CN105400652B - The method for quickly preparing artificial distiller's yeast using the functional microorganism flora of high yield butyric acid and caproic acid - Google Patents
The method for quickly preparing artificial distiller's yeast using the functional microorganism flora of high yield butyric acid and caproic acid Download PDFInfo
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Abstract
The present invention relates to the methods that Clostridium production butyric acid, caproic acid functional microorganism flora is used quickly to prepare artificial distiller's yeast.Belong to brewing technical field, the technical problems to be solved by the invention are to provide a kind of method that artificial distiller's yeast is quickly prepared using functional microorganism flora, and the aromatic Chinese spirit of high-quality can be produced using the pit of the artificial distiller's yeast.Present invention specific implementation includes the following steps：A, the culture of functional microorganism microflora fermentation liquid；B, pit mud heap fermentation is just trained 20~25 days, temperature control is at 25~35 DEG C；C, artificial distiller's yeast heap fermentation 30~35 days, you can obtain finished product.The method of the present invention protects quality pit mud resource, and can be quickly obtained the artificial distiller's yeast of high quality, improves the quality for the white wine newly stored by the use of functional microorganism microflora fermentation liquid as one of main inoculum.The present invention provides a kind of new scheme for the preparation of new pit pit mud, has broad application prospects.
Description
Technical field
The invention belongs to brewing technical fields, and in particular to make the functional microorganism flora using high yield butyric acid and caproic acid fast
The method that speed prepares artificial distiller's yeast.
Background technology
During Production of Luzhou-flavor Liquor, pit is both round, and pit mud therein is brewing functional microorganism again
One of main source.Microorganism in pit mud is mostly anaerobic bacteria, be metabolized generate butyric acid, caproic acid and with ethyl alcohol generation
Aliphatic ester is the main body flavor substance of aromatic Chinese spirit, therefore the sour flavor ability of production of pit mud directly decides the matter of pit
Amount, it is final to influence white wine vinosity and liquor body style.Production acid function microorganism in pit mud is that process is tamed and dociled all the year round in process of production
Change, be progressively enriched with, therefore the saying that there is industry to have " centurial aged pit has gone out wine ".
With the expansion of China's liquor industry scale, each wine enterprise has all built a considerable amount of pits, but in pit
Functional microorganism is not overnight to be formed, thus artificial distiller's yeast to be prepared into restriction high-quality white wine yield increased
Key constraints.At present, the microorganism during prepared by artificial distiller's yeast is mainly derived from the old cellar for storing things age quality pit mud of inoculation, by
In the scarcity of quality pit mud, the inoculum concentration of pit mud is limited, finally substantially prolongs the preparation time and quality of artificial distiller's yeast.
Invention content
The technical problems to be solved by the invention are to provide a kind of functional microorganism with efficiently production butyric acid, caproic acid ability
Bacteria group culture, the quick preparation applied to artificial distiller's yeast.
The present invention provides a kind of methods that artificial distiller's yeast is prepared using functional microorganism flora.The method of the present invention include with
Lower step：
A, butyric acid, caproic acid microorganism are produced with semi-synthetic yellow water culture medium difference anaerobic fermentation culture Clostridium,
Then culture solution is mixed into obtain pit mud production butyric acid, caproic acid functional microorganism microflora fermentation liquid；
B, by the 1 of pH value 5.0:The mixed liquor of 1 yellow water and water, functional microorganism microflora fermentation liquid, quality pit mud are according to matter
Measure ratio 4~5：1：1 ratio mixing, is made suspension；
C, according to mass ratio 9~12：6：1 yellow mud, pit sealing mud and koji powder mixing add in suspension made from step b, control
The mixing water content of matter is made between 45~50%；
D, the pit mud for completing mix is stacked, and claps tight, sealing anaerobic fermentation 20~25 days, first training pit mud；
E, by the 1 of pH value 5.0:The mixed liquor of 1 yellow water and water, functional microorganism microflora fermentation liquid are according to mass ratio 4~5：1
It is uniformly mixed；
F, according to mass ratio 30~40：20：1 yellow mud, just training pit mud and koji powder mixing are added in and are hanged made from step e
Pit mud mixture is made in liquid, mixing, and control pit mud mixes water content of matter between 45~50%；
G, the pit mud mixture other side for obtaining step f, claps tight, is sealed by fermentation 30~35 days, artificial distiller's yeast.
Wherein, the functional microorganism bacterium of the high yield butyric acid described in above method step a and caproic acid is clostridium butyricum, junket butyric acid
At least one of clostridium or clostridium klebsi.
Wherein, the formula of the semi-synthetic yellow water culture medium described in above method step a is：4~6g/L of sodium acetate, sulfuric acid
0.3~0.7g/L of ammonium, 0.2~0.6g/L of dipotassium hydrogen phosphate, 0.1~0.3g/L of magnesium sulfate, 0.5~1.5g/L of yeast extract, half Guang
Propylhomoserin 0.3~0.7g/L of hydrochloride is dissolved in water and yellow water and respectively accounts in the dilution of half by volume, pH value for 5.5~
6.5, sterilizing, thus obtaining the product.
Wherein, the Clostridium production butyric acid described in above method step a, caproic acid microorganism inoculum concentration for 5~
10%, 33~37 DEG C of fermentation temperature, the time of anaerobic fermentation culture is 7 days.
Wherein, the fermentation of pit mud pile core is controlled during the sealing anaerobic fermentation of the first training pit mud described in above method step d
Temperature is between 25~35 DEG C.
Wherein, controlled during pit mud mixture described in above method step g is sealed by fermentation pile core fermentation temperature 25~
Between 35 DEG C.
Wherein, the zymotic fluid equal proportion that each bacterial strain obtains after fermenting respectively in above method step a is mixed to get mixing hair
Zymotic fluid.
It is understood that the quality pit mud described in above method step b be derived from it is existing by production practices
The old of high-quality white wine can be produced by, which proving, stores pit mud.Preferably, quality pit mud is stored always from 100 years ages of cellar for storing things or more
Pit mud.
The present invention is additionally provided as the artificial distiller's yeast prepared by the above method simultaneously.
The beneficial effects of the present invention are：The method of the present invention is by the use of functional microorganism microflora fermentation liquid as main inoculum
One of, the initial content of functional microorganism in artificial distiller's yeast is increased, while with the use of 1 ︰, 1 yellow waters that pH value is 5 as people
The Major Nutrient source of microorganism in work pit mud, additionally establishes unique pit mud preparation method, is prepared so as to substantially reduce
The time of artificial distiller's yeast, it is only necessary to can obtain good artificial distiller's yeast in 50~60 days.The method of the present invention reduces high-quality simultaneously
The inoculum concentration of pit mud, 1 part of quality pit mud can obtain 60~70 parts of artificial distiller's yeasts, protect quality pit mud resource, and can be quick
The artificial distiller's yeast of high quality is obtained, improves the quality for the white wine newly stored.The present invention provides one for the preparation of new pit pit mud
The new scheme of kind, has broad application prospects.
Figure of description
Respectively take 3 parts of cellar for storing things bed muds, after extracting total DNA, bacterial flora in pit mud is parsed using high-flux sequence in Fig. 1,4 pits
Structure is fallen, to the result of 12 sample principal component analyses (PCA).
Specific embodiment
The specific embodiment of the present invention is described further with reference to embodiment, is not therefore limited the invention to
In the embodiment described range.
The present invention uses functional microorganism microflora fermentation liquid that can increase artificial distiller's yeast in this way as one of main inoculum
The initial content of middle functional microorganism, and substantially reduce the time for preparing artificial distiller's yeast, it is only necessary to 50~60 days are i.e. available
Good artificial distiller's yeast.It is at least one of clostridium butyricum, clostridium tyrobutyricum or clostridium klebsi it is preferable to use microorganism.
In addition, invention also uses 1 ︰, 1 yellow waters that pH value is 5, (1 ︰, 1 yellow waters in the present invention refer to yellow water and water
Yellow water after isometric mixed liquor, i.e. proportional diluted) Major Nutrient source as microorganism in artificial distiller's yeast, it is main using wherein
The carbon sources such as the lower alcohol being rich in and short chain fatty acids the study found that these carbon sources can quickly be utilized by cellar mud microorganisms, accelerate micro-
The growth and breeding of biology.And the present invention creatively the pH value of yellow water culture medium is adjusted to from general 3.5 or so it is weakly acidic
5.0, it can keep acidic environment that varied bacteria growing and peracid is inhibited to inhibit functional microorganism activity in this way.Meanwhile according to this
Concentration of alcohol can be down to 2.5% or so using dilution, relieve high concentration ethanol to target of the present invention by the early-stage study of invention
Microorganism and other fermentation beneficial microorganisms inhibit.Above various measures combine, and have reached reduction quality pit mud
Inoculum concentration, protect quality pit mud resource, while shorten preparation time, artificial distiller's yeast made from raising.
Specific the method for the present invention can carry out according to the following steps：
A, the culture of functional microorganism flora
In semi-synthetic yellow water culture medium, (formula of semi-synthetic yellow water culture medium is respectively for production butyric acid, caproic acid functional microorganism
4~6g/L of sodium acetate, 0.3~0.7g/L of ammonium sulfate, 0.2~0.6g/L of dipotassium hydrogen phosphate, 0.1~0.3g/L of magnesium sulfate, yeast
0.5~1.5g/L of cream, 0.3~0.7g/L of cysteine hydrochloride, water and yellow water respectively account for half by volume, pH value for 5.5~
6.5, sterilizing, thus obtaining the product) in cultivate, inoculum concentration 5~10%, 33~37 DEG C of temperature, Anaerobic culturel 7 days.Then by culture solution equal proportion
It mixes up to pit mud production butyric acid, caproic acid functional microorganism microflora fermentation liquid.
B, the 1 of pH value 5.0:1 yellow water, functional microorganism microflora fermentation liquid, quality pit mud are according to mass ratio 4~5：1：1
Ratio mixes, and suspension is made.
C, blocky yellow mud and pit sealing mud are respectively crushed into the particle of grain size 1~3cm sizes, according to mass ratio 9~12：6：
1 yellow mud, pit sealing mud and koji powder mixing.
D, suspension made from step b is added in material heap made from step c, control material heap moisture content 45~50% it
Between.
E, the pit mud heap for completing mix coning, is clapped tight, is sealed by fermentation 20~25 days, first training pit mud.Period
Pit mud pile core fermentation temperature is controlled between 25~35 DEG C.
F, take adjust pH value to 5.0 1：1 yellow water, functional microorganism microflora fermentation liquid are according to mass ratio 4~5：1 mixing is equal
It is even.
G, blocky yellow mud and just, training pit mud is respectively crushed into the particles of grain size 1~3cm sizes, according to mass ratio 30~
40：20：1 yellow mud, just training pit mud and koji powder mixing.
H, suspension made from step f is added in material heap made from step g, control material heap moisture content 45~50% it
Between.
I, by the pit mud heap machined coning, clap tight, be sealed by fermentation 30~35 days, artificial distiller's yeast.Period
Pit mud pile core fermentation temperature is controlled between 25~35 DEG C.
Those skilled in the art may be used classification and expand when functional microorganism zymotic fluid is largely prepared in a steps
The mode of culture can finally carry out interval shake culture in fermentation tank or closed container.These are all people in the art
Member is easy to implement according to the prior art.In this field, inoculum concentration refer to seed liquor volume and inoculation after system body
The ratio between product.
In a steps and f steps, in order to which cellar mud microorganisms can be grown as early as possible, yellow water temperature preferably control 30~40 DEG C it
Between.
In b step, quality pit mud should derive from out the high pit of wine quality, preferably select 100 years cellars for storing things pit more than age.
The yellow mud of middle selection preferably selects the farmland soil layer of 10~30cm of depth in step c and g steps, and wherein phosphorus, potassium and humus enriches,
Be conducive to the growth of cellar mud microorganisms.And in step c and g steps, koji powder selects commercially available conventional yeast.Further to carry
High pit mud quality, preferably uses daqu of middle temperature.
Artificial distiller's yeast prepared by the above method of the present invention is a kind of outstanding artificial distiller's yeast product, can be used for high quality liquor
In fermentation process.
More specific description is carried out to the method for the present invention with embodiment below.
Embodiment one prepares artificial distiller's yeast using the method for the present invention
1st, bacterial strain clostridium butyricum (Clostridium butyricum, the preserving number of tool production butyric acid ability are chosen：ATCC
And clostridium tyrobutyricum (Clostridium tyrobutyricum, preserving number 19398)：ATCC 25755) and with production caproic acid
Bacterial strain clostridium klebsi (Clostridium kluyveri, the preserving number of ability：ATCC 8527) exist respectively in 50L fermentation tanks
Semi-synthetic yellow water inoculation of medium fermentation, the formula of semi-synthetic yellow water culture medium used is sodium acetate 5g g/L, ammonium sulfate
0.5g g/L, dipotassium hydrogen phosphate 0.4g g/L, magnesium sulfate 0.2g g/L, yeast extract 1g g/L, cysteine hydrochloride 0.5g g/
L, add 1：1 sterile yellow water (with water diluted yellow water in equal volume) adjusts pH value as 6.0,115 DEG C of sterilizing 20min to 1000mL.Fermentation
Canned liquid measure 20L, inoculum concentration 10%, 35 DEG C of temperature, anaerobic fermentation 7 days mix the culture solution equal proportion of each strain to obtain the final product
Functional microorganism microflora fermentation liquid.
2nd, 60kg centurial aged pit pit muds, 60kg functional microorganism microflora fermentation liquid are taken, 300kg pH value is the 1 of 5.0：1 is yellow
Water (with water diluted yellow water in equal volume), stirs into suspension.
3rd, the pit sealing mud 300kg crushed is taken, crop field yellow mud 450kg, daqu of middle temperature 50kg are uniformly mixed three, contain
Water about 25%.
4th, step 1 suspension is added in solid material heap, plus, be sufficiently mixed on one side on one side, make grogs, thalline, starter powder mixing
Uniformly, water content about 50%.
5th, material heap heap is next, is sealed by fermentation 25 days, between temperature control 25~35, obtains just training pit mud 1100kg, contain
Water about 46%.
6th, 2200kg yellow mud (water content about 20%), 55kg starter powders, 200kg functional microorganism floras just are added in training pit mud
Zymotic fluid, 800kg pH value are 5.0 1 ︰, 1 yellow waters.
7th, material heap heap is next, is sealed by fermentation 35 days, between temperature control 25~35, obtains artificial distiller's yeast about 4000kg,
Water content about 45%.
8th, artificial distiller's yeast, input grain grain fermentation production wine are applied in newly-built pit.
Using the pit (artificial aged pit) of the artificial distiller's yeast of the method for the present invention preparation, new cellar for storing things, (yellow mud, cellar for storing things bed mud and envelope are stored
Mud mixes), 30 years pits, 50 years pits, under the conditions of similary raw material and technique productions, produce aromatic Chinese spirit, compare
Main body fragrance content of material in white wine, the results are shown in Table 1.3 parts of cellar for storing things bed muds are respectively taken in 4 pits respectively, extract total DNA
Afterwards, bacterial quorum sensing in pit mud is parsed using high-flux sequence, (PCA) is analyzed to 12 sample principal components, as a result such as Fig. 1
It is shown.
1 each pit of table goes out main body fragrance content of material (unit in wine：g/L)
| Ethyl hexanoate | Ethyl butyrate | Ethyl acetate | Caproic acid | Butyric acid | Acetic acid | |
| New cellar for storing things | 1.90 | 0.24 | 1.22 | 0.41 | 0.10 | 0.27 |
| Artificial aged pit | 2.27 | 0.35 | 1.39 | 0.63 | 0.13 | 0.30 |
| It stores within 30 years | 2.35 | 0.38 | 1.53 | 0.65 | 0.18 | 0.33 |
| It stores within 50 years | 3.55 | 0.42 | 2.08 | 0.78 | 0.17 | 0.48 |
As can be seen from Table 1, the aromatic Chinese spirit quality that the artificial aged pit of the artificial distiller's yeast prepared using the present invention is produced
New cellar for storing things is apparently higher than, main aroma content still had certain gap close to 30 years pits with 50 years pits.And by the poly- of Fig. 1
Alanysis result will also realize that, the sample from 4 pits is polymerized to 3 classes in PCA figures, pit mud in artificial aged pit and 30 years pits
Bacterium composition is close, differs greatly with what new cellar for storing things and 50 years were stored.In addition, the every 1 part of quality pit mud of this method can obtain 60~70 parts
Artificial distiller's yeast.Compared with 1 part of quality pit mud output, 7~8 parts of people disclosed in the prior art, such as 102181342 B of patent document CN
Work pit mud, the inoculum concentration of quality pit mud reduce nearly 90%, protect quality pit mud resource.
Claims (5)
1. the method for quickly preparing artificial distiller's yeast using the functional microorganism flora of high yield butyric acid and caproic acid, it is characterised in that including
Following steps：
A, butyric acid, caproic acid microorganism are produced with semi-synthetic yellow water culture medium difference anaerobic fermentation culture Clostridium, then
Culture solution is mixed into obtain pit mud production butyric acid, caproic acid functional microorganism microflora fermentation liquid；The function of the high yield butyric acid and caproic acid is micro-
Biological bacteria is at least one of clostridium butyricum, clostridium tyrobutyricum or clostridium klebsi；Described Clostridium production butyric acid,
The inoculum concentration of caproic acid microorganism is 5~10%, 33~37 DEG C of fermentation temperature, and the time of anaerobic fermentation culture is 7 days；
B, by the 1 of pH value 5.0:The mixed liquor of 1 yellow water and water, functional microorganism microflora fermentation liquid, quality pit mud are according to mass ratio 4
~5：1：1 ratio mixing, is made suspension；
C, according to mass ratio 9~12：6：1 yellow mud, pit sealing mud and koji powder mixing, add in suspension made from step b, and control should
Water content of matter is mixed between 45~50%；
D, the pit mud for completing mix is stacked, and claps tight, sealing anaerobic fermentation 20~25 days, first training pit mud；
E, by the 1 of pH value 5.0:The mixed liquor of 1 yellow water and water, functional microorganism microflora fermentation liquid are according to mass ratio 4~5：1 mixing
Suspension uniformly is made；
F, according to mass ratio 30~40：20：1 yellow mud, just training pit mud and koji powder mixing, add in suspension made from step e, mix
Even that pit mud mixture is made, control pit mud mixes water content of matter between 45~50%；
G, the pit mud mixture obtained step f is stacked, and claps tight, is sealed by fermentation 30~35 days, artificial distiller's yeast.
2. according to the method described in claim 1, it is characterized in that the formula of the semi-synthetic yellow water culture medium described in step a is：
4~6g/L of sodium acetate, 0.3~0.7g/L of ammonium sulfate, 0.2~0.6g/L of dipotassium hydrogen phosphate, 0.1~0.3g/L of magnesium sulfate, yeast
0.5~1.5g/L of cream, 0.3~0.7g/L of cysteine hydrochloride are dissolved in water and respectively account for the dilute of a meromict by volume with yellow water
It releases in liquid, pH value is 5.5~6.5, sterilizing, thus obtaining the product.
3. according to the method described in claim 1, it is characterized in that：The sealing anaerobic fermentation of first training pit mud described in step d
Period controls pit mud pile core fermentation temperature between 25~35 DEG C.
4. according to the method described in claim 1, it is characterized in that：The pit mud mixture of step g controls heap during being sealed by fermentation
Heart fermentation temperature is controlled at 25~35 DEG C.
5. as the artificial distiller's yeast prepared by Claims 1 to 4 any one of them method.
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| CN106085741B (en) * | 2016-08-30 | 2019-05-21 | 花冠集团酿酒股份有限公司 | A kind of Luzhou-flavor liquo band pit mud nutrient fluid cultural method |
| CN108570384B (en) * | 2018-06-12 | 2021-06-04 | 安徽省金裕皖酒业有限公司 | Production process of cellar mud-free strong-flavor liquor |
| CN111500507A (en) * | 2020-05-28 | 2020-08-07 | 江南大学 | Pit mud flora enrichment method based on oligoculture and application thereof |
| CN112410263B (en) * | 2020-12-08 | 2022-03-29 | 北京工商大学 | New strain of ancient Liangbacillus and application thereof |
| CN113831978B (en) * | 2021-10-21 | 2023-05-02 | 四川省宜宾高洲酒业有限责任公司 | Artificial pit mud and preparation, maintenance and pit-outside production method thereof |
| CN116083191B (en) * | 2022-09-07 | 2024-04-30 | 江南大学 | Method for reducing and controlling lactic acid accumulation in fermentation process of Luzhou-flavor liquor by utilizing chemotaxis of clostridium |
| CN115747128A (en) * | 2022-11-10 | 2023-03-07 | 泸州老窖股份有限公司 | Method for directionally culturing pit mud functional microorganisms |
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| CN101074419A (en) * | 2006-05-18 | 2007-11-21 | 姚云 | Technology for culturing hexylic-acid bacterium by artificial aged pit |
| CN102181342B (en) * | 2011-03-08 | 2014-02-12 | 泸州品创科技有限公司 | Artificial pit mud and preparation method thereof |
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