CN110804576A - Preparation method of composite caproic acid bacterial liquid - Google Patents
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Abstract
The invention discloses a preparation method of composite caproic acid bacterial liquid, which comprises the following steps: s1, selecting 0.95-1.05L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, calcium carbonate and 95% ethanol. The invention relates to the technical field of wine brewing. According to the preparation method of the compound caproic acid bacterial liquid, a specific culture medium formula is selected preferably, the bacterial liquid cultured in the traditional pure caproic acid bacterial liquid culture mode and the bacterial liquid cultured in the pit mud enrichment culture mode are mixed in a certain proportion by using the formula, the defect that the structure of a strain flora cultured in the traditional caproic acid bacterial liquid culture mode is single is overcome, and the defect that the growth advantage of the strain caproic acid bacteria cultured in the pit mud enrichment culture mode is unstable is overcome, so that the problems of degradation, type reduction and quantity reduction of wine-making microorganisms in pit mud are solved by using the cultured compound caproic acid bacterial liquid, the quality of produced wine is greatly improved, the aging of the pit mud in a new production workshop is accelerated, and the quality of the pit mud is improved.
Description
Technical Field
The invention relates to the technical field of wine brewing, in particular to a preparation method of a compound caproic acid bacterial liquid.
Background
Caproic acid bacteria are very important acid-producing microorganisms in the production process of the strong aromatic Chinese spirits, and caproic acid generated by the metabolism of the caproic acid bacteria and alcohol generated by the fermentation of Daqu produce ethyl caproate which is the main aroma component of the strong aromatic Chinese spirits. The caproic acid bacteria culture solution can be widely applied to aspects of cellar filling maintenance, artificial cellar mud culture, esterified liquid production and the like of the strong aromatic Daqu liquor, and the aim of improving the quality of the strong aromatic Daqu liquor is fulfilled. Wherein: the manmade pit mud functional caproic acid bacterial liquid is widely used for cultivation of manmade pit mud and daily maintenance of pit.
The traditional culture mode of the caproic acid bacteria liquid is mostly pure caproic acid bacteria culture or cellar mud enrichment culture, and the pure caproic acid bacteria liquid culture has the advantages of fast strain growth, good acid production capacity and simpler culture conditions. The method has the advantages that the pure caproic acid bacterium liquid flora is single in structure, not beneficial to long-term maintenance of a pit and aging of pit mud in a new production workshop, long in culture period, high in requirements for operator operation skills and site hardware conditions, and high in production cost. The enrichment culture of pit mud has the advantages of simple production operation process, less requirements on production hardware conditions and rich structure of caproic acid bacteria liquid flora. The method has the defects that the growth advantage of caproic acid bacteria liquid cultured by enriching pit mud is unstable, the caproic acid yield is low, the culture period is long, the caproic acid bacteria liquid is easy to infect infectious microbes, the quality of the bacteria liquid is unstable, and meanwhile, the aging and hardening of pit mud are easy to occur when the components such as calcium carbonate with large use amount enter a pit in the traditional culture mode.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a preparation method of a compound caproic acid bacterial liquid, which solves the problems of longer culture period, easy infectious microbes and unstable bacterial liquid quality.
In order to achieve the purpose, the invention is realized by the following technical scheme: the preparation method of the composite caproic acid bacterial liquid comprises the following steps:
s1, selecting 0.95-1.05L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, calcium carbonate and 95% ethanol, placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate and the yeast extract in a container for mixing, placing the mixed product in an autoclave for sterilization treatment, and adding the 95% ethanol and the calcium carbonate into the container to form a liquid culture medium;
s2, selecting 1000L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, vinasse, Daqu powder and old cellar mud, and placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the vinasse, the Daqu powder and the old cellar mud in another container for mixing to form a composite caproic acid bacteria liquid culture medium;
s3, selecting caproic acid bacteria which are screened from the old cellar pool and have vigorous growth and strong acid production capacity, inoculating the caproic acid bacteria into a liquid culture medium, and culturing to obtain a caproic acid bacteria seed liquid;
s4, selecting high-quality pit mud, adding the pit mud into 0.05L of sterilized liquid culture medium, performing constant-temperature water bath, cooling, inoculating the pit mud into 0.95L of sterilized liquid culture medium, and culturing to obtain 1L of pit mud enriched liquid;
s5, proportionally mixing the caproic acid bacteria seed liquid in the S3 and the cellar mud compound liquid in the S4, inoculating the mixture into a triangular flask of a 1L liquid culture medium, wherein the inoculation amount is 10-15%, and performing standing culture to obtain a first-level seed liquid;
s6, cleaning and pre-sterilizing a Kaschin jar, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting a primary seed solution with vigorous growth and good acid and gas production capacity, inoculating 10-15% of the primary seed solution into the Kaschin jar, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as a secondary seed solution;
s7, cleaning and pre-eliminating a stainless steel vat, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting secondary seed liquid with vigorous growth and good acid and gas production capacity, inoculating 10% -15% of the secondary seed liquid into the stainless steel fermentation vat, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as tertiary seed liquid;
s8, adding 1000L of water into a stainless steel pot, introducing steam, heating to 40-45 ℃, adding a caproic acid bacteria liquid culture medium material according to a proportion, inoculating 10-15% of three-stage seed liquid, stirring uniformly, introducing steam, heating to 80-85 ℃, keeping for 30min, cooling to 45-50 ℃, taking a 1000L container as a fermentation tank, pumping into 1000L of the fermentation tank, adding 25L of 95% (v/v) alcohol, regulating the pH value to 7.0 by NaOH, and standing and culturing at 32-35 ℃ for 5-10d for use;
the composite caproic acid bacterial liquid cultured and produced by the method has stable quality, strong caproic acid producing capacity and rich flora structure types by taking caproic acid bacteria as dominant strains, simplifies production operation, reduces production steps, shortens production period, and greatly enlarges culture amount by continuous graded culture, thereby improving production efficiency and saving production cost.
Further, the content of the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the calcium carbonate and the ethanol in the S1 are 4-6g of the anhydrous sodium acetate, 0.4-0.6g of the ammonium sulfate, 0.3-0.4g of the monopotassium phosphate, 0.1-0.2g of the magnesium sulfate, 2-3g of the yeast extract, 4-6g of the calcium carbonate and 20-25g of 95% ethanol.
Further, the internal temperature of the autoclave in S1 was 120-130 ℃, and the sterilization time was 20 min.
Further, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the vinasse, the Daqu powder and the old cellar mud in the S2 comprise 4000-6000g of anhydrous sodium acetate, 400-600g of ammonium sulfate, 300-400g of monopotassium phosphate, 100-200g of magnesium sulfate, 3000g of yeast extract, 90000g of vinasse, 60000g of Daqu powder and 5000g of old cellar mud.
Further, the culture environment in the S3 is 32-35 ℃, and the culture time is 5-10 d.
Further, the temperature of the thermostatic water bath in the S3 is 85-90 ℃, and the time of the thermostatic water bath is 20 min.
Further, the culture environment in the S4 is 32-35 ℃, and the culture time is 5-10 d.
Further, the ratio in S5 is 4: 6, the time of static culture is 5-10d, and the temperature of the time of static culture is 32-35 ℃.
Further, the formula ratio of the liquid culture medium in the S5 is as follows: 1L of water, 5g of anhydrous sodium acetate, 0.5g of ammonium sulfate, 0.4g of monopotassium phosphate, 0.2g of magnesium sulfate, 3g of yeast extract, 5g of calcium carbonate and 0.025L of 95% ethanol, wherein the liquid culture medium in the S6 has the formula ratio: 10L of water, 50g of anhydrous sodium acetate, 5g of ammonium sulfate, 4g of monopotassium phosphate, 2g of magnesium sulfate, 30g of yeast extract, 50g of calcium carbonate and 0.25L of 95% ethanol, wherein the liquid culture medium in the S7 comprises the following formula: 100L of water, 500g of anhydrous sodium acetate, 50g of ammonium sulfate, 40g of monopotassium phosphate, 20g of magnesium sulfate, 300g of yeast extract, 500g of calcium carbonate and 2.5L of 95% ethanol.
Further, the proportion of the complex caproic acid bacteria liquid culture medium in the S8 is 1000L of water, 5000g of sodium acetate trihydrate, 500g of ammonium sulfate, 1100g of monopotassium phosphate, 250g of magnesium sulfate, 3000g of yeast extract, 25L of 95% ethanol, 90000g of vinasse, 50000g of Daqu powder and 5000g of old cellar mud.
Compared with the prior art, the invention has the beneficial effects that:
according to the preparation method of the compound caproic acid bacterial liquid, a specific culture medium formula is selected preferably, the bacterial liquid cultured in the traditional pure caproic acid bacterial liquid culture mode and the bacterial liquid cultured in the pit mud enrichment culture mode are mixed in a certain proportion by using the formula, the defect that the structure of a strain flora cultured in the traditional caproic acid bacterial liquid culture mode is single is overcome, and the defect that the growth advantage of the strain caproic acid bacteria cultured in the pit mud enrichment culture mode is unstable is overcome, so that the problems of wine-making microorganism degradation, type reduction and quantity reduction in pit mud are solved by using the cultured compound caproic acid bacterial liquid, the quality of produced wine is improved, the aging of the pit mud in a new production workshop is accelerated, and the quality of the pit mud is improved.
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FIG. 1 is a flow chart of the preparation method of the compound caproic acid bacterial liquid of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1, the present invention provides a technical solution: the preparation method of the composite caproic acid bacterial liquid comprises the following steps:
s1, selecting 0.95-1.05L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, calcium carbonate and 95% ethanol, placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract and the calcium carbonate in a container for mixing, placing a mixed product in an autoclave for sterilization treatment, and adding 95% ethanol into the container to form a liquid culture medium;
s2, selecting 1000L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, vinasse, Daqu powder and old cellar mud, and placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the vinasse, the Daqu powder and the old cellar mud in another container for mixing to form a composite caproic acid bacteria liquid culture medium;
s3, selecting caproic acid bacteria which are screened from the old cellar pool and have vigorous growth and strong acid production capacity, inoculating the caproic acid bacteria into a liquid culture medium, and culturing to obtain a caproic acid bacteria seed liquid;
s4, selecting high-quality pit mud, adding the pit mud into 0.05L of sterilized liquid culture medium, performing constant-temperature water bath, cooling, inoculating the pit mud into 0.95L of sterilized liquid culture medium, and culturing to obtain 1L of pit mud enriched liquid;
s5, proportionally mixing the caproic acid bacteria seed liquid in the S3 and the cellar mud compound liquid in the S4, inoculating the mixture into a triangular flask of a 1L liquid culture medium, wherein the inoculation amount is 10-15%, and performing standing culture to obtain a first-level seed liquid;
s6, cleaning and pre-sterilizing a Kaschin jar, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting a primary seed solution with vigorous growth and good acid and gas production capacity, inoculating 10-15% of the primary seed solution into the Kaschin jar, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as a secondary seed solution;
s7, cleaning and pre-eliminating a stainless steel vat, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting secondary seed liquid with vigorous growth and good acid and gas production capacity, inoculating 10% -15% of the secondary seed liquid into the stainless steel fermentation vat, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as tertiary seed liquid;
s8, adding 1000L of water into a stainless steel pot, introducing steam, heating to 40-45 ℃, adding a caproic acid bacteria liquid culture medium material according to a proportion, inoculating 10-15% of three-stage seed liquid, stirring uniformly, introducing steam, heating to 80-85 ℃, keeping for 30min, cooling to 45-50 ℃, taking a 1000L container as a fermentation tank, pumping into 1000L of the fermentation tank, adding 25L of 95% (v/v) alcohol, regulating the pH value to 7.0 by NaOH, and standing and culturing at 32-35 ℃ for 5-10d for use;
the composite caproic acid bacterial liquid cultured and produced by the method has stable quality, takes caproic acid bacteria as dominant bacteria, has strong caproic acid production capacity, has rich flora structure types, simplifies production operation, reduces production steps, shortens production period, and greatly enlarges culture amount by continuous graded culture, thereby improving production efficiency and saving production cost;
the culture medium formula of the composite caproic acid bacteria liquid removes calcium carbonate, and is added with vinasse, yeast and aged pit mud.
In the step of culturing the composite caproic acid bacterial liquid, adding all materials into a stainless steel pot, and after the preparation is finished, extracting supernatant to a large container with the constant volume of 1000L for culture to achieve the aim of removing redundant impurities.
The components of the S1 include anhydrous sodium acetate 4-6g, ammonium sulfate 0.4-0.6g, potassium dihydrogen phosphate 0.3-0.4g, magnesium sulfate 0.1-0.2g, yeast extract 2-3g, calcium carbonate 4-6g and 95% ethanol 20-25 g.
The internal temperature of the autoclave in S1 was 120-.
The contents of anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, distiller 'S grains, Daqu powder and aged pit mud in the S2 are 4000-6000g of anhydrous sodium acetate, 400-600g of ammonium sulfate, 300-400g of monopotassium phosphate, 200g of magnesium sulfate 100-90 g, 3000g of yeast extract, 90000g of distiller' S grains, 60000g of Daqu powder and 4000-5000g of aged pit mud.
The culture environment in the S3 is 32-35 ℃, and the culture time is 5-10 days.
The temperature of the constant-temperature water bath in the S3 is 85-90 ℃, and the time of the constant-temperature water bath is 20 min.
The culture environment in the S4 is 32-35 ℃, and the culture time is 5-10 days.
The ratio in S5 is 4: 6, the time of static culture is 5-10d, and the temperature of the time of static culture is 32-35 ℃.
The formula proportion of the liquid culture medium in the S5 is as follows: 1L of water, 5g of anhydrous sodium acetate, 0.5g of ammonium sulfate, 0.4g of monopotassium phosphate, 0.2g of magnesium sulfate, 3g of yeast extract, 5g of calcium carbonate and 0.025L of 95% ethanol, wherein the liquid culture medium in the S6 has the formula ratio: 10L of water, 50g of anhydrous sodium acetate, 5g of ammonium sulfate, 4g of monopotassium phosphate, 2g of magnesium sulfate, 30g of yeast extract, 50g of calcium carbonate and 0.25L of 95% ethanol, wherein the liquid culture medium in the S7 comprises the following formula: 100L of water, 500g of anhydrous sodium acetate, 50g of ammonium sulfate, 40g of monopotassium phosphate, 20g of magnesium sulfate, 300g of yeast extract, 500g of calcium carbonate and 2.5L of 95% ethanol;
water, anhydrous sodium acetate, ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate, yeast extract, calcium carbonate (separately sterilized) in the liquid medium formulations of S5, S6 and S7, 95% ethanol (separately added without sterilization) in the liquid medium formulations of S5, S6 and S7, and all were sterilized at 121 ℃ for 20min after fusion.
The proportion of the composite caproic acid bacteria liquid culture medium in the S8 is 1000L of water, 5000g of sodium acetate trihydrate, 500g of ammonium sulfate, 1100g of monopotassium phosphate, 250g of magnesium sulfate, 3000g of yeast extract, 25L of 95% ethanol, 90000g of vinasse, 50000g of Daqu powder and 5000g of old cellar mud.
When the culture medium works, S1, 0.95-1.05L of water, anhydrous sodium acetate, ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate, yeast extract, calcium carbonate and 95% ethanol are selected, the water, the anhydrous sodium acetate, the ammonium sulfate, the potassium dihydrogen phosphate, the magnesium sulfate, the yeast extract and the calcium carbonate are placed in a container for mixing, then the mixed product is placed in an autoclave for sterilization treatment, and then the 95% ethanol is added into the container to form a liquid culture medium;
s2, selecting 1000L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, vinasse, Daqu powder and old cellar mud, and placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the vinasse, the Daqu powder and the old cellar mud in another container for mixing to form a composite caproic acid bacteria liquid culture medium;
s3, selecting caproic acid bacteria which are screened from the old cellar pool and have vigorous growth and strong acid production capacity, inoculating the caproic acid bacteria into a liquid culture medium, and culturing to obtain a caproic acid bacteria seed liquid;
s4, selecting high-quality pit mud, adding the pit mud into 0.05L of sterilized liquid culture medium, performing constant-temperature water bath, cooling, inoculating the pit mud into 0.95L of sterilized liquid culture medium, and culturing to obtain 1L of pit mud enriched liquid;
s5, proportionally mixing the caproic acid bacteria seed liquid in the S3 and the cellar mud compound liquid in the S4, inoculating the mixture into a triangular flask of a 1L liquid culture medium, wherein the inoculation amount is 10-15%, and performing standing culture to obtain a first-level seed liquid;
s6, cleaning and pre-sterilizing a Kaschin jar, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting a primary seed solution with vigorous growth and good acid and gas production capacity, inoculating 10-15% of the primary seed solution into the Kaschin jar, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as a secondary seed solution;
s7, cleaning and pre-eliminating a stainless steel vat, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting secondary seed liquid with vigorous growth and good acid and gas production capacity, inoculating 10% -15% of the secondary seed liquid into the stainless steel fermentation vat, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as tertiary seed liquid;
s8, adding 1000L of water into a stainless steel pot, introducing steam, heating to 40-45 ℃, adding caproic acid bacteria liquid culture medium materials according to a proportion, inoculating 10-15% of three-stage seed liquid, stirring uniformly, introducing steam, heating to 80-85 ℃, keeping for 30min, cooling to 45-50 ℃, taking a 1000L container as a fermentation tank, pumping into 1000L of the fermentation tank, adding 25L of 95% (v/v) alcohol, adjusting the pH value to 7.0 by NaOH, and standing and culturing at 32-35 ℃ for 5-10 d.
The caproic acid bacteria compound liquid is used for culturing the new cellar mud, and a comparison experiment is carried out with the new cellar mud which independently uses the caproic acid bacteria culture liquid, and the comparison result is as follows:
in the table, the 1# and 2# cellar mud uses caproic acid bacteria compound liquid as culture liquid, the 3# and 4# cellar mud uses caproic acid bacteria liquid as culture liquid, and the culture conditions of the two new cellar mud are consistent except for the strains. The inspection time of the pit mud is 45 days of fermentation culture.
The caproic acid bacteria compound liquid is used for maintaining a cellar pool, and is compared with the cellar pool which independently uses a caproic acid bacteria culture solution, and the comparison result is as follows:
in the table, the 1# and 2# pit mud takes caproic acid bacteria compound liquid as maintenance liquid, the 3# and 4# pit mud takes caproic acid bacteria liquid as maintenance liquid, and the fermented grains, the fermentation time, the distillation, the inspection and other production processes and inspection modes of the two pit ponds are the same;
according to the actual production results, the caproic acid bacteria composite liquid is used for carrying out new pit mud culture and pit maintenance, the pit mud culture quality is effectively improved, the quality of base wine is improved, and the effect quality is obviously superior to that of a single caproic acid bacteria culture liquid.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. The preparation method of the composite caproic acid bacterial liquid is characterized by comprising the following steps of:
s1, selecting 0.95-1.05L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, calcium carbonate and 95% ethanol, placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate and the yeast extract in a container for mixing, placing the mixed product in an autoclave for sterilization treatment, and adding the 95% ethanol and the calcium carbonate into the container to form a liquid culture medium;
s2, selecting 1000L of water, anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, vinasse, Daqu powder and old cellar mud, and placing the water, the anhydrous sodium acetate, the ammonium sulfate, the monopotassium phosphate, the magnesium sulfate, the yeast extract, the vinasse, the Daqu powder and the old cellar mud in another container for mixing to form a composite caproic acid bacteria liquid culture medium;
s3, selecting caproic acid bacteria which are screened from the old cellar pool and have vigorous growth and strong acid production capacity, inoculating the caproic acid bacteria into a liquid culture medium, and culturing to obtain a caproic acid bacteria seed liquid;
s4, selecting high-quality pit mud, adding the pit mud into 0.05L of sterilized liquid culture medium, performing constant-temperature water bath, cooling, inoculating the pit mud into 0.95L of sterilized liquid culture medium, and culturing to obtain 1L of pit mud enriched liquid;
s5, proportionally mixing the caproic acid bacteria seed liquid in the S3 and the cellar mud compound liquid in the S4, inoculating the mixture into a triangular flask of a 1L liquid culture medium, wherein the inoculation amount is 10-15%, and performing standing culture to obtain a first-level seed liquid;
s6, cleaning and pre-sterilizing a Kaschin jar, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting a primary seed solution with vigorous growth and good acid and gas production capacity, inoculating 10-15% of the primary seed solution into the Kaschin jar, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as a secondary seed solution;
s7, cleaning and pre-eliminating a stainless steel vat, adding materials according to the formula proportion of a liquid culture medium, introducing steam for sterilization, boiling, keeping for 20min, cooling, selecting secondary seed liquid with vigorous growth and good acid and gas production capacity, inoculating 10% -15% of the secondary seed liquid into the stainless steel fermentation vat, adding alcohol and calcium carbonate together, and performing standing culture at 32-35 ℃ for 5-10d to be used as tertiary seed liquid;
s8, adding 1000L of water into a stainless steel pot, introducing steam, heating to 40-45 ℃, adding caproic acid bacteria liquid culture medium materials according to a proportion, inoculating 10-15% of three-stage seed liquid, stirring uniformly, introducing steam, heating to 80-85 ℃, keeping for 30min, cooling to 45-50 ℃, taking a 1000L container as a fermentation tank, pumping into 1000L of the fermentation tank, adding 25L of 95% (v/v) alcohol, adjusting the pH value to 7.0 by NaOH, and standing and culturing at 32-35 ℃ for 5-10 d.
2. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the components of the S1 include anhydrous sodium acetate 4-6g, ammonium sulfate 0.4-0.6g, potassium dihydrogen phosphate 0.3-0.4g, magnesium sulfate 0.1-0.2g, yeast extract 2-3g, calcium carbonate 4-6g and 95% ethanol 20-25 g.
3. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the internal temperature of the autoclave in S1 was 120-.
4. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the contents of anhydrous sodium acetate, ammonium sulfate, monopotassium phosphate, magnesium sulfate, yeast extract, distiller 'S grains, Daqu powder and aged pit mud in the S2 are 4000-6000g of anhydrous sodium acetate, 400-600g of ammonium sulfate, 300-400g of monopotassium phosphate, 200g of magnesium sulfate 100-90 g, 3000g of yeast extract, 90000g of distiller' S grains, 60000g of Daqu powder and 4000-5000g of aged pit mud.
5. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the culture environment in the S3 is 32-35 ℃, and the culture time is 5-10 days.
6. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the temperature of the constant-temperature water bath in the S3 is 85-90 ℃, and the time of the constant-temperature water bath is 20 min.
7. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the culture environment in the S4 is 32-35 ℃, and the culture time is 5-10 days.
8. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the ratio in S5 is 4: 6, the time of static culture is 5-10d, and the temperature of the time of static culture is 32-35 ℃.
9. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the formula proportion of the liquid culture medium in the S5 is as follows: 1L of water, 5g of anhydrous sodium acetate, 0.5g of ammonium sulfate, 0.4g of monopotassium phosphate, 0.2g of magnesium sulfate, 3g of yeast extract, 5g of calcium carbonate and 0.025L of 95% ethanol, wherein the liquid culture medium in the S6 has the formula ratio: 10L of water, 50g of anhydrous sodium acetate, 5g of ammonium sulfate, 4g of monopotassium phosphate, 2g of magnesium sulfate, 30g of yeast extract, 50g of calcium carbonate and 0.25L of 95% ethanol, wherein the liquid culture medium in the S7 comprises the following formula: 100L of water, 500g of anhydrous sodium acetate, 50g of ammonium sulfate, 40g of monopotassium phosphate, 20g of magnesium sulfate, 300g of yeast extract, 500g of calcium carbonate and 2.5L of 95% ethanol.
10. The method for preparing a compound caproic acid bacterial liquid according to claim 1, which is characterized by comprising the following steps: the proportion of the composite caproic acid bacteria liquid culture medium in the S8 is 1000L of water, 5000g of sodium acetate trihydrate, 500g of ammonium sulfate, 1100g of monopotassium phosphate, 250g of magnesium sulfate, 3000g of yeast extract, 25L of 95% ethanol, 90000g of vinasse, 50000g of Daqu powder and 5000g of old cellar mud.
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| CN111440685A (en) * | 2020-04-14 | 2020-07-24 | 河南省宋河酒业股份有限公司 | Pit mud maintenance liquid and preparation method and application thereof |
| CN111500507A (en) * | 2020-05-28 | 2020-08-07 | 江南大学 | Pit mud flora enrichment method based on oligoculture and application thereof |
| CN111778181A (en) * | 2020-06-19 | 2020-10-16 | 保定五合窖酒业有限公司 | Method for preparing pit mud caproic acid bacterial liquid |
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