CN105400652A - Method for rapidly preparing man-made pit mud through functional microbial group of high-yield butyric acid and caproic acid - Google Patents
Method for rapidly preparing man-made pit mud through functional microbial group of high-yield butyric acid and caproic acid Download PDFInfo
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Abstract
The invention relates to a method for rapidly preparing man-made pit mud through a functional microbial group of butyric acid and caproic acid produced through clostridium, belongs to the technical field of liquor brewing and aims to achieve the technical purpose that the method for rapidly preparing man-made pit mud through the functional microbial group is provided. A pit applying the man-made pit mud can produce high-quality strong-flavor baijiu. The method includes the following specific steps that 1, functional microbial group fermentation liquid is cultured; 2, preliminarily-cultured pit mud is piled and fermented for 20-25 days, and the temperature is conducted to 25-35 DEG C; 3, man-made pit mud is piled and fermented for 30-35 days, and a finished product can be obtained. According to the method, the functional microbial group fermentation liquid serves as one of the most important inoculums, the high-quality pit mud resource is protected, high-quality man-made pit can be rapidly obtained, and the quality of baijiu of a new pit is improved. A new scheme is provided for preparation of the pit mud of a new pit pool, and wide application prospect is achieved.
Description
Technical field
The invention belongs to brewing technical field, be specifically related to make the functional microorganism flora utilizing high yield butyric acid and caproic acid prepare the method for artificial distiller's yeast fast.
Background technology
In Production of Luzhou-flavor Liquor process, Jiao Chi is fermenting container, and cellar for storing things mud is wherein again one of main source of brewing functional microorganism.Microorganism in the mud of cellar for storing things mostly is anerobe, its metabolism produce butyric acid, caproic acid and be the main body flavour substances of aromatic Chinese spirit with the fatty acid ester that ethanol generates, the product acid flavor ability of therefore storing mud directly decides the quality of Jiao Chi, finally affects white wine vinosity and liquor body style.Product acid function microorganism in the mud of cellar for storing things is in process of production through domestication, enrichment gradually all the year round, therefore has industry to have the saying of " centurial aged pit has gone out wine ".
Along with the expansion of China's liquor industry scale, each wine enterprise has all built a considerable amount of Jiao Chi, but the functional microorganism in pond, cellar for storing things is not overnight formed, therefore the key constraints being prepared into the increase of restriction high-quality white wine output of artificial distiller's yeast.At present, the artificial distiller's yeast microorganism of preparing in process is mainly derived from the old cellar for storing things quality pit mud in age of inoculation, due to the scarcity of quality pit mud, limits the inoculum size of cellar for storing things mud, finally substantially prolongs preparation time and the quality of artificial distiller's yeast.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind ofly has the functional microorganism bacteria group culture efficiently producing butyric acid, caproic acid ability, is applied to the quick preparation of artificial distiller's yeast.
The invention provides a kind of method utilizing functional microorganism flora to prepare artificial distiller's yeast.The inventive method comprises the following steps:
A, with semi-synthetic yellow water substratum respectively anaerobically fermenting cultivate Clostridium and produce butyric acid, caproic acid microorganism, then nutrient solution is mixed to get cellar for storing things mud and produces butyric acid, caproic acid functional microorganism flora fermented liquid;
B, the mixed solution of the 1:1 yellow water of pH value 5.0 and water, functional microorganism flora fermented liquid, quality pit mud to be mixed according to the ratio of mass ratio 4 ~ 5:1:1, make suspension;
C, according to mass ratio 9 ~ 12:6:1 the mixing of yellow mud, pit sealing mud and koji powder, add the suspension that step b is obtained, control this mixture water ratio between 45 ~ 50%;
D, the cellar for storing things dust completed by mix are put, and clap tight, sealing anaerobically fermenting 20 ~ 25 days, first training cellar for storing things mud;
E, the mixed solution of the 1:1 yellow water of pH value 5.0 and water, functional microorganism flora fermented liquid to be mixed according to mass ratio 4 ~ 5:1;
F, according to mass ratio 30 ~ 40:20:1 yellow mud, just training cellar for storing things mud and koji powder mixing, add the suspension that step e is obtained, cellar for storing things mud mixture is made in mixing, controls cellar for storing things mud mixture water ratio between 45 ~ 50%;
G, the cellar for storing things mud mixture the other side obtained by step f, clap tight, sealing and fermenting 30 ~ 35 days, artificial distiller's yeast.
Wherein, the high yield butyric acid described in aforesaid method step a and the functional microorganism bacterium of caproic acid are at least one in clostridium butylicum, clostridium tyrobutyricum or kirschner clostridium.
Wherein, the formula of the semi-synthetic yellow water substratum described in aforesaid method step a is: sodium acetate 4 ~ 6g/L, ammonium sulfate 0.3 ~ 0.7g/L, dipotassium hydrogen phosphate 0.2 ~ 0.6g/L, magnesium sulfate 0.1 ~ 0.3g/L, yeast extract paste 0.5 ~ 1.5g/L, cysteine hydrochloride 0.3 ~ 0.7g/L, be dissolved in water and yellow water respectively accounts in the diluent of half by volume, pH value is 5.5 ~ 6.5, sterilizing and get final product.
Wherein, the Clostridium described in aforesaid method step a produces butyric acid, the inoculum size of caproic acid microorganism is 5 ~ 10%, and leavening temperature 33 ~ 37 DEG C, the time that anaerobically fermenting is cultivated is 7 days.
Wherein, the first training cellar for storing things mud described in aforesaid method steps d sealing anaerobically fermenting during control cellar for storing things dust heart leavening temperature between 25 ~ 35 DEG C.
Wherein, pile core leavening temperature is controlled between 25 ~ 35 DEG C during the cellar for storing things mud mixture sealing and fermenting described in aforesaid method step g.
Wherein, the fermented liquid equal proportion that in aforesaid method step a, each bacterial strain obtains after fermenting respectively is mixed to get mixed fermentation liquid.
Be understandable that, the quality pit mud described in aforesaid method step b derives from existing cellar for storing things, the old cellar for storing things mud proving to produce high-quality white wine through production practice.Preferably, quality pit mud is the cellar for storing things mud deriving from old cellar for storing things in more than 100 years age, cellar for storing things.
The present invention additionally provides the artificial distiller's yeast prepared by aforesaid method simultaneously.
Beneficial effect of the present invention is: the inventive method utilizes functional microorganism flora fermented liquid as one of main inoculum, add the initial content of functional microorganism in artificial distiller's yeast, be simultaneously that 1 ︰ 1 yellow water of 5 is as the Major Nutrient source of microorganism in artificial distiller's yeast with the use of pH value, establish unique pit mud Preparation Method in addition, thus substantially reduce the time of preparing artificial distiller's yeast, only need the artificial distiller's yeast that can obtain high-quality for 50 ~ 60 days.The inventive method reduces the inoculum size of quality pit mud simultaneously, and 1 part of quality pit mud can obtain 60 ~ 70 parts of artificial distiller's yeasts, protects quality pit mud resource, and can obtain high-quality artificial distiller's yeast fast, improves the quality of the white wine at new cellar for storing things.The present invention is that the preparation of cellar for storing things, Xin Jiao pond mud provides a kind of new scheme, has broad application prospects.
Figure of description
Respectively get 3 parts of cellar for storing things bed muds in Fig. 1, pond, 4 cellars for storing things, after extracting STb gene, utilize high-flux sequence to resolve bacterial quorum sensing in the mud of cellar for storing things, 12 sample principal component are analyzed to the result of (PCA).
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described further, does not therefore limit the present invention in described scope of embodiments.
Using function microorganism species fermented liquid of the present invention is as one of main inoculum, the initial content of functional microorganism in artificial distiller's yeast can be increased like this, and substantially reduce the time of preparing artificial distiller's yeast, only need the artificial distiller's yeast that can obtain high-quality for 50 ~ 60 days.Preferred use microorganism is clostridium butylicum, at least one in clostridium tyrobutyricum or kirschner clostridium.
In addition, invention also uses pH value be 51 ︰ 1 yellow water (1 ︰ 1 yellow water in the present invention refers to the equal-volume mixed solution of yellow water and water, namely the yellow water after proportional diluted) as the Major Nutrient source of microorganism in artificial distiller's yeast, the carbon sources such as the lower alcohol that main utilization is wherein rich in and short chain fatty acid, research finds, these carbon sources can be utilized fast by cellar for storing things mud microorganism, accelerate microbial growth breeding.And the present invention is creatively adjusted to weakly acidic 5.0 the pH value of yellow water substratum from general about 3.5, sour environment can be kept like this to suppress varied bacteria growing, prevent again peracid inhibit feature microorganism active.Meanwhile, according to early-stage Study of the present invention, utilize dilution alcohol concn can be down to about 2.5%, relieve high concentration ethanol and target microorganism of the present invention and other fermentation beneficial microorganisms are suppressed.Above all measures combine, and reach the inoculum size reducing quality pit mud, and protection quality pit mud resource, shortens preparation time simultaneously, the artificial distiller's yeast that raising is obtained.
Concrete the inventive method can be carried out according to the following steps:
The cultivation of a, functional microorganism flora
Produce butyric acid, (formula of semi-synthetic yellow water substratum is sodium acetate 4 ~ 6g/L, ammonium sulfate 0.3 ~ 0.7g/L, dipotassium hydrogen phosphate 0.2 ~ 0.6g/L, magnesium sulfate 0.1 ~ 0.3g/L, yeast extract paste 0.5 ~ 1.5g/L, cysteine hydrochloride 0.3 ~ 0.7g/L to caproic acid functional microorganism at semi-synthetic yellow water substratum respectively, water and yellow water respectively account for half by volume, pH value is 5.5 ~ 6.5, sterilizing and get final product) in cultivate, inoculum size 5 ~ 10%, temperature 33 ~ 37 DEG C, Anaerobic culturel 7 days.Then nutrient solution equal proportion is mixed and obtain cellar for storing things mud and produce butyric acid, caproic acid functional microorganism flora fermented liquid.
The 1:1 yellow water of b, pH value 5.0, functional microorganism flora fermented liquid, quality pit mud mix according to the ratio of mass ratio 4 ~ 5:1:1, make suspension.
C, block yellow mud and pit sealing mud are ground into respectively the particle of particle diameter 1 ~ 3cm size, according to mass ratio 9 ~ 12:6:1, yellow mud, pit sealing mud and koji powder are mixed.
D, the suspension that step b obtains added the obtained bank of step c, control bank water ratio between 45 ~ 50%.
E, the cellar for storing things dust that completed by mix coning, are clapped tight, sealing and fermenting 20 ~ 25 days, first training cellar for storing things mud.Period controls cellar for storing things dust heart leavening temperature between 25 ~ 35 DEG C.
F, the 1:1 yellow water getting adjust pH to 5.0, functional microorganism flora fermented liquid mix according to mass ratio 4 ~ 5:1.
G, block yellow mud and just training cellar for storing things mud be ground into the particle of particle diameter 1 ~ 3cm size respectively, according to mass ratio 30 ~ 40:20:1 yellow mud, just training cellar for storing things mud and koji powder mixing.
H, the suspension that step f obtains added the obtained bank of step g, control bank water ratio between 45 ~ 50%.
I, by the cellar for storing things dust that machines coning, clap tight, sealing and fermenting 30 ~ 35 days, artificial distiller's yeast.Period controls cellar for storing things dust heart leavening temperature between 25 ~ 35 DEG C.
Those skilled in the art's functional microorganism fermented liquid in a step can adopt the mode of classification enlarged culturing when preparing in a large number, finally can carry out interval in fermentor tank or encloses container and shake cultivation.These are all that those skilled in the art easily realize according to prior art.In this area, inoculum size refers to the ratio of the volume of seed liquor and the volume of postvaccinal system.
In a step and f step, can grow as early as possible to store mud microorganism, yellow water temperature should control between 30 ~ 40 DEG C.
In b step, quality pit mud should derive from out the high Jiao Chi of wine quality, preferably selects the Jiao Chi storing age more than for 100 years.The yellow mud selected in step c and g step should select the farmland soil layer of the degree of depth 10 ~ 30cm, and wherein phosphorus, potassium and soil ulmin are abundant, is conducive to cellar for storing things mud microbial growth.And in step c and g step, koji powder selects commercially available conventional Daqu.For improving pit mud quality further, preferably use daqu of middle temperature.
Artificial distiller's yeast prepared by aforesaid method of the present invention is a kind of outstanding artificial distiller's yeast product, can be used in the fermenting process of high quality liquor.
By embodiment, more specific description is carried out to the inventive method below.
Embodiment one, employing the inventive method prepare artificial distiller's yeast
1, choose the bacterial strain clostridium butylicum (Clostridiumbutyricum that tool produces butyric acid ability, preserving number: ATCC19398) and clostridium tyrobutyricum (Clostridiumtyrobutyricum, preserving number: ATCC25755) and there is the bacterial strain kirschner clostridium (Clostridiumkluyveri producing caproic acid ability, preserving number: ATCC8527) ferment at semi-synthetic yellow water inoculation of medium respectively in 50L fermentor tank, the formula of the semi-synthetic yellow water substratum used is sodium acetate 5gg/L, ammonium sulfate 0.5gg/L, dipotassium hydrogen phosphate 0.4gg/L, magnesium sulfate 0.2gg/L, yeast extract paste 1gg/L, cysteine hydrochloride 0.5gg/L, add the aseptic yellow water of 1:1 (yellow water with the dilution of water equal-volume) to 1000mL, adjust pH is 6.0, 115 DEG C of sterilizing 20min.Fermentor tank liquid amount 20L, inoculum size 10%, temperature 35 DEG C, anaerobically fermenting 7 days, mixes the nutrient solution equal proportion of each bacterial classification and obtains functional microorganism flora fermented liquid.
2, get 60kg centurial aged pit cellar for storing things mud, 60kg functional microorganism flora fermented liquid, 300kgpH value is the 1:1 yellow water (yellow water with the dilution of water equal-volume) of 5.0, stirs into suspension.
3, get the pit sealing mud 300kg crushed, land for growing field crops yellow mud 450kg, daqu of middle temperature 50kg, mixes three, water content about 25%.
4, step 1 suspension is added solids stockpile, add, fully mix on one side, make grogs, thalline, bent powder mixes, water content about 50%.
5, bank is piled tightly, sealing and fermenting 25 days, and temperature controls between 25 ~ 35, obtains just training and stores mud 1100kg, water content about 46%.
6, just training is stored in mud and is added 2200kg yellow mud (water content about 20%), and the bent powder of 55kg, 200kg functional microorganism flora fermented liquid, 800kgpH value is 1 ︰ 1 yellow water of 5.0.
7, bank is piled tightly, sealing and fermenting 35 days, and temperature controls, between 25 ~ 35, to obtain artificial distiller's yeast and be about 4000kg, water content about 45%.
8, in pond, newly-built cellar for storing things, apply artificial distiller's yeast, drop into the fermentation of grain grain and produce wine.
The Jiao Chi (artificial aged pit) of the artificial distiller's yeast adopting the inventive method to prepare, new cellar for storing things (yellow mud, cellar for storing things bed mud and pit sealing mud mix), 30 years Jiao Chi, 50 years Jiao Chi, under same raw material and explained hereafter condition, produce aromatic Chinese spirit, relatively main body fragrance substances content in white wine, result is as shown in table 1.In pond, 4 cellars for storing things, respectively get 3 parts of cellar for storing things bed muds respectively, after extracting STb gene, utilize high-flux sequence to resolve bacterial quorum sensing in the mud of cellar for storing things, analyze (PCA) to 12 sample principal component, result as shown in Figure 1.
The each Jiao Chi of table 1 goes out main body fragrance substances content (unit: g/L) in wine
| 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 |
| 30 years cellars for storing things | 2.35 | 0.38 | 1.53 | 0.65 | 0.18 | 0.33 |
| 50 years cellars for storing things | 3.55 | 0.42 | 2.08 | 0.78 | 0.17 | 0.48 |
As can be seen from Table 1, the aromatic Chinese spirit quality of the artificial aged pit production of the artificial distiller's yeast adopting the present invention to prepare is apparently higher than Xin Jiao, and main aroma content, close to 30 years Jiao Chi, still has certain gap with 50 years Jiao Chi.And from Fig. 1 cluster analysis result also, the sample from 4 Jiao Chi is polymerized to 3 classes in PCA figure, artificial aged pit and store the bacterium of storing mud in pond for 30 years and form close, and newly to store and 50 years store differs greatly.In addition, the every 1 part of quality pit mud of present method can obtain 60 ~ 70 parts of artificial distiller's yeasts.Comparatively prior art, such as 7 ~ 8 parts of artificial distiller's yeasts of 1 part of quality pit mud output disclosed in patent documentation CN102181342B, the inoculum size of quality pit mud reduces nearly 90%, protects quality pit mud resource.
Claims (7)
1. utilize the functional microorganism flora of high yield butyric acid and caproic acid to prepare the method for artificial distiller's yeast fast, it is characterized in that comprising the following steps:
A, with semi-synthetic yellow water substratum respectively anaerobically fermenting cultivate Clostridium and produce butyric acid, caproic acid microorganism, then nutrient solution is mixed to get cellar for storing things mud and produces butyric acid, caproic acid functional microorganism flora fermented liquid;
B, the mixed solution of the 1:1 yellow water of pH value 5.0 and water, functional microorganism flora fermented liquid, quality pit mud to be mixed according to the ratio of mass ratio 4 ~ 5:1:1, make suspension;
C, according to mass ratio 9 ~ 12:6:1 the mixing of yellow mud, pit sealing mud and koji powder, add the suspension that step b is obtained, control this mixture water ratio between 45 ~ 50%;
D, the cellar for storing things dust completed by mix are put, and clap tight, sealing anaerobically fermenting 20 ~ 25 days, first training cellar for storing things mud;
E, the mixed solution of the 1:1 yellow water of pH value 5.0 and water, functional microorganism flora fermented liquid mixed according to mass ratio 4 ~ 5:1 and makes suspension;
F, according to mass ratio 30 ~ 40:20:1 yellow mud, just training cellar for storing things mud and koji powder mixing, add the suspension that step e is obtained, cellar for storing things mud mixture is made in mixing, controls cellar for storing things mud mixture water ratio between 45 ~ 50%;
G, the cellar for storing things mud mixture the other side obtained by step f, clap tight, sealing and fermenting 30 ~ 35 days, artificial distiller's yeast.
2. method according to claim 1, is characterized in that: the functional microorganism bacterium of high yield butyric acid and caproic acid described in step a is at least one in clostridium butylicum, clostridium tyrobutyricum or kirschner clostridium.
3. method according to claim 1, it is characterized in that the formula of the semi-synthetic yellow water substratum described in step a is: sodium acetate 4 ~ 6g/L, ammonium sulfate 0.3 ~ 0.7g/L, dipotassium hydrogen phosphate 0.2 ~ 0.6g/L, magnesium sulfate 0.1 ~ 0.3g/L, yeast extract paste 0.5 ~ 1.5g/L, cysteine hydrochloride 0.3 ~ 0.7g/L, water-solublely respectively to account for by volume in the diluent of a meromict with yellow water, pH value is 5.5 ~ 6.5, sterilizing and get final product.
4. method according to claim 1, is characterized in that: the Clostridium described in step a produces butyric acid, the inoculum size of caproic acid microorganism is 5 ~ 10%, and leavening temperature 33 ~ 37 DEG C, the time that anaerobically fermenting is cultivated is 7 days.
5. method according to claim 1, is characterized in that: the first training cellar for storing things mud described in steps d sealing anaerobically fermenting during control cellar for storing things dust heart leavening temperature between 25 ~ 35 DEG C.
6. method according to claim 1, is characterized in that: control pile core leavening temperature during the cellar for storing things mud mixture sealing and fermenting of step g and control at 25 ~ 35 DEG C.
7. the artificial distiller's yeast prepared by the method described in any one of claim 1 ~ 6.
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| CN106085741A (en) * | 2016-08-30 | 2016-11-09 | 花冠集团酿酒股份有限公司 | A kind of aromatic Chinese spirit band pit mud nutrient fluid cultural method |
| CN108570384A (en) * | 2018-06-12 | 2018-09-25 | 安徽省金裕皖酒业有限公司 | A kind of production technology of no pit mud aromatic Chinese spirit |
| CN111500507A (en) * | 2020-05-28 | 2020-08-07 | 江南大学 | Pit mud flora enrichment method based on oligoculture and application thereof |
| CN112410263A (en) * | 2020-12-08 | 2021-02-26 | 北京工商大学 | New strain of ancient Liangbacillus and application thereof |
| CN113831978A (en) * | 2021-10-21 | 2021-12-24 | 四川省宜宾高洲酒业有限责任公司 | Artificial pit mud and preparation, maintenance and ex-pit production method thereof |
| 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 |
| CN116083191A (en) * | 2022-09-07 | 2023-05-09 | 江南大学 | Method for reducing and controlling lactic acid accumulation in fermentation process of Luzhou-flavor liquor by utilizing chemotaxis of clostridium |
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| CN106085741A (en) * | 2016-08-30 | 2016-11-09 | 花冠集团酿酒股份有限公司 | A kind of aromatic Chinese spirit band pit mud nutrient fluid cultural method |
| CN108570384A (en) * | 2018-06-12 | 2018-09-25 | 安徽省金裕皖酒业有限公司 | A kind of production technology of no pit mud aromatic Chinese spirit |
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| CN111500507A (en) * | 2020-05-28 | 2020-08-07 | 江南大学 | Pit mud flora enrichment method based on oligoculture and application thereof |
| CN112410263A (en) * | 2020-12-08 | 2021-02-26 | 北京工商大学 | New strain of ancient Liangbacillus and application thereof |
| CN113831978A (en) * | 2021-10-21 | 2021-12-24 | 四川省宜宾高洲酒业有限责任公司 | Artificial pit mud and preparation, maintenance and ex-pit production method 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 |
| CN116083191A (en) * | 2022-09-07 | 2023-05-09 | 江南大学 | Method for reducing and controlling lactic acid accumulation in fermentation process of Luzhou-flavor liquor by utilizing chemotaxis of clostridium |
| 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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