CN108865567A - A kind of preparation method and low alcohol bubbling applejack of low alcohol bubbling applejack - Google Patents
A kind of preparation method and low alcohol bubbling applejack of low alcohol bubbling applejack Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of low alcohol bubbling applejack and low alcohol bubbling applejack, the applejack alcoholic strength for using method of the invention to be prepared reaches low alcohol standard, CO for 1.8%vol2(20 DEG C) of pressure are 0.45MPa, reach fizz standard；The preparation method includes the fermentation preparation of apple former wine being carried out using zymophyte and cider as raw material, then the part ethyl alcohol in apple former wine is removed to obtain low alcohol applejack, and saccharomyces cerevisiae is finally recycled to carry out the secondary fermentation of low alcohol applejack；The zymophyte is S. cervisiae and candidiasis of being addicted to drink.Low alcohol bubbling applejack can be brewed using zymotechnique disclosed above, the aroma substance type for the low alcohol bubbling applejack brewed using the technique of saccharomyces cerevisiae and secondary fermentation after be addicted to drink Candida mixed fermentation and dealcoholysis is enriched, organoleptic indicator is good, golden yellow color, clear, mellow in taste, it is a kind of low alcohol fruit wine of nutrient health that bubble is fine and smooth persistently.
Description
Technical field
The invention belongs to applejack brewing technology fields, and in particular to the preparation method of a kind of low alcohol bubbling applejack and low
Alcohol alcoholic foamed apple wine.
Background technique
Producing low alcohol and the method without alcohol applejack mainly has three classes, focuses on the three phases of production process respectively, i.e.,
Before fermentation, among fermentation and after fermentation.Fermented pretreatment mainly reduces finished wine by fermentable sugars content in reduction matrix
Precision, including add glucose oxidase, use immature fruit fermentation etc.；The generation that intermediate treatment of fermenting passes through reduction ethyl alcohol
Alcoholic strength is controlled, including uses the methods of Non-Saccharomyces or Yeast engineering bacteria, stuck fermentation；Fermentation post-processing passes through physics
Method removes ethyl alcohol in wine, mainly there is non-film method such as rotating cone scapus method；Embrane method includes reverse osmosis, nanofiltration, infiltration extraction
It takes, pervaporation and osmotic distillation etc., low alcohol that these methods obtain and partially light without the generally existing taste of alcohol applejack, wine body is not
It is full.
The production method of fizz mainly has fermentation method i.e. " champagne method ", bulk fermentation method and transfer in conventional bottles at present
Method.The fragrance of applejack is the important indicator for evaluating applejack quality, but the above method mostly all generates the fragrance component of wine
Negative effect, causing must fragrance deficiency.
Summary of the invention
For the defects in the prior art and insufficient, the object of the present invention is to provide a kind of preparations of low alcohol bubbling applejack
Method and low alcohol bubbling applejack overcome the disadvantage that low alcohol aroma of cider is insufficient, mouthfeel is partially light and wine body is not full, obtain one
It kind gives off a strong fragrance, the low alcohol bubbling applejack that mouthfeel is excellent.
For this purpose, the technical scheme is that：
A kind of preparation method of low alcohol bubbling applejack, the preparation method include using zymophyte and cider as raw material
The fermentation preparation of apple former wine is carried out, then the part ethyl alcohol in apple former wine is removed to obtain low alcohol applejack, is finally recycled
Saccharomyces cerevisiae carries out the secondary fermentation of low alcohol applejack；
The zymophyte is S. cervisiae and candidiasis of being addicted to drink.
Optionally, in terms of the volume of cider, the inoculum concentration of fermentation bacteria seed solution is 4%；Inoculative proportion is saccharomyces cerevisiae
Bacterium：It is addicted to drink candidiasis=1:3；
The preparation of the fermentation bacteria seed solution includes：Zymophyte is inoculated in 28 DEG C, 120rpm shaking table in YPD culture medium
Culture activates twice for 24 hours, and the zymophyte after activation is inoculated in cider with 5% inoculum concentration, 28 DEG C, the training of 120rpm shaking table
It supports；
The shaking table culture time of S. cervisiae is 18h, and the shaking table culture time for candidiasis of being addicted to drink is for 24 hours.
Optionally, in the fermentation preparation of the apple former wine：Fermentation temperature is 18 DEG C, fermentation time 7d, cider
Soluble solid content be 12~14 ° of Brix, the pH of cider is 3.9 or so.
Optionally, the ethyl alcohol removing in the apple former wine uses cryogenic vacuum distillating method.
Optionally, the vacuum degree of the cryogenic vacuum distillating method is 0.08~0.09MPa, and vapo(u)rizing temperature is 35 DEG C,
Distillation time is 5min.
Optionally, secondary fermentation is carried out again after also carrying out moisturizing to low alcohol applejack, mend sugar and supplement potassium metabisulfite,
The magnitude of recruitment of water is the volume reduction of low alcohol applejack after removing ethyl alcohol, and sugared magnitude of recruitment is the low alcohol guaranteed after removing ethyl alcohol
Sugared content in applejack is 25~30g/L, and the magnitude of recruitment of potassium metabisulfite is 90mg/L.
Optionally, the temperature of the secondary fermentation is 15 DEG C, and saleratus additive amount is 1g/L, fermentation time 18d.
Optionally, in terms of the volume of low alcohol applejack for removing ethyl alcohol, the seed liquor of the S. cervisiae in secondary fermentation
Inoculum concentration is 6%；
The preparation method of the seed liquor of saccharomyces cerevisiae in secondary fermentation includes：Saccharomyces cerevisiae is cultivated by the effective YPD of glycerol
After base activated for two generations, with the cider after the access sterilizing of 5% inoculum concentration, 120rpm is cultivated for 24 hours at 28 DEG C；Obtained bacterium solution with
In dealcoholysis applejack after the access sterilizing of 8% inoculum concentration, at 28 DEG C after 120rpm culture 36h to obtain the final product.
A kind of low alcohol bubbling applejack, the low alcohol bubbling applejack are prepared using the preparation method.
Compared with prior art, advantages of the present invention is：
This method is by approach of the biological method in conjunction with physical method, while applejack alcoholic strength is effectively reduced,
Most of aroma substance is remained, and secondary fermentation not only brings the bubble for enriching exquisiteness and tasty and refreshing sense to wine body, it is more raw
At a part of new fragrance, the fragrance system of low alcohol applejack is enriched, organoleptic indicator is good, golden yellow color, clear, wine
The mellow coordination of body, mouthfeel are soft.
Detailed description of the invention
Fig. 1 is influence of the different fermentations mode to applejack alcoholic strength；
Fig. 2 is influence of the different fermentations temperature to applejack；
Fig. 3 is influence of the different vaccination ratio to applejack alcoholic strength；
Fig. 4 is influence of the different vaccination amount to applejack alcoholic strength；
Fig. 5 is influence of the different liquid amounts to applejack alcoholic strength；
Fig. 6 is the influence of fermentation temperature and inoculum concentration to applejack alcoholic strength；
Fig. 7 is the influence of inoculative proportion and inoculum concentration to applejack alcoholic strength；
Fig. 8 is influence of the vapo(u)rizing temperature to applejack ethanol content；
Fig. 9 is influence of the distillation time to applejack ethanol content；
Figure 10 is applejack (b) and low alcohol bubbling applejack (c) GC-MS total ion current after low alcohol applejack (a), dealcoholysis
Figure；
The present invention is illustrated below in conjunction with specification drawings and specific embodiments.
Specific embodiment
The present invention by saccharomyces cerevisiae and is addicted to drink using fresh apple juice as raw material and Candida while accessing fresh squeezing apple clear juice
The middle fermentation for carrying out apple former wine, then cryogenic vacuum distillation removing ethyl alcohol is carried out to obtained apple former wine and obtains low alcohol apple
Wine；Using the low alcohol applejack after dealcoholysis as base liquor, inoculation saccharomyces cerevisiae carries out secondary fermentation after adjusting sugar to obtain the final product.It measures and divides
Physical and chemical index, fragrance component before and after analysis distillation dealcoholysis and after secondary fermentation.
Being addicted to drink Candida can be using the ethyl alcohol that saccharomyces cerevisiae generates without sugar fermentation, using saccharomyces cerevisiae and false silk of being addicted to drink
The mode of yeast for fermentation can reduce the alcoholic strength of applejack, while the fragrance system of abundant applejack；Under vacuum conditions,
Ethyl alcohol boiling point reduces, therefore can remove the ethyl alcohol in applejack under lower vapo(u)rizing temperature, and what cider fermentation generated
Fermented material all retains, and obtains low alcohol applejack；Secondary fermentation in bottle is carried out using the low alcohol applejack that dealcoholysis obtains to generate
The carbon dioxide gas of new fruit wine fragrance and carbon dioxide gas, generation dissolves in wine, and wine is made to have tasty and refreshing sense.
Saccharomyces cerevisiae used in the present invention selects WLS21 saccharomyces cerevisiae strain (document " the dedicated ferment of Wang Lin pine applejack
Female building and fermentation dynamics research [D] Xibei Univ. of Agricultural & Forest Science & Technology, 21# bacterial strain disclosed in 2007 "), SJ03 is addicted to drink vacation
Silk barms (document " and Song Jingjing, Yuan Yahong, Liu Bin, Wang Huxuan, Cen Tao .2015. ethyl alcohol using bacterial strain separation identification and
Application modern food science and technology in low alcohol applejack, (6):SJ03 bacterial strain disclosed in 254-258. ") and WF41 wine brewing ferment
(" northwest research [D] the agriculture and forestry science and technology that Wang Li prestige protoplast fusion constructs flavored type kiwifruit wine yeast is big for document for female strain
It learns, 41# bacterial strain disclosed in 2004. ").
Research process of the invention is as follows：Below unless otherwise specified, percentage composition is mass percentage.
One, the preparation of apple clear juice
Select abundant mature, non-rot red fuji apple, cleaning, stoning and stripping and slicing be placed on containing 0.05%Vc and
Color protection 20min is stood in 1.5% citric acid, takes out juicing.Residue is discarded, the weighting sulfurous acid of 70mg/L is added into fruit juice
Potassium stands 15min, pectase is added according to the ratio of 450mg/L in fruit juice, 45 DEG C of water-bath 2h, it is clear that vacuum filtration obtains apple
Juice saves backup at -20 DEG C.
Two, prepared by yeast starter liquid
By two saccharomycetes being preserved in glycerol tube (WLS21 saccharomyces cerevisiae strain be addicted to drink with SJ03 candida strain)
In the YPD culture medium (1% yeast extract, 2% glucose, 2% peptone) activation (28 DEG C of temperature, shaking speed 120r/
Min, the time is for 24 hours) after two generations, it is inoculated in cider (105 DEG C of sterilizing 15min), 28 DEG C of temperature, is shaken respectively with 5% inoculum concentration
Bed revolving speed 120r/min, WLS21 S. cervisiae cultivates 18h, and SJ03 is addicted to drink candidiasis culture for 24 hours, respectively obtains growth
Vigorous WLS21 S. cervisiae seed liquor and SJ03 are addicted to drink candidiasis seed liquid.
Three, low alcohol applejack mixed fungus fermentation process optimization
(1) selection of fermentation method
The WLS21 S. cervisiae seed liquor and SJ03 that prepare are addicted to drink candidiasis seed liquid with 8% inoculum concentration
In cider after being inoculated in sterilizing respectively, 21 DEG C of standing for fermentation measure the soluble solid content of fermentation liquid, until can daily
Stop fermentation when dissolubility solid content is no longer substantially reduced, 6000r/min is centrifuged 10min and removes thallus, obtained low alcohol apple
Fruit wine is measured for alcoholic strength.
Fermentation method：
1. WLS21 and SJ03 single bacterium are fermented；
2. WLS21 and SJ03 are inoculated with mixed fermentation simultaneously；
3. being first inoculated with WLS21, SJ03 mixed fermentation is inoculated with after 2d, 3d；
4. being first inoculated with SJ03, WLS21 mixed fermentation is inoculated with after 2d, 3d.
It chooses the minimum fermentation method of alcoholic strength and carries out follow-up test.As a result as shown in Figure 1.
By Fig. 1, it can be concluded that, when being addicted to drink the fermentation of Candida SJ03 single bacterium, the alcoholic strength of applejack is 0.2%vol,
It is essentially close to 0, i.e., it is believed that SJ03 single bacterium hardly generates ethyl alcohol when fermenting.In the fermentation of saccharomyces cerevisiae WLS21 single bacterium, apple
The alcoholic strength of fruit wine is 6.1%vol, and the alcoholic strength of applejack, that is, use saccharomyces cerevisiae when being significantly higher than the two mixed fermentation
WLS21 and the fermentation method for Candida SJ03 mixed fermentation of being addicted to drink have obvious effect in terms of the alcoholic strength for reducing applejack.
In terms of strain inoculated sequence, saccharomyces cerevisiae WLS21 and the Candida SJ03 while when being inoculated with of being addicted to drink, the alcoholic strength of applejack is most
It is low；In addition, first be inoculated with SJ03 when, than being first inoculated with WLS21 in the case where applejack alcoholic strength it is low；In inoculation time interval side
Face, two plants of bacterium are separated by 2d or 3d inoculation, and the alcoholic strength of applejack is without significant difference.Therefore, it selects saccharomyces cerevisiae WLS21 and is addicted to drink
Candida SJ03 is inoculated with simultaneously, and the minimum applejack of alcoholic strength can be obtained, and follow-up test uses this vaccination ways.
(2) mixed fungus fermentation process conditions single factor experiment
1. influence of the fermentation temperature to alcoholic strength
Cider soluble solid content after sterilizing is 13 ° of Brix, pH 3.94, with 8% inoculum concentration, inoculative proportion
WLS21：SJ03=1：1, liquid amount 150mL, fermentation temperature are respectively 15 DEG C, 18 DEG C, 21 DEG C, 24 DEG C, 27 DEG C of standing for fermentation.Often
It weighs to fermentation liquid band bottle, and fermentation ends are thought when its mass change is lower than 0.2g/d, and 7000g centrifugation 10min goes degerming
Body measures applejack alcoholic strength.Parallel 2 times.As a result as shown in Figure 2.
By Fig. 2, it can be concluded that, with the raising of fermentation temperature, alcoholic strength shows raised trend after first reducing.In temperature
When degree is 24 DEG C, applejack alcoholic strength is minimum, is 5.6%vol；When temperature is 27 DEG C, applejack alcoholic strength highest is
6.6%vol.This may be since within the scope of 15 DEG C -24 DEG C, as the temperature rises, the Candida SJ03 that is addicted to drink is metabolized second
The vigor of alcohol gradually increases, and the efficiency of degradation ethyl alcohol improves, so that alcoholic strength is increased with temperature and reduced；And work as fermentation temperature
When reaching 27 DEG C, metabolic activity is suppressed, the decline of ethanol degradation ability, and the alcohol generated in fermentation process cannot be timely
Degradation, therefore alcoholic strength increases instead.To sum up, 18 DEG C, 21 DEG C and 24 DEG C these three levels is selected to carry out response surface optimization test.
2. influence of the inoculative proportion to alcoholic strength
Cider soluble solid content after sterilizing is 13 ° of Brix, pH 3.94, with 8% inoculum concentration, liquid amount
150mL, inoculative proportion SJ03：WLS21 is respectively 0.5:1,1:1,2:1,3:1,21 DEG C of standing for fermentation.Daily to fermentation liquid band bottle
Fermentation ends are thought in weighing when its mass change is lower than 0.2g/d, and 7000g is centrifuged 10min and removes thallus, measure applejack wine
Precision, parallel 2 times.As a result as shown in Figure 3.
By Fig. 3, it can be concluded that, with the reduction of fermentation ratio, that is, the inoculative proportion for the Candida SJ03 that is addicted to drink increases, apple
The alcoholic strength of fruit wine shows the trend being gradually reduced.This explanation increases Candida of being addicted to drink in the identical situation of inoculum concentration
The relative scale of SJ03, can effectively it is degradable fermented during generate ethyl alcohol, thus reach reduce applejack alcoholic strength work
With；And the relative scale for the Candida SJ03 that is addicted to drink is bigger, it is more significant to the drop alcohol effect of applejack.To sum up, WLS21 is selected：
SJ03=1：1,1：2 and 1：3 this 3 horizontal progress response surface optimization tests.
3. influence of the inoculum concentration to alcoholic strength
Cider soluble solid content after sterilizing is 13 ° of Brix, pH 3.94, with inoculative proportion SJ03：WLS21
=1：1, liquid amount 150mL, inoculum concentration are respectively 4%, 6%, 8%, 10% inoculation, 21 DEG C of standing for fermentation.Daily to fermentation liquid
Band bottle is weighed, and fermentation ends are thought when its mass change is lower than 0.2g/d, and 7000g is centrifuged 10min and removes thallus, measures apple
Wine alcoholic strength, parallel 2 times.As a result as shown in Figure 4.
By Fig. 4, it can be concluded that, when inoculum concentration is 4%, alcoholic strength is minimum, is 5.2%vol；When inoculum concentration is 10%, alcohol
Highest is spent, is 6.6%vol, and with the increase of inoculum concentration, the alcoholic strength of applejack is gradually risen, this may be to connect due to working as
One timing of kind ratio, with the increase of inoculum concentration, the biomass of saccharomyces cerevisiae WLS21 increases, although being addicted to drink Candida
The biomass of SJ03 is also increasing, but the vigor of WLS21 ratio SJ03 is strong, therefore WLS21 may generate centainly SJ03
Inhibiting effect, cause SJ03 that can not show enough drop alcohol abilities, though increase inoculum concentration, apple can not be effectively reduced
The alcoholic strength of wine.To sum up, these three levels of inoculum concentration selection 4%, 6% and 8% carry out response surface optimization test.
4. influence of the liquid amount to alcoholic strength
Cider soluble solid content after sterilizing is 13 ° of Brix, pH 3.94, with 8% inoculum concentration, inoculative proportion
WLS21：SJ03=1：1,250mL conical flask liquid amount is respectively 100,150 and 200mL, 21 DEG C of standing for fermentation.Daily to fermentation
Fermentation ends are thought in the weighing of liquid band bottle when its mass change is lower than 0.2g/d, and 7000g is centrifuged 10min and removes thallus, measure apple
Fruit wine alcoholic strength, parallel 2 times.As a result as shown in Figure 5.
By Fig. 5 it can be concluded that, when 250mL conical flask liquid amount is respectively 100,150 and 200mL, the alcoholic strength of applejack
Respectively 5.5%vol, 5.7%vol and 5.8%vol.Alcoholic strength peak and minimum difference are only 0.3%vol, illustrate this
Factor influences less applejack alcoholic strength, therefore directly selected liquid amount is 150mL.
(3) mixed fungus fermentation process conditions response surface optimization
1. being made wine according to the result of single factor experiment and Box-Behnken design principle using experimental design software optimization
Yeast and Non-Saccharomyces mixed fungus fermentation make the technique of low alcohol applejack, using alcoholic strength as response, to influence zymotechnique
Factor carry out 3 factor, 3 horizontal centre Combination Design, to result carry out response surface analysis, response surface design factor level code
Table is shown in Table 1.
1 factor level coding schedule of table
According to BBD response surface design principle, design point, and each factor are selected using three factor center combination designs
3 levels are designated as to optimize fermentation parameter.Influence for hydrolysis and fermentation technological parameter to low alcohol applejack quality, using more
First linear regression method is fitted test data, generates second-order response surface regression equation.Box-Behnken experimental design scheme and test
As a result such as table 2, regression model variance analysis table such as table 3.
2 BBD experimental design scheme of table and result
3 regression model variance analysis table of table
Note：*p<0.05 indicates significant difference, * * p<0.01 indicates extremely significant.
According to Design Expert V8.06 software the results of analysis of variance, tentatively establish low alcohol applejack alcoholic strength with
Regression equation between three fermentation temperature, inoculative proportion and inoculum concentration factors is as follows：
Y=4.68+0.19X1+0.038X2+0.075X3+0.15X1X2-0.48X1X3-0.57X2X3-0.47X1 2+
0.24X2 2-0.39X3 2
Wherein X1It is the encoded radio of fermentation temperature, X2It is the encoded radio of inoculative proportion, X3It is the encoded radio of inoculum concentration.By table 3
It is found that the quadratic regression model p value built less than 0.01, illustrates that the model is significant, and lose quasi- item p value and be greater than 0.05, loses quasi- item
It is not significant, illustrate that built secondary model fitting degree is higher, the error during testing is smaller.It can also be seen that in table
Determine coefficients R2Coefficient AdjR is determined with adjusting2Coefficient is respectively 0.9326 and 0.8460.The result shows that factor is to test result
It has a significant impact, the relationship between factor variation and test result is true and reliable.Meanwhile the low coefficient of variation (C.V.%=
4.88) show model precision with higher and reliability.By table 3 it can also be seen that in the regression model, AC, BC,
A2、C2For extremely significant factor (p<0.01), A is significant factors (p<0.05), B, C, AB, B2For not significant factor.
2. response surface analysis
3D surface chart in response surface analysis and contour map being capable of reciprocations between intuitive reaction factor.According to
DesignExpert V8.06 obtains the regression equation of low alcohol cider fermentation technique, factor fermentation temperature and factor inoculum concentration it
Between reciprocation it is as shown in Figure 6.
By Fig. 6, it can be concluded that, the timing of fermentation temperature one, alcoholic strength is with after being gradually increased and show and first increase of inoculum concentration
Reduced trend；When the timing of inoculum concentration one, applejack alcoholic strength is with after being gradually increased and also show and first increase of fermentation temperature
Reduced trend.
The reciprocation of inoculative proportion and inoculum concentration is as shown in Figure 7.By Fig. 7 it can be concluded that, when the timing of inoculative proportion one, apple
The alcoholic strength of fruit wine shows the trend of first increases and then decreases with being gradually increased for inoculum concentration, this is because suitable inoculation
Amount so that the Candida SJ03 that is addicted to drink is capable of forming certain biomass advantage without being inhibited by saccharomyces cerevisiae WLS21, thus
Its ethyl alcohol of degrading is played, reduces the effect of applejack alcoholic strength, inoculum concentration is too high or too low, cannot all reach this effect.
3. verifying
The low alcohol applejack Optimal technique process that software Design Expert V8.06 is provided is 18.56 DEG C of fermentation temperature,
Inoculative proportion WLS21：SJ03=1:3, inoculum concentration 4%.Under this optimal conditions, prediction can be obtained alcoholic strength and obtain for 3.2%vol
Applejack.For the ease of industrial operation, verification test is at 18 DEG C of fermentation temperature, inoculative proportion WLS21：SJ03=1:3, inoculation
Implement under conditions of amount 4%, obtain the applejack that alcoholic strength is 3.3%vol, more coincide with model predication value, compared to control
The applejack alcoholic strength of group alcoholic strength 6.1%vol, process conditions brewing reduce 2.8%vol, the low alcohol apple made
The analysis of fruit wine physical and chemical index is as shown in table 4.
The low alcohol applejack physical and chemical index of table 4
As shown in Table 4, low alcohol applejack alcoholic strength is 3.3%vol, meets national regulations.Therefore, it is obtained using the method
Low alcohol apple wine brewage technology parameter is feasible, there is certain values in practical applications.
Four, cryogenic vacuum distillation technique determines
(1) selection of cryogenic vacuum dealcoholysis temperature
Using cryogenic vacuum distillation equipment, it will heat up to 20 DEG C of low alcohol applejack graduated cylinder and measure 20mL, respectively at 20
DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, 45 DEG C, cryogenic vacuum distillation dealcoholysis test is carried out at 50 DEG C, control distilling flask revolving speed is protected
It holds constant, starts timing when whole system vacuum degree reaches 0.08MPa, distillation stops timing after carrying out 5min, sample is removed,
20 DEG C of water-bath 30min supply volume to 20mL with ultrapure water.Every group of test is repeated twice.
Obtained sample uses potassium dichromate oxidation colorimetric method after producing distillate, absorbance is measured at 600nm, by standard
Curve obtains the alcohol content of sample.As a result as shown in Figure 8.
By Fig. 8, it can be concluded that, with the raising of vapo(u)rizing temperature, the ethanol content in sample is in the trend being gradually reduced.It is former
Ethanol content is 18.17mg/mL in wine, and under vacuum conditions, 25 DEG C of whens can remove part ethyl alcohol；When temperature reaches 35 DEG C
When, ethanol content is only 5.17mg/mL in sample, and ethyl alcohol removal efficiency is 71.5%.Even if hereafter temperature improves again, ethyl alcohol removing
The raising of rate is not also significant, it is contemplated that while removing ethyl alcohol, temperature is higher, and the loss of fragrance component is also got over for vacuum distillation
Greatly, therefore, select 35 DEG C for vapo(u)rizing temperature.
(2) selection of cryogenic vacuum dealcoholysis time
Using cryogenic vacuum distillation equipment, 20 DEG C of low alcohol applejack is measured into 20mL with graduated cylinder and carries out cryogenic vacuum distillation
Dealcoholysis test, distilling flask revolving speed are kept constant, and bath temperature is 35 DEG C, are opened when whole system vacuum degree reaches 0.08MPa
Beginning timing, respectively handle 2min, 3min, 4min, 5min, 6min, after remove sample, 20 DEG C of water-bath 30min use ultrapure water
Volume is supplied to 20mL.Every group of test is repeated 2 times.Ethanol content measuring method is same as above.As a result as shown in Figure 9.
By Fig. 9, it can be concluded that, the initial ethanol content of sample is 14.66mg/mL, with the extension of distillation time, in sample
Ethanol content gradually decreases, and reduces rate and reduce at any time.When distillation time reaches 5min, ethanol content is in sample
3.54mg/mL, ethyl alcohol removal efficiency reaches 75.9%, even if hereafter distillation time extends again, ethyl alcohol removal efficiency is not also significantly improved,
In view of under vacuum condition, distillation time is longer, product fragrance loss is bigger, therefore, selects distillation time for 5min.
Five, reverse osmosis dealcoholysis technique determines
It is tested using domestic nanofiltration membrane (Jinan Rong De Water Management Equipment Ltd.).
(1) determination of reverse osmosis time
Film is measured in reverse osmosis process from starting to work into the 1h time, the membrane flux every 10min changes, and determines film
The time point tended towards stability.
(2) determination of enrichment factor
During carrying out dealcoholysis to former wine using film, film is measured in enrichment factor (remaining reduction of feed volume/raw material liquid measure)
Extract loss rate when being 0.5,0.6,0.7,0.8,0.9 determines that film water supplement is to original volume in reverse osmosis process respectively
Technique.Extract loss rate is extract content in unit volume permeate.
(3) determination of operating pressure
Measure ethyl alcohol flux and extract flux and ethyl alcohol of the film when pressure is 0.6,0.7,0.8,0.9,1.0MPa
Transmitance and extract transmitance determine the Optimum Operation pressure of film.
By the technical study to film, show that the best reverse osmosis process of the film is：Reverse osmosis time 40min, concentration system
Number is 0.7, film pressure 1.6MPa.
Six, the determination of dealcoholysis mode
The reason of the dealcoholysis applejack obtained under the conditions of preferable to above-mentioned cryogenic vacuum distillation technique and reverse osmosis dealcoholysis technique
Change index and Aromatic Matter Contents are measured analysis, as shown in table 5 and table 6.
5 dealcoholysis applejack physical and chemical index of table
As shown in Table 5, cryogenic vacuum distillation technique generates big shadow to the soluble solid, total reducing sugar and total acid of applejack
It rings less, and reverse osmosis dealcoholysis technique is then affected to it；Influence of two kinds of techniques to applejack alcoholic strength is unobvious.It is comprehensive
It closes analysis and show that the low alcohol applejack wine body that cryogenic vacuum distillation technique obtains is full, mellow coordination.
6 sample Aromatic Matter Contents (mg/L) of table
It can be obtained by table 6, the applejack after applejack and reverse osmosis dealcoholysis after cryogenic vacuum distills is fragrant compared with former wine
Gas substance has certain loss.54 kinds of aroma substances, total content 7.092mg/L are detected in former wine sample altogether.Its Higher Alcohols
16 kinds of substance, total content 1.997mg/L；1 kind of phenolic substances, total content 0.013mg/L；2 kinds of aldehyde material, total content
For 0.119mg/L；9 kinds of acid, total content 1.327mg/L；26 kinds of Ester, total content 3.636mg/L.Through low
27 kinds of aroma substance, total content 3.041mg/L are detected altogether in applejack sample after temperature vacuum distillation, than sample before distilling
27 kinds are reduced, total content reduces 3.456mg/L, reduces 48.7%.After distillation in sample, 8 kinds of higher alcohols substance, total content
For 0.405mg/L；1 kind of phenolic substances, total content 0.045mg/L；7 kinds of acid, total content 0.717mg/L；Esters
11 kinds of substance, total content 1.874mg/L.Applejack after reverse osmosis dealcoholysis detects 13 kinds of aroma substance altogether, wherein alcohols 3
Kind, total content 0.172mg/L；1 kind of phenols, total content 0.017mg/L；3 kinds of acids, total content 0.433mg/L；Esters
6 kinds, total content 1.581mg/L.
In conclusion carrying out dealcoholysis to applejack using cryogenic vacuum distillation, obtained applejack is in terms of fragrance and instead
It permeates obtained applejack to compare, substance classes and content are all higher.
This technology determines that cryogenic vacuum distillation technique is got well than reverse osmosis dealcoholysis technique, obtained applejack wine body is full,
Mellow coordination and fragrance loss are less.
Seven, prepared by secondary fermentation seed liquor
The optimal bacterial strain of secondary fermentation that screening obtains is activated twice in YPDL culture medium, each inoculum concentration 5%, 28 DEG C
Lower 120r/min culture is for 24 hours；Obtained bacterium solution accesses the cider after sterilizing with 5% inoculum concentration, and 120r/min is cultivated at 28 DEG C
24h；Obtained bacterium solution is accessed in the dealcoholysis applejack distilled through cryogenic vacuum after sterilizing with 8% inoculum concentration, at 28 DEG C
120r/min cultivates 36h, takes out spare.
Eight, secondary fermentation condition optimizing
Saccharomyces cerevisiae WF41 is selected, selection saleratus additive amount (g/L), inoculum concentration (%), fermentation time (d) are to investigate
Factor is tested using software Minitab16.0 design 3 factor, 3 horizontal quadrature, is selected most using sensory evaluation scores value as inspection target
Excellent process conditions, as shown in table 7.
7 orthogonal test factor level table of table
Select 250mL brown pressure resistance beer bottle tested, per it is bottled enter 200mL 4.2.1 in dealcoholysis applejack, be
Prevent living contaminants, add 90mg/L potassium metabisulfite, add sugar to 25g/L, then according to testing program in table 5 into
Row test is left to ferment at 15 DEG C after gland sealing.Corkage sampling after fermentation carries out subjective appreciation.
It is reported according to trial test result and pertinent literature, selects bacterial strain WF41, chosen saleratus additive amount (g/L), connect
Kind amount (%), fermentation time (d) use software Minitab16.0 using the sensory evaluation scores value of each processing as index for investigation factor
Three factors-levels orthogonal test is designed, and carries out range analysis.Test result is as shown in table 8.
8 secondary fermentation orthogonal test table of table
It is obtained by range analysis, the factor for influencing low alcohol bubbling applejack sensory evaluation scores descending is followed successively by B>A>C,
That is saleratus additive amount>Inoculum concentration>Fermentation time, so finally determine inoculum concentration 6%, saleratus additive amount 1.0g/L,
Fermentation time 18d.
Verification test is carried out under obtained secondary fermentation Optimal technique process, obtained low alcohol bubbling applejack sense organ is commented
Score value is 80.4, and the clarification of wine body, aroma of pure, have fizz should have mouthfeel；Alcoholic strength is 1.8%vol, CO2Pressure (20
DEG C) it is 0.45MPa.
Applejack physical and chemical index is as shown in table 9 after secondary fermentation.
Physical and chemical index before and after 9 applejack secondary fermentation of table
Nine, fragrance component measures
1. fragrance component is enriched with：The enrichment of fragrance component is carried out using Headspace solid phase microextractiom (HS-SPME).It pipettes
1.5g sodium chloride and a certain amount of inner mark solution is added in 15mL sample bottle in 5mL applejack sample, on magnetic stirring apparatus,
45 DEG C of balance 10min, then sample bottle, headspace absorption 35min are inserted into 100 μm of PDMS extracting heads that aging process is crossed.
2. fragrance component GC-MS is detected：Will adsorb fragrance component PDMS fiber head insertion gas chromatographic sample introduction mouth, 250
5min is parsed at DEG C.The chromatographic column used is DB-WAX fused-silica capillary column (30m*0.25mm*0.25 μm).Shunting mode
Not shunt, chromatographic column temperature program is 40 DEG C of initial temperature, keeps 5min, rises to 120 DEG C with the heating rate of 3 DEG C/min,
230 DEG C are risen to the heating rate of 8 DEG C/min again, keeps 10min, carrier gas He, flow velocity 0.8mL/min.Mass Spectrometry Conditions：EI
Ionization source, electron energy 70eV, ion source temperature are 200 DEG C, scanning range 33-450AMU, and filament flow is 0.25mA,
Detector voltage is 350V.
The fragrance of applejack and low alcohol bubbling applejack measures total ion current figure such as Figure 10 institute after low alcohol applejack, dealcoholysis
Showing, a figure in Figure 10 is low alcohol applejack, and b is applejack after dealcoholysis, and c is the low alcohol bubbling applejack that the present invention is prepared,
Aroma substance and content are as shown in table 10.
10 sample Aromatic Matter Contents (mg/L) of table
By the analysis to applejack, low alcohol bubbling aroma of cider substance after low alcohol applejack, dealcoholysis it is found that secondary hair
Aroma substance total amount after ferment significantly improves, and is mainly reflected in the raising of higher alcohol and Ester content.Illustrate secondary fermentation
The Aromatic Matter Contents that can effectively improve applejack after dealcoholysis, fragrance loss caused by making up because of distillation increase low alcohol bubbling
The aroma substance of applejack.
Embodiment 1：
(1) preparation of apple clear juice
Abundant mature, non-rot red fuji apple is selected, clear water cleaning, stoning and stripping and slicing are placed on containing 0.05%
Color protection 20min is stood in Vc and 1.5% citric acid, takes out juicing.Residue is discarded, the weighting sulfurous of 70mg/L is added into fruit juice
Sour potassium stands 15min, pectase, 45 DEG C of water-bath 2h is added according to the ratio of 450mg/L in fruit juice, vacuum filtration obtains apple
Juice saves backup at -20 DEG C.
(2) prepared by yeast starter liquid
By two saccharomycetes being preserved in glycerol tube respectively at YPD culture medium (1% yeast extract, 2% glucose, 2%
Peptone) in activation (28 DEG C of temperature, shaking speed 120r/min, the time is for 24 hours) after two generations, be inoculated in respectively with 5% inoculum concentration
In cider (105 DEG C of sterilizing 15min), 28 DEG C of temperature, shaking speed 120r/min, WLS21 cultivate 18h, and SJ03 is cultivated for 24 hours,
Obtain eugonic seed liquor.
(3) mixed fungus fermentation
105 DEG C of sterilizing 15min of cider, in terms of cider volume, with 4% inoculum concentration, inoculative proportion WLS21：SJ03=
1:3 inoculation yeasts, 18 DEG C of fermentation 7d, 5000rpm centrifugation 10min remove thallus, obtain low alcohol applejack.
(4) cryogenic vacuum dealcoholysis technique
The low alcohol applejack that (3) are obtained carries out cryogenic vacuum using Rotary Evaporators and distills dealcoholysis, vacuum degree 0.08
~0.09MPa, vapo(u)rizing temperature are 35 DEG C, distillation time 5min.Original volume is complemented to pure water after the completion of distillation.
(5) base liquor is deployed
The low alcohol applejack that (4) are obtained adds sugar to 25g/L, adds potassium metabisulfite 90mg/L, adds bicarbonate
Potassium 1g/L.
(6) prepared by yeast starter liquid
Saccharomyces cerevisiae WF41 is activated twice in YPDL culture medium by glycerol tube, each inoculum concentration 5%, at 28 DEG C
120r/min is cultivated for 24 hours；Obtained bacterium solution accesses the cider after sterilizing with 5% inoculum concentration, and 120r/min is cultivated at 28 DEG C
24h；Obtained bacterium solution is in the dealcoholysis applejack (5) after the access sterilizing of 8% inoculum concentration, 120r/min cultivates 36h at 28 DEG C, takes
It is spare out.
(7) secondary fermentation
Deployed base liquor is packed into 250mL pressure resistance bottle, per bottled 200mL.Saccharomyces cerevisiae WF41 is inoculated with 6% inoculum concentration
Seed liquor, after gland sealing, 15 DEG C of ferment at constant temperature 18d.Body is rotated by a certain angle daily during fermentation until close to vertical
Handstand state after fermentation freezes bottleneck in liquid nitrogen, opens scum, supplements a certain amount of low alcohol applejack, and gland is close
Envelope, obtains finished product.
The CO for the low alcohol bubbling applejack that the embodiment obtains2Pressure is 0.45MPa (20 DEG C), alcoholic strength 1.8%
vol.37 kinds of aroma substance, total content 6.113mg/L are detected altogether, than improving 101.1% before fermentation.Wherein, alcohols object
Quality inspection goes out 9 kinds, total content 1.395mg/L, improves 244.4% than total content 0.405mg/L before fermenting；Phenolic substances is examined altogether
4 kinds out, 3 kinds, total content 0.035mg/L are increased compared with first a kind of fermentation, compared with the preceding total content 0.045mg/L that ferments
It remains basically stable；2 kinds of aldehyde material, total content 0.006mg/L；Two kinds of acid, total content 0.109mg/L, with fermentation
Preceding 0.717mg/L, which is compared, reduces 84.8%；Ester detects 20 kinds, total content 4.568mg/L altogether, before fermentation
1.874mg/L compared to improving 143.8%.Low alcohol bubbling applejack organoleptic quality with higher：In golden yellow, clarification is saturating
Bright, fragrance is coordinated, is strong, without peculiar smell, the typicalness with applejack.
Claims (9)
1. a kind of preparation method of low alcohol bubbling applejack, which is characterized in that the preparation method includes with zymophyte and apple
Fruit juice is the fermentation preparation that raw material carries out apple former wine, then the part ethyl alcohol in apple former wine is removed to obtain low alcohol applejack,
Saccharomyces cerevisiae is finally recycled to carry out the secondary fermentation of low alcohol applejack；
The zymophyte is S. cervisiae and candidiasis of being addicted to drink.
2. the preparation method of low alcohol applejack as described in claim 1, which is characterized in that in terms of the volume of cider, fermentation
The inoculum concentration of bacterium seed liquor is 4%；Inoculative proportion is S. cervisiae：It is addicted to drink candidiasis=1:3；
The preparation of the fermentation bacteria seed solution includes：Zymophyte is inoculated in 28 DEG C, 120rpm shaking table culture in YPD culture medium
It activates twice, the zymophyte after activation is inoculated in cider with 5% inoculum concentration, 28 DEG C, 120rpm shaking table culture for 24 hours；
The shaking table culture time of S. cervisiae is 18h, and the shaking table culture time for candidiasis of being addicted to drink is for 24 hours.
3. the preparation method of low alcohol applejack as claimed in claim 1 or 2, which is characterized in that the hair of the apple former wine
In ferment preparation：Fermentation temperature is 18 DEG C, fermentation time 7d, and the soluble solid content of cider is 12~14 ° of Brix, apple
The pH of fruit juice is 3.9 or so.
4. the preparation method of low alcohol applejack as claimed in claim 1 or 2, which is characterized in that in the apple former wine
Ethyl alcohol removing uses cryogenic vacuum distillating method.
5. the preparation method of low alcohol applejack as claimed in claim 4, which is characterized in that the cryogenic vacuum distillating method
Vacuum degree be 0.08~0.09MPa, vapo(u)rizing temperature be 35 DEG C, distillation time 5min.
6. the preparation method of low alcohol bubbling applejack as claimed in claim 1 or 2, which is characterized in that also to low alcohol applejack
Moisturizing is carried out, sugar is mended and carries out secondary fermentation again after supplementing potassium metabisulfite, the magnitude of recruitment of water is low alcohol apple after removing ethyl alcohol
The volume reduction of fruit wine, sugared magnitude of recruitment are that the sugared content in the low alcohol applejack after guaranteeing removing ethyl alcohol is 25~30g/L,
The magnitude of recruitment of potassium metabisulfite is 90mg/L.
7. the preparation method of low alcohol bubbling applejack as claimed in claim 1 or 2, which is characterized in that the secondary fermentation
Temperature be 15 DEG C, saleratus additive amount be 1g/L, fermentation time 18d.
8. the preparation method of low alcohol bubbling applejack as claimed in claim 1 or 2, which is characterized in that remove the low of ethyl alcohol
The stereometer of alcohol applejack, the seed liquor inoculum concentration of the S. cervisiae in secondary fermentation are 6%；
The preparation method of the seed liquor of saccharomyces cerevisiae in secondary fermentation includes：Saccharomyces cerevisiae is living by the effective YPD culture medium of glycerol
After changing for two generations, with the cider after the access sterilizing of 5% inoculum concentration, 120rpm is cultivated for 24 hours at 28 DEG C；Obtained bacterium solution connects with 8%
In dealcoholysis applejack after kind amount access sterilizing, at 28 DEG C after 120rpm culture 36h to obtain the final product.
9. a kind of low alcohol bubbling applejack, which is characterized in that the low alcohol bubbling applejack uses any power of claim 1-8
Benefit requires the preparation method to be prepared.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112961746A (en) * | 2021-03-09 | 2021-06-15 | 四川大学 | Brewing method for improving flavor characteristics of cider |
| CN113025455A (en) * | 2021-03-12 | 2021-06-25 | 四川大学 | Two-stage fermentation method of cider |
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| CN1079988A (en) * | 1993-05-10 | 1993-12-29 | 大连轻工业学院 | Non-alcoholic fruit wine and method for making |
| CN103060150A (en) * | 2013-01-06 | 2013-04-24 | 西北农林科技大学 | Production method of low-alcohol cider |
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| CN1079988A (en) * | 1993-05-10 | 1993-12-29 | 大连轻工业学院 | Non-alcoholic fruit wine and method for making |
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| CN112961746A (en) * | 2021-03-09 | 2021-06-15 | 四川大学 | Brewing method for improving flavor characteristics of cider |
| CN113025455A (en) * | 2021-03-12 | 2021-06-25 | 四川大学 | Two-stage fermentation method of cider |
| CN113025455B (en) * | 2021-03-12 | 2023-08-08 | 四川大学 | Two-stage fermentation method of cider |
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