7, the content of 8-dihydro-β-ionol:
7,8-dihydro-β-ionol content (mg/L)=7,8-dihydro-β-ionol quality (mg)/0.1 μ L
Detection draws this embodiment fermented product and contains 65.2mg/L7,8-dihydro-β-ionol.
Adopt conventional perfuming process program that the fermented product of present embodiment preparation is sprayed on the pipe tobacco, balance 48 hours in climatic chamber is processed cigarette again and is propped up then, carries out sensory evaluating smoking's evaluation.
Described sensory evaluating smoking's authentication method is that five cigarettes of magnificent precious essence and flavoring agent company are formed evaluation group with the perfumer by our company; Adopt the method for secretly commenting, carry out grade estimation according to fragrance matter, amount, harmony, assorted gas, stimulation, strength and six index grade standards of pleasant impression that our company sets up.
Confirm 7,8-dihydro-β-ionol has increases unique this hay-scented effect of tobacco, significantly strengthens the fragrance matter of cigarette, reduces assorted gas simultaneously, and pleasant impression is comfortable clean.
Embodiment 2: preparation has the fermented product of VT fragrance
Preparation process is following:
A, seed culture
It is yeast extract 5.0g/L that seed culture medium is formed, NaCl 30.0g/L, glucose 0.2g/L, peptone 10.0g/L, liquid microelement 1.0mL/L; Liquid microelement preparation: H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L, all the other are deionized water,
Place culturing bottle to the said seed culture medium of 100mL, stir into the liquid medium of homogeneous.With the NaOH aqueous solution pH value of this seed culture medium is adjusted to 7.3; Add to 150mL with deionized water; The cotton plug mouth, the 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes inserts with said seed culture medium volumeter 2% again and separates starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340; At room temperature cultivated then 110 hours, and obtained seed liquor;
B, shake flask fermentation
Replace glucose as carbon source with xenthophylls, its substratum is formed: yeast extract 5.2g/L, NaCl30.0g/L, peptone 8.5g/L, xenthophylls 195.0mg/L, liquid microelement 1.0mL/L; Liquid microelement preparation: H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L;
Be added to said shake flask fermentation substratum 400mL in the culturing bottle; With the NaOH aqueous solution pH value of this fermention medium is adjusted to 7.2; The 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes then at room temperature inserts in steps A again) seed culture medium that obtains, inoculum size is in said shake flask fermentation culture volume 10%; In the 60rpm shaking table, cultivate 105h down for 29 ℃, obtain fermented liquid in temperature;
See photolysis for fear of xenthophylls (carrotenoid), culturing bottle is used the aluminium foil shading.
C, separation
With step B) fermented liquid that obtains carries out centrifugal (cf-5400g) and separates 20min, again in its supernatant adding with 1: 1 organic solvent (organic solvent is acetone, ether and sherwood oil mixture, and its ratio is by volume 1: 1: 1) of said supernatant volumeter; Under 20 ℃ of temperature, carry out liquid-liquid extraction 20min; Separate then and contain 7, the organic phase of 8-dihydro-β-ionol, it volatilizes again; Obtain having the fermented product of VT fragrance, its fermented product carries out following analysis.
D, product analysis:
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately the fermented product 1.0mg of present embodiment preparation is settled to 100 μ L with methylene dichloride, obtains sample to be analyzed; Accurately draw 0.1 its sample of μ L again and carry out gas-matter coupling analysis according to condition same as described above;
Detection draws this embodiment fermented product and contains 66.3mg/L7,8-dihydro-β-ionol.
Adopt embodiment 1 described grade estimation method, confirm 7,8-dihydro-β-ionol has increases unique this hay-scented effect of tobacco, significantly strengthens the fragrance matter of cigarette, reduces assorted gas simultaneously, and pleasant impression is comfortable clean.
Embodiment 3: preparation has the fermented product of VT fragrance
Preparation process is following:
A, seed culture
It is yeast extract 5.0g/L that seed culture medium is formed, NaCl 30.0g/L, glucose 0.2g/L, peptone 10.0g/L, liquid microelement 1.0mL/L; Liquid microelement preparation: H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L, all the other are deionized water,
Place culturing bottle to the said seed culture medium of 100mL, stir into the liquid medium of homogeneous.With the NaOH aqueous solution pH value of this seed culture medium is adjusted to 7.3; Add to 150mL with deionized water; The cotton plug mouth, the 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes inserts with said seed culture medium volumeter 3% again and separates starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340; At room temperature cultivated then 85 hours, and obtained seed liquor;
B, shake flask fermentation
Replace glucose as carbon source with xenthophylls, its substratum is formed: yeast extract 5.2g/L, NaCl30.0g/L, peptone 8.5g/L, xenthophylls 195.0mg/L, liquid microelement 1.0mL/L; Liquid microelement preparation: H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L;
Be added to said shake flask fermentation substratum 500mL in the culturing bottle; With the NaOH aqueous solution pH value of this fermention medium is adjusted to 7.2; The 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes then at room temperature inserts in steps A again) seed culture medium that obtains, inoculum size is in said shake flask fermentation culture volume 10%; In the 50rpm shaking table, cultivate 120h down for 32 ℃, obtain fermented liquid in temperature;
See photolysis for fear of xenthophylls (carrotenoid), culturing bottle is used the aluminium foil shading.
C, separation
With step B) fermented liquid that obtains carries out centrifugal (cf-5400g) and separates 20min, again in its supernatant adding with 1: 1 organic solvent (organic solvent is acetone, ether and sherwood oil mixture, and its ratio is by volume 1: 1: 1) of said supernatant volumeter; Extract 8min down for 20 ℃ in temperature; Separate then and contain 7, the organic phase of 8-dihydro-β-ionol, it volatilizes again; Obtain having the fermented product of VT fragrance, its fermented product carries out following analysis.
D, product analysis:
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing the fermented product 1.0mg of this embodiment preparation, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 its sample of μ L again and carry out gas-matter coupling analysis according to condition same as described above;
Detection draws this embodiment fermented product and contains 66.3mg/L7,8-dihydro-β-ionol.
Adopt embodiment 1 described grade estimation method, confirm 7,8-dihydro-β-ionol has increases unique this hay-scented effect of tobacco, significantly strengthens the fragrance matter of cigarette, reduces assorted gas simultaneously, and pleasant impression is comfortable clean.
Embodiment 4: preparation has the fermented product of VT fragrance
Preparation process is following:
A, seed culture
It is yeast extract 5.0g/L that seed culture medium is formed, NaCl 30.0g/L, glucose 0.2g/L, peptone 10.0g/L, liquid microelement 1.0mL/L; Liquid microelement preparation: H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O 0.04g/L, all the other are deionized water,
Place culturing bottle to the said seed culture medium of 80mL, stir into the liquid medium of homogeneous.With the NaOH aqueous solution pH value of this seed culture medium is adjusted to 7.3; Add to 100mL with deionized water; The cotton plug mouth, the 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes inserts with said seed culture medium volumeter 5% again and separates starch based genus bacillus (Paenibacillus amylolyticus) BM-B-340; At room temperature cultivated then 80 hours, and obtained seed liquor;
B, shake flask fermentation
Replace glucose as carbon source with xenthophylls, it is yeast extract 5.5g/L, NaCl 30.0g/L, peptone 10.0g/L, xenthophylls 190.0mg/L, liquid microelement 1.0mL/L that its fermention medium is formed; It is H that liquid microelement is formed
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O 0.03g/L, ZnCl
20.02g/L, CoCl
26H
2O0.04g/L;
Be added to said shake flask fermentation substratum 300mL in the culturing bottle; With the NaOH aqueous solution pH value of this fermention medium is adjusted to 7.2; The 15min that under the condition of 121 ℃ of 15psig and temperature, sterilizes then at room temperature inserts in steps A again) seed culture medium that obtains, inoculum size is in said shake flask fermentation culture volume 10%; In the 80rpm shaking table, cultivate 85h down for 28 ℃, obtain fermented liquid in temperature;
See photolysis for fear of xenthophylls (carrotenoid), culturing bottle is used the aluminium foil shading.
C, separation
Step B) fermented liquid that obtains carries out centrifugal (cf-5400g) separates 20min, in its supernatant, adds with 1: 1 organic solvent (organic solvent is acetone, ether and sherwood oil mixture, and its ratio is by volume 1: 1: 1) of said supernatant volumeter again; Extract 20min down for 20 ℃ in temperature; Separate then and contain 7, the organic phase of 8-dihydro-β-ionol, it volatilizes again; Obtain having the fermented product of VT fragrance, its fermented product carries out following analysis.
D, product analysis
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing the fermented product 1.0mg of this embodiment preparation, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 its sample of μ L again and carry out gas-matter coupling analysis according to condition same as described above;
Detection draws this embodiment fermented product and contains 66.2mg/L7,8-dihydro-β-ionol.
Adopt embodiment 1 described grade estimation method, confirm 7,8-dihydro-β-ionol has increases unique this hay-scented effect of tobacco, significantly strengthens the fragrance matter of cigarette, reduces assorted gas simultaneously, and pleasant impression is comfortable clean.
Claims (8)
1. working method with fermented product of VT fragrance is characterized in that described method may further comprise the steps:
A, seed culture
Use yeast extract, NaCl, glucose, peptone, liquid microelement and deionized water preparation to contain the seed culture medium of yeast extract 4-6g/L, NaCl 25-32g/L, glucose 0.1-0.4g/L, peptone 8-15g/L, liquid microelement 1.0mL/L; Described liquid microelement is to contain H
3BO
33.0g/L, MgSO
41.8g/L, FeSO
41.36g/L, C
4H
4KNaO
64H
2O 1.77g/L, CuCl
22H
2O0.03g/L, ZnCl
20.02g/L, CoCl
26H
2The aqueous solution of O 0.04g/L;
Be added to said seed culture medium in the culturing bottle; With mineral alkali the pH value of this seed culture medium is adjusted to weakly alkaline; Sterilization then; Insert again with said seed culture medium volumeter 0.1-5% and separate starch based genus bacillus Paenibacillus amylolyticus BM-B-340, at room temperature cultivated then 80-120 hour, obtain seed liquor;
B, shake flask fermentation
Use yeast extract, NaCl, peptone, xenthophylls, above-mentioned liquid microelement and deionized water preparation to contain the shake flask fermentation substratum of yeast extract 5.0-5.5g/L, NaCl 25-32g/L, peptone 8.5-12.0g/L, xenthophylls 180-200mg/L, liquid microelement 1.0mL/L;
Be added to said shake flask fermentation substratum in the culturing bottle; With mineral alkali the pH value of this shake flask fermentation substratum is adjusted to weakly alkaline; Sterilization at room temperature inserts the seed culture medium that obtains in steps A more then, and inoculum size is in said shake flask fermentation culture volume 5-10%; In the 50-80rpm shaking table, cultivate 80-120h down, obtain fermented liquid at temperature 28-34 ℃;
C, separation
The fermented liquid that step B is obtained carries out spinning, in its supernatant, adds with 1: 1 organic solvent of said supernatant volumeter, at room temperature carries out oscillation extraction, separates the organic phase that obtains and volatilizes, and obtains having the fermented product of VT fragrance.
2. method according to claim 1 is characterized in that described mineral alkali is NaOH, KOH or Ca (OH)
2
3. method according to claim 1 is characterized in that in steps A and B said weakly alkaline pH value is 7.2-7.4.
4. method according to claim 1 is characterized in that among the step C, said organic solvent is selected from normal hexane, chloroform, acetone, ether, tetracol phenixin and/or sherwood oil.
5. method according to claim 1 is characterized in that among the step C, and it is nitrogen flushing method, vacuum decompression method or distillation method that organic phase volatilizes.
6. the fermented product with VT fragrance that obtains according to the said method of each claim among the claim 1-5 is characterized in that it contains 60.0-70.0mg/L 7,8-dihydro-β-ionol.
7. the fermented product with VT fragrance according to claim 6 is characterized in that 7, and 8-dihydro-β-ionol is to adopt the following analysis method to measure:
Accurately draw 0.1 μ L and contain 7, the dichloromethane solution of 8-dihydro-β-ionol carries out gas-matter coupling and analyzes;
HP 5890II type analytical conditions for gas chromatography is: quartz capillary column 20-M, column length 30m, thickness 1 μ m; Heating schedule: 50 ℃ keep 5min, are warming up to 200 ℃ with 4 ℃/min of heat-up rate, under this temperature, keep 25min; 200 ℃ of injector temperatures;
HP 5945A type mass spectroscopy condition is: EI source, ionization voltage 70eV, 200 ℃ of ion source temperatures, 200 ℃ of interface temperature, detector voltage 1KV, gamut scanning;
Then, accurately take by weighing the said product 1.0mg of claim 6, be settled to 100 μ L, obtain sample to be analyzed with methylene dichloride; Accurately draw 0.1 μ L sample and carry out gas-matter coupling analysis according to condition same as described above;
These are measured the result and obtain in the steps A in the sample solution 7 through computes, the content of 8-dihydro-β-ionol:
7,8-dihydro-β-ionol content (mg/L)=7,8-dihydro-β-ionol quality (mg)/0.1 μ L.
8. the purposes with fermented product of VT fragrance as tobacco aromatics using according to claim 6.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498539A (en) * | 2014-11-19 | 2015-04-08 | 上海应用技术学院 | Method for preparing 2-phenylethyl alcohol |
| CN108732265A (en) * | 2018-04-18 | 2018-11-02 | 湖南农业大学 | A method of four kinds of Monoterpenes of Quantitative detection from yeast fermentation broth |
| CN109298082A (en) * | 2018-02-27 | 2019-02-01 | 广东省农业科学院茶叶研究所 | The identification method of artificial essence is added in a kind of tealeaves |
| CN109998153A (en) * | 2019-03-22 | 2019-07-12 | 河南中烟工业有限责任公司 | A kind of tobacco juice for electronic smoke prepared using microbial fermentation tobacco leaf |
| CN110122920A (en) * | 2019-03-22 | 2019-08-16 | 河南中烟工业有限责任公司 | A method of utilizing microbial flora fermentation process cigar tobacco |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102140484A (en) * | 2010-12-16 | 2011-08-03 | 郑州奥利实业有限公司 | Synthesis method of 3-oxo-alpha-ionol esters |
-
2011
- 2011-12-06 CN CN2011104001404A patent/CN102492729A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102140484A (en) * | 2010-12-16 | 2011-08-03 | 郑州奥利实业有限公司 | Synthesis method of 3-oxo-alpha-ionol esters |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498539A (en) * | 2014-11-19 | 2015-04-08 | 上海应用技术学院 | Method for preparing 2-phenylethyl alcohol |
| CN109298082A (en) * | 2018-02-27 | 2019-02-01 | 广东省农业科学院茶叶研究所 | The identification method of artificial essence is added in a kind of tealeaves |
| CN109298082B (en) * | 2018-02-27 | 2021-07-02 | 广东省农业科学院茶叶研究所 | Method for identifying artificial essence added in tea |
| CN108732265A (en) * | 2018-04-18 | 2018-11-02 | 湖南农业大学 | A method of four kinds of Monoterpenes of Quantitative detection from yeast fermentation broth |
| CN108732265B (en) * | 2018-04-18 | 2021-12-28 | 湖南农业大学 | Method for rapidly and quantitatively detecting four monoterpene substances from yeast fermentation liquor |
| CN109998153A (en) * | 2019-03-22 | 2019-07-12 | 河南中烟工业有限责任公司 | A kind of tobacco juice for electronic smoke prepared using microbial fermentation tobacco leaf |
| CN110122920A (en) * | 2019-03-22 | 2019-08-16 | 河南中烟工业有限责任公司 | A method of utilizing microbial flora fermentation process cigar tobacco |
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