阅读说明：本技术 谢瓦散囊菌在白酒窖池窖泥微生物复壮中的应用及方法 (Application and method of eurotium scherzeri in microbial rejuvenation of pit mud of white spirit cellar ) 是由 蔡鹏飞 白秀彬 信春晖 石鲁博 许玲 夏晓波 于 2019-09-06 设计创作，主要内容包括：本发明属于生物技术领域,涉及谢瓦散囊菌在白酒窖池窖泥微生物复壮中的应用及方法。采用白酒固态发酵中产生的黄水、酒尾为主要发酵底物,加入灭菌的软化水,含谢瓦散囊菌大曲和酯化红曲,搅拌均匀,通入空气,30-35℃发酵25-45天所得含有谢瓦散囊菌的发酵液接入到窖池窖泥或粮醅中发酵,可提高窖泥中厌氧微生物数量,有效实现窖池窖泥微生物的复壮；同时还可改善窖池窖泥的风味,改善糟醅的发酵环境,进一步提高国井集团浓香白酒的质量,提升白酒的风味,可酿造一种兼具茶香、果香风味的国酝香白酒。(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.)

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.

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.

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.

Drawings

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.