CN108624518A - Yeast strain and its application - Google Patents
Yeast strain and its application Download PDFInfo
- Publication number
- CN108624518A CN108624518A CN201810461991.1A CN201810461991A CN108624518A CN 108624518 A CN108624518 A CN 108624518A CN 201810461991 A CN201810461991 A CN 201810461991A CN 108624518 A CN108624518 A CN 108624518A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- wine
- content
- delbrueckii
- yeast strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
Abstract
The present invention provides a kind of yeast strain, which is CGMCC NO:15430, Classification And Nomenclature, which is the triumphant formula of Dell, spore torula (Torulaspora delbrueckii).The experimental results showed that the bacterial strain has stronger ethanol tolerance, SO2Tolerance and high sugared tolerance, it has fermentability in the zymotic fluid of a concentration of 200~400g/L of sugar, and the bacterial strain has the characteristic of low yield volatile acid and high yield glycerine, in making sweet white and ice-wine, the yeast strain and saccharomyces cerevisiae mixed fermentation can be reduced fruit wine volatilization acid yield, improve fruit wine glycerol content and assign ice wine more fragrance of a flower, fruit wine fragrance complexity is increased.Yeast strain ferments ability provided by the invention is strong, fermenting property is excellent, has important value in promoting the application of fruit wine quality.
Description
Technical field
The invention belongs to microorganisms technical field more particularly to a kind of yeast strain and its applications.
Background technology
All the time, saccharomyces cerevisiae is the main bacteria seed of wine fermentation, fruit wine a big chunk zymophyte that we produce
Strain all uses saccharomyces cerevisiae.In recent years, have researcher's discovery, the aroma substance that Non-Saccharomyces generate account for the total fragrance of grape wine at
The 90% of part.Wherein, it is a kind of relevant with grape wine that the triumphant formula of Dell, which has spore torula (Torulaspora delbrueckii),
Non-Saccharomyces belong to true yeast (phorozoon of Candida colliculosa candida colliculosas).Its cell is small, nearly circle
Shape (6.5 5.5 μm of μ m), the bacterium colony on WLN culture mediums is in cream color with light green, protrusion spherical in shape, and table
Face is smooth opaque.T.delbrueckii has 9~11Mb genomes, is distributed on 8 chromosomes.Generally, it is considered that
T.delbrueckii fermentabilities are relatively low, SO2Tolerance is low, during the fermentation, SO2Addition can reduction by a relatively large margin its
Maximum biomass.T.delbrueckii can influence Portugal by influencing some compounds (content as increased bata-phenethyl alcohol)
The fragrance of grape wine.It can also influence the generation of volatility Ester, such as make acetic acid isoamyl fat, C6-C10Under content of fatty acid
Drop.
The higher fruit wine of the sugar contents such as semi-sweet grape wine, sweet wine, due to easily drinking, it is comfortable, in good taste the features such as
It is well received by consumers.This fruit wine is fermented using the higher fruit juice of sugar content, according to production requirement, specified residual
Fermentation is terminated in the case of sugar amount, for example, ice wine is fermented using sugar content more than 35 ° of Brix ice grape juices, when wine degree reaches 9
~14% (v/v) when residual sugar is more than 125g/L, terminates fermentation.Currently, the fermentation of this fruit wine mostly uses saccharomyces cerevisiae, sending out
When ferment is carried out to a certain extent, fermentation is terminated.This fermentation process needs to monitor sugar content variation always during the fermentation, and
Fermentation is terminated when suitable.But in actual production, often due to ice grape juice sugar content is higher, fermentation time is long, fermentation
Often volatile acid content is higher for the ice wine of formation, and not easy to control, and this ice wine to be fermented with single culture, flavor are complicated
Degree is inadequate, tends not to constitute a excellent grape wine.In addition, glycerol content is also critically important one of influence ice Quality of Liquors
Factor, higher glycerol content can largely promote the mouthfeel of ice wine.And there are studies have shown that Non-Saccharomyces that can carry
The quality of grape wine is risen, but currently, China is seldom to the research of native country Non-Saccharomyces, is applied to the non-wine brewing ferment of brewing fruit wine
Female strain is less.
Therefore, for the quality of the higher fruit wine of sugar contents such as raising ice wine, good, fermentability that there is an urgent need for a kind of high sugared tolerances
By force, low yield volatile acid, high yield glycerine and the Non-Saccharomyces with excellent fermentation character.
Invention content
Good, fermentability that the purpose of the present invention is to provide a kind of high sugared tolerances is by force, production volatile acid content is low, produces glycerine
Content is high and has the yeast strain of excellent fermentation character.
To achieve the above object, the present invention provides a kind of yeast strain, and deposit number is CGMCC NO:15430, classification
Being named as the triumphant formula of Dell has spore torula (Torulaspora delbrueckii).Yeast strain CEC R12, the deposit date is
On 03 08th, 2018, depositary institution was entitled：China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Specifically, the areas bacterial strain 26s D1-D2 have such as SEQ ID:Nucleotide sequence shown in 2.
Specifically, which there is fermentability, the bacterial strain can drop in the zymotic fluid of a concentration of 200~400g/L of sugar
Volatile acid content in low fruit wine, the bacterial strain can enhance the fragrance complexity of fruit wine.
The present invention also provides a kind of application of above-mentioned bacterial strains in reducing fruit wine volatile acid.Specifically, the fruit wine is sweet tea
Type grape wine or ice wine.
The present invention also provides a kind of methods reducing fruit wine volatile acid, by above-mentioned bacterial strains and saccharomyces cerevisiae mixed fermentation fruit juice
Raw material.Specifically, fruit juice raw material is shrimps and crabs juice or ice grape juice.
Specifically, above-mentioned bacterial strains and the saccharomyces cerevisiae are pressed 1:1~20:1 inoculative proportion mixed fermentation fruit juice raw material.
The present invention also provides a kind of methods of enhancing fruit wine fragrance, and above-mentioned yeast strain is taken to mix hair with other yeast strains
Ferment fruit juice raw material.
The present invention also provides a kind of methods improving fruit wine glycerol production, and above-mentioned bacterial strain is taken to be mixed with other yeast strains
Fermented juice raw material.
The present invention also provides a kind of application of above-mentioned bacterial strain in mixed fermentation fruit wine.Fruit wine may include grape wine, Mi
A variety of fruit wine such as monkey peach wine, blueberry wine.
The present invention also provides application of the above-mentioned bacterial strains in high sugared fermented product.
The present invention also provides a kind of active dry yeasts, are made by above-mentioned bacterial strains.
The present invention also provides a kind of methods that screening can reduce the bacterial strain of fruit wine volatile acid, and it is former material to take above-mentioned bacterial strains
Material.
Torulaspora delbrueckii R12 provided by the invention show that bacterial strain R12 can in physiological property research
With in alcoholic strength, the SO of 14% (v/v)2It is grown in the environment that content is 600mg/L, sugar content is 400g/L.In fermentation character
It is shown in research, volatile acid contents of the bacterial strain R12 in the zymotic fluid of 3 kinds of different sugar concentration is minimum.In a concentration of 200g/L of sugar
It is high with glycerol content is produced in the zymotic fluid of 300g/L.
Bacterial strain R12 provided by the invention is shown in the research of mixed fermentation sweet white wine, while inoculation fermentation is mixed
The fermentation rate for closing fermentation is more than the mixed fermentation of interval 5d inoculations, and in the two ways of mixed fermentation, volatile acid contains
Amount is reduced as the triumphant formula of Dell has the increase of spore torula R12 ratios, and glycerol content has spore torula R12 with the triumphant formula of Dell
Ratio increase and increase.It is obtained by sensory evaluation while being inoculated with (20:1) intervals ＞ 5d inoculations (20:1) intervals ＞ 5d is inoculated with
(1:1) ＞ is inoculated with (1 simultaneously:1).In conclusion the triumphant formula of Dell has spore torula R12 with saccharomyces cerevisiae by 20:1 inoculative proportion
The quality of sweet white wine can be improved by being inoculated with mixed fermentation simultaneously.
Bacterial strain R12 provided by the invention shows in the research of mixed fermentation ice grape juice, T.delbrueckii R12 and
The volatile acid content that the intervals S.cerevisiae 5d is inoculated with mixed fermentation is minimum, is 0.42g/L.Its volatile acid yield is also most simultaneously
It is low, it is 0.001g/L.Although being inoculated with the volatile acid content of mixed fermentation simultaneously higher than the mixed fermentation of interval 5d inoculations, it is less than
R12 and the respective independent fermentations of S.cerevisiae.The result shows that either in a manner of inoculation simultaneously or spaced inoculations, hair
Ferment terminates the volatile acid content of wine and yield is all reduced with the increase of R12 ratios, this illustrates that R12 and S.cerevisiae is mixed
Volatile acid content can be reduced by closing fermentation.On the other hand, at the same be inoculated with mixed fermentation production amounts of glycerol it is most, illustrate R12 and
The mixed fermentation that S.cerevisiae is inoculated with simultaneously can improve glycerol production.In addition, R12 can improve volatility object in ice wine
The content of matter can especially improve the content of diethyl succinate, benzyl carbinol and rose oxide in ice wine, and it is more to assign ice wine
The fragrance of a flower, improve ice wine aroma complexity.
In conclusion bacterial strain R12 provided by the invention has stronger ethanol tolerance, SO2Tolerance and high sugar are resistant to
Property, there is fermentability in the zymotic fluid of a concentration of 200~400g/L of sugar, and the bacterial strain has low yield volatile acid and high yield
The yeast strain and saccharomyces cerevisiae mixed fermentation can be reduced fruit wine by the characteristic of glycerine in brewing sweet white wine and ice wine
Volatilization acid yield improves fruit wine glycerol content and improves the content of diethyl succinate, benzyl carbinol and rose oxide in ice wine, assigns
The more fragrance of a flower of ice wine are given, the complexity of ice wine aroma is increased.Yeast strain ferments ability provided by the invention is strong, fermenting property
It is excellent, there is important value in promoting the application of fruit wine quality.
The bacterial strain CEC R12 of the present invention, the deposit date is on 03 08th, 2018, deposit number was CGMCC NO:
15430, Classification And Nomenclature, which is the triumphant formula of Dell, spore torula (Torulaspora delbrueckii), and depositary institution is entitled：In
State's Microbiological Culture Collection administration committee common micro-organisms center, collection address are：BeiChen West Road, Chaoyang District, BeiJing City 1
Number No. 3 Institute of Microorganism, Academia Sinica of institute.
Description of the drawings
Fig. 1 is the growthform figure of bacterial strain R85 and bacterial strain R12；
Fig. 2 is bacterial strain R85, R12 growth curve chart；
Fig. 3 is CO in bacterial strain R85, R12 fermentation process2Weight loss rate figure；
Fig. 4 is bacterial strain R85, R12 ethanol tolerance measurement result figure；
Fig. 5 is bacterial strain R85, R12SO2Tolerance measurement result figure；
Fig. 6 is bacterial strain R85, R12 high sugared tolerance measurement result figures；
Fig. 7 is bacterial strain R85, R12 flocculability measurement result figure；
Fig. 8 is bacterial strain R85, R12 leaven line chart in 200g/L sugar concentration；
Fig. 9 is bacterial strain R85, R12 leaven line chart in 300g/L sugar concentration；
Figure 10 is bacterial strain R85, R12 leaven line chart in 400g/L sugar concentration；
Figure 11 is bacterial strain R12 and S.cerevisiaeNX11424 mixed fermentation sweet white wine leaven line charts；
Figure 12 is bacterial strain R12 and S.cerevisiaeNX11424 mixed fermentation ice wine fermentation curve graphs；
Figure 13 is the variation of bacterial strain R12 and the total group's bacterium colony of yeast during NX11424 mixed fermentations；
Figure 14 is bacterial strain R12 and key point Flora dynamics during NX11424 mixed fermentations.
Specific implementation mode
For the purpose of the present invention, technical solution and advantageous effect is better described, below in conjunction with attached drawing and specific implementation
The invention will be further described for example.It should be noted that following the methods of implementing are explained further to what the present invention was done
It is bright, it should not be taken as limitation of the present invention.If material used in the embodiment of the present invention, reagent all can be from without specified otherwise
Commercial sources obtain.
The triumphant formula of 1 Dell of embodiment has spore torula (Torulaspora delbrueckii) R12 physiological properties
1.1 bacterium source
The triumphant formula of Dell has spore torula (Torulaspora delbrueckii) R12, R85, Gansu Qilian brewery.Bacterial strain
R12 and R85 is isolated from Gansu Qilian brewery grape wine natural fermentation process, belongs to China native country yeast.Wherein, bacterial strain R85
The areas 26s D1-D2 have such as SEQ ID:Nucleotide sequence shown in 1, the areas bacterial strain R1226s D1-D2 have such as SEQ ID:2 institutes
The nucleotide sequence shown.In addition, growthform figures of the bacterial strain R85 and R12 on WLN culture mediums is as shown in Figure 1, wherein A is bacterium
The growthform figure of strain R85, B are the growthform figure of bacterial strain R12.
1.2 experiment contents and method
1.2.1 the drafting of growth curve
Bacterial strain Torulaspora delbrueckii R12, the R85 that -20 DEG C are preserved are in YPD fluid nutrient mediums, and 28
DEG C, under the conditions of 150r/min, activation is for 24 hours.Using turbidimetry, activated bacterial strain is taken to be inoculated in 40mL by 10% inoculum concentration
In YPD fluid nutrient mediums, 28 DEG C of stationary cultures, if 3 repetitions.A sample is taken per 2h, absorbance is surveyed at OD600.
1.2.2 fermenting power is tested
It by the T.delbrueckii after activation, is inoculated into liquid YPD medium by 2% inoculum concentration, 28 DEG C of cultures, with
It is not inoculated with as a contrast, if 3 repetitions.Per periodically weighing for 24 hours and recording data in fermentation process, fermentation flask weight loss is calculated
(i.e. CO2Burst size).According to CO2Burst size understand yeast fermenting power
1.2.3 ethanol tolerance
YPD fluid nutrient mediums are added in test tube and Du Shi pipes are added, and absolute ethyl alcohol adjustment is separately added into after sterilizing just
Beginning alcohol concentration is 4%, 6%, 8%, 10%, 12%, 14% (v/v), activated bacterial strain is inoculated with by inoculum concentration 2%, 28
DEG C culture, if 3 repetitions.Per its aerogenesis situation of 4h time recordings after culture 15h, OD600 measures absorbance A after 3d.
1.2.4 SO2Tolerance
YPD culture mediums are added in test tube and Du Shi pipes are added, and K is added after sterilizing2S2O5(in superclean bench through purple
Outer sterilization 30min), adjust initial SO2A concentration of 50mg/L, 100mg/L, 150mg/L, 200mg/L, 250mg/L, 300mg/L,
Bacterial strain after activation is inoculated with by 350mg/L, 400mg/L, 450mg/L, 500mg/L, 550mg/L, 600mg/L by inoculum concentration 2%,
28 DEG C of cultures, if 3 repetitions.Per its aerogenesis situation of 4h time recordings after culture 15h, OD600 measures absorbance A after 3d.According to
Aerogenesis situation and bacterial concentration understand yeast SO2Tolerance.
1.2.5 high sugared tolerance
Du Shi pipes are added in addition YPD culture mediums (Initial sugar concentration 200g/L, 300g/L, 400g/L) in test tube, go out
Bacterium.Bacterial strain after activation is inoculated with by inoculum concentration 2%, 28 DEG C of cultures, if 3 repetitions.OD600 measures absorbance A after 3d.According to
Aerogenesis situation and bacterial concentration understand the high sugared tolerance of yeast.
1.2.6 yeast flocculence measures
Bacterial strain is seeded to by 10% inoculum concentration in YPD fluid nutrient mediums, is fermented, until growth stationary phase mid-term, centrifugation are received
Collect somatic cells, spends flocculation buffer solution (50mmol/L sodium citrates, 5mmol/L EDTA, pH value 3.0) and sterile water washing 2
Time, it is suspended in flocculation buffer solution (50mmol/L sodium citrates, 20mmol/LCaCl2), 30 DEG C are placed in 50mL shaking flasks, 100r/
Min cultivates 2h.The cell suspending liquid of 5mL vertically stands 30min in 10mL test tubes, and 350 μ L samples are accurately drawn under concave meniscus
Product measure its OD value at wavelength 600nm, are repeated 3 times, and each bacterial strain does 3 parallel tests.The calculation formula of flocculation value：
Flo=B/A × 100%
In formula：The cell OD values being suspended in before A expression shaking flask cultures in flocculation buffer solution, B indicate cell flocculation sedimentation
OD values after 30min, Flo indicate flocculation value.
1.3 results and analysis
1.3.1 the drafting of growth curve
The results are shown in Figure 2 for the growth curve of bacterial strain R85 and R12, figure it is seen that bacterial strain R85 and R12 is being inoculated with
Almost enter growth logarithmic phase after 8h simultaneously, the wherein growth of R12 is very fast, and R85 growths are slower.They after being inoculated with 16h almost
Enter growth stationary phase simultaneously.
1.3.2 fermenting power measures
It can be obtained by Fig. 3 fermentation rates curve, the fermentation rate variation of two saccharomycetes is consistent, and fermentation is entered after 48h
Stationary phase.In the preceding 48h of fermentation, the fermentation rate of R85 is always more than R12, the basic phase of fermentation rate of two saccharomycetes after 48h
Deng.
1.3.3 ethanol tolerance
From table 1, it can be seen that the size of initial alcohol concentration can directly affect the speed of yeast fermentation starting.With
The increase of alcohol concentration, CO2Burst size gradually decrease, fermentation starting slows down gradually.As shown in figure 4, with alcohol concentration
Increase, saccharomycete thalline quantity gradually decreases, while being influenced on the growth and breeding of saccharomycete increasing.When alcohol concentration is
8%, 10% and 12% when, saccharomycete can be grown but reproduction speed is had nothing in common with each other.Wherein, the growth rate of R85 is faster than R12.
When alcohol concentration is 14%, although seriously inhibiting the growth of two saccharomycete, the two can still be grown.The result shows that
Two plants of T.delbrueckii resistance to 14% (v/v) alcohol.It can thus be seen that fermentability and bacterial strain are just to the patience of alcohol
It is relevant.
The ethanol tolerance of 1 T.delbrueckii of table
Note：Bubble-free is represented ,+bubble accounts for Du Shi pipes 1/3 is represented, ++ bubble accounts for Du Shi pipes 2/3 is represented, +++ it represents
Bubble is managed full of Du Shi.
1.3.4 SO2Tolerance
From table 2 it can be seen that initial SO2Concentration can directly affect the speed of fermentation starting.With SO2Concentration
Increase, CO2Burst size gradually decrease, fermentation starting slows down gradually.As shown in figure 5, working as SO2A concentration of 100mg/L, 150mg/
L, when 200mg/L, 250mg/L, 300mg/L, 350mg/L, 400mg/L and 450mg/L, the equal growth and breeding of two saccharomycetes but numerous
Speed is grown to have nothing in common with each other.Work as SO2When a concentration of 500mg/L, 550mg/L and 600mg/L, although the growth of the two is by serious
Inhibit, but still can grow.The result shows that two plants of T.delbrueckii are in SO2It is raw in the case of a concentration of 600mg/L
It is long.Therefore, in Fermentation of Grape Wine, 600mg/L SO2Additive amount will not be to the growth of T.delbrueckii R12 and numerous
It grows and generates strong inhibiting effect.
The SO of 2 T.delbrueckii of table2Tolerance
Note：Bubble-free is represented ,+bubble accounts for Du Shi pipes 1/3 is represented, ++ bubble accounts for Du Shi pipes 2/3 is represented, +++ it represents
Bubble is managed full of Du Shi.
1.3.5 high sugared tolerance
Shown in Fig. 6, two plants of T.delbrueckii can be grown the case where sugar content is 400g/L, and upgrowth situation
More preferable to be grown in the environment of 200g/L and 300g/L than content, wherein R85 growing states are better than R12, in 400g/L environment,
R12 and R85 growing states are not much different.
1.3.6 flocculability
In general, during the fermented soy of grape wine, dispersion increases and fast deposition was made wine in standardization
Yeast strain is required in journey.When yeast has good flocculability vinous fermentation liquid clarifying speed can accelerate, drop
The energy expenditure of low yeast separation, while preventing yeast cells because being suspended in zymotic fluid for a long time, make aqtocytolysis and influences
The flavor of grape wine.In saccharomycete flocculability is evaluated, when Flo be 70%~100%, belong to low flocculability；Flo is
30%~70%, belong to middle flocculability：Flo is 0%~30%, belongs to high flocculability.As shown in Figure 7, two plants
T.delbrueckii's has middle flocculability, and without low flocculability, the wherein flocculability of R12 is preferable.
The triumphant formula of 2 Dell of embodiment have spore torula (Torulaspora delbrueckii) R12 fermentation characters study and its
Application in fermentation
There are spore torula R85 and R12 to carry out fermentation character research using Triple M simulation juice the triumphant formula of above-mentioned Dell,
The formula of Triple M simulation juice is：
ergo stock：95% ethyl alcohol of 12.5mL Tween80,37.5mL, 0.125g ergosterols, solution A：375mL
Add 100g glucose, 100g fructose, 4mL ergo stock that deionized water is added after dissolving and adds to 500mL in deionized water；
Solution B：Add 6g L (+) tartaric acid, 3g L (-) malic acid, 0.5g citric acids in 250mL deionized waters；Solution C：250mL go from
1.7g YNB, 2g acid hydrolyzed caseins, 6mg inositols, 0.2g anhydrous calcium chlorides, L-arginine 0.8g, L- dried meat ammonia are added in sub- water
Sour 1g/L, DL-tryptophan 0.1g/L, ammonium phosphate 1g.A, after B, C mixing, with 4mol/L potassium hydroxide tune pH to 3.25,0.22 μm
Membrane filtration degerming, it is now with the current.
Processing one：Triple M simulate juice stoste；
Processing two：Add 50g glucose, 50g fructose respectively in Triple M simulation juice stostes；
Processing three：Add 100g glucose, 100g fructose respectively in Triple M simulation juice stostes.
Fermentation physical and chemical index and glycerol content are measured after fermentation, physical and chemical index includes alcoholic strength % (v/v),
Remaining reduced sugar, Titrable acid, volatile acid, pH, assay method is with reference to grape and wine trials technical operation specification.Glycerol content
It measures and uses glycerine kit, be purchased from Megazyme glycerol GK.
2.1 T.delbrueckii fermentation rates
As shown in figure 8, two plants of T.delbrueckii are in the zymotic fluid of a concentration of 200g/L of sugar, fermentation time is long, fermentation
Rate is slow, and their fermentation rate no significant difference.It is slightly larger than R12,720h in the fermentation rate of the preceding 720h of fermentation, R85
Afterwards, the fermentation rate of R12 is slightly larger than R85.As shown in figure 9, zymotic fluids of two plants of T.delbrueckii in a concentration of 300g/L of sugar
In, fermentation time is longer, and fermentation rate is slower.729h before fermentation, the fermentation rate no significant difference of the two.R85 after 729h
Fermentation rate is more than the fermentation rate of R12.As shown in Figure 10, fermentations of two plants of T.delbrueckii in a concentration of 400g/L of sugar
In liquid, fermentation rate is slow, and fermentation time is long.And fermentation rate no significant difference.
2.2 basic physical and chemical indexes
When table 3 is a concentration of 200g/L of sugar, two plants of main physical and chemical indexes of T.delbrueckii fermentations.It can be with from the table
Find out, two plants of T.delbrueckii are under the identical environment of a concentration of 200g/L of sugar, the wine degree, the total acid that are generated through alcoholic fermentation
With pH without significant difference.There were significant differences with R85 on production volatile acid content by R12, and the production volatile acid content of R12 is minimum.R85 is sent out
The wine degree of ferment is about 11.7% (v/v), and the wine degree ratio R85 of R12 fermentations is more slightly lower, and 11.51%.Two plants of T.delbruecki fermentations
Residual sugar content be below 2g/L, reach the requirement of dryness grape wine.
Two plants of T.delbrueckii fermentation physical and chemical indexes when 3 a concentration of 200g/L of sugar of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in T.
When table 4 is a concentration of 300g/L of sugar, two plants of main physical and chemical indexes of T.delbrueckii fermentations.It can be with from the table
Find out, two plants of T.delbrueckii are under the identical environment of a concentration of 300g/L of sugar, and the volatile acid generated through alcoholic fermentation is without aobvious
Write difference.There were significant differences with R12 in residual sugar content by R85.There were significant differences with R85 on total acid content and pH by R12.Two plants
Saccharomycete generates in fermentation and is respectively present significant difference on wine degree.R85 fermentation production wine degree is up to 17.1%.Two plants
The residual sugar content of T.delbruecki fermentations is all higher than 12g/L, reaches the requirement of semi-sweet grape wine.The production volatile acid of R12 contains
It measures minimum.
Two plants of T.delbrueckii fermentation physical and chemical indexes when 4 a concentration of 300g/L of sugar of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in T.
When table 5 is a concentration of 400g/L of sugar, two plants of main physical and chemical indexes of T.delbrueckii fermentations.It can be with from the table
Find out, under the identical environment of a concentration of 400g/L of sugar, alcoholic fermentation terminates later total acid, waves two plants of T.delbrueckii
Content of turning sour and pH are without significant difference.The residual sugar content of R85 is substantially less than R12, correspondingly, R12 (5.8%) fermented wine sample
Alcoholic strength is less than R85 (6.75%), and the residual sugar content of two plants of T.delbruecki fermentations is all higher than 45g/L, reaches sweet tea type grape
The requirement of wine.
Two plants of T.delbrueckii fermentation physical and chemical indexes when 5 a concentration of 400g/L of sugar of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in T.
The measurement of 2.3 glycerol contents
As shown in table 6, in the zymotic fluid of a concentration of 200g/L of sugar two plants of T.delbrueckii production glycerol content difference
Not significantly.In Fermentation of Grape Wine, glycerine is converted by carbon source, and yeast strain, culture medium and fermentation condition etc. are no
With the yield that can all influence glycerine.It is a concentration of that R12 and R85 glycerol productions in the zymotic fluid of a concentration of 300g/L of sugar are more than sugar
200g/L.As it can be seen that the zymotic fluid low compared to sugared concentration, the production glycerol content of high glucose concentration zymotic fluid are higher.
Glycerol content (g/L) under 6 bacterial strain R12, R85 different sugar concentration of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in T.
Volatile acid contents of the R12 in the zymotic fluid of 3 kinds of different sugar concentration is minimum.In the sugared concentration of 200g/L and 300g/L
It is high that glycerol content is produced in zymotic fluid.In residual sugar, wine degree, total acid and pH this 4 physical and chemical indexes, two saccharomycetes are substantially without significantly
Difference.
Influence of the 3 T.delbrueckii and S.cerevisiae mixed fermentations of embodiment to sweet white wine quality
There are spore torula R12 and Saccharomyces Cerevisiae in S accharomyces cerevisiae NX11424 to carry out the triumphant formula of Dell
Mixed fermentation grape juice, wherein saccharomyces cerevisiae NX11424 are stored in Xibei Univ. of Agricultural ＆ Forest Science ＆ Technology's microbial strains holding room, grape
Kind：Chardonnay；Plucking time：In September, 2015；Grape juice physicochemical property：Total reducing sugar 239g/L, total acid 6.6g/L, pH 3.7.
It ferments by sweet white small container zymotechnics, total inoculum concentration 5 × 10 of fermenting every time5(blood cell plate counts cells/mL
Method), standing for fermentation, 18 DEG C of fermentation temperature, if two are parallel.By surveying proportion indirect monitoring fermentation process, hair in fermentation process
Basic physical and chemical index and volatile matter content are measured after ferment.Fermentation inoculative proportion design is shown in Table 7.
7 mixed fermentation experimental design table of table
Process route：Picking fruit → cooling → squeezing grape juice, 60mg/LSO2→ standing is clarified → connects barms, pure
Grape juice ferments before (18 DEG C) → reduced sugar >=45g/L, terminates fermentation and (adds SO24 DEG C of low-temperature treatments simultaneously).
Per proportion of survey for 24 hours in fermentation process.Each processing is repeated to survey 3 times and is averaged.After fermentation, reason is measured
Change index, glycerol content and organic acid, physical and chemical index is measured to carry out referring in particular to grape and wine trials technical operation specification,
Separation Depuration Craft uses glycerine kit, organic acidity test to use liquid chromatography.
Organic acidity test：Chromatographic condition：Mobile phase：0.005N H2SO4(HPLC grades aqueous solution)；Chromatographic column：
PHenomenex Rezex ROA organic acid analysis columns (150mm × 7.8mm i.d, 8 μm)；Column temperature：Room temperature；Leacheate：
0.005mol/L sulfuric acid solutions；Flow velocity：0.5mL/min；Sampling volume：20μL.Detector：UV@210nm.
Sample analysis：After wine samples are suitably diluted after 0.45 μm of membrane filtration, sample detection.
The measurement of volatile materials：Using solid-phase microextraction method to the volatile matter content in grape wine
It is measured.
A, extraction conditions
8mL sample solutions are pipetted in ml headspace bottles of the 15mL equipped with magnetic stirring apparatus, 2.0g NaCl and 5 μ L internal standards are added
Solution after adsorbing 40min with 100 μ l PDMS extracting heads under the conditions of 45 DEG C, is analyzed into GC-MS.
B, GC-MS analysis conditions
Chromatographic condition：Shunting mode is not shunt, and chromatographic column is DB-Wax (30m × 0.25mm × 0.25 μm).Program liter
Wen Wei：40 DEG C of holding 3min, rise to 120 DEG C, then rise to 230 with the heating rate of 8 DEG C/min with the heating rate of 5 DEG C/min
DEG C, keep 10min.Gas is He, and volume flow 1mL/min, injector temperature is 250 DEG C.
Mass Spectrometry Conditions are：EI+ ionization sources, electron energy 70eV, filament flow are 0.20mA.Detector voltage is
350V, scanning range 33-450AMU, ion source temperature are 200 DEG C.
C, fragrance component qualitative method
Data Processing in Experiment is completed by Xcalibur software systems, unknown compound be computed machine examination rope simultaneously with
NISTlibrary (107k compounds) and Wiley library (320k compounds, version 6.0) match.
It is just reported more than the qualification result of 800 (maximum values 1000) through working as matching degree and purity.
Sensory quality assessment analysis is carried out according to Wine standard (GB 15037-2006), mainly from appearance, perfume (or spice)
Gas, mouthfeel, four aspect of typicalness are evaluated.15 people of taste panel, it is for statistical analysis to tasting result.
3.1 fermentation diagram
As seen from Figure 11, the temperature during entire fermentation is stablized at 14~16 DEG C, meets the temperature of sweet white fermentation.Together
When inoculation fermentation (T1, T2) fermentation rate variation tendency it is identical, and difference unobvious, illustrate T.delbrueckii have compared with
Strong fermentability can be used as and start yeast.
The fermentation rate of mixed fermentation (T1, T2) is inoculated with simultaneously in 4 kinds of fermented types integrally than being spaced 5d inoculation fermentations
(J1, J2) mixed fermentation rate is big.It is inoculated with T.delbrueckii and S.cerevisiae, the inoculation of T.delbrueckii simultaneously
The inhibition no significant difference to S.cerevisiae is measured, interval 5d is inoculated with T.delbrueckii and S.cerevisiae, it is possible to find
There is difference to the inhibition of S.cerevisiae in the inoculum concentration of T.delbrueckii, and the T.delbrueckii the how right
The inhibition of S.cerevisiae is more apparent.This explanation, T.delbrueckii can be with S.cerevisiae in Non-Saccharomyces
Syntrophism, and T.delbrueckii also has stronger fermentability, can be improved with S.cerevisiae mixed fermentations
The quality of grape wine.
The physicochemical characteristics of 3.2 fermented wines are analyzed
8 are shown in Table to the physical and chemical index analysis result of fermented wine.
(1) volatile acid content
As can be seen from Table 8, relative to T1 and T2, the volatile acid content of J1 and J2 are low, respectively 0.63g/L and
The volatile acid content highest of 0.75g/L, T2 are 1.06g/L.Illustrate that volatile acid can be reduced by the way of indirect inoculation fermentation
The generation of content, this be perhaps because relative at the same be inoculated with, in the mode being inoculated with indirectly, the thalline of T.delbrueckii
Number is more than S.cerevisiae, and the fermentation of T.delbrueckii accounts for leading.In two groups of different fermentation process of J1, J2 and T1, T2
In, the volatile acid content of J1 is less than J2, and the volatile acid content of T1 is less than T2, this explanation in T.delbrueckii and
During S.cerevisiae mixed fermentations, T.delbrueckii not will produce excessive volatilization in vinous fermentation
Acid, but be conducive to the reduction of volatile acid content.
(2) alcohol content and residual sugar content
The wine degree of 4 sections of fermented wines has reached the set goal, has met Portugal all in the left and right 10% (v/v) in this experiment
The requirement of grape wine standard.
The residual sugar content of 4 sections of fermented wines of this experiment meets the requirement of sweet wine all in 45g/L or more.It can from table 8
To find out, the alcohol content of J1 and J2 are both greater than T1 and T2, and residual sugar content is more than T1 and T2.This illustrates the fermentation being inoculated with indirectly
Power is more than while being inoculated with, and is perhaps because while being deposited between T.delbrueckii and S.cerevisiae in the zymotic fluid of inoculation
In stronger relationship of vying each other, the effect mutually inhibited.It is can be found that when indirect 5d is followed by S.cerevisiae this
Inhibiting effect has significantly reduced.
The physical and chemical index of the fermentation sweet white wine of table 8
Note：Different letters is represented examines (P ＜ 0.05) significant difference in Duncan
(3) organic acid content
The final mouthfeel and stability of grape wine have important relationship with organic acid, it is also to influence grape wine organoleptic attribute
One of important indicator.As can be seen from Table 8, either more than inoculation simultaneously or indirect inoculation T.delbrueckii contents
The content of citric acid and tartaric acid is relatively more in zymotic fluid.Zymotic fluid in being inoculated at the same time more than T.delbrueckii contents
The content of middle malic acid and succinic acid is relatively more, but apple in the zymotic fluid in inoculation more than T.delbrueckii contents indirectly
The content of acid and succinic acid is relatively fewer.As it can be seen that in the mixed fermentation of T.delbrueckii and S.cerevisiae,
T.delbrueckii contents are to influence the principal element of citric acid and tartaric acid content, vaccination ways substantially to citric acid and
Tartaric acid content does not influence.And the content and vaccination ways of T.delbrueckii have the content of malic acid and succinic acid
It influences.
(4) glycerine
The content of glycerine can influence the sweet taste of grape wine, wine body and richness.As can be seen from Table 8,4 sections of fermented wines
In, while the glycerol content of mixed fermentation and inoculation mixed fermentation indirectly is inoculated with there are significant difference, while two kinds be inoculated with are not
For glycerol content with processing there is also significant difference, the difference between two processing being inoculated with indirectly is not notable.The glycerine of T1 contains
Amount is more than T2, and the glycerol content of J1 is more than J2.Illustrate, the number of T.delbrueckii is positively correlated with glycerol production.
3.3 volatile materials measure
The most important fragrance source of white wine is the aroma substance that it is formed during the fermentation, includes mainly advanced
The difference of the type and content of alcohol, ester and acid, especially ester can bring different fruit flavors to white wine, as known from Table 9, no
Influence with vaccination ways to different material is different.
The volatile matter content of the fermentation sweet white wine of table 9
Different letters is represented examines (P ＜ 0.05) significant difference, ND in Duncan：It can't detect
It can be obtained by table 9, there is 3 kinds of alcohol, 4 kinds of esters and a kind of acid in Ta.Wherein, ethyl acetate content highest, 86.49 μ g/L.Second
Acetoacetic ester has the fragrance of banana.There are 9 kinds of volatile materials in Tb.Wherein alcohols has 4 kinds, and esters have 4 kinds, and acids has a kind,
The content highest of middle Kui acetoacetic ester is 11.34 μ g/L, assigns grape wine fruity and aroma fragrance.Tb contains other 3 sections of wine samples
No 2- nonyl alcohols.Alcohols in Ja has 4 kinds, and esters have 4 kinds and a kind of acid, and wherein ethyl acetate content is highest, is
20.16μg/L.There are 3 kinds of alcohol, 4 kinds of esters and a kind of acid in Jb, is equally ethyl acetate content highest, 14.08 μ g/L.From table 9 also
It can be seen that (E)-blatter alcohol can only detect in Ja and Jb, this perhaps because vaccination ways are different, to (E) -3- oneself
The influence that alkene -1- alcohol generates is also different.
3.4 sensory evaluation
Wine sample after alcoholic fermentation is store by one month bottle and is stablized, and sensory evaluation is carried out.Pass through sense organ group product
Mirror, the scoring event of the fermented wine of 4 sections of mixed fermentation is shown in Table 10, it will be seen that the highest scoring of T1 from table, 90.33.
T2 scores are minimum, and 71.33.Whole score is T1 ＞ J1 ＞ J2 ＞ T2.
Sense organ taste's result of the different mixed fermentation grape wine of table 10
Note：100 points of systems：Perfect ＞ 86；Very well：81-86；Good 71-80；Generally：50-70；It is bad：＜ 50.
In the mixed fermentation of T.delbrueckii and S.cerevisiae, from fermentation rate, indirect inoculation fermentation
Fermentation rate ratio simultaneously inoculation fermentation it is small, fermentation time is long.This is in the fermenting power for the bacterial strain being first added, the time of spaced inoculations
There is relationship.From the physical and chemical index of fermented wine, it is known that, as T.delbruecki R12 ratios increase, production volatile acid content drop
Low, glycerol content increases.Sensory evaluation is carried out to 4 sections of wine samples, obtained whole sense organ quality good or not sequence is T1 ＞ J1 ＞ J2
＞ T2.
The application of embodiment 4T.delbruecki and S.cerevisiae mixed fermentation ice grape juice
T.delbruecki R12 and S.cerevisiae NX11424 are subjected to ice grape juice fermentation, grape variety：Prestige
Dai Er；Grape physical and chemical index：Total sugar content 398.7g/L, total acid 10.19g/L, pH 3.14；Grape juice processing：Add 250mg/
L(NH4)2SO4Nitrogen source is supplemented, while 600mg/L DMDC being added to be sterilized, stands 6h or more.
The target residual sugar of ice wine is set as 50g/L, is made by the small container zymotechnics of sweet white wine, and fermentation every time always connects
Kind amount 1 × 106Cells/mL (blood cell plate counting method), 18 DEG C, 150r/min of fermentation temperature, if three parallel.In fermentation process
By surveying reduced sugar monitor fermentation process, and WLN colony counting methods are used, (fermentation starting, sugar consumption are sampled in fermentation key point
1/2, before sugar consumption 3/4, termination fermentation) saccharomycete is counted, basic physical and chemical index and volatility are measured after fermentation
Content of material.Inoculative proportion design of fermenting is as shown in table 11.Simultaneously be respectively adopted T.delbrueckii individually fermentation and
Individually fermentation compares S.cerevisiae.
11 mixed fermentation experimental design table of table
4.1 fermentation diagram
As shown in figure 12, the rate that S.cerevisiae individually ferments individually is fermented more than T.delbrueckii.Indirect
In the fermentation of 5d inoculations, when residual sugar content is 90g/L or so in zymotic fluid, residual sugar content does not change within 6d, then
It is considered as fermentation to terminate.Illustrate the mixed fermentation carried out in such a way that indirect 5d is inoculated in ice grape juice, target wine can not be completed
The fermentation of sample.The fermentation rate variation tendency of indirect inoculation fermentation (Ja, Jb and Jc) is identical, almost without difference, and connects simultaneously
Kind, T.delbrueckii individually ferment and
S.cerevisiae individually compare by fermentation, and the fermentation rate of spaced inoculations is slow, and fermentation time is long.This in high sugar
In environment, S.cerevisiae is accessed after accessing T.delbrueckii 5d, T.delbrueckii contents are more at this time, seriously
Inhibit the growth of S.cerevisiae related.
The 3 groups of experiments that can be obtained, while be inoculated with by Figure 12, the fermentation time of Tc is most short, Ta longests, and fermentation rate is
Tc ＞ Tb ＞ Ta.Compared with saccharomyces cerevisiae individually ferments, to be all slower than saccharomyces cerevisiae independent for the fermentation rate of 3 groups of inoculation fermentations simultaneously
Fermentation.In the mixed fermentation that this explanation is inoculated at the same time, with increasing for saccharomyces cerevisiae, the fermentation rate of mixed fermentation becomes larger.
Have spore torula individually compared with fermentation with the triumphant formula of Dell, Tc ＞ R ＞ Tb ＞ Ta, this be perhaps because in T.delbrueckii and
S.cerevisiae inoculative proportions are 20:1 and 10:The mixed fermentation of 1 inoculation simultaneously, there is no because of saccharomyces cerevisiae for mixed fermentation
Addition and promote their fermentation rate, they become a kind of competition instead, and the relationship mutually inhibited reduces mutual hair
Ferment rate.Here the content of saccharomyces cerevisiae is less, and the triumphant formula of Dell has the alcoholic fermentation of spore torula to occupy an leading position.When wine brewing ferment
Female content increase to Dell triumphant formula have spore torula it is equal when (Tc), saccharomyces cerevisiae content increases, competitiveness enhancing, this
When saccharomyces cerevisiae alcoholic fermentation occupy an leading position, along with saccharomyces cerevisiae fermenting power be more than the triumphant formula of Dell have spore torula,
Then Tc ＞ R.
The dynamic change of saccharomycete quantity in 4.2 fermentation process
By 4 key points to fermenting (when fermentation starting, sugar consumption 1/2, sugar consumption 3/4 and before terminating fermentation) into
Row sampling, WLN coated plates count result, draw Figure 13.As shown in Figure 13, before sugar consumption amount 1/2, two primary yeasts
Yeast total amount is being continuously increased always, and after sugar consumption amount is more than 1/2, saccharomycete total bacteria count is also increasing, but it is slow to increase trend
Slowly, when sugar consumption is close to 3/4, saccharomycete total bacteria count drastically declines.In general, as saccharomyces cerevisiae flora number ratio increases
Greatly, total yeast count in mixed fermentation also increases, this is consistent with their fermentation diagram trend.
As can be known from Fig. 14, at the same time in 3 processing of inoculation mixed fermentation, 3/4 from fermentation starting to sugar consumption,
The bacterium number of T.delbrueckii is more than S.cerevisiae, and before fermentation terminates, the bacterium number of S.cerevisiae is more than
T.delbrueckii.This is perhaps because in high saccharide ring border, the growth ability of T.delbrueckii is more than
S.cerevisiae, but as the progress of fermentation, alcohol are continuously generated, the bacterium number of 2 primary yeasts constantly declines, due to
The ethanol tolerance of T.delbrueckii is without S.cerevisiae high, therefore the fall off rate of T.delbrueckii is more than wine
Wine.Appear in the phenomenon that bacterium number of fermentation later stage S.cerevisiae is more than T.delbrueckii.
The physicochemical characteristics of 4.3 ice wine are analyzed
The physicochemical characteristics analysis of ice wine is as shown in table 12.
(1) volatile acid content
As can be seen from Table 11, by 20：The T.delbrueckii and S.cerevisiae of 1 inoculative proportion spaced inoculations
(Ja) mixed fermentation volatile acid content is minimum, is 0.42g/L, and S.cerevisiae individually ferment (N) highest, for 1.57g/L.
In T.delbrueckii and S.cerevisiae mixed fermentations, the volatile acid content that spaced inoculations mixed fermentation generates is significantly low
It is inoculated with mixed fermentation and two plants of bacterium are individually fermented in simultaneously.And at the same time in the processing of inoculation and spaced inoculations mixed fermentation,
With increasing for S.cerevisiae, volatile acid content increases.It is inoculated with simultaneously in T.delbrueckii and S.cerevisiae mixed
Close fermentation, the independent threes of individually fermenting with T.delbrueckii of S.cerevisiae compare, it can be seen that with
S.cerevisiae's increases, and volatile acid content increases.With increasing for T.delbrueckii, volatile acid content reduces.
The volatile acid that T.delbrueckii individually ferments is above T.delbrueckii and S.cerevisiae spaced inoculations mixing hair
Ferment.It can obtain, spaced inoculations ferment according to a certain percentage by T.delbrueckii and S.cerevisiae, can effectively drop
The generation of low volatile acid.
The basic physical and chemical index of 12 ice wine of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in Duncan
It can be seen that from volatile acid yield by 20:1、10:1 inoculative proportion spaced inoculations (Ja, Jb)
T.delbrueckii and S.cerevisiae volatile acids yield is minimum, is 0.001g/L, and S.cerevisiae individually ferments (N)
Maximum is 0.0042g/L.It is similar to volatile acid content, in T.delbrueckii and S.cerevisiae mixed fermentations, interval
The volatile acid yield of inoculation mixed fermentation is significantly less than while being inoculated with the fermentation of mixing.And inoculation and spaced inoculations are mixed at the same time
In the processing for closing fermentation, with increasing for S.cerevisiae, volatile acid yield increases.In T.delbrueckii and
S.cerevisiae is inoculated with mixed fermentation simultaneously, the independent threes of individually fermenting with T.delbrueckii of S.cerevisiae compare,
As can be seen that increasing with S.cerevisiae, volatile acid yield increases.With increasing for T.delbrueckii, volatile acid
Yield reduces.The volatile acid yield that T.delbrueckii individually ferments is all higher than between T.delbrueckii and S.cerevisiae
Every inoculation mixed fermentation.By the comparison of the volatile acid yield to 8 kinds of fermented wines, further prove that the triumphant formula of Dell has spore torula
Mixed fermentation can effectively reduce the volatilization acid yield in ice grape juice fermentation process in a manner of spaced inoculations with saccharomyces cerevisiae.
(2) wine degree and residual sugar content
As shown in Table 12, the wine degree of 8 sections of fermented wines is all 12.15% or more, and saccharomyces cerevisiae individually ferments the alcohol of generation
Highest, up to 19.16%.Followed by the triumphant formula of Dell have spore torula individually ferment generation alcoholic strength be 18.86%.Here it is spaced
The wine degree that inoculation mixed fermentation generates is below while being inoculated with, consistent with fermentation diagram trend.The hair of Tc in fermentation diagram
Ferment power is more than R, but the alcohol producing amount of R is more than Tc, this may be because the triumphant formula of Dell has spore torula and wine brewing in Tc fermentations
Yeast is vied each other, is inhibited, and the generation of alcohol is reduced.It is inoculated with that mixed fermentation, saccharomyces cerevisiae individually ferment and Dell is triumphant simultaneously
Formula has spore torula remaining residual sugar amount of individually fermenting to be reduced with the raising of wine degree, at 3 of spaced inoculations mixed fermentation
Reason is also such.
(3) organic acid content
As can be seen from Table 13, in addition to Jb and Jc, all significant difference of each citric acid content of other 6 sections fermentations.8
Jb citric acid contents highest in money fermented wine, Tb contents are minimum.Between two processing of Jb and Tb, in addition to vaccination ways are different,
His condition is all identical, and it is apparent to generate citric acid content difference.Ja and Ta, Jc and Tc are also such.Table 13 can also be seen that
In the case that same ratio is inoculated with, while the tartaric acid, malic acid, citric acid content that are inoculated with mixed fermentation are below spaced inoculations
Mixed fermentation.Wherein, the tartaric acid of Jb and malic acid content all highests.Therefore, the number of organic acid content and connecing for saccharomycete
Whether kind mode has direct relationship, it is still necessary to further research.
Organic acid and glycerol content in 13 ice wine of table
Note：Different letters is represented examines (P ＜ 0.05) significant difference in Duncan.
(4) glycerol content
As can be seen from Table 13, it while being inoculated with the glycerol production of mixed fermentation and is both greater than spaced inoculations mixed fermentation,
More than saccharomyces cerevisiae individually and the triumphant formula of Dell there is spore torula individually to ferment.It is inoculated with Ta ＞ Tb ＞ in mixed fermentation at the same time
Tc, spaced inoculations mixed fermentation are also such.This explanation is in mixed fermentation, the content of Dell's yeast and the generation positive of glycerine
It closes.
The volatile matter content of 4.4 ice wine is analyzed
It is fixed altogether in the ice wine of the pure fermentations of T.delbrueckii R12 and S.cerevisiae NX11424 and mixed fermentation
Measure 3 kinds of 45 kinds of important aroma substances (table 14), 20 kinds of esters, 15 kinds of alcohols, 4 kinds of acids, 3 kinds of aldoketones and terpenes.No
Volatile materials total content with mixed fermentation is 605.23~1376.32mg/L.Ta fermentation ice wine in fragrance content most
Height is higher than control strain N (S.cerevisiae NX11424), is secondly Tb, and followed by Ja, fermentation purer than saccharomyces cerevisiae is fragrant
The high 281.21mg/L of gas total content has independent ferment of spore torula R12 to be higher by 411.95mg/L than Dell.It can be seen that simultaneously
Inoculation and T.delbrueckii R12 and S.cerevisiae NX11424 mixed fermentations can improve the fragrance of ice wine, and be inoculated with
Ratio is 20:It is most that the ice wine that ferments when 1 generates aroma substance.
The content of 14 T.delbrueckii and S.cerevisiae mixed fermentation ice wine volatile materials of table
As can be seen from Table 14, the esters of Tb and Ta mixed fermentations, alcohols and aldoketones content are above saccharomyces cerevisiae, acids is low
In saccharomyces cerevisiae, illustrate the quality that there is spore torula in Dell and saccharomyces cerevisiae mixed fermentation can improve ice wine.In Ester,
The content for the diethyl succinate that S.cerevisiae NX11424 individually ferment is minimum, and T.delbrueckii R12 are individually sent out
The content highest of ferment illustrates the content that the triumphant formula of Dell has spore torula that can improve diethyl succinate in ice wine, succinic acid two
Ethyl ester can assign the fruit such as grape wine muskmelon perfume.In alcohols material, in addition to Tc, S.cerevisiae NX11424 are individually sent out
The benzyl carbinol content of ferment is less than the ice wine of other vaccination ways fermentation, illustrates that the triumphant formula of Dell has spore torula R12 in most cases
The content of benzyl carbinol in ice wine can be improved down.Benzyl carbinol has typical rose fragrance, can assign ice wine more fragrance of a flower.
In terpenoid substance, in addition to Tc, the content for the rose oxide that S.cerevisiae NX11424 individually ferment connects less than other
The ice wine that kind mode is fermented, illustrates the content that the triumphant formula of Dell has spore torula R12 that can improve rose oxide in ice wine.Aoxidize rose
It is rare that there is Rose Essentielle and fresh spiceleaf fragrance.Comprehensive explanation, it is more that the triumphant formula of Dell has spore circle ferment R12 that can assign ice wine
Fragrance of a flower fragrance, especially rose fragrance.
Comprehensive analysis results show the content that the triumphant formula of Dell has spore torula R12 that can improve volatile materials in ice wine,
The content of diethyl succinate, benzyl carbinol and rose oxide in ice wine can be especially improved, the more fragrance of a flower of ice wine is assigned, carries
The complexity of high ice wine aroma.
Bacterial strain R12 provided by the invention shows that bacterial strain R12 can be in the alcohol of 14% (v/v) in physiological property research
Degree, SO2It is grown in the environment that content is 600mg/L, sugar content is 400g/L.It is shown in fermentation character research, bacterial strain R12 exists
Volatile acid content in the zymotic fluid of 3 kinds of different sugar concentration is minimum.It is produced in the zymotic fluid of a concentration of 200g/L and 300g/L of sugar
Glycerol content is high.And the fragrance ratio R85 of R12 simulation fermented wines is more prominent.
Bacterial strain R12 provided by the invention shows in the research in mixed fermentation sweet white wine, while inoculation fermentation
The fermentation rate of mixed fermentation is more than the mixed fermentation of interval 5d inoculations, and in the two ways of mixed fermentation, volatile acid
Content is reduced as the triumphant formula of Dell has the increase of spore torula R12 ratios, and glycerol content has spore torula with the triumphant formula of Dell
The ratio of R12 increases and increases.It is obtained by sensory evaluation while being inoculated with (20:1) intervals ＞ 5d inoculations (20:1) intervals ＞ 5d
Inoculation (1:1) ＞ is inoculated with (1 simultaneously:1).In conclusion the triumphant formula of Dell has spore torula R12 with saccharomyces cerevisiae by 20:1 inoculation
The quality of sweet white wine can be improved to ratio by being inoculated with mixed fermentation simultaneously.
Bacterial strain R12 provided by the invention shows in the research of mixed fermentation ice grape juice, T.delbrueckii R12 and
The volatile acid content that the intervals S.cerevisiae 5d is inoculated with mixed fermentation is minimum, is 0.42g/L.Its volatile acid yield is also most simultaneously
It is low, it is 0.001g/L.Although being inoculated with the volatile acid content of mixed fermentation simultaneously higher than the mixed fermentation of interval 5d inoculations, it is less than
R12 and S.cerevisiae respectively individually ferments.The result shows that either in a manner of inoculation simultaneously or spaced inoculations, hair
Ferment terminates the volatile acid content of wine and volatile acid yield is all reduced with the increase of R12 ratios.This illustrate R12 and
S.cerevisiae mixed fermentations can reduce volatile acid content.On the other hand, while the production amounts of glycerol of mixed fermentation is inoculated with most
It is more.Illustrate R12 and S.cerevisiae while the mixed fermentation of inoculation can improve glycerol production.In addition, R12 can improve ice
The content of volatile materials in wine, can especially improve the content of diethyl succinate, benzyl carbinol and rose oxide in ice wine,
The more fragrance of a flower of ice wine are assigned, the complexity of ice wine aroma is increased.
It should be noted that bacterial strain R12 provided by the invention can also be used for the brewing of other fruit wine, and bacterial strain R12 is being sent out
Have the characteristics that low yield volatile acid, high yield glycerine and fragrance protrude in ferment fruit wine, can be used to the quality for improving fruit wine.In addition, this
The bacterial strain R12 provided is provided there is stronger fermentability under the conditions of high sugar, therefore be also applied for fermentation bread, cake etc. it
His fermented product.In actual use, bacterial strain R12 can be made into active dry yeast, in order to store and produce.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than is protected to the present invention
The limitation of range is protected, although being explained in detail to the present invention with reference to preferred embodiment, those skilled in the art should
Understand, technical scheme of the present invention can be modified or replaced equivalently, without departing from the essence of technical solution of the present invention
And range.
Sequence table
<110>Xibei Univ. of Agricultural ＆ Forest Science ＆ Technology
<120>Yeast strain and its application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 589
<212> DNA
<213>The triumphant formula of Dell has spore torula (Torulaspora delbrueckii)
<400> 1
ccatcggtat gccttagtac ggcgagtgaa gcggcaaaag ctcaaatttg aaatctggta 60
ccttcggtgc ccgagttgta atttgtagaa ggtaactttg gggctggtcc ttgtctatgt 120
tccttggaac aggacgtcat agagggtgag aatcccgtgt ggcgaggatc ccagttcttt 180
gtaaagtgct ttcgaagagt cgagttgttt gggaatgcag ctctaagtgg gtggtaaatt 240
ccatctaaag ctaaatattg gcgagagacc gatagcgaac aagtacagtg atggaaagat 300
gaaaagaact ttgaaaagag agtgaaaaag tacgtgaaat tgttgaaagg gaagggcatt 360
tgatcagaca tggtgttttg cgccctctgc tccttgtggg tgggggaatc tcgcagctca 420
ctgggccagc atcagttttg gcggcaggat aaatctgcag gaatgtagct tgcctcggta 480
agtgttatat cctgtagaaa tactgccagc tgggactgag gactgcgact ttacgtcaag 540
gatgctggca taatggttat atgccgcccg tctgaaaacc cgggaacaa 589
<210> 2
<211> 589
<212> DNA
<213>The triumphant formula of Dell has spore torula (Torulaspora delbrueckii)
<400> 2
tcatcggtat gcttagtacg gcgagtgagc ggcaaaagct caaatttgaa atctggtacc 60
ttcggtgccc gagttgtaat ttgtagaagg taactttggg gctggtcctt gtctatgttc 120
cttggaacag gacgtcatag agggtgagaa tcccgtgtgg cgaggatccc agttctttgt 180
aaagtgcttt cgaagagtcg agttgtttgg gaatgcagct ctaagtgggt ggtaaattcc 240
atctaaagct aaatattggc gagagaccga tagcgaacaa gtacagtgat ggaaagatga 300
aaagaacttt gaaaagagag tgaaaaagta cgtgaaattg ttgaaaggga agggcatttg 360
atcagacatg gtgttttgcg ccctctgctc cttgtgggtg ggggaatctc gcagctcact 420
gggccagcat cagttttggc ggcaggataa atctgcagga atgtagcttg cctcggtaag 480
tgttatatcc tgtagaaata ctgccagctg ggactgagga ctgcgacttt acgtcaagga 540
tgctggcata atggttatat gccgcccgtc ttaaaaaccg gaacaaaac 589
Claims (10)
1. a kind of yeast strain, which is characterized in that its deposit number is CGMCC NO:15430, Classification And Nomenclature, which is the triumphant formula of Dell, to be had
Spore torula (Torulaspora delbrueckii).
2. yeast strain as described in claim 1, which is characterized in that the yeast strain is in a concentration of 200~400g/L of sugar
Zymotic fluid in have fermentability.
3. a kind of method reducing fruit wine volatile acid, which is characterized in that take such as claim 1 to 2 any one of them saccharomycete
Strain and saccharomyces cerevisiae mixed fermentation fruit juice raw material.
4. the method for reducing fruit wine volatile acid as claimed in claim 3, which is characterized in that will be such as any one of claim 1 to 2
The yeast strain presses 1 with the saccharomyces cerevisiae:1~20:1 inoculative proportion mixed fermentation fruit juice raw material.
5. a kind of method of enhancing fruit wine fragrance, which is characterized in that take such as claim 1 to 2 any one of them yeast strain
With other yeast strain mixed fermentation fruit juice raw materials.
6. a kind of method improving fruit wine glycerol production, which is characterized in that take such as claim 1 to 2 any one of them yeast
Bacterial strain and other yeast strain mixed fermentation fruit juice raw materials.
7. such as application of claim 1 to 2 any one of them yeast strain in mixed fermentation fruit wine.
8. such as application of claim 1 to 2 any one of them yeast strain in high sugared fermented product.
9. a kind of active dry yeast, which is characterized in that be made by such as claim 1 to 2 any one of them yeast strain.
10. a kind of method that screening can reduce the bacterial strain of fruit wine volatile acid, which is characterized in that take as claim 1 to 2 is any
Yeast strain described in is raw material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810461991.1A CN108624518A (en) | 2018-05-15 | 2018-05-15 | Yeast strain and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810461991.1A CN108624518A (en) | 2018-05-15 | 2018-05-15 | Yeast strain and its application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108624518A true CN108624518A (en) | 2018-10-09 |
Family
ID=63693326
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810461991.1A Pending CN108624518A (en) | 2018-05-15 | 2018-05-15 | Yeast strain and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108624518A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110093231A (en) * | 2019-05-07 | 2019-08-06 | 甘肃省农业科学院农产品贮藏加工研究所 | A kind of preparation method of applejack |
| CN110804523A (en) * | 2019-11-25 | 2020-02-18 | 四川大学 | Method for producing high-quality mulberry wine by fermentation |
| CN110951632A (en) * | 2019-12-31 | 2020-04-03 | 江南大学 | Torulaspora delbrueckii producing α -farnesene and fermentation method thereof |
| CN113667611A (en) * | 2021-07-13 | 2021-11-19 | 西北农林科技大学 | Heat-resistant Kluyveromyces strain and application thereof |
| CN114540148A (en) * | 2020-11-26 | 2022-05-27 | 天津市农业科学院 | Brewing process of dragon fruit wine with low volatile acid content |
| CN115948210A (en) * | 2023-02-28 | 2023-04-11 | 北京燕京啤酒股份有限公司 | Preparation method of honey-flavor low-alcohol beer |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016140966A1 (en) * | 2015-03-04 | 2016-09-09 | Danisco Us Inc | Process for production of bio-alcohol |
| CN106987531A (en) * | 2017-05-05 | 2017-07-28 | 湖南农业大学 | One plant of grape wine laminating adhesive spore yeast and its application in monoterpene production |
-
2018
- 2018-05-15 CN CN201810461991.1A patent/CN108624518A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016140966A1 (en) * | 2015-03-04 | 2016-09-09 | Danisco Us Inc | Process for production of bio-alcohol |
| CN106987531A (en) * | 2017-05-05 | 2017-07-28 | 湖南农业大学 | One plant of grape wine laminating adhesive spore yeast and its application in monoterpene production |
Non-Patent Citations (5)
| Title |
|---|
| MARINA BELY 等: "Impact of mixed Torulaspora delbrueckii-Saccharomyces cerevisiae culture on high-sugar fermentation", 《INT J FOOD MICROBIOL.》 * |
| PHILIPPE RENAULT 等: "Enhanced 3-Sulfanylhexan-1-ol Production in Sequential Mixed Fermentation with Torulaspora delbrueckii/Saccharomyces cerevisiae Reveals a Situation of Synergistic Interaction between Two Industrial Strains", 《FRONT MICROBIOL.》 * |
| PHILIPPE RENAULT 等: "Increase of fruity aroma during mixed T. delbrueckii/S. cerevisiae wine fermentation is linked to specific esters enhancement", 《INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY》 * |
| SANTIAGO BENITO: "The impact of Torulaspora delbrueckii yeast in winemaking", 《APPL MICROBIOL BIOTECHNOL》 * |
| 秦伟帅: "香格里拉本土葡萄酒酵母的筛选及其胁迫耐受性分析", 《中外葡萄与葡萄酒》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110093231A (en) * | 2019-05-07 | 2019-08-06 | 甘肃省农业科学院农产品贮藏加工研究所 | A kind of preparation method of applejack |
| CN110804523A (en) * | 2019-11-25 | 2020-02-18 | 四川大学 | Method for producing high-quality mulberry wine by fermentation |
| CN110804523B (en) * | 2019-11-25 | 2021-08-20 | 四川大学 | Method for producing high-quality mulberry wine by fermentation |
| CN110951632A (en) * | 2019-12-31 | 2020-04-03 | 江南大学 | Torulaspora delbrueckii producing α -farnesene and fermentation method thereof |
| CN110951632B (en) * | 2019-12-31 | 2022-04-29 | 江南大学 | Torulopsis delkesii producing alpha-farnesene and fermentation method thereof |
| CN114540148A (en) * | 2020-11-26 | 2022-05-27 | 天津市农业科学院 | Brewing process of dragon fruit wine with low volatile acid content |
| CN113667611A (en) * | 2021-07-13 | 2021-11-19 | 西北农林科技大学 | Heat-resistant Kluyveromyces strain and application thereof |
| CN113667611B (en) * | 2021-07-13 | 2023-10-13 | 西北农林科技大学 | Heat-resistant Kluyveromyces strain and application thereof |
| CN115948210A (en) * | 2023-02-28 | 2023-04-11 | 北京燕京啤酒股份有限公司 | Preparation method of honey-flavor low-alcohol beer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108624518A (en) | Yeast strain and its application | |
| CN101792719B (en) | Saccharomyces cerevisiae and application thereof in wine brewing | |
| CN106916758A (en) | A kind of Hansenula yeast and its application in wine production | |
| CN108239608B (en) | Torulaspora delbrueckii and application thereof in wine brewing | |
| CN109988720B (en) | Yeast ZB412 and application thereof | |
| CN108251318A (en) | A kind of Dell is furnished with spore torula and its application in Wine Aroma quality is improved | |
| CN105802865B (en) | One plant height fermentation activity and the fragrant characteristic of production ice brewer yeast outstanding and its application | |
| CN109182156B (en) | Saccharomyces cerevisiae suitable for brewing red-core pitaya wine and application thereof | |
| CN101838615B (en) | Saccharomyces cerevisiae and application thereof in reducing acidity in process of producing wine | |
| CN108559713A (en) | A kind of saccharomyces cerevisiae and its application | |
| CN109988685A (en) | The production method of fruity and fragrance of a flower fragrance in a kind of raising grape wine | |
| CN108179120A (en) | One Accharomyces cerevisiae and its application | |
| CN112553091A (en) | Non-saccharomyces cerevisiae and fermentation method for increasing fragrance of blueberry fruit wine by using same | |
| CN110343625B (en) | Saccharomyces cerevisiae strain and application thereof | |
| CN108179119A (en) | A kind of zymotechnique for improving ice-wine flavouring essence quality using Non-Saccharomyces | |
| CN108384728B (en) | Saccharomyces cerevisiae and application thereof | |
| CN108865910B (en) | Saccharomyces cerevisiae, screening method thereof and application of saccharomyces cerevisiae in blueberry red wine fermentation | |
| CN109929766A (en) | A kind of Crewe dimension saccharomyces lactis CVE-LT1 and its application in wine production | |
| CN111621430B (en) | Saccharomyces cerevisiae suitable for brewing yellow peach fruit wine and application thereof | |
| CN116515654B (en) | Saccharomyces cerevisiae and application thereof in longan wine brewing | |
| CN108410745A (en) | One Accharomyces cerevisiae and its application in wine production | |
| CN105296369A (en) | Ageing-type saccharomyces cerevisiae and its use in wine making | |
| CN108342333B (en) | Yeast strain and application thereof | |
| CN113667611B (en) | Heat-resistant Kluyveromyces strain and application thereof | |
| CN109852555A (en) | A kind of Acid-tolerant yeasts and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181009 |
|
| RJ01 | Rejection of invention patent application after publication |