CN109628253A - A kind of method and its application preparing biological esterification liquid using Aspergillus niger liquid state fermentation - Google Patents
A kind of method and its application preparing biological esterification liquid using Aspergillus niger liquid state fermentation Download PDFInfo
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Abstract
The invention discloses a kind of methods for preparing biological esterification liquid using Aspergillus niger liquid state fermentation, the following steps are included: 1, the preservation of bacteria strain of aspergillus niger activated, single bacterium colony after selecting activation is transferred to culture in the first culture medium and obtains the seed liquor of aspergillus niger, then seed liquor is transferred to and is spread cultivation to obtain fermentation of Aspergillus niger liquid in the fermentor containing the second culture medium；2, the preservation of bacteria strain JZZ of caproic acid bacteria is inoculated into third culture medium, Anaerobic culturel 48-360h under the conditions of 25-43 DEG C, so that caproic acid bacteria ferments to obtain caproic acid bacteria bacterium solution；3, fermentation mixed liquor is formed after mixing fermentation of Aspergillus niger liquid with esterification mixture material, the pH value for the mixed liquor that ferments is 3.0~5.0, and biological esterification liquid finally is made using liquid fermentation method.The present invention also promotes esterification using the compound enzyme that Aspergillus niger liquid state fermentation generates, improves the esterifying efficiency of liquid state fermentation fermentation system on the basis ofs adding caproic acid bacteria solution, esterified red yeast, yeast, pit mud etc..
Description
Technical field
The present invention relates to brewed spirit field more particularly to a kind of biological esterification liquid is prepared using Aspergillus niger liquid state fermentation
Method and its application.
Background technique
In conventional solid-state Luzhou-flavor liquo fermentation process, ethyl hexanoate is generated slowly, increases aroma in production, is led to
The technical measures such as " fermentation of reflux liquor ", " fermentation of folder mud ", " double bottom " can be often taken to improve containing for the esters Studies of The Aromatic Substances such as ethyl hexanoate
Amount, but these technical measures, in actual production there is also such as fermentation period is long, utilization rate of equipment and installations is low, the great high, base liquor of grain
The problems such as different miscellaneous taste is larger, distillation yield is low, female groove is second-rate.The generally existing own ester content of acid of Production of Luzhou-flavor Liquor is small at present
In the phenomenon that ethyl lactate, oneself cream are not protruded than numerous imbalances, giving off a strong fragrance style.
Chinese invention patent application CN106987500A discloses a kind of preparation method of esterifying liquid and its in wine brewing
Using: 52%w/w yellow water, 29%w/w liquor tailing, 10%w/w residual water, the compound caproic acid bacteria solution of 4%w/w, 2%w/w esterification is red
Song, 2%w/w high temperature Daqu, 1%w/w pit mud mix, and adjustment pH value is 3.5-3.8, then cultivate under the conditions of 32-35 DEG C
28-35d.The ester that the preparation method is mainly generated using the microbial metabolisms such as compound caproic acid bacteria solution, esterified red yeast, yeast, pit mud
Change enzyme, to promote esterification to generate the Ester based on ethyl hexanoate, but there are the esterifying efficiencies of esterifying liquid for this method
Lowly, the problems such as esterification cannot be promoted well.
Therefore, those skilled in the art, which is dedicated to developing, a kind of prepares biological esterification liquid using Aspergillus niger liquid state fermentation
Method is also answered using what Aspergillus niger liquid state fermentation generated on the basis ofs adding caproic acid bacteria solution, esterified red yeast, yeast, pit mud etc.
Synthase class promotes the esterification of fermentation system, improves the esterifying efficiency of liquid state fermentation system, is advantageously implemented wine brewing by-product
The comprehensive utilization of object.
Summary of the invention
In view of the above drawbacks of the prior art, the technical problem to be solved by the present invention is to how utilize microbial enzyme method
Improve the esterifying efficiency of liquid state fermentation system.
To achieve the above object, the present invention provides a kind of sides that biological esterification liquid is prepared using Aspergillus niger liquid state fermentation
Method, which comprises the following steps:
The preservation of bacteria strain of aspergillus niger is inoculated on solid-state slant medium and activates by step 1, bacterium colony after activation
Choosing colony is transferred in the first culture medium in group, and is placed in shaking table constant temperature incubation and is obtained the seed liquor of aspergillus niger, then will kind
Sub- liquid is transferred in the second culture medium, and fermentor spreads cultivation to obtain fermentation of Aspergillus niger liquid；
Step 2 activates the preservation of bacteria strain JZZ of caproic acid bacteria, the caproic acid bacteria colony inoculation after selecting activation to third
In culture medium, Anaerobic culturel 48-360h under the conditions of 25-43 DEG C, so that caproic acid bacteria ferments to obtain caproic acid bacteria bacterium solution, by caproic acid bacteria
Bacterium solution and yellow serofluid, liquor tailing, edible alcohol, yeast, fermented grain and pit mud are mixed to get esterification mixture material；
Step 3 forms fermentation mixed liquor, adjustment after mixing the fermentation of Aspergillus niger liquid in step 1 with esterification mixture material
The pH value of fermentation mixed liquor is 3.0-5.0, and biological esterification liquid finally is made using liquid fermentation method.
Further, esterification mixture material includes yellow serofluid 20-30 parts, 30-50 parts of liquor tailing, 5-10 parts of edible alcohol, greatly
It is 4-6 parts bent, 4-6 parts of fermented grain, 4-6 parts of pit mud, 1-3 parts and 1-3 parts of caproic acid bacteria bacterium solution of esterified red yeast.
Further, the mixed proportion of fermentation of Aspergillus niger liquid and esterification mixture material is 1:3~5, the temperature of liquid state fermentation
It is 30-35 DEG C, time 30-40d.
Further, constant temperature is 30 DEG C in step 1, and the revolving speed of shaking table is 120r/min, obtains the incubation time of seed liquor
For 48-72h, the temperature to spread cultivation is 30 DEG C -40 DEG C, time 48-72h, and the revolving speed of fermentor is 120r/min, and ventilatory capacity is
0.2m3/h。
Further, the temperature of Anaerobic culturel is 37 DEG C in step 2, time 192h.
Further, the first culture medium includes wheat bran 10-30g/L, glucose 20-30g/L, ammonium sulfate 10-20g/L, phosphorus
Sour hydrogen dipotassium 1-3g/L, epsom salt 0.5-1g/L, the pH value 4.0-6.0 of the first culture medium；Second culture medium includes wheat bran
40-50g/L, ammonium sulfate 10-30g/L, sodium chloride 1-4g/L, epsom salt 0.5-3g/L, the pH value 4.0- of the second culture medium
6.0。
Further, third culture medium includes sodium acetate 5.0g/L, yeast extract 1.0g/L, epsom salt 0.2g/
L, dipotassium hydrogen phosphate 0.4g/L, ammonium sulfate 0.5g/L, and in third culture medium be added third culture volume 2% second
Alcohol, the pH value of third culture medium are 7.0.
The present invention also provides a kind of methods of biological esterification liquid using Aspergillus niger liquid state fermentation preparation in Luzhou-flavor base
Application in wine production, which is characterized in that carry out biological esterification liquid in fermented grain fermentation process to fill cellar operation or and fermented grain
It carries out mixing and is distilled to get Luzhou-flavor base liquor.
Further, cellar operation is filled to comprise the following steps:
1, Fermented grains feeding pond is sealed by fermentation 15d；
2, the surrounding of the pond head mud of pit is excavated to expose fermented grain；
3, cellar was filled 1 time along pit surrounding with biological esterification liquid 15kg at interval of 1 day, each filling cellar terminates again at once will be exposed
Fermented grain covering pond head mud；
4, continuous to fill cellar 15d, then seal pit and fermented grain is allowed to continue the 30d that ferments, Luzhou-flavor base liquor is obtained to go out unstrained spirits distillation.
Further, the preservation of bacteria strain JZZ of caproic acid bacteria, it is commonly micro- to be preserved in China Committee for Culture Collection of Microorganisms
Bio-Centers, preservation date: 2015-12-29, deposit number: CGMCCNo.11948.
Compared with prior art, the present invention is also sharp on the basis ofs adding caproic acid bacteria solution, esterified red yeast, yeast, pit mud etc.
Promote esterification with the compound enzyme that Aspergillus niger liquid state fermentation generates, improves the esterifying efficiency of liquid state fermentation fermentation system；
Present invention application fills cellar mode, biological esterification liquid can be applied in fermentation process, has and promotes alcohol esterification
Effect；
The present invention can be such that base liquor quality improves rapidly with mixed fermented grain of steaming, and realize that common white spirit is converted to quality liquor, prominent
The main body fragrant and wine body of Luzhou-flavor liquo ethyl hexanoate are coordinated.
It is described further below with reference to technical effect of the attached drawing to design of the invention, specific structure and generation, with
It is fully understood from the purpose of the present invention, feature and effect.
Detailed description of the invention
Fig. 1 is the process that biological esterification liquid is prepared using Aspergillus niger liquid state fermentation of a preferred embodiment of the invention
Figure.
Specific embodiment
Multiple preferred embodiments of the invention are introduced below with reference to Figure of description, keep its technology contents more clear and just
In understanding.The present invention can be emerged from by many various forms of embodiments, and protection scope of the present invention not only limits
The embodiment that Yu Wenzhong is mentioned.
Herein presented caproic acid bacteria JZZ should be construed as a kind of strain of caproic acid bacteria, be preserved in Chinese microorganism strain
Preservation administration committee common micro-organisms center, preservation date: 2015-12-29, deposit number: CGMCCNo.11948；r/min
It is the unit of revolving speed, means rev/min；H is chronomere, means hour；D is the unit of time, means number of days；
m3The distance that/h is passed through within the unit time for finger gas liquid point, indicates cubic meter per hour；G/L is mass concentration list
Position；Kg is mass unit；%w/w is weight percent.
Biological esterification liquid is exactly to utilize microbial technique and fermentation engineering, using enzyme esterification method, by yellow serofluid,
In tail water etc. organic acid substance and alcohols material be converted under the action of lipase the multiple fragrance based on ethyl hexanoate at
The fragrant ester mixed liquor divided.
As shown in Figure 1, in the present embodiment, the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation, including it is following
Step:
Step 1 prepares fermentation of Aspergillus niger liquid: the preservation of bacteria strain of aspergillus niger is inoculated into potato dextrose agar
Solid-state slant medium on activated, select single bacterium colony in bacterium colony group after activation and be transferred to comprising the first culture medium
Shaking flask in, shaking flask is placed in shaking table culture, cultivation temperature is 30 DEG C, and the revolving speed of shaking table is 120r/min, incubation time 48-
72h (preferably 58h) obtains the seed liquor of aspergillus niger, then the seed liquor of 3%-5% mass fraction is taken to be transferred to containing the second training
In the fermentor for supporting base, it is preferred that the seed liquor of 5% mass fraction is taken to be transferred in the fermentor containing the second culture medium；Kind
Sub- liquid is in the fermenter by spreading cultivation to obtain fermentation of Aspergillus niger liquid, wherein and the temperature to spread cultivation is 30 DEG C -40 DEG C (preferably 30 DEG C),
The revolving speed of fermentor is 120r/min, and the ventilatory capacity of fermentor is 0.2m3/ h, fermentation time is 48- to seed liquor in the fermenter
72h (preferably 58h).
Step 2 prepares caproic acid fermented liquid: the preservation of bacteria strain JZZ of caproic acid bacteria being inoculated into third culture medium, in 25-
Anaerobic culturel 48-360h under the conditions of 43 DEG C, so that caproic acid bacteria ferments to obtain caproic acid bacteria bacterium solution；Preferably, the temperature of Anaerobic culturel
It is 37 DEG C, the time of Anaerobic culturel is 192h, by caproic acid bacteria bacterium solution and yellow serofluid, liquor tailing, edible alcohol, yeast, fermented grain and cellar
Mud is mixed to get esterification mixture material.
Step 3 prepares biological esterification liquid: by step 1 fermentation of Aspergillus niger liquid and esterification mixture material be proportionally
1:5 forms fermentation mixed liquor after being mixed, fermentation mixed liquor is formed after mixing, wherein esterification mixture material is yellow serofluid 20-
30 parts, 30-50 parts of liquor tailing, 5-10 parts of edible alcohol, 4-6 parts of yeast, 4-6 parts of fermented grain, 4-6 parts of pit mud, 1-3 parts of esterified red yeast and
1-3 parts of caproic acid bacteria bacterium solution, be 3.0~5.0 using the pH value that hydrochloric acid adjusts fermentation mixed liquor, will finally be sent out using liquid fermentation method
Ferment mixed liquor ferments 40d at 30-35 DEG C.
The first culture medium in the present embodiment includes wheat bran 10-30g/L, glucose 20-30g/L, ammonium sulfate 10-20g/L,
Potassium dihydrogen phosphate 1-3g/L, epsom salt 0.5-1g/L, the pH value 4.0-6.0 of the first culture medium；Second culture medium includes bran
Skin 40-50g/L, ammonium sulfate 10-30g/L, sodium chloride 1-4g/L, epsom salt 0.5-3g/L, the pH value of the second culture medium
4.0-6.0；Third culture medium includes sodium acetate 5.0g/L, yeast extract 1.0g/L, epsom salt 0.2g/L, phosphoric acid hydrogen two
Potassium 0.4g/L, ammonium sulfate 0.5g/L, and the ethyl alcohol of its volume 2% is added in third culture medium, utilize wine brewing yellow serofluid will
PH is adjusted to 7.0,121 DEG C of sterilizing 20min.
In the present embodiment, yellow serofluid is the by-product in Luzhou-flavor liquo brewing process, is dark-brown glue transparency liquid,
Appearance is thick, the flavor and fragrance substances such as alcohols rich in, acids, aldehydes, esters.It is also rich in organic in yellow serofluid
Sour especially lactic acid, acetic acid, caproic acid, butyric acid content is most abundant, is the optimal material for producing esterifying liquid.
Composition analysis result is shown in Table 1,
Constituent analysis testing result in 1 yellow serofluid of table
In the present embodiment, mature biological esterification liquid has the strong compound fragrance based on ethyl hexanoate, and appearance is in
It is yellow.The ingredient of mature biological esterification liquid is detected, the results are shown in Table shown in 2,
The analysis detection result of the mature esterifying liquid of table 2
In the present embodiment, mature biological esterification liquid is carried out to fill cellar operation, Luzhou-flavor during grain unstrained spirits is sealed by fermentation
Pit grain unstrained spirits enters pond and is sealed by fermentation 15d, the surrounding of pit pond head mud is excavated to expose fermented grain, at interval of 1d obtained by step c
Mature esterifying liquid 15kg fills cellar 1 time along pit surrounding, and filling cellar every time terminates again at once by exposed fermented grain covering pond head mud, continuously
Cellar 15d is filled, then seals and continues the 30d that ferments, Luzhou-flavor base liquor is taken out in unstrained spirits distillation out.To the Luzhou-flavor base obtained by filling cellar operation
Wine carries out sensory judgments and constituent analysis with base liquor is compareed, and the Luzhou-flavor base liquor cellar aroma flavoring in the present embodiment is strong, and wine body is plentiful, alcohol
Thickness, smell coordination, tail is neter, and compares base liquor cellar aroma flavoring and protrude, and wine body is pure and mild, and more plentiful, mellow, tail is net, the results are shown in Table 3,
The mature esterifying liquid of table 3 carries out filling the Luzhou-flavor base liquor constituent analysis testing result produced after the operation of cellar
In other embodiments, bottom pot is added in mature biological esterification liquid before fermented grain loaded steamer, then fermented grain uploading in rice steamer is carried out
String, which steams, obtains Luzhou-flavor base liquor, or mature biological esterification liquid is splashed in fermented grain and turning to be uniformly mixed and carry out mixed steam and obtain
Luzhou-flavor base liquor.
The preferred embodiment of the present invention has been described in detail above.It should be appreciated that the ordinary skill of this field is without wound
The property made labour, which according to the present invention can conceive, makes many modifications and variations.Therefore, all technician in the art
Pass through the available technology of logical analysis, reasoning, or a limited experiment on the basis of existing technology under this invention's idea
Scheme, all should be within the scope of protection determined by the claims.
Claims (10)
1. a kind of method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation, which comprises the following steps:
The preservation of bacteria strain of aspergillus niger is inoculated on solid-state slant medium and activates by step 1, in bacterium colony group after activation
Choosing colony is transferred in the first culture medium, and is placed in shaking table constant temperature incubation and is obtained the seed liquor of the aspergillus niger, then by institute
It states seed liquor to be transferred in the second culture medium, fermentor spreads cultivation to obtain fermentation of Aspergillus niger liquid；
Step 2 activates the preservation of bacteria strain JZZ of caproic acid bacteria, the caproic acid bacteria colony inoculation after selecting activation to third culture
In base, Anaerobic culturel 48-360h under the conditions of 25-43 DEG C, so that the caproic acid bacteria ferments to obtain caproic acid bacteria bacterium solution, by caproic acid bacteria
Bacterium solution and yellow serofluid, liquor tailing, edible alcohol, yeast, fermented grain and pit mud are mixed to get esterification mixture material；
Step 3 forms fermentation mixed liquor after mixing the fermentation of Aspergillus niger liquid in step 1 with esterification mixture material, described in adjustment
The pH value of fermentation mixed liquor is 3.0-5.0, and biological esterification liquid finally is made using liquid fermentation method.
2. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that described
Esterification mixture material include yellow serofluid 20-30 parts, 30-50 parts of liquor tailing, 5-10 parts of edible alcohol, 4-6 parts of yeast, 4-6 parts of fermented grain,
4-6 parts of pit mud, 1-3 parts of 1-3 parts of esterified red yeast and the caproic acid bacteria bacterium solution.
3. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation stated such as claim 1, which is characterized in that described black
The mixed proportion of Aspergillus fermentation broth and the esterification mixture material is 1:3~5, and the temperature of liquid state fermentation is 30-35 DEG C, and the time is
30-40d。
4. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that step
Constant temperature is 30 DEG C in 1, and the revolving speed of shaking table is 120r/min, and the incubation time for obtaining seed liquor is 48-72h, and the temperature to spread cultivation is
30 DEG C -40 DEG C, time 48-72h, the revolving speed of fermentor is 120r/min, ventilatory capacity 0.2m3/h。
5. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that step
The temperature of Anaerobic culturel is 37 DEG C in 2, time 192h.
6. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that described
First culture medium includes wheat bran 10-30g/L, glucose 20-30g/L, ammonium sulfate 10-20g/L, dipotassium hydrogen phosphate 1-3g/L, and seven
Water magnesium sulfate 0.5-1g/L, the pH value 4.0-6.0 of first culture medium；Second culture medium includes wheat bran 40-50g/L,
Ammonium sulfate 10-30g/L, sodium chloride 1-4g/L, epsom salt 0.5-3g/L, the pH value 4.0-6.0 of second culture medium.
7. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that described
Third culture medium includes sodium acetate 5.0g/L, yeast extract 1.0g/L, epsom salt 0.2g/L, dipotassium hydrogen phosphate 0.4g/
L, ammonium sulfate 0.5g/L, and the ethyl alcohol of the third culture volume 2%, the third is added in the third culture medium
The pH value of culture medium is 7.0.
8. a kind of method of the biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1 preparation is in Luzhou-flavor base
Wine production in application, which is characterized in that by the biological esterification liquid carried out in fermented grain fermentation process fill cellar operation or with
Fermented grain carries out mixing and is distilled to get the Luzhou-flavor base liquor.
9. raw in Luzhou-flavor base liquor using the method for the biological esterification liquid of Aspergillus niger liquid state fermentation preparation as claimed in claim 8
Application in production, which is characterized in that filling cellar operation comprises the following steps:
1, the Fermented grains feeding pond is sealed by fermentation 15d；
2, the surrounding of the pond head mud of the pit is excavated to expose fermented grain；
3, cellar was filled 1 time along the pit surrounding with the biological esterification liquid 15kg at interval of 1 day, filling cellar every time terminates at once will again
The exposed fermented grain covers the pond head mud；
4, continuous to fill cellar 15d, then seal pit and the fermented grain is allowed to continue the 30d that ferments, the Luzhou-flavor base is obtained to go out unstrained spirits distillation
Wine.
10. the method for preparing biological esterification liquid using Aspergillus niger liquid state fermentation as described in claim 1, which is characterized in that institute
The preservation of bacteria strain JZZ for stating caproic acid bacteria, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation is compiled
Number: CGMCC No.11948.
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|---|---|---|---|---|
| CN110527635A (en) * | 2019-09-29 | 2019-12-03 | 北京工商大学 | One plant of aspergillus niger cultural method and its catalyzing and synthesizing the application in liquor flavor ester |
| CN110652748A (en) * | 2019-10-22 | 2020-01-07 | 江南大学 | Method for separating and enriching flavor substances of white spirit from yellow seriflux for brewing wine |
| CN114806969A (en) * | 2022-06-06 | 2022-07-29 | 江苏汤沟两相和酒业有限公司 | Acinetobacter pitteus, esterification liquid, method for treating yellow serofluid and application |
| CN114806969B (en) * | 2022-06-06 | 2023-09-15 | 江苏汤沟两相和酒业有限公司 | Acinetobacter petite, esterification liquid, method for treating yellow serofluid and application |
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