CN109722343A - A kind of flavor and fragrance prepared by fermentation of Debaryomyces hansenii - Google Patents
A kind of flavor and fragrance prepared by fermentation of Debaryomyces hansenii Download PDFInfo
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- 230000004151 fermentation Effects 0.000 title claims abstract description 87
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- 239000000796 flavoring agent Substances 0.000 title claims abstract description 48
- 235000019634 flavors Nutrition 0.000 title claims abstract description 48
- 241000235036 Debaryomyces hansenii Species 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 30
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Landscapes
- Fats And Perfumes (AREA)
Abstract
The invention discloses a kind of flavors and fragrancess being prepared using the inferior Dbaly yeast fermentation of the Chinese, belong to technical field of microbial fermentation.The method is fermented by being seeded to the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) in the fermentation medium containing sophora flower etc., is assigned the sophora flower fragrance of the slightly fresh and sweet taste of tunning, has been obtained a kind of flavors and fragrances of pleasant aroma；The method simple process, raw material sources are abundant, low in cost, are applicable to industrial production；Also, the flavors and fragrances being prepared using the method is in addition to pleasant aroma, the active constituent rich in sophora flower and yeast, with anti-oxidant, anti-inflammatory and other effects and securely and reliably, can be widely applied to the fields such as food, drug, cosmetics, tobacco.
Description
Technical field
The present invention relates to a kind of flavors and fragrancess being prepared using the inferior Dbaly yeast fermentation of the Chinese, belong to microbial fermentation
Technical field.
Background technique
For now, flavors and fragrances industry has played extremely crucial effect in national life production, by
Step becomes an important industry closely related with people's lives.Also, spices and essence industrial market is wide, dosage is big, in China
It deserves to be called for rising sun-industry.For objectively, relatively stable development is had been obtained in spices and essence industry in China's at present, with
The development level of other mating industries is adapted, such as medical industry, food industry, environmental protection industry and beverage industry.In turn
It sees, the development of above-mentioned mating industry also can generate crucial facilitation to the development of spices and essence industry, effectively promotion fragrance
The development and growth of essence industry.In terms of comprehensive, flavors and fragrances industry in China's gradually develops towards the direction of Basic national topographic map,
Important role has been played in national economy.
And in flavors and fragrances, natural perfume material is with the flower, leaf, branch, root, skin, natural gum of spice berry in nature, complete
Grass and fruit are raw material, a kind of native compound produced through steam distillation, squeezing method, absorption process or solvent extraction.Benefit
Fragrance such as butyric acid, diacetyl, benzaldehyde produced with biotechnologys such as fermentation process etc. also all belongs to natural perfume material.This kind of fragrance
Since quality is higher, is gradually paid attention to and developed in edible essence.
Currently, preparing the history of natural perfume material and not very long using fermentation technique, still, fermentation method prepares natural perfume material tool
It has the advantage that (1) reaction safety, can be carried out under room temperature, normal pressure, it is lower to production site and equipment requirement；(2) environment friend
Good, the three wastes formed after product separation are few；(3) highly-safe, entire production process is required without miscellaneous bacteria, a degree of to reduce
Security risk；(4) overcome for a long time from animal and plant as natural perfume material sole source existing active constituent content
Low, the defects of separation is difficult, climate and animal and plant harm influence, above-mentioned advantage makes fermentation method prepare natural perfume material more
Requirement of the people to environmental protection, health is adapted to, powerful vitality and huge market potential are shown.
Therefore, natural perfume material is prepared using Fermentation Engineering and have become a kind of development trend, not only to perfume industry sheet
Body, meanwhile, the development of other industries associated with it can all be had a huge impact.
The inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) is a kind of aroma-producing yeast, due to that can produce when its fermentation
Raw thin fragrance and this fragrance are sweet tea ester perfume (or spice) slightly aroma, therefore, the inferior Dbaly yeast (Debaryomyces of the Chinese
Hansenii) it is widely used in such as preparation of sausage, sausage, meat pie, halogen meat food, is sausage, sausage, meat pie, halogen
The food such as meat increase special aroma and flavor.
But also just because of when the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) ferments the fragrance that generates compared with
Light and this fragrance is sweet tea ester perfume (or spice) slightly aroma, so that the application limit of the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii)
It makes in the preparation of food, currently, there is no natural with the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) fermenting and producing
Flavors and fragrances, this is undoubtedly to the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) in the development of flavors and fragrances industry
And the development of the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) application itself causes serious obstruction.
Therefore, it is badly in need of finding one kind with the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) as fermentation raw material system
It is standby to obtain the method for the good flavors and fragrances of the flavouring essence qualities such as aroma quality, aroma strength and fragrance remaining time to promote Han Xundeba
Sharp yeast (Debaryomyces hansenii) in the development of flavors and fragrances industry, meanwhile, expand the inferior Dbaly yeast of the Chinese
(Debaryomyces hansenii) application range itself.
Summary of the invention
[technical problem]
It is that fragrance is prepared in fermentation raw material that one kind, which is provided, with the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii)
The method of the good flavors and fragrances of the flavouring essence qualities such as quality, aroma strength and fragrance remaining time.
[technical solution]
To solve the above problems, the present invention provides a kind of sides using the inferior Dbaly yeast fermentation preparation flavors and fragrances of the Chinese
Method, the method are that the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) is seeded to fermentation medium to ferment
Afterwards, obtained fermentation material is extracted, obtains flavors and fragrances；The ingredient of the fermentation medium includes flower；The flower includes
One of sophora flower, chrysanthemum, Radix Paeoniae Alba, lavender, rose or sweet osmanthus are a variety of.
In one embodiment of the invention, the flower is sophora flower.
In one embodiment of the invention, the ingredient of the fermentation medium includes sophora flower, glucose and water.
In one embodiment of the invention, the mass ratio of the sophora flower, glucose and water is (45~65): (3~
5): (30~50).
In one embodiment of the invention, the mass ratio of the sophora flower, glucose and water is 50:4:46.
In one embodiment of the invention, the temperature of the fermentation is 25~30 DEG C, the time is 7~10d.
In one embodiment of the invention, the inferior Dbaly yeast of the Chinese is seeded to fermentation medium in the form of seed liquor
In ferment；The volume of the inferior Dbaly yeast seed liquor of the Chinese accounts for the 2~10% of fermentation liquid total volume, the fermentation liquid by
Seed liquor and fermentation medium composition；The bacteria concentration of the inferior Dbaly yeast seed liquor of the Chinese is 1 × 108~1 × 1010CFU/
mL；The inferior Dbaly yeast seed liquor of the Chinese is obtained by the way that the inferior Dbaly yeast of the Chinese to be seeded to cultivate in seed culture medium
's.
The present invention also provides a kind of flavors and fragrances being prepared using the inferior Dbaly yeast fermentation of the Chinese, the essence is fragrant
The material application above method is prepared, and has certain fragrance.
The present invention also provides the products for containing above-mentioned flavors and fragrances.
In one embodiment of the invention, the product can be tobacco, cosmetics, food or drug.
[beneficial effect]:
(1) method of the invention by by the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii) be seeded to containing
It ferments in the fermentation medium of sophora flower etc., assigns the sophora flower fragrance of the slightly fresh and sweet taste of tunning, obtained a kind of fragrance
Pleasant flavors and fragrances；
(2) the pleasant aroma specific manifestation of the flavors and fragrances obtained using the inferior Dbaly yeast fermentation sophora flower of the Chinese are as follows: with the Chinese
The fermentation material that inferior Dbaly yeast individually ferments is compared, the extracting solution obtained using the inferior Dbaly yeast fermentation sophora flower of the Chinese
In, phenol, à-pine tar alcohol content dramatically increase；Some content higher building matter are newly increased, such as: mushroom alcohol, cis- α, α -5- front three
Base -5- vinyl tetrahydrofuran -2- methanol, trans- α trans- α, α -5- trimethyl -5- vinyl tetrahydro -2- furancarbinol, virtue
Camphor tree alcohol.
(3) sophora flower is rich in vitamin and several mineral materials, has clearing heat and detoxicating, cool blood moistening lung, blood pressure lowering, pre- anti-stroke
Effect；Yeast can produce active substances, have certain anti-oxidant, anti-inflammatory, it is antitumor, adjust the physiological functions such as immune,
Therefore, the flavors and fragrances being prepared using method of the invention can also have sophora flower and yeast in addition to pleasant aroma simultaneously
Beneficial functional；
(4) method of the invention has not only expanded flavors and fragrances resource, and the flavors and fragrances safety obtained is higher, can be with
It is widely used in the fields such as food, tobacco, cosmetics, medical treatment, assigns product feature fragrance, note and feature functionality；
(5) method and process of the invention is simple, raw material sources are abundant, low in cost, is suitable for industrial production.
Detailed description of the invention
Fig. 1 is flavors and fragrances A in embodiment 10Total ion current map；
Fig. 2 is flavors and fragrances A in embodiment 11Total ion current map.
Specific embodiment
The present invention will be further elaborated combined with specific embodiments below.
The inferior Dbaly yeast of the Chinese in following embodiments is purchased from China General Microbiological collection, and deposit number is
CGMCC 2.1335；Sophora flower involved in following embodiments is purchased from the Wuxi market of farm produce；The purchase of pipe tobacco involved in following embodiments
(the inferior Dbaly yeast CGMCC 2.1335 of the above-mentioned bacterial strains Chinese can be commercially available, no for cigarette industry Co., Ltd from Sichuan
It needs to carry out the preservation for proprietary program).
Culture medium involved in following embodiments is as follows:
YPD seed culture medium (m/v): peptone 2%, yeast powder 1%, glucose 2% (115 DEG C of sterilizing 20min).
Detection method involved in following embodiments is as follows:
Flavors and fragrances solid phase microextraction-makings analysis method:
It is analyzed using the triple level four bars gas chromatograph-mass spectrometers of TSQ8000.
Sample pretreatment: taking the fermentation material for preparing that 30~95% ethyl alcohol are added according to the volume ratio of 1:3~4, in 150~
2~4h is extracted under conditions of 200rpm, obtains extract；Extract is centrifuged 10min under conditions of 10000rpm, is obtained
Clear liquid；Supernatant is rotated under conditions of 40mbar, 40 DEG C, obtains extracting solution；Add after extracting solution is diluted to suitable concentration
Enter to 20mL ml headspace bottle, be added internal standard sec-n-octyl alcohol (final concentration 8.2ppb), total volume 3mL, NaCl is added to its saturation, then surveys
It is fixed；
Headspace solid-phase microextraction condition: then extracting head is extracted in GC injection port aging to no miscellaneous peak at 60 DEG C
30min；The volatile compound being adsorbed desorbs 1min at 260 DEG C of GC injection port, pours GC chromatographic column；Start instrument simultaneously
Acquire data；Extracting head: DVB/CAR/PMDS；
GC conditions: chromatographic column HP-5；Carrier gas: helium, 1.0mLmin-1；Temperature programming condition: 40 DEG C of maintenances
1min, 3 DEG C of min-1180 DEG C are warming up to, 15 DEG C of min-1230 DEG C are warming up to, 3min is maintained；
Mass Spectrometry Conditions: 250 DEG C of interface temperature；Source temperature: 300 DEG C；150 DEG C of level four bars temperature；Ionize electricity 70eV；From
Sub-ization mode: EI+；35~350amu of mass scan range m/z.
Data processing: sample carries out Headspace-solid phase microextraction-gas chromatography-mass spectrum (HS-SPME-GC- by above-mentioned condition
MS it) analyzes, carries out retrieval according to NIST standard mass spectrum search library and compare identification, the relative amount of each component is calculated by internal standard method.
Tobacco sensory evaluation:
Tobacco aesthetic quality is influenced referring to National Standard Method to be evaluated (YC/T 138-1998).
Toner sense organ, physical and chemical evaluation:
Being evaluated (QB/T 2660-2004) to the sense organ of toner, physical and chemical quality influence referring to National Standard Method, (evaluation is marked
1) standard is shown in Table.
1 toner sense organ of table, physical and chemical evaluation criterion
Cell concentration detection method:
It takes yeast starter liquid 1mL in cuvette, measures OD600, normal saline dilution seed liquor is then used, is 25 with specification
× 16 blood counting chamber counts yeast count, calculates cell concentration；
Wherein, the cell number in every mL=(number of cells/80 counted in blood counting chamber) × 400 × 10000 ×
Extension rate.
Embodiment 1: the inferior Dbaly yeast+sophora flower of the Chinese
Specific step is as follows:
(1) ring is taken to be connected to YPD seed culture medium (115 DEG C of sterilizing 20min) yeast strain saved at 4 DEG C, 30 DEG C,
Revolving speed 200rmin-1Under conditions of cultivate 2d, obtain activating solution；
(2) it takes 80mLYPD seed culture medium to be placed in 250mL triangular flask (115 DEG C of sterilizing 20min), 200 μ L (1) is taken to obtain
To activating solution be seeded in YPD seed culture medium, 30 DEG C of Yu Wendu, revolving speed 200rmin-1Under conditions of cultivate 2d, obtain
Seed liquor；At this point, the cell concentration in seed liquor is about 9 × 109CFU/mL；
(3) 100g fermentation medium (105 DEG C of sterilizing 20min) is taken to be placed in 250mL shaking flask, by seed liquor according to table 2
Parameter, which is seeded in fermentation medium, ferments, and obtains fermentation material A0、A1、A2、A3；Wherein, A0Group is blank control group.
Using sec-n-octyl alcohol as internal standard (final concentration 8.2ppb), by fermentation material A0、A1、A2、A3According to the micro- extraction of flavors and fragrances solid phase
Take-makings analysis method in the analysis method recorded analyzed, wherein fermentation material A0、A1Analysis the results are shown in Table 3-4 and figure
1-2。
To fermentation material A0、A1、A2、A3It carries out smelling perfume (or spice), the results are shown in Table 5.
By table 3-4 and Fig. 1-2 it is found that fermentation material A1Compared with blank control group fermentation material A0Substance classes obviously enrich, content
It changes greatly, wherein phenol, à-pine tar alcohol content dramatically increase, the objects such as 3- methyl-1-butanol, 2-methyl-1-butene alcohol, benzyl carbinol
Matter content is remarkably decreased；Some content higher building matter are newly increased, such as: mushroom alcohol, cis- α, α -5- trimethyl -5- vinyl four
Hydrogenate furans -2- methanol, trans- α trans- α, α -5- trimethyl -5- vinyl tetrahydro -2- furancarbinol, linalool.As it can be seen that through
It crosses volatile species, content in fermentation material obtained by the inferior Dbaly yeast of the sophora flower fermentation Chinese all to have a distinct increment, flavouring essence quality
Variation is obvious.
As shown in Table 5, fermentation material A1、A2、A3Good fragrance is all had, can be widely applied to food, tobacco, makeup
The fields such as product, medical treatment assign product feature fragrance and note.
2 fermentation material A of table0、A1、A2、A3Fermentation parameter
3 fermentation material A of table0Material base analyze result
4 fermentation material A of table1Material base analyze result
5 fermentation material A of table0、A1、A2、A3Smell fragrant result
| Group | Fragrance |
| Fermentation material A0 | Ester fragrance, slightly aroma sweet taste |
| Fermentation material A1 | The peculiar fragrance of sophora flower |
| Fermentation material A2 | The peculiar fragrance of sophora flower, slightly faint scent |
| Fermentation material A3 | Sophora flower is clear soft fragrant and sweet |
Embodiment 2: the inferior Dbaly yeast+Radix Paeoniae Alba of the Chinese
Specific step is as follows:
Sophora flower in embodiment 1 is substituted for Radix Paeoniae Alba, repeats A in embodiment 11The experiment of group, obtains fermentation material A4.It is right
Obtained fermentation material A4It carries out smelling perfume (or spice), the results are shown in Table 6.
Embodiment 3: the inferior Dbaly yeast+lavender of the Chinese
Specific step is as follows:
Sophora flower in embodiment 1 is substituted for lavender, repeats A in embodiment 11The experiment of group, obtains fermentation material A5.It is right
Obtained fermentation material A5It carries out smelling perfume (or spice), the results are shown in Table 6.
Embodiment 4: the inferior Dbaly yeast+sweet osmanthus of the Chinese
Specific step is as follows:
Sophora flower in embodiment 1 is substituted for sweet osmanthus, repeats A in embodiment 11The experiment of group, obtains fermentation material A6.To
The fermentation material A arrived6It carries out smelling perfume (or spice), the results are shown in Table 6.
Embodiment 5: the inferior Dbaly yeast+rose of the Chinese
Specific step is as follows:
Sophora flower in embodiment 1 is substituted for rose, repeats A in embodiment 11The experiment of group, obtains fermentation material A7.It is right
Obtained fermentation material A7It carries out smelling perfume (or spice), the results are shown in Table 6.
Embodiment 6: the inferior Dbaly yeast+chrysanthemum of the Chinese
Specific step is as follows:
Sophora flower in embodiment 1 is substituted for chrysanthemum, repeats A in embodiment 11The experiment of group, obtains fermentation material A8.To
The fermentation material A arrived8It carries out smelling perfume (or spice), the results are shown in Table 6.
As shown in Table 6, fermentation material A4、A5、A6、A7、A8Flavouring essence quality is poor, is not suitable for as flavors and fragrances.
6 fermentation material A of table4、A5、A6、A7、A8Smell fragrant result
| Group | Fragrance |
| Fermentation material A4 | The weak Radix Paeoniae Alba feature fragrance of a flower, cigarette sweet taste with irritating |
| Fermentation material A5 | The distinctive radix aucklandiae note of lavender, slightly smell |
| Fermentation material A6 | Sweet osmanthus characteristic perfume is weaker, slightly sweet taste |
| Fermentation material A7 | The light distinctive fragrance of a flower of rose and fragrant and sweet, slightly fresh gas |
| Fermentation material A8 | The thin peculiar fragrance of chrysanthemum, it is fragrant and sweet with stronger sauce |
Embodiment 4: the preparation of tobacco
Specific step is as follows:
1, it extracts
The fermentation material A that embodiment 1 is obtained1It is carried out according to the Parameter Conditions of scheme one, scheme two, scheme three, scheme four
It extracts, obtains flavors and fragrances A, B, C, D；Wherein, the extraction conditions of each scheme are as follows:
Scheme one: being added 30% ethyl alcohol (v/v) by 1:3 volume ratio in fermentation material, and 150rpm extracts 2h, extract
10000rpm is centrifuged 10min and obtains supernatant, this supernatant is flavors and fragrances；
Scheme two: being added 50% ethyl alcohol (v/v) by 1:4 volume ratio in fermentation material, and 200rpm extracts 2h, extract
10000rpm is centrifuged 10min and obtains supernatant, this supernatant is flavors and fragrances；
Scheme three: being added 40% ethyl alcohol (v/v) by 1:3 volume ratio in fermentation material, and 150rpm extracts 3h, extract
10000rpm is centrifuged 10min and obtains supernatant, this supernatant is flavors and fragrances；
Scheme four: being added 95% ethyl alcohol (v/v) by 1:3 volume ratio in fermentation material, and 150rpm extracts 3h, extract
10000rpm is centrifuged 10min and obtains supernatant, this supernatant is flavors and fragrances；
2, it detects
Pipe tobacco is added according to the additive amount ratio of pipe tobacco table 7 in flavors and fragrances A, B, C, D for being prepared, sense organ is carried out and comments
Fixed, evaluation result is shown in Table 7.
As shown in Table 7, flavors and fragrances A, B, C may be applicable to tobacco flavoring, have good effect；Flavors and fragrances D effect
Fruit is bad, is not suitable for tobacco flavoring.
7 tobacco Analyses Methods for Sensory Evaluation Results of table
Note: "+" is positive-effect；"-" is negative effect；" ± " is that effect is unobvious.
Embodiment 5: the preparation of toner
Specific step is as follows:
Obtained flavors and fragrances A, B, C, D are extracted according to the recipe configuration of table 12 into toner by 4 kinds of embodiment, are felt
Official's evaluation, evaluation result are shown in Table 8.
As shown in Table 8, flavors and fragrances A, B, C, D may be applicable to toner flavoring, have good effect.
12 toner formula of table
13 toner Analyses Methods for Sensory Evaluation Results of table
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not intended to limit the invention, any to be familiar with this skill
The people of art can do various change and modification, therefore protection model of the invention without departing from the spirit and scope of the present invention
Enclosing subject to the definition of the claims.
Claims (10)
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| CN109837137A (en) * | 2019-03-20 | 2019-06-04 | 四川中烟工业有限责任公司 | The method for preparing essence spice for cigarette using the inferior Dbaly yeast of the Chinese and flower fermentation |
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| CN113355249A (en) * | 2021-07-15 | 2021-09-07 | 四川中烟工业有限责任公司 | Debaryomyces hansenii strain and application thereof |
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