CN108315270A - Table Grape Spirit saccharomyces cerevisiae and its application - Google Patents
Table Grape Spirit saccharomyces cerevisiae and its application Download PDFInfo
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- CN108315270A CN108315270A CN201810007943.5A CN201810007943A CN108315270A CN 108315270 A CN108315270 A CN 108315270A CN 201810007943 A CN201810007943 A CN 201810007943A CN 108315270 A CN108315270 A CN 108315270A
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- 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
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
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- 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
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C11/00—Fermentation processes for beer
- C12C11/02—Pitching yeast
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- 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
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
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- 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
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- 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
- C12G2200/00—Special features
- C12G2200/05—Use of particular microorganisms in the preparation of wine
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Abstract
The present invention relates to a kind of Table Grape Spirit saccharomyces cerevisiae (Saccharomyces cerevisiae) and its applications, belong to field of microorganism engineering.The Table Grape Spirit saccharomyces cerevisiae isolated is named as Miao 2.1609, is suitable for the brewing of wine spirit.Table Grape Spirit saccharomyces cerevisiae Miao 2.1609 carries out preservation by China Microbiological preservation administration committee's common micro-organisms center (abbreviation CGMCC), address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the deposit date is on October 11st, 2017, deposit number was CGMCC NO.14779.
Description
Technical field
The present invention relates to a kind of Wine-making yeast and its applications, belong to field of microorganism engineering.
Background technology
In brewing industry, saccharomycete sucks the monosaccharide such as glucose, fructose, mannose into the cell, in the condition of anaerobic
Under, by the effect of interior enzyme, monosaccharide is decomposed into carbon dioxide and alcohol, this effect is fermented.But the ferment of variety classes, source
Mother is different to the fermentability of same liquor-making raw material, and content, the mouthfeel of the alcohol of fermentation also have prodigious difference.And because making
The difference of liquor raw material is more easy to influence the yield and quality of product.It is well known that the raw material grape of production red wine exists with Table Grape
There is larger difference on ingredient, such as produces Table Grape Spirit using the saccharomycete of production red wine, it will certainly be great
The yield and quality of product is influenced and reduced, therefore, is found a kind of specifically for being that brewing materials are used to produce using Table Grape
The yeast strain of Spirit, it appears particularly important.
The production strain of present making fruit wine, general using the active dry yeast for brewing grape red wine, some also adds
Enter common grain wine koji fermentation, the barms for production lack specificity, cause uva fragrance that grape wine should have,
Flavor is not pure.Disinfection, the filling mode of production after directly filtering are generally taken in the processing of zymotic fluid.Zymotic fluid is merely through filtering
With the processing of disinfection, complicated component (active ingredient and metabolic waste are simultaneously deposited) and alcohol content in filtrate are low, and (10% is left
It is right), the safety of product and shelf-life are limited apparent.
Invention content
In view of the above technical problems, the object of the present invention is to provide a kind of bacterium for distilling liquor brewing suitable for Table Grape
Strain.The bacterial strain can also be used for the brewing of grape red wine and other fruit wine or beer.
To achieve the goals above, the present invention provides the following technical solutions：
A kind of wine spirit saccharomyces cerevisiae (Saccharomyces cerevisiae) Miao 2.1609, in China Microbiological
The common micro-organisms center preservation of culture presevation administration committee, preservation registration number are CGMCC No.14779.
The basic physiological feature of wine spirit saccharomyces cerevisiae Miao 2.1609 of the present invention：It observes under the microscope, cell ovum
It is round；Bacterium colony is rounded, milky, neat in edge, protrusion, surface wettability, sticky；Unicellular is 8-12 × 11-17 microns；It is suitable
Suitable cultivation temperature is：29~31 DEG C, suitable culture pH value is：5.5~6.5.
The present invention also provides a kind of saccharomyces cerevisiae preparation, the active constituent of the preparation is aforementioned saccharomyces cerevisiae
(Saccharomyces cerevisiae) Miao 2.1609.
The present invention also provides a kind of above-mentioned wine spirit saccharomyces cerevisiae Miao 2.1609 answering in wine production
With.
The present invention also provides a kind of above-mentioned wine spirit saccharomyces cerevisiae Miaos 2.1609 in grape red wine or other fruits
Application in the brewing of wine or beer.
The beneficial effects of the invention are as follows：Table Grape Spirit saccharomyces cerevisiae Miao 2.1609 provided solves fresh food Portugal
Because using Commercial active dry ferment of the common fruit wine yeast bacterium as brewed grape wine in grape distillation wine brewing process, lead to wine body not
Coordinate, the grape flavor fragrance of wine is not pure and the low problem of yield rate.Use the fresh food Portugal of the saccharomyces cerevisiae production of the present invention
Grape flavor in grape distilled spirit is strong, quiet and tastefully laid out, and wine body is mellow, hangs that cup is apparent, and not top after people's drink, there are few bad physiology is anti-
It answers, amino acid, vitamin containing there are many, there is higher nutritive value, long shelf-life are drunk in right amount beneficial to health.
Description of the drawings
Fig. 1 is 2.1609 tablet of saccharomyces cerevisiae Miao of the present invention separation figure.
Fig. 2 is 2.1609 test tube transposing figure of saccharomyces cerevisiae Miao of the present invention.
Fig. 3 is the lower 640 times of observation charts of 2.1609 microscope of saccharomyces cerevisiae Miao of the present invention.
Specific implementation mode
Separation, the purification of 1 grape wild yeasts bacterium of embodiment
Multiplying culture 48 hours, acquisition have the grape of more individual under the conditions of 29~31 DEG C after grape is crushed
Wild yeasts Multiplying culture liquid.By obtained Multiplying culture liquid by 10 times, 20 times, 40 times, 80 times, 160 times, 320 times, 640 times
Seven concentration gradients are diluted with sterile distilled water, are then applied on tablet Selective agar medium that (tablet Selective agar medium is matched respectively
Side：4~5 ° of B é fresh grape pulps 24.5%, 4 ° of filtrate, 24.5%, 4 ° of B é brewer's worts, 24.5%, 4 ° of B é murphy juices of B é soybean juices
24.5%, agar 2%), the culture solution of each diluted concentration gradient makees 4 tablets, is isolated using " plate streak " tens of
The single cell colonies of a grape wild yeasts bacterium.According to the colonial morphology of the single cell colonies of wild yeasts bacterium, unicellular individual
Form, growing way etc., selecting five healthy and strong pure bacterial strains to distinguish transposing to 50ml test tubes becomes first class inoculum, obtains 6 plants of wild ferment
Female pure bacterial strain, is respectively designated as the 1st, 2,3,4,5, No. 6 grape wild yeasts.It is tablet Selective agar medium as shown in Fig. 1,2
Upper to screen obtained grape wild yeasts bacterium, bacterium colony is rounded, milky, neat in edge, protrusion.By the wild ferment of obtained grape
Female bacterium is placed under 640 power microscopes and observes, as shown in Fig. 3, it is seen that cell is oval.
The performance measurement small-scale test of the 2 pure bacterial strain of grape wild yeasts bacterium of embodiment
1 test objective
Verify six isolated grape wild yeasts bacteria strain fermentabilities and production wine effect.
2 test materials and method
New 18 kilograms of fresh grape (kind sunlight rose) obtains 14.0 kilograms of grape slurries after removal base of fruit, stalk and mashing,
It is divided into 7 test samples, each test sample dosage is 2.0 kilograms.Wherein 6 test samples are for 6 plants of Portugals in embodiment 1
Grape wild yeasts are as experimental group, and in addition 1 sample is lived for commercial goods Italy " English receive Supreme Being Si " (Enartis) grape wine
Property dry ferment is as a contrast.10.0 grams of fresh wheat bran yeast, inoculum concentration 0.5% are inoculated with into each grape slurries test sample
(the fresh wheat bran yeast being added in experimental group is respectively the 1st in embodiment 1,2,3,4,5, No. 6 grape wild yeasts are prepared into
It arrives, the fresh wheat bran yeast in control group is that " English receive Supreme Being Si " wine active dry yeast is prepared).It mixes well and is placed on
25 DEG C ferment 15 days.
Fresh wheat bran yeast preparation method：1) into embodiment 1, the 50ml equipped with grape wild yeasts bacterium first class inoculum is tried
30ml sterile waters are added in pipe, liquid is sucked out for use after jiggling.It 2) will be after 100 grams of grape slurries, 100 grams of wheat bran mixings
115 DEG C sterilize 30 minutes, and taking-up is cooled to room temperature, and the grape saccharomycete liquid 5ml that step 1) obtains, 28~30 DEG C of cultures are added
60~72h, (therebetween in 24 hours, 8 hours each shaking flasks 1 time) obtain fresh wheat bran yeast.Control " English receive Supreme Being Si " grape wine
The fresh wheat bran yeast of yeast is also made in this way.
3 test results
It will be distilled after the fermentation 25 days of 25 DEG C of above-mentioned 7 test samples, every plant of grape wild yeasts have stronger production
Wine ability.Alcoholic strength is measured on the basis of distilling out 200 milliliters of equivalent wine liquids respectively, measures the 1st, 2,3,4,5, No. 6 grape
The alcoholic strength of white wine made from wild yeasts and " English receive Supreme Being Si " wine yeast is followed successively by： 48、45、46、49、47、51、50
(control), it is seen that No. 6 grape wild yeasts bacterial strain is closest with the fermentability for compareing " English receive Supreme Being Si " wine yeast.
It organizes related personnel to carry out " blind to comment " organoleptic indicators such as fragrance, the flavor of wine liquid simultaneously, unanimously thinks No. 6 wild strain
It is most characteristic and be substantially better than control strain " English receive Supreme Being Si ".
The performance measurement medium-sized test of the 3. pure bacterial strain of grape wild yeasts bacterium (Miao 2.1609) of embodiment
1 test objective
In being carried out on the basis of small-scale test with No. 6 grape wild yeasts bacterium (Miao 2.1609) for detaching, filtering out
Type is tested, further to verify the performance (color and luster, flavor, liquor-producing ability etc.) of the wild yeasts bacteria strain.
2 test materials and method
No. 6 bacterial strain (Miao in Example 2 by being selected in the 1st, 2,3,4,5, No. 66 plants of grape wild yeasts bacterial strains
2.1609) fresh wheat bran yeast 2.5 kilograms (inoculum concentrations 0.5%), 50 kilograms of the grape slurries being prepared, after mixing well often
(September part) is fermented 25 days under the conditions of temperature.It is compared simultaneously with the equally processing of " English receive Supreme Being Si " (Enartis) wine yeast.
3 test results
After fermenting 15 days under room temperature, No. 6 wild yeasts bacterial strain (Miao 2.1609) obtains 53 degree of 7 kilograms of distilled spirits and (goes out
Product rate 14.0%), control strain obtains 53 degree of distilled spirits 6.5 kilograms (yield rates 13.0%), and liquor-producing ability is better than control bacterium
Strain " English receive Supreme Being Si ".
The identification to related organoleptic indicator has been organized simultaneously, it is obtained with No. 6 bacterial strains (Miao 2.1609) of grape wild yeasts
Distilled spirit wine liquid color clarification is transparent, grape flavor is strong, quiet and tastefully laid out, good taste, and excitation that entrance is stingless, wine body is mellow, hangs
Cup is apparent, and not top after people's drink, there are few bad physiological reactions, hence it is evident that is better than control strain.
The performance measurement trial production experiment of the 4 pure bacterial strain of grape wild yeasts bacterium of embodiment
1 test objective
Further fermentability in production of verification separation, the obtained No. 6 grape wild yeasts bacteria strain of selection and
Produce wine effect.
2 test materials and method
(1) 2.1609 grape yeast strain two level of Miao, three-level seed are made
The pure culture of the bacterial strain is made into 2.5 kilograms of wheat bran yeast secondary seeds by preceding method.
5 kilograms of fresh grape liquid are prepared, uniform with 6 kilograms of fresh wheat bran mixs, 100 DEG C of normal pressures maintain sterilize for 60 minutes, from
It is so cooled to room temperature, enters seeding tank and be inoculated with 2.5 kilograms of wheat bran yeast secondary seeds, 25 DEG C of intervals shake tank and lead to oxygen, culture 60~72
Hour, obtain 12.5 kilograms of wheat bran yeast three-level seeds.
(2) it is inoculated with
12.5 kilograms of No. 6 grape wild yeasts bacterium are accessed in the stainless steel fermentation tank equipped with 1000 kilograms of fresh grape liquid
The wheat bran yeast three-level seed of bacterial strain, inoculum concentration 1.25%.
(3) it ferments
First three day maintains 25~28 DEG C of Multiplying cultures and intermittent stirring (stirring daily three times, 30 minutes every time), to promote
It is proliferated into somatic cells, it is ensured that enough thalline quantities needed for fermentation.22~23 DEG C are maintained later to be left to ferment 15~20 days.
(4) physical and chemical index of zymotic fluid is detected
The physical and chemical indexes such as alcoholic strength, pH, the reduced sugar of zymotic fluid are detected, to determine fermentation termination.
10% or more alcohol content, pH3~3.5,0.4 gram/100 milliliters or less of reduced sugar (residual sugar) can terminate fermentation.
(reducing sugar test uses ferropotassium cyanide method；Alcoholometry uses hydrometer method；Acidity assaying uses pH meter).
(5) it filters, distill
It is voluntarily filtered using fermentation tank ontology gravity flow method.Filtrate takes wine by the method distilled twice.
Distillating method is as follows：
A, it distills for the first time：Zymotic fluid is distilled at 90~92 DEG C, distillation is terminated when alcoholic strength is 5 degree；
B, it distills for second：It is to distill obtained first wine for the first time to distill object, and vapo(u)rizing temperature is 90~92 DEG C, is taken point
The method that section takes wine：It is divided into three sections：First segment is " chieftain's wine ", contains more fusel oil in wine body, and taste is more bitter, amount
It is few, the white wine that alcoholic strength measuring device at liquid shows 70~76 degree is collected out, is mainly used for allocating wine；Second segment is " center
Wine " is the main body of Spirit, and amount is big, and for alcoholic strength generally between 58~70 degree, taste is more pure, needs to be added a certain amount of
Distilled water be configured to finished wine；Third section is " original plasm wine ", and amount is less, and alcoholic strength is between 53~58 degree, because it has not
The characteristics of can directly being drunk with allotment (blending), therefore it is called " original plasm wine ".It can be terminated when alcohol concentration is less than 53 degree
Distillation, extraction raffinate participate in next pot of distillation, and second pot of tail wine for connecing first pot, third pot connects second pot of tail wine, and so on (i.e.
Cycle distillation).
(6) semi-finished product physical and chemical index detects：Measure the alcoholic strength of each cylinder semi-finished product wine one by one with alcoholometer.By each cylinder
Quantity, alcohol concentration, production time, deposit position of semi-finished product wine etc. make eye-catching mark and accurate recording.
(7) store sth. in a cellar ageing：Object is semi-finished product wine.Wine cellar requires to clean clean and tidy, constant temperature and humidity；Wine vat requirement is closed, old
It is more than the moon to make time at least three.
Using strain Italy " English receive Supreme Being Si " (Enartis) commodity wine active dry yeast reference while production
The above method prepares wine spirit, and inoculum concentration is 20 grams/100 liters (2/10000ths), inventory, fermentation condition, filtering, distillation
It is identical as above-mentioned processing.
3 test results
It handles (No. 6 pure bacterial strain, that is, Miao 2.1609 of grape wild yeasts bacterium)：53 degree of distilled spirits 135,000 of comprehensive conversion production
Gram, yield rate 13.5%.Wine liquid color clarification is transparent, and grape flavor is strong, quiet and tastefully laid out, good taste, entrance is stingless excitation, wine
Body is mellow, hangs that cup is apparent, and not top after people's drink, there are few bad physiological reactions.
Control (Italy " English receive Supreme Being Si " (Enartis) commodity wine active dry yeast：Comprehensive 53 degree of steamings of conversion production
Evaporate 128 kilograms of white wine, yield rate 12.8%.Wine liquid color clarification is transparent, grape fragrance is thin, taste is purer, entrance thorn
Excitation is weak, and wine body is more mellow, and it is more apparent to hang cup.
Two compare, and voluntarily selection and breeding No. 6 grape wild yeasts bacteria strain Miao 2.1609 is better than control strain Italy
" English receive Supreme Being Si " (Enartis).According to lab scale, pilot scale, trial production properties synthesis measuring as a result, it is believed that Miao
2.1609 have apparent advantage in the production of Table Grape distilled spirit.
By No. 6 wild yeasts bacterial strain (Miao 2.1609), China Microbiological preservation administration committee is sent in October, 2017
Common micro-organisms center (abbreviation CGMCC) carries out preservation, and address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation date
For on October 11st, 2017, deposit number was CGMCC NO.14779, and classify entitled Saccharomyces Cerevisiae in S accharomyces
cerevisiae。
Claims (5)
1. a kind of wine spirit saccharomyces cerevisiae (Saccharomyces cerevisiae) Miao 2.1609, in China Microbiological bacterium
The common micro-organisms center preservation of kind preservation administration committee, preservation registration number are CGMCC No.14779.
2. wine spirit saccharomyces cerevisiae according to claim 1, it is characterised in that：Cell oval；Bacterium colony is rounded,
Milky, neat in edge, protrusion are surface wettability, sticky；Unicellular is 8-12 × 11-17 microns；Suitable cultivation temperature is：
29~31 DEG C；Suitable culture pH value is：5.5~6.5.
3. a kind of saccharomyces cerevisiae preparation, it is characterised in that：The active constituent of the preparation is saccharomyces cerevisiae described in claim 1
(Saccharomyces cerevisiae) Miao 2.1609.
4. application of wine spirit saccharomyces cerevisiae Miao 2.1609 as described in claim 1 in wine production.
5. wine spirit saccharomyces cerevisiae Miao 2.1609 as described in claim 1 is in grape red wine or other fruit wine or beer
Application in brewing.
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2018
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2008034337A1 (en) * | 2006-09-05 | 2008-03-27 | Longji Wei | Excrement and urine treatment apparatus for an ecological flush-free toilet |
| CN101701195A (en) * | 2009-11-25 | 2010-05-05 | 天津科技大学 | Yeast specially used for brewing rose-flavor grape wine and application thereof to grape wine brewing |
| CN103305431A (en) * | 2012-03-16 | 2013-09-18 | 湖北智敏农业有限公司 | Saccharomyces cerevisiae and application thereof in brewing of plum wine |
| CN103215195A (en) * | 2012-12-17 | 2013-07-24 | 青岛蔚蓝生物集团有限公司 | Saccharomyces cerevisiae and application of the same in dry red wine brewing |
| CN104371936A (en) * | 2014-11-25 | 2015-02-25 | 重庆市南岸区广阳镇回龙枇杷种植观光园 | Saccharomyces cerevisiae and application thereof |
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