CN111925975A - Functional yeast liquid and functional bacteria and preparation method thereof - Google Patents
Functional yeast liquid and functional bacteria and preparation method thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/32—Nature of the water, waste water, sewage or sludge to be treated from the food or foodstuff industry, e.g. brewery waste waters
- C02F2103/325—Nature of the water, waste water, sewage or sludge to be treated from the food or foodstuff industry, e.g. brewery waste waters from processes relating to the production of wine products
Abstract
The invention discloses a yeast functional bacterial liquid, a functional bacterium and a preparation method thereof, and particularly relates to the technical field of biological fermentation. A preparation method of yeast functional bacteria liquid comprises the following steps: adding 1 part of yellow water into 5-7 parts of water, adding Daqu into the diluted yellow water, adjusting the pH to 5.0-7.0, and performing anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution; transferring the seed liquid into yellow water according to the mass ratio of 10-50%; repeating for 4-6 times to obtain functional bacterial liquid containing acid-producing bacteria. A preparation method of a yeast functional bacterium comprises the following steps: adding ethanol or lactic acid into the functional bacterial liquid, and blowing off oxygen by nitrogen for 20-30 min; transferring the fermented bacterial liquid into a pasteurization culture medium, blowing off oxygen by nitrogen for 20-30 min, and then carrying out anaerobic culture at 30 ℃ for 7-15 d; diluting and coating the fermented bacteria liquid, and separating and screening out functional bacteria. The technical scheme of the invention solves the problem of development and utilization of functional microorganisms in the yeast for making hard liquor and can be used for development and research of functional bacteria.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a yeast functional bacterial liquid, a functional bacterium and a preparation method thereof.
Background
The Daqu is a microecological product simultaneously containing a microbial strain, a microbial enzyme system and a compound yeast aroma substance, and is an important wine production agent and aroma generation agent in the brewing process of the Daqu wine. The Daqu serves as a nutrient carrier and has a brewing effect by selectively enriching microorganisms in the environment. The organisms have diversity and unity, and the functional microorganisms in the yeast are not only used for brewing, but also can play an irreplaceable role in pharmacy and human intestinal tracts. Therefore, finding a proper method for preparing the functional microorganisms in the Daqu has important significance for recognizing and developing the microorganisms and further improving the quality of the Daqu wine.
In the solid-state fermentation process of the white spirit, water generated by microbial metabolism and water which is not utilized in the fermented grains seep from the fermentation materials to the bottom of the pit to form yellow water. The yellow water contains organic acid, yeast extract, soluble starch, reducing sugar, ethanol, amino acid, etc. In the fermentation process of the Luzhou-flavor liquor by taking a pit as a fermentation container, the bottom fermented grains containing the Daqu powder are soaked in yellow water, so that microorganisms in the bottom fermented grains adapt to the environment and play a role. Therefore, it is feasible to prepare acid-producing bacteria in koji by using yellow water as a natural culture medium.
Disclosure of Invention
The invention aims to provide a yeast functional bacterial liquid, a functional bacterium and a preparation method thereof, so as to solve the problem of development and utilization of functional microorganisms in yeast.
In order to achieve the purpose, the technical scheme of the invention is as follows: a preparation method of yeast functional bacteria liquid comprises the following steps:
s102, adding 1 part of yellow water into 5-7 parts of water, crushing Daqu, uniformly mixing, adding the crushed Daqu into diluted yellow water according to the mass ratio of 1% -5%, adjusting the pH value to 5.0-7.0, and performing anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution;
s104, transferring the seed solution of the S102 into yellow water with the same dilution ratio as that in the step S102 according to the mass ratio of 10-50%, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
and S106, repeating the step S104 for 6 times to obtain the functional bacterial liquid containing the acid-producing bacteria group.
The working principle of the scheme is as follows: the waste water (such as yellow water) generated in the brewing process contains a large amount of lactic acid and a small amount of ethanol. In the process of brewing the Daqu liquor, the Daqu is used as an important starter and contains abundant functional microorganisms. After the steps of S102-S106, the microorganisms in the yeast are domesticated and enriched continuously, so that the yeast functional bacterial liquid capable of metabolizing lactic acid or ethanol can be prepared.
Further, the yeast functional bacterial liquid can be applied to white spirit brewing and substance conversion of brewing wastewater.
The other technical scheme of the invention is as follows: a preparation method of a yeast functional bacterium comprises the following steps:
s108, adding ethanol or lactic acid into the functional bacterial liquid obtained in the S106, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
s110, transferring the bacterial liquid obtained in the step S108 to a pasteurization medium with ethanol or lactic acid as a unique carbon source according to an inoculation ratio of 10-30%, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 7-15 d;
s112, diluting and coating the bacterial liquid obtained in the step S110 in a pasteurization culture medium, and carrying out anaerobic culture at the temperature of 30 ℃ for 7-15 days; and (3) selecting a single colony on the plate, placing the single colony in a liquid pasteurization culture medium, carrying out anaerobic culture at the temperature of 30 ℃ for 7-15 d, and detecting the organic acid in the bacterial liquid by adopting a gas chromatography-mass spectrometer so as to screen the acid-producing strain.
Further, the pasteur culture medium is prepared by the following method: 1-3 parts of yeast extract, 0.1-0.5 part of peptone, 0.5-1.0 part of NaCl and 0.2-0.5 part of MgSO4·7H20. 0.5-1.2 parts of K2HPO40.1 to 0.5 part of FeSO40.1 to 0.5 parts of MnSO4、20～30Parts of lactic acid, and 1000 parts of distilled water. Adjusting the pH value to 5.0-7.0 by using 5mol/L NaOH, sterilizing at 115-121 ℃ for 20-30 min, cooling the culture medium, and adding 5-10 parts by weight of sterile CaCO into the culture medium3。
6. Further, the pasteur culture medium is prepared by the following method: 1-3 parts of yeast extract, 0.1-0.5 part of peptone, 0.5-1.0 part of NaCl and 0.2-0.5 part of MgSO4·7H20. 0.5-1.2 parts of K2HPO40.1 to 0.5 part of FeSO40.1 to 0.5 parts of MnSO4And 1000 parts of distilled water. Adjusting the pH value to 5.0-7.0 by using 5mol/LNaOH, sterilizing at 115-121 ℃ for 20-30 min, cooling the culture medium to normal temperature, and adding 15-20 parts by weight of ethanol and 5-10 parts by weight of sterile CaCO3。
Further, 15-20 parts by weight of agar powder is added into the Pasteur culture medium, so that the solid culture medium is prepared.
Through the steps, the microorganisms for metabolizing lactic acid or ethanol in the yeast functional bacteria liquid can be domesticated and enriched again. Yellow water is a natural culture medium which is rich in nutrients but complex, and substrates and products metabolized by the yeast functional microorganisms can be further illustrated by a pasteur culture medium containing lactic acid or ethanol as a sole carbon source.
Further, the yeast functional strain is applied to the processes of liquor brewing and brewing wastewater treatment.
Compared with the prior art, the beneficial effect of this scheme: the acid-producing functional bacteria in the yeast are domesticated and enriched for many times by using yellow water, so that the yellow water can be converted into functional bacteria liquid with recycling value. By adding ethanol or lactic acid into the fermentation liquor again, functional bacteria capable of metabolizing lactic acid or ethanol can form dominant flora in the fermentation environment, so that the aim of enriching the microorganisms beneficial to liquor brewing is directly fulfilled. The method of the scheme can not only rapidly obtain acid-producing functional bacteria in the yeast, but also has good practical application value for the obtained flora and single bacteria.
Detailed Description
The present invention will be described in further detail below by way of specific embodiments:
example 1:
a preparation method of yeast functional bacteria liquid comprises the following steps:
s102, adding 1 part of yellow water into 5 parts of water, crushing Daqu, uniformly mixing, adding the crushed Daqu into diluted yellow water according to the mass ratio of 1%, adjusting the pH value to 5.0, and carrying out anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution;
s104, transferring the seed solution of the S102 into yellow water with the same dilution ratio as that in the step S102 according to the mass ratio of 10%, adjusting the pH value to 5.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
and S106, repeating the step S104 for 4 times to obtain the yeast functional bacterial liquid containing the acid-producing bacteria group. Detecting the functional bacterial liquid by a gas chromatography-mass spectrometer, and comparing the content of the organic acid in the yellow water after dilution according to parts, wherein the content is shown in table 1:
table 1: comparing the content of organic acid in diluted yellow water with that in fermented yellow water
| Acetic acid (g/L) | Propionic acid (g/L) | Butyric acid (g/L) | Hexanoic acid (g/L) | Lactic acid (g/L) | |
| Diluted yellowWater (W) | 0.62 | 0.08 | 0.27 | 0.32 | 27.51 |
| Fermented yellow water | 2.28 | 0.17 | 2.55 | 1.18 | 10.51 |
As can be seen from table 1, the flora in the yeast for making hard liquor plays a huge acid-producing role compared to the diluted yellow water, wherein acetic acid is increased by 2.68 times, propionic acid is increased by 1.13 times, butyric acid is increased by 8.44 times, caproic acid is increased by 2.69 times, and lactic acid is decreased by 0.62 times. Therefore, under the condition, the yellow water can enrich acid-producing bacteria in the yeast and has a good acid-producing effect.
A preparation method of a yeast functional bacterium comprises the following steps:
s108, adding 15g/L ethanol or 20g/L lactic acid into the functional bacterial liquid of S106, adjusting the pH to 5.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
s110, transferring the bacterial liquid of the S108 into a pasteurization culture medium with ethanol or lactic acid as a unique carbon source according to the inoculation ratio of 30%, adjusting the pH to 5.0, blowing oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 7-15 d.
The pasteur culture medium is prepared by the following method: mixing yeast extract 3 parts, peptone 0.1 part, NaCl 0.5 part, and MgSO 0.2 part4·7H20. 1.0 part of K2HPO40.2 part of FeSO40.1 part of MnSO420 parts of lactic acid and 1000 parts of distilled waterAnd (6) mixing. Adjusting the pH value to 5.0 by using NaOH with the concentration of 5mol/L, sterilizing at the temperature of 115-121 ℃ for 20-30 min, and adding 5 parts by weight of CaCO after the culture medium is cooled to normal temperature3. If ethanol is used to replace lactic acid to prepare the Pasteur culture medium, 15 parts by weight of ethanol and 5 parts by weight of CaCO are added after the culture medium is cooled to normal temperature3. If a solid pasteur medium is to be prepared, 15 parts by weight of agar powder are additionally added to the above mixed solution.
The bacteria liquid is detected by a gas chromatography-mass spectrometer, and the content of organic acid before fermentation and after fermentation is shown in the following table 2:
table 2: comparison of organic acid content in functional bacteria liquid before and after fermentation
As can be seen from Table 2, the functional bacterial liquid after the fermentation of the yellow water contains functional bacteria which utilize ethanol and lactic acid and can convert ethanol or lactic acid into acetic acid, propionic acid, butyric acid and caproic acid again. The functional bacteria can form dominant flora in the fermentation environment by adding ethanol or lactic acid, and a foundation is laid for the separation and screening of acid-producing bacteria in the next step.
S112, diluting the bacterial liquid of the S110, coating the diluted bacterial liquid in a pasteurization culture medium, and carrying out anaerobic culture at the temperature of 30 ℃ for 7-15 d; and (2) selecting a single colony on the plate, placing the single colony in a liquid pasteurization culture medium, carrying out anaerobic culture at 30 ℃ for 7-15 days, detecting organic acid in the bacterial liquid by adopting a gas chromatography-mass spectrometer, and screening 3 acid-producing bacteria which utilize ethanol or lactic acid, wherein 2 acid-producing bacteria which can utilize lactic acid to produce propionic acid and butyric acid and 1 acid-producing bacteria which utilize ethanol to produce butyric acid.
Example 2
A preparation method of yeast functional bacteria liquid comprises the following steps:
s102, adding 1 part of yellow water into 7 parts of water, crushing Daqu, uniformly mixing, adding the crushed Daqu into diluted yellow water according to the mass ratio of 2%, adjusting the pH value to 7.0, and carrying out anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution;
s104, transferring the seed solution of the S102 into yellow water with the same dilution ratio as that in the step S102 according to the mass percentage of 50%, adjusting the pH to 7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
and S106, repeating the step S104 for 5 times to obtain the yeast functional bacterial liquid containing the acid-producing bacteria group. Detecting the functional bacterial liquid by a gas chromatography-mass spectrometer, and comparing the content of the organic acid in the yellow water after dilution according to parts, wherein the content is shown in table 3:
table 3: comparing the content of organic acid in the diluted yellow water with that in the fermented yellow water
| Acetic acid (g/L) | Propionic acid (g/L) | Butyric acid (g/L) | Hexanoic acid (g/L) | Lactic acid (g/L) | |
| Diluted yellow water | 0.44 | 0.06 | 0.19 | 0.23 | 19.65 |
| Fermented yellow water | 1.29 | 1.39 | 5.22 | 1.86 | 6.96 |
As can be seen from Table 3, the contents of acetic acid, propionic acid, butyric acid and hexanoic acid in the yellow water after the fermentation is completed are all increased, and the lactic acid is obviously reduced. Wherein, acetic acid is increased by 1.93 times, propionic acid is increased by 22.17 times, butyric acid is increased by 26.47 times, caproic acid is increased by 7.09 times, and lactic acid is decreased by 0.65 times. Therefore, under the condition, the microorganisms in the yeast are domesticated by yellow water, and the microorganisms with acid-producing function are enriched.
A preparation method of a yeast functional bacterium comprises the following steps:
s108, adding 20g/L ethanol or 30g/L lactic acid into the functional bacterial liquid of S106, adjusting the pH to 7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
s110, transferring the bacterial liquid of the S108 into a pasteurization culture medium with ethanol or lactic acid as a unique carbon source according to the inoculation ratio of 20%, adjusting the pH to 7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 7-15 d.
The pasteur culture medium is prepared by the following method: 2 parts of yeast extract, 0.5 part of peptone, 1.0 part of NaCl and 0.4 part of MgSO4·7H20. 0.5 part of K2HPO40.1 part of FeSO40.5 part of MnSO430 parts of lactic acid, and 1000 parts of distilled water. Adjusting the pH value to 7.0 by using NaOH with the concentration of 5mol/L, sterilizing at the temperature of 115-121 ℃ for 20-30 min, and adding 10 parts by weight of CaCO after the culture medium is cooled to normal temperature3. If ethanol is used to replace lactic acid to prepare the Pasteur culture medium, 20 parts by weight of ethanol and 10 parts by weight of CaCO are added after the culture medium is cooled to normal temperature3. If the solid pasteur medium is to be prepared, it is necessary to mix the solutions20 parts by weight of agar powder is additionally added into the liquid.
The ratio of the organic acid content before fermentation to the organic acid content after fermentation is shown in table 4, when the bacterial liquid is detected by a gas chromatography-mass spectrometer:
table 4: comparison of organic acid content in functional bacteria liquid before and after fermentation
S112, performing coating culture on the functional bacterial liquid obtained in the step S110 according to a gradient dilution method, and performing anaerobic culture at the temperature of 30 ℃ for 7-15 days; and (2) selecting a single colony on the plate, placing the single colony in a liquid culture medium taking ethanol or lactic acid as a carbon source, carrying out anaerobic culture at 30 ℃ for 10-15 d, detecting the fermented bacterial liquid by a gas chromatography-mass spectrometer, and screening out 4 acid-producing bacteria using ethanol or lactic acid, wherein 3 acid-producing bacteria capable of producing acetic acid and butyric acid by using lactic acid and 1 acid-producing bacteria capable of producing butyric acid by using ethanol.
Example 3:
a preparation method of yeast functional bacteria liquid comprises the following steps:
s102, adding 1 part of yellow water into 6 parts of water, crushing Daqu, uniformly mixing, adding the crushed Daqu into diluted yellow water according to the mass ratio of 5%, adjusting the pH value to 6.0, and carrying out anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution;
s104, transferring the seed solution of the S102 into yellow water with the same dilution ratio as that in the step S102 according to the mass ratio of 30%, adjusting the pH to 6.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
and S106, repeating the step S104 6 times to obtain the yeast functional bacterial liquid containing the acid-producing bacteria group. Detecting the functional bacterial liquid by a gas chromatography-mass spectrometer, and comparing the content of the organic acid in the yellow water after dilution according to parts, wherein the content is shown in table 5:
table 5: comparing the content of organic acid in the diluted yellow water with that in the fermented yellow water
| Acetic acid (g/L) | Propionic acid (g/L) | Butyric acid (g/L) | Hexanoic acid (g/L) | Lactic acid (g/L) | |
| Diluted yellow water | 0.52 | 0.06 | 0.23 | 0.27 | 22.93 |
| Fermented yellow water | 6.95 | 1.66 | 8.64 | 5.38 | 5.82 |
As can be seen from table 5, under this condition, acetic acid increased 12.37 times, propionic acid increased 26.67 times, butyric acid increased 36.56 times, caproic acid increased 18.93 times, and lactic acid decreased 0.75 times. Therefore, the yellow water can enrich acid-producing flora in the yeast, has good acid-producing effect, is reflected in 'increasing already and reducing milk', and can change the yellow water into things of value.
A preparation method of a yeast functional bacterium comprises the following steps:
s108, adding 18g/L ethanol or 25g/L lactic acid into the functional bacterial liquid of S106, adjusting the pH to 6.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
s110, transferring the bacterial liquid obtained in the step S108 to a pasteur culture medium which takes ethanol or lactic acid as a unique carbon source according to an inoculation ratio of 10%, adjusting the pH to 6.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 7-15 d.
The pasteur culture medium is prepared by the following method: 1 part of yeast extract, 0.3 part of peptone, 0.8 part of NaCl and 0.5 part of MgSO4·7H20. 1.2 parts of K2HPO40.5 part of FeSO40.3 part of MnSO425 parts of lactic acid, and 1000 parts of distilled water. Adjusting the pH value to 6.0 by using NaOH with the concentration of 5mol/L, sterilizing at the temperature of 115-121 ℃ for 20-30 min, and adding 8 parts by weight of CaCO after the culture medium is cooled to normal temperature3. If ethanol is used to replace lactic acid to prepare the Pasteur culture medium, 18 parts by weight of ethanol and 8 parts by weight of CaCO are added after the culture medium is cooled to normal temperature3. If a solid pasteur medium is to be prepared, 18 parts by weight of agar powder are additionally added to the above mixed solution.
The bacteria liquid is detected by a gas chromatography-mass spectrometer, and the content of organic acid before fermentation is compared with that after fermentation, as shown in table 6:
table 6: comparison of organic acid content in fermentation broth before and after addition of ethanol or lactic acid
As can be seen from Table 6, under these conditions, the contents of acetic acid, propionic acid, butyric acid and caproic acid continued to increase after ethanol or lactic acid was added to the functional bacterial liquid, and it can be seen that the functional bacterial liquid contains a strain producing an organic acid by ethanol or lactic acid. The functional bacteria can form dominant flora in the fermentation environment by adding ethanol and lactic acid, and a foundation is laid for the separation and screening of acid-producing bacteria in the next step.
S112, performing coating culture on the functional bacterial liquid of the S110 according to a gradient dilution method, and performing anaerobic culture at 30 ℃ for 7-15 days; and (3) selecting the single colony on the plate, and carrying out anaerobic culture for 10-15 days at 30 ℃ in a liquid culture medium taking ethanol or lactic acid as a carbon source. And (3) detecting the fermented bacterial liquid by using a gas chromatography-mass spectrometer, and screening 10 acid-producing bacteria which utilize ethanol or lactic acid, wherein 7 acid-producing bacteria which can utilize lactic acid to produce acetic acid, butyric acid and caproic acid, and 3 acid-producing bacteria which utilize ethanol to produce butyric acid and caproic acid.
The foregoing are merely examples of the present invention and common general knowledge of known specific structures and/or features of the schemes has not been described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (6)
1. A preparation method of yeast functional bacteria liquid is characterized by comprising the following steps: the method comprises the following steps:
s102, adding 1 part of yellow water into 5-7 parts of water, crushing and uniformly mixing Daqu, adding the Daqu into diluted yellow water according to the mass ratio of 1% -5%, adjusting the pH value to 5.0-7.0, and carrying out anaerobic culture at 30 ℃ for 10-20 days to obtain a seed solution;
s104, transferring the seed solution of the S102 into yellow water with the same dilution ratio as that in the step S102 according to the mass ratio of 10-50%, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
and S106, repeating the step S104 for 4-6 times to obtain the functional bacterial liquid containing acid-producing bacteria.
2. Application of Daqu functional bacteria liquid in brewing white spirit and converting substances of brewing wastewater is provided.
3. A preparation method of a yeast functional bacterium is characterized by comprising the following steps: the method comprises the following steps:
s108, adding ethanol or lactic acid into the functional bacterial liquid obtained in the S106, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 10-20 d;
s110, transferring the bacterial liquid obtained in the step S108 to a pasteurization medium with ethanol or lactic acid as a unique carbon source according to an inoculation ratio of 10-30%, adjusting the pH to 5.0-7.0, blowing off oxygen by nitrogen for 20-30 min, and carrying out anaerobic culture at 30 ℃ for 7-15 d;
s112, diluting and coating the bacterial liquid obtained in the step S110 in a pasteurization culture medium, and carrying out anaerobic culture at the temperature of 30 ℃ for 7-15 days; and (3) selecting a single colony on the plate, placing the single colony in a liquid pasteurization culture medium, carrying out anaerobic culture at the temperature of 30 ℃ for 7-15 d, and detecting the organic acid in the bacterial liquid by adopting a gas chromatography-mass spectrometer so as to screen the acid-producing strain.
4. The method for preparing a functional yeast for making hard liquor according to claim 3, wherein the method comprises the following steps: the pasteur culture medium is prepared by the following method: 1-3 parts of yeast extract, 0.1-0.5 part of peptone, 0.5-1.0 part of NaCl and 0.2-0.5 part of MgSO4·7H20. 0.5-1.2 parts of K2HPO40.1 to 0.5 part of FeSO40.1 to 0.5 parts of MnSO420-30 parts of lactic acid and 1000 parts of distilled water. Adjusting the pH value to 5.0-7.0 by 5mol/LNaOH, sterilizing at 115-121 ℃ for 20-30 min, cooling the culture medium to normal temperature, and adding 5-10 parts by weight of sterile CaCO3。
5. The method for preparing a functional yeast for making hard liquor according to claim 3, wherein the method comprises the following steps: the pasteur culture medium is prepared by the following method: 1-3 parts of yeast extract, 0.1-0.5 part of peptone, 0.5-1.0 part of NaCl and 0.2-0.5 part of MgSO4·7H20. 0.5-1.2 parts of K2HPO40.1 to 0.5 part of FeSO40.1 to 0.5 parts of MnSO4Mixing with 1000 parts of distilled water, adjusting the pH value to 5.0-7.0 by using 5mol/L NaOH, sterilizing at 115-121 ℃ for 20-30 min, cooling the culture medium to normal temperature, adding 15-20 parts of ethanol and 5-10 parts of sterile CaCO3。
6. The method for producing a functional koji mold according to any one of claims 4 or 5, wherein: and adding 15-20 parts by weight of agar powder into the Pasteur culture medium to obtain the solid culture medium.
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