CN110591852A - Application and method of eurotium scherzeri in microbial rejuvenation of pit mud of white spirit cellar - Google Patents
Application and method of eurotium scherzeri in microbial rejuvenation of pit mud of white spirit cellar Download PDFInfo
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Abstract
The invention belongs to the technical field of biology, and relates to application of eurotium scherzerianum in microbial rejuvenation of pit mud of a white spirit pit and a method thereof. Yellow water and feints generated in the solid state fermentation of white spirit are used as main fermentation substrates, sterilized softened water containing eupatorium adenophorum yeast and esterified red yeast rice is added, the mixture is uniformly stirred, air is introduced, and fermentation liquor containing eupatorium adenophorum is obtained by fermenting for 25 to 45 days at the temperature of between 30 and 35 ℃ and is inoculated into pit mud or fermented grains of a pit for fermentation, so that the number of anaerobic microorganisms in the pit mud can be increased, and the rejuvenation of the pit mud microorganisms of the pit is effectively realized; meanwhile, the flavor of pit mud in a pit can be improved, the fermentation environment of fermented grains is improved, the quality of the Gujing group strong-flavor liquor is further improved, the flavor of the liquor is improved, and the Gujing-flavor liquor with tea flavor and fruit flavor can be brewed.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to application of eurotium scherzerianum in microbial rejuvenation of pit mud of a white spirit pit and a method thereof.
Background
In the process of brewing white spirit, the types and the quantity of beneficial microorganisms in the pit and pit mud have important influence on the acquisition of high-quality wine. However, the aging of the pit and the degradation of pit mud microorganisms are problems which need to be solved urgently in the white spirit industry at present. At present, white spirit manufacturers mostly use supplementary inorganic salt and caproic acid bacteria to age and rejuvenate pit mud microorganisms, but the effect is very little.
The Chinese invention patent application (CN106085741A) provides a culture method of a pit mud culture solution with a strong aromatic Chinese spirit, wherein the pit mud culture solution prepared by utilizing various components such as high-quality old pit mud, an extraction mixture, Daqu, yellow clay, growth factors, wine-making byproducts and the like is rich in beneficial wine-making microorganisms and nutritional components suitable for brewing the Chinese spirit. The pit mud culture solution is added in the pit mud making, pit maintenance and pit protection and pit rejuvenation, so that beneficial brewing microorganisms can rapidly grow and propagate in the pit mud and pit, and the deterioration of the pit mud is delayed.
In the prior art, the Shewanella is reported to exist in strong-flavor, Maotai-flavor and fen-flavor liquor Daqu. In the invention, a plurality of Ascomycetes Shewanella (such as strain CICC41587(M9), strain M6 and the like) are obtained by screening in the previous research, and the application of the Ascomyces Shewanella in the production of white spirit is intensively researched, and in the patent application (CN10967855A), the strain CICC41587(M9) is enhanced in the production of Daqu in white spirit in a bacterial powder form, and the Daqu has a tea flavor after being mature.
Disclosure of Invention
The invention mainly aims to provide a new application of eurotium scherzeri in the rejuvenation of pit mud microorganisms of a white spirit pit, and provide a rejuvenation method, so that the rejuvenation of the pit mud microorganisms of the pit is effectively realized, and the quality of the strong aromatic white spirit is improved.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention aims to provide application of eurotium scherzeri in rejuvenation of microorganisms in pit mud of a white spirit pit.
In the above application, preferably, the fermentation liquid containing the Eurotium scherzeylanicum is applied to the rejuvenation of microorganisms in pit mud of a white spirit cellar.
In the above application, preferably, the Eurotium ascheri is the strain Eurotium ascheri CICC41587(M9) (Eurotium chevalieri) with the preservation number of CGMCC No. 14623.
The second purpose of the invention is to provide a method for rejuvenating microorganisms in pit mud of a white spirit pit by using eurotium aschersonii, which is to introduce fermentation liquor containing the eurotium aschersonii into pit mud or fermented grains for fermentation.
In the method, preferably, the inoculation amount of the fermentation liquor is 1-10% of the weight of pit mud or grain; further preferably, the inoculation amount of the fermentation liquor is 1-5% of the weight of pit mud or grain.
In the above method, preferably, the culture method of the fermentation broth containing eurotium shikawari comprises: adopting yellow water and feints generated in solid state fermentation of Chinese liquor as main fermentation substrates, adding sterilized softened water containing Ascomyces sieboldii Daqu and esterified red yeast, stirring, introducing air, and fermenting at 30-35 deg.C for 25-45 days.
In the above method, preferably, the mass ratio of yellow water, feints, softened water, eurotium schlegelii containing Daqu and esterified red yeast rice is 30-40:40-50:15-20:1-5: 0.1-1; further preferably, the mass ratio of the yellow water, the feints, the softened water, the eurotium schlegelii containing Daqu and the esterified red yeast rice is 39:41:17-19:1-2: 0.4-0.6.
In the above method, preferably, the yellow water is extracted on the same day, has pH of 4.0-4.5, and bacillus number of 0.7-1 × 108Per ml; the standard alcohol content of the feints is 20-21 degrees; the softened water is the slurry water for blending the white spirit.
In the above method, preferably, the eudesmium-containing yeast is prepared by a direct preparation method or an enhanced method of eudesmium.
The direct preparation method comprises the following steps: the time containing the eurotium scherzerianum Daqu is 30 days after the solar term of Qingming or 30 days after the solar term of late autumn; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, sieving the powder by a 30-40-mesh sieve, and finely dividing the powder to 40-50 percent of the total weight; adding water and stirring, wherein the water addition amount is 40-50% of the weight of the yeast material; mechanically treading the koji, wrapping into koji blocks, putting into a room for koji setting, scattering a layer of fresh rice hulls with the thickness of 2-3cm on the ground, placing the koji blocks on the ground, wherein the gaps between the koji blocks are 2-3cm, and preserving the heat of the koji blocks by using dry straws with the thickness of 10-15 cm; turning over the koji when the temperature of the koji blocks reaches 58-60 ℃, and turning over the koji for 2 times; closing the house 10-15 days after the secondary turning of the yeast, continuously culturing the yeast blocks for 7-10 days after the temperature of the yeast blocks approaches the room temperature, keeping the moisture of the yeast blocks to 10% -15%, and putting the yeast blocks out of the house and storing the yeast blocks in a warehouse, wherein the yeast cores have obvious yellow bacterial plaque of aschersonia aleyrodis.
In the preparation process of the method, 30 days after the solar term of Qingming or 30 days after the solar term of late autumn are the optimal culture period, and the Daqu containing golden yellow bacterial plaques can hardly be produced at other times; in addition, after the temperature of the koji block is close to room temperature, the koji block needs to be cultured for 7-10 days to produce the Daqu with the koji core having the obvious golden yellow bacterial plaque of aschersonia aleurida.
The eustachian holangium reinforcing method includes the following steps:
(1) preparing xiewasan bursin fungus powder: inoculating eurotium scherzeri into a 4% glucose MEB liquid medium, and culturing at 28 ℃ for 5-7 d to obtain an activated culture solution; mixing bran and distilled water according to a mass ratio of 1: 1-5, sterilizing at 121 ℃ for 40min, and sterilizing once again at night according to the same conditions to obtain a bran culture medium; inoculating the prepared activation culture solution into a bran culture medium according to the proportion of 1: 4-10 ml/g, and culturing for 10-14 days at 28 ℃; transferring the bran culture into a rotary evaporator rotary bottle of 20L, and vacuum drying in water bath at 40 deg.C for 26 h; crushing for 30 seconds by using a miniature plant crusher, and sieving by using a 20-mesh sieve to obtain aschersonia aleyrodis powder;
(2) eustachian eurotium reinforcing yeast: dissolving the eurotium aschersonii powder prepared in the step (1) in distilled water containing 2-3% of sodium chloride according to the mass-volume ratio of 10-15: 1g/L to obtain a stirring solution; grinding wheat into slices, grinding wheat kernels into powder, sieving with a 30-40 mesh sieve, finely dividing by 40-50% of the total weight, adding stirring liquid, and stirring, wherein the adding amount of the stirring liquid is 40-50% of the weight of the yeast material; mechanically treading the koji, wrapping into koji blocks, putting into a room for koji setting, scattering a layer of fresh rice hulls with the thickness of 2-3cm on the ground, placing the koji blocks on the ground, wherein the gaps between the koji blocks are 2-3cm, and preserving the heat of the koji blocks by using dry straws with the thickness of 10-15 cm; turning over the koji when the temperature of the koji blocks reaches 58-60 ℃, and turning over the koji for 2 times; closing the house 10-15 days after the secondary turning over of the yeast, keeping the moisture of the yeast blocks to 14% -15% after the temperature of the yeast blocks approaches the room temperature, taking the yeast blocks out of the house and warehousing the yeast blocks, wherein the yeast cores have obvious yellow bacterial plaques of aschersonia aleyrodis.
In the above method, preferably, in the preparation method of the eupatorium adenophorum strengthened yeast, the eupatorium adenophorum is CICC41587(M9) (Eurotium chevalieri) with the collection number of CGMCC No. 14623.
Compared with the prior art, the invention has the following beneficial effects:
in the process of researching the rejuvenation of the pit mud microorganisms of the white spirit pit, the inventor unexpectedly discovers that the rejuvenation effect of the fermentation liquor obtained by mixing and fermenting the golden flower yeast (namely the yeast containing golden yellow bacterial plaque prepared 30 days after the solar term of Qingming or 30 days after the solar term of autumn) with yellow water, wine tails and esterified red yeast is obviously better than that of the yeast of the general pouring pit by the microorganism detection center of the Chinese food fermentation institute. Further, after the yeast is strengthened by the eurotium scherzeri CICC41587(M9) obtained through earlier stage research and screening, the rejuvenation effect of the obtained fermentation liquor on the white wine cellar pit mud microorganisms is further improved.
The invention further enriches the utilization value of the Eurotium ascheri in the white spirit production fermentation industry, and provides possibility for large-scale application of the bacterial strain of Eurotium ascheri CICC41587(M9) (Eurotium chevalieri) in the white spirit fermentation process.
Mixing and fermenting the yeast for making hard liquor containing the eupatorium Chinese, yellow water, feints, esterified red yeast rice and softened water, wherein the obtained fermentation liquor has the mixed flavor of tea aroma, fruit aroma and sweet aroma and is inoculated into pit mud of a pit, so that the number of anaerobic microorganisms in the pit mud can be increased, and the rejuvenation of the microorganisms in the pit mud of the pit can be effectively realized; meanwhile, the flavor of pit mud in a pit can be improved, the fermentation environment of fermented grains is improved, the quality of the Gujing group strong-flavor liquor is further improved, the flavor of the liquor is improved, and the Gujing-flavor liquor with tea flavor and fruit flavor can be brewed.
The method has simple steps, easily obtained and cheap materials, low investment cost and easy popularization and application.
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FIG. 1 is a flow chart of the method for culturing a fermentation broth containing Ascomyces sieboldii according to the present invention.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, and/or combinations thereof, unless the context clearly indicates otherwise.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
The Eurotium ascheri strain CICC41587(M9) used in the invention is obtained by early screening and is disclosed in the patent application (CN 109679855A). The strain is preserved in 2017, 09 and 14 days to China general microbiological culture Collection center (China Beijing China academy of sciences microbial research institute), and the preservation number is as follows: CGMCC No.14623, taxonomic name: eurotium scherrieri (Eurotium chevalieri). The esterified red yeast rice can be purchased from commercial sources, has red or light red color and no peculiar smell, and the esterification force is more than or equal to 23 mg/g.100 h.
Example 1 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps:
390 kg of yellow water, 410 kg of feints and 170 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 10 kg of crushed eupatorium-containing yeast and 4 kg of crushed esterified red yeast are added, air is introduced for 10 minutes, the mixture is uniformly stirred, and the fermentation is carried out for 30 days at the temperature of 32 ℃.
After the fermentation is finished, the color of the fermentation liquor is brownish red, and the fermentation liquor has tea aroma, flower and fruit aroma and ester sweet taste, sampling, heating and distilling are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.1538g/L of ethyl acetate, 0.2192g/L of ethyl butyrate, 7.1152g/L of ethyl lactate and 1.6357g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The preparation method of the Eurotium sieboldii containing Daqu by adopting a direct preparation method comprises the following steps: the preparation time of the eustachyosis-containing Daqu is 30 days after the Ming season or 30 days after the early autumn season; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, and sieving the powder by a 40-mesh sieve, wherein the fine powder accounts for 40 percent of the total weight; adding water and mixing, wherein the water adding amount is 40% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of about 15cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 58 ℃, and turning over the koji for 2 times; closing the house 15 days after the secondary turning of the yeast, continuously culturing the yeast blocks for 7 days after the temperature of the yeast blocks approaches the room temperature, keeping the moisture of the yeast blocks to 15 percent, and taking the yeast blocks out of the house and warehousing the yeast blocks for storage, wherein the yeast blocks have obvious yellow bacterial plaques of aschersonia aleyrodis.
Example 2 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 390 kg of yellow water, 410 kg of feints and 180 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 5 kg of crushed Daqu containing the Eurotium schlegelii CICC41587(M9) and 5 kg of crushed esterified red yeast rice are added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 30 days at the temperature of 32 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, the fermentation liquor has more obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.1918g/L of ethyl acetate, 0.2852g/L of ethyl butyrate, 7.7480g/L of ethyl lactate and 1.9259g/L of ethyl caproate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The preparation method of the Eurotium sieboldii containing Daqu by adopting a direct preparation method comprises the following steps: the preparation time of the eustachyosis-containing Daqu is 30 days after the Ming season or 30 days after the early autumn season; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, and sieving the powder by a 40-mesh sieve, wherein the fine powder accounts for 40 percent of the total weight; adding water and mixing, wherein the water adding amount is 42% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 12cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 60 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, continuously culturing for 8 days after the temperature of the yeast block approaches to the room temperature, keeping the moisture of the yeast block to 12 percent, and taking the yeast block out of the house and warehousing the yeast block for storage, wherein the yeast block has obvious yellow bacterial plaque of aschersonia aleyrodis.
Example 3 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 390 kg of yellow water, 410 kg of feints and 190 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 20 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 6 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 30 days at the temperature of 32 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.1978g/L of ethyl acetate, 0.2913g/L of ethyl butyrate, 7.7516g/L of ethyl lactate and 1.9548g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The preparation method of the Eurotium sieboldii containing Daqu by adopting a direct preparation method comprises the following steps: the preparation time of the eustachyosis-containing Daqu is 30 days after the Ming season or 30 days after the early autumn season; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, and sieving the powder by a 40-mesh sieve, wherein the fine powder accounts for 40 percent of the total weight; adding water and mixing, wherein the water adding amount is 45% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 10cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 59 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, continuously culturing for 8 days after the temperature of the yeast block approaches to the room temperature, keeping the moisture of the yeast block to 10 percent, and taking the yeast block out of the house and warehousing the yeast block for storage, wherein the yeast block has obvious yellow bacterial plaque of aschersonia aleyrodis.
Example 4 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 300 kg of yellow water, 400 kg of feints and 150 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 10 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 1 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 45 days at the temperature of 30 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.1813g/L of ethyl acetate, 0.2176g/L of ethyl butyrate, 7.1368g/L of ethyl lactate and 1.6391g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The preparation method of the Eurotium sieboldii containing Daqu by adopting a direct preparation method comprises the following steps: the preparation time of the eustachyosis-containing Daqu is 30 days after the Ming season or 30 days after the early autumn season; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, and sieving the powder by a 40-mesh sieve, wherein the fine powder accounts for 40 percent of the total weight; adding water and mixing, wherein the water adding amount is 45% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 10cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 59 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, continuously culturing for 8 days after the temperature of the yeast block approaches to the room temperature, keeping the moisture of the yeast block to 10 percent, and taking the yeast block out of the house and warehousing the yeast block for storage, wherein the yeast block has obvious yellow bacterial plaque of aschersonia aleyrodis.
Example 5 culture method of fermentation broth containing Ascomyces sieboldii
The method comprises the following specific operation steps: 400 kg of yellow water, 500 kg of feints and 200 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 50 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 10 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 25 days at the temperature of 35 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.2243g/L of ethyl acetate, 0.3247g/L of ethyl butyrate, 9.2371g/L of ethyl lactate and 2.1753g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The preparation method of the eurotium scherzeri-containing Daqu is the same as that of example 4.
Example 6 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 400 kg of yellow water, 500 kg of feints and 200 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 50 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 10 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 25 days at the temperature of 35 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.2461g/L of ethyl acetate, 0.3584g/L of ethyl butyrate, 9.9238g/L of ethyl lactate and 2.6185g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
Adopts a eustachian holangium reinforcing method, which comprises the following steps:
(1) preparing xiewasan bursin fungus powder: inoculating Eurotium scherzerianum CICC41587(M9) (Eurotium chevalieri) into 4% glucose MEB liquid culture medium, and culturing at 28 deg.C for 7d to obtain activated culture solution; mixing bran and distilled water according to a mass ratio of 1:1, sterilizing at 121 ℃ for 40min, and sterilizing once again at the same condition overnight to obtain a bran culture medium; inoculating the prepared activation culture solution into a bran culture medium according to the proportion of 1:4ml/g, and culturing for 14d at 28 ℃; transferring the bran culture into a rotary evaporator rotary bottle of 20L, and vacuum drying in water bath at 40 deg.C for 26 h; crushing for 30 seconds by using a miniature plant crusher, and sieving by using a 20-mesh sieve to obtain aschersonia aleyrodis powder;
(2) eustachian eurotium reinforcing yeast: dissolving the eurotium aschersonii powder prepared in the step (1) in distilled water containing 2% of sodium chloride according to the mass volume ratio of 10:1g/L to obtain a stirring solution; grinding wheat into slices, grinding wheat cores into powder, sieving with a 40-mesh sieve, finely dividing to 45% of the total weight, adding stirring liquid, and stirring, wherein the adding amount of the stirring liquid is 45% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 10cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 59 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, keeping the moisture of the yeast blocks to 14 percent after the temperature of the yeast blocks approaches to the room temperature, taking the yeast blocks out of the house and warehousing the yeast blocks, wherein the yeast blocks have obvious yellow bacterial plaque of aschersonia aleyrodis.
Example 7 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 400 kg of yellow water, 500 kg of feints and 200 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 50 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 10 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 25 days at the temperature of 35 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.2302g/L of ethyl acetate, 0.3356g/L of ethyl butyrate, 9.7562g/L of ethyl lactate and 2.4257g/L of ethyl caproate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
Adopts a eustachian holangium reinforcing method, which comprises the following steps:
(1) preparing xiewasan bursin fungus powder: inoculating Eurotium scherzerianum CICC41587(M9) (Eurotium chevalieri) into 4% glucose MEB liquid culture medium, and culturing at 28 deg.C for 5d to obtain activated culture solution; mixing bran and distilled water according to a mass ratio of 1:5, sterilizing at 121 ℃ for 40min, and sterilizing once again at the same condition overnight to obtain a bran culture medium; inoculating the prepared activation culture solution into a bran culture medium according to the proportion of 1:10ml/g, and culturing for 10 days at 28 ℃; transferring the bran culture into a rotary evaporator rotary bottle of 20L, and vacuum drying in water bath at 40 deg.C for 26 h; crushing for 30 seconds by using a miniature plant crusher, and sieving by using a 20-mesh sieve to obtain aschersonia aleyrodis powder;
(2) eustachian eurotium reinforcing yeast: dissolving the eurotium aschersonii powder prepared in the step (1) in distilled water containing 2% of sodium chloride according to the mass volume ratio of 10:1g/L to obtain a stirring solution; grinding wheat into slices, grinding wheat cores into powder, sieving with a 40-mesh sieve, finely dividing to 45% of the total weight, adding stirring liquid, and stirring, wherein the adding amount of the stirring liquid is 45% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 10cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 59 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, keeping the moisture of the yeast blocks to 14 percent after the temperature of the yeast blocks approaches to the room temperature, taking the yeast blocks out of the house and warehousing the yeast blocks, wherein the yeast blocks have obvious yellow bacterial plaque of aschersonia aleyrodis.
Example 8 culture method of fermentation broth containing Eurotium scherzeylanicum
The method comprises the following specific operation steps: 400 kg of yellow water, 500 kg of feints and 200 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 50 kg of crushed Daqu containing the aschersonia alexiva CICC41587(M9) is added, 10 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 25 days at the temperature of 35 ℃.
After fermentation is finished, the color of fermentation liquor is brownish red, and the fermentation liquor has obvious tea aroma, flower and fruit aroma and ester sweet taste, sampling and heating distillation are carried out, 100ml of distillate is obtained by steaming every 500ml of fermentation liquor, and the contents of ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are as follows after analysis by a gas chromatograph (GC-6890A Lunan instrument): 0.2612g/L of ethyl acetate, 0.3978g/L of ethyl butyrate, 9.8124g/L of ethyl lactate and 2.5493g/L of ethyl hexanoate.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
Adopts a eustachian holangium reinforcing method, which comprises the following steps:
(1) preparing xiewasan bursin fungus powder: inoculating Eurotium scherzerianum CICC41587(M9) (Eurotium chevalieri) into 4% glucose MEB liquid culture medium, and culturing at 28 deg.C for 5d to obtain activated culture solution; mixing bran and distilled water according to a mass ratio of 1:5, sterilizing at 121 ℃ for 40min, and sterilizing once again at the same condition overnight to obtain a bran culture medium; inoculating the prepared activation culture solution into a bran culture medium according to the proportion of 1:10ml/g, and culturing for 10 days at 28 ℃; transferring the bran culture into a rotary evaporator rotary bottle of 20L, and vacuum drying in water bath at 40 deg.C for 26 h; crushing for 30 seconds by using a miniature plant crusher, and sieving by using a 20-mesh sieve to obtain aschersonia aleyrodis powder;
(2) eustachian eurotium reinforcing yeast: dissolving the eurotium aschersonii powder prepared in the step (1) into distilled water containing 2% of sodium chloride according to the mass-volume ratio of 15:1g/L to obtain a stirring solution; grinding wheat into slices, grinding wheat cores into powder, sieving with a 40-mesh sieve, finely dividing to 45% of the total weight, adding stirring liquid, and stirring, wherein the adding amount of the stirring liquid is 45% of the weight of the yeast material; mechanically treading the yeast to make the yeast block into a package yeast shape, wherein the yeast block is 33cm long, 20cm wide, 8cm high and 2cm high; cleaning the koji house, ventilating and airing for 3 days before putting the koji house into the house, then scattering a layer of fresh rice husks with the thickness of 2-3cm on the ground, putting the koji blocks in a shape of a Chinese character of a finished product from north to south, putting the koji blocks in a gap of 2-3cm, putting 2 layers in total, after finishing yeast installation, preserving heat by using dry straws with the thickness of 10cm on the koji blocks, closing doors and windows, preserving heat and preserving moisture; turning over the koji when the temperature of the koji blocks reaches 59 ℃, and turning over the koji for 2 times; closing the house after 10 days after the secondary turning over of the yeast, keeping the moisture of the yeast blocks to 14 percent after the temperature of the yeast blocks approaches to the room temperature, taking the yeast blocks out of the house and warehousing the yeast blocks, wherein the yeast blocks have obvious yellow bacterial plaque of aschersonia aleyrodis.
Comparative example 1 culture method of fermentation broth
The method comprises the following specific operation steps: 400 kg of yellow water, 500 kg of feints and 200 kg of softened water are pumped into a fermentation pottery jar with the capacity of 1000 liters, 50 kg of crushed Daqu is added, 10 kg of crushed esterified red yeast rice is added, air is introduced for 10 minutes, the mixture is stirred uniformly, and the mixture is fermented for 25 days at the temperature of 35 ℃.
The yellow water is fresh yellow water extracted on the same day, and has brown yellow color, transparency, sour and astringent taste, sweet taste, suspension silk, pH value of about 4.0, and bacillus amount of 0.7-1.0 × 108One per ml. The used feints have a standard alcohol content of 20-21 °, the used softened water is the slurry water used for blending the white spirit, the used esterified red yeast rice is commercially available, the color is red or light red, no peculiar smell exists, and the esterification force is more than or equal to 23 mg/g.100 h.
The used yeast is a general well-wrenching yeast.
Example 9 microbial rejuvenation of Artificial pit mud
Respectively culturing by using the methods of examples 5 and 8 and comparative example 1 to obtain fermentation liquor 1, fermentation liquor 2 and fermentation liquor 3, respectively adding the three fermentation liquors into newly prepared artificial pit mud according to the proportion of 3% of the weight of the pit mud (respectively obtaining pit mud 1, pit mud 2 and pit mud 3), uniformly mixing by using a grab bucket, sealing, fermenting for 30 days at room temperature, simultaneously sampling and testing with control pit mud without any fermentation liquor, wherein the test results are shown in the following table 1:
TABLE 1
As can be seen from table 1, compared with the control pit mud sample, the pit mud samples 1 and 2 have fragrant smell, tea fragrance and flower and fruit fragrance, and have the smell of mature old pit mud in the fermentation pit, so that the amounts of moisture, ammonium nitrogen and anaerobic bacteria are obviously increased, the pH value is slightly reduced, the amount of aerobic bacteria is obviously reduced, and the effect of rejuvenating the pit mud microorganisms is achieved; compared with pit mud 3, the pit mud 1 and pit mud 2 samples have tea aroma and flower and fruit aroma, the amount of water, ammonium nitrogen and anaerobic bacteria is also obviously increased, the amount of aerobic bacteria is obviously reduced, the rejuvenation effect of the fermentation liquor obtained by culturing in the embodiments 5 and 8 on pit mud microorganisms is obviously more obvious, and meanwhile, the rejuvenation effect of the fermentation liquor obtained by culturing in the embodiment 8 on pit mud microorganisms is better than that of the fermentation liquor obtained in the embodiment 5. Therefore, the Eurotium scherzeylanicum can be beneficial to the rejuvenation of white spirit pit mud microorganisms.
Example 10 rejuvenation of microorganisms in pit mud of wine cellar
The cellar wall cellar mud that will make wine the cellar for storing things pond has ageing phenomenon shovels out, and the sample chemical examination utilizes the zymotic fluid of embodiment 8 preparation, adds the zymotic fluid according to the proportion of cellar mud weight 3% in the cellar wall cellar for storing things mud, and the manual work is stepped on evenly, fills back in the cellar for storing things pond cellar for storing things wall, and floating is smooth, and the grain unstrained spirits goes into the cellar for storing things, produces wine fermentation, and after 60 days of fermentation, it evaporates wine to go out the cellar for storing things, and cellar wall cellar mud sample chemical examination, the chemical examination result is shown in:
TABLE 2
As shown in the table 2, compared with the aged pit mud sample, the test pit mud sample has tea fragrance, flower and fruit fragrance and mature old pit mud smell, the amounts of water, ammonium nitrogen and anaerobic bacteria are obviously increased, the PH value is slightly reduced, the amount of aerobic bacteria is obviously reduced, and the effect of rejuvenating the pit mud microorganisms is achieved.
Example 11 rejuvenation of microbial fermentation in fermented grains
And (3) adding the fermentation liquor prepared in the example 8 into fermented grains in a cellar according to 1% of the weight of grains added into the cellar, performing solid state fermentation for 60 days, taking out the cellar, and steaming the wine, wherein the fermentation liquor is added into a bottom pot according to 100 kg/steamer during wine steaming.
Selecting a wine sample of which the reference pit and the test pit are steamed for the same time, and analyzing the wine sample by a gas chromatograph (GC-6890A Lunan instrument) to obtain an analysis result shown in the following 3:
TABLE 3
From the table 3, compared with the reference pit wine sample, the wine sample of the test pit has tea aroma and fruit aroma, is sweet in taste and has a clean aftertaste, the contents of main ester substances such as ethyl acetate, ethyl butyrate, ethyl lactate and ethyl caproate are increased, the ratio of the ethyl acetate to the ethyl caproate is reduced from 2.1520 to 1.7191, the relative content of the ethyl caproate is increased, the quality of the raw wine is improved, and the Guotungang fragrant white wine with the tea aroma and the fruit aroma is obtained.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. Application of Ascomycetes Shewanella in microorganism rejuvenation of pit mud of a white spirit pit.
2. The application of the fermentation liquor containing the eurotium scherzeri as claimed in claim 1, wherein the fermentation liquor containing the eurotium scherzeri is applied to rejuvenation of microorganisms in pit mud of a white spirit pit.
3. The use according to claim 1 or 2, wherein the Eurotium ascheri is the strain Eurotium ascheri CICC41587(M9) (Eurotium chevalieri) with a accession number CGMCC No. 14623.
4. A method for rejuvenating microorganisms in pit mud of white spirit by using Ascomyces ascheri is characterized in that fermentation liquor containing Ascomyces ascheri is inoculated into pit mud or fermented grains for fermentation.
5. The method according to claim 4, wherein the inoculation amount of the fermentation liquor is 1% -10% of the weight of pit mud or grain charge; preferably, the inoculation amount of the fermentation liquor is 1-5% of the weight of pit mud or grain.
6. The method of claim 4, wherein the culture of the fermentation broth comprising Ascomyces sieboldii comprises: adopting yellow water and feints generated in solid state fermentation of Chinese liquor as main fermentation substrates, adding sterilized softened water containing Ascomyces sieboldii Daqu and esterified red yeast, stirring, introducing air, and fermenting at 30-35 deg.C for 25-45 days.
7. The method according to claim 6, wherein the mass ratio of yellow water, feints, softened water, eurotium schlegelii containing Daqu and esterified red yeast rice is 30-40:40-50:15-20:1-5: 0.1-1; preferably, the mass ratio of yellow water, feints, softened water, the eurotium schwannioides containing Daqu and the esterified red yeast rice is 39:41:17-19:1-2: 0.4-0.6.
8. The method according to claim 6, wherein the yellow water is extracted on the same day, has pH of 4.0-4.5, and bacillus number of 0.7-1 x 108Per ml; the standard alcohol content of the feints is 20-21 degrees; the softened water is the slurry water for blending the white spirit.
9. The method of claim 6, wherein the Daqu containing Ascomyces sieboldii is prepared by a direct preparation method or by an Ascomyces sieboldii enhanced Daqu;
the direct preparation method comprises the following steps: the time containing the eurotium scherzerianum Daqu is 30 days after the solar term of Qingming or 30 days after the solar term of late autumn; taking wheat as a starter propagation raw material, grinding the wheat into slices, grinding wheat cores into powder, sieving the powder by a 30-40-mesh sieve, and finely dividing the powder to 40-50 percent of the total weight; adding water and stirring, wherein the water addition amount is 40-50% of the weight of the yeast material; mechanically treading the koji, wrapping into koji blocks, putting into a room for koji setting, scattering a layer of fresh rice hulls with the thickness of 2-3cm on the ground, placing the koji blocks on the ground, wherein the gaps between the koji blocks are 2-3cm, and preserving the heat of the koji blocks by using dry straws with the thickness of 10-15 cm; turning over the koji when the temperature of the koji blocks reaches 58-60 ℃, and turning over the koji for 2 times; closing the house 10-15 days after the secondary turning of the yeast, continuously culturing the yeast blocks for 7-10 days after the temperature of the yeast blocks approaches the room temperature, keeping the moisture of the yeast blocks to 10% -15%, and enabling the yeast cores to have obvious golden yellow bacterial plaques of aschersonia aleyrodis, and taking the yeast blocks out of the house and warehousing for storage;
the preparation method of the eustachian holboensis reinforced Daqu comprises the following steps:
(1) preparing xiewasan bursin fungus powder: inoculating eurotium scherzeri into a 4% glucose MEB liquid medium, and culturing at 28 ℃ for 5-7 d to obtain an activated culture solution; mixing bran and distilled water according to a mass ratio of 1: 1-5, sterilizing at 121 ℃ for 40min, and sterilizing once again at night according to the same conditions to obtain a bran culture medium; inoculating the prepared activation culture solution into a bran culture medium according to the proportion of 1: 4-10 ml/g, and culturing for 10-14 days at 28 ℃; transferring the bran culture into a rotary evaporator rotary bottle of 20L, and vacuum drying in water bath at 40 deg.C for 26 h; crushing for 30 seconds by using a miniature plant crusher, and sieving by using a 20-mesh sieve to obtain aschersonia aleyrodis powder;
(2) eustachian eurotium reinforcing yeast: dissolving the eurotium aschersonii powder prepared in the step (1) in distilled water containing 2-3% of sodium chloride according to the mass-volume ratio of 10-15: 1g/L to obtain a stirring solution; grinding wheat into slices, grinding wheat kernels into powder, sieving with a 30-40 mesh sieve, finely dividing by 40-50% of the total weight, adding stirring liquid, and stirring, wherein the adding amount of the stirring liquid is 40-50% of the weight of the yeast material; mechanically treading the koji, wrapping into koji blocks, putting into a room for koji setting, scattering a layer of fresh rice hulls with the thickness of 2-3cm on the ground, placing the koji blocks on the ground, wherein the gaps between the koji blocks are 2-3cm, and preserving the heat of the koji blocks by using dry straws with the thickness of 10-15 cm; turning over the koji when the temperature of the koji blocks reaches 58-60 ℃, and turning over the koji for 2 times; closing the house 10-15 days after the secondary turning over of the yeast, keeping the moisture of the yeast blocks to 14% -15% after the temperature of the yeast blocks approaches the room temperature, taking the yeast blocks out of the house and warehousing the yeast blocks, wherein the yeast cores have obvious yellow bacterial plaques of aschersonia aleyrodis.
10. The method as claimed in claim 9, wherein the reinforced koji of Ascomyces Shewanella is prepared by using the strain Ascomyces Shewanella CICC41587(M9) (Eurotium chevalieri) with a preservation number of CGMCCNo.14623.
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