CN111196967A - White spirit pit mud containing esterified liquid and preparation process and application thereof - Google Patents
White spirit pit mud containing esterified liquid and preparation process and application thereof Download PDFInfo
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- CN111196967A CN111196967A CN202010238403.5A CN202010238403A CN111196967A CN 111196967 A CN111196967 A CN 111196967A CN 202010238403 A CN202010238403 A CN 202010238403A CN 111196967 A CN111196967 A CN 111196967A
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- 239000007788 liquid Substances 0.000 title claims abstract description 113
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 claims abstract description 118
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 86
- 239000000843 powder Substances 0.000 claims abstract description 52
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 49
- 241000894006 Bacteria Species 0.000 claims abstract description 44
- 238000005886 esterification reaction Methods 0.000 claims abstract description 27
- 230000032050 esterification Effects 0.000 claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 claims abstract description 24
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims abstract description 18
- 244000046052 Phaseolus vulgaris Species 0.000 claims abstract description 18
- 239000003337 fertilizer Substances 0.000 claims abstract description 17
- 239000003415 peat Substances 0.000 claims abstract description 17
- 210000000988 bone and bone Anatomy 0.000 claims abstract description 16
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 16
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 16
- 239000002994 raw material Substances 0.000 claims abstract description 10
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 9
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims description 71
- 238000012216 screening Methods 0.000 claims description 49
- 238000012258 culturing Methods 0.000 claims description 42
- 239000004033 plastic Substances 0.000 claims description 35
- 238000002156 mixing Methods 0.000 claims description 24
- 235000013339 cereals Nutrition 0.000 claims description 23
- 238000012360 testing method Methods 0.000 claims description 23
- 238000011081 inoculation Methods 0.000 claims description 22
- 229940041514 candida albicans extract Drugs 0.000 claims description 21
- 239000012138 yeast extract Substances 0.000 claims description 21
- 239000000203 mixture Substances 0.000 claims description 18
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 229920002472 Starch Polymers 0.000 claims description 14
- 230000003321 amplification Effects 0.000 claims description 14
- 239000008367 deionised water Substances 0.000 claims description 14
- 229910021641 deionized water Inorganic materials 0.000 claims description 14
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 14
- 238000007789 sealing Methods 0.000 claims description 14
- 239000001632 sodium acetate Substances 0.000 claims description 14
- 235000017281 sodium acetate Nutrition 0.000 claims description 14
- 239000008107 starch Substances 0.000 claims description 14
- 235000019698 starch Nutrition 0.000 claims description 14
- 230000001954 sterilising effect Effects 0.000 claims description 14
- 238000003756 stirring Methods 0.000 claims description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 12
- 239000012535 impurity Substances 0.000 claims description 10
- 238000000926 separation method Methods 0.000 claims description 10
- 239000002985 plastic film Substances 0.000 claims description 9
- 229920006255 plastic film Polymers 0.000 claims description 9
- 229940026314 red yeast rice Drugs 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 8
- 238000002474 experimental method Methods 0.000 claims description 8
- 239000007789 gas Substances 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 108091005804 Peptidases Proteins 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 239000004365 Protease Substances 0.000 claims description 7
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 7
- 235000019347 bone phosphate Nutrition 0.000 claims description 7
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 7
- 239000011248 coating agent Substances 0.000 claims description 7
- 238000000576 coating method Methods 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 230000007613 environmental effect Effects 0.000 claims description 7
- 239000000284 extract Substances 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 7
- 239000011780 sodium chloride Substances 0.000 claims description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims description 7
- 230000009286 beneficial effect Effects 0.000 abstract description 10
- 230000032683 aging Effects 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 description 13
- 238000000034 method Methods 0.000 description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 239000002253 acid Substances 0.000 description 10
- 239000000796 flavoring agent Substances 0.000 description 9
- 235000019634 flavors Nutrition 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 239000003205 fragrance Substances 0.000 description 8
- 150000002148 esters Chemical class 0.000 description 7
- SHZIWNPUGXLXDT-UHFFFAOYSA-N ethyl hexanoate Chemical compound CCCCCC(=O)OCC SHZIWNPUGXLXDT-UHFFFAOYSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 238000010298 pulverizing process Methods 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 4
- 125000003118 aryl group Chemical group 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 238000002955 isolation Methods 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- 230000001953 sensory effect Effects 0.000 description 4
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 238000013124 brewing process Methods 0.000 description 3
- 239000004568 cement Substances 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 239000003864 humus Substances 0.000 description 3
- 230000002045 lasting effect Effects 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 2
- 235000017491 Bambusa tulda Nutrition 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000228347 Monascus <ascomycete fungus> Species 0.000 description 2
- 244000082204 Phyllostachys viridis Species 0.000 description 2
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 2
- 239000011425 bamboo Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 238000011514 vinification Methods 0.000 description 2
- HNAGHMKIPMKKBB-UHFFFAOYSA-N 1-benzylpyrrolidine-3-carboxamide Chemical compound C1C(C(=O)N)CCN1CC1=CC=CC=C1 HNAGHMKIPMKKBB-UHFFFAOYSA-N 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 244000007853 Sarothamnus scoparius Species 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000005587 bubbling Effects 0.000 description 1
- OBNCKNCVKJNDBV-UHFFFAOYSA-N butanoic acid ethyl ester Natural products CCCC(=O)OCC OBNCKNCVKJNDBV-UHFFFAOYSA-N 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229940116333 ethyl lactate Drugs 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a white spirit cellar mud containing esterified liquid and a preparation process and application thereof, wherein the white spirit cellar mud is prepared from the following raw materials: 10-15 kg of Daqu powder, 10-15 kg of bean cake powder, 55-65 kg of wet distiller's grains and 0.1-0.15 m of pit skin mud30.03-0.08 m of aged cellar mud31-1.6 kg of peat, 0.2-0.8 kg of micro-fertilizer, 0.2-0.8 kg of organic bone, 0.4-0.9kg of monopotassium phosphate, 15-25 kg of alcohol, 130-150L of caproic acid bacteria tertiary seed liquid, 90-110 kg of esterification liquid, 200-300 kg of cool boiled water and 0.8-1.2 m of loess3. The pit mud of the invention shortens the aging period of the pit mud, is beneficial to improving the quality of white spirit and improving the production efficiency, and can be widely applied to the enlargement of the pit pool of the wine enterprise and the improvement of the pit mud.
Description
Technical Field
The invention belongs to the technical field of wine brewing, and particularly relates to white spirit pit mud containing esterified liquid and a preparation process and application thereof.
Background
In the process of brewing white spirit, the quality of pit mud is a key factor for determining the quality of the white spirit, and the key factor is that a large number of pit mud functional bacteria such as caproic acid bacteria, methane bacteria, butyric acid bacteria and lactic acid bacteria are inhabited in the pit mud, and the functional bacteria metabolize a large number of flavor substances of the white spirit by using brewing raw materials in a pit, so that the style of the white spirit is determined. Along with the expanding production of white spirit enterprises, the number of the cellar pools also needs to be correspondingly increased, and the increase of the cellar pools inevitably leads to the increase of the demand of high-quality old cellar mud. Generally, pit mud is naturally aged through uninterrupted grain feeding, wine production and aroma generation, and pit mud microorganisms need to be enriched and domesticated for a long time, so that the types and the number of brewing beneficial microorganisms reach certain values, and the process at least needs more than 10 years, so that the development of famous wine enterprises is seriously restricted. Therefore, how to manufacture high-quality artificial pit mud, shorten the aging period of the pit mud, and improve the quality of the white spirit becomes a technical key for the white spirit production enterprises and white spirit researchers to fight against.
Chinese patent publication No. CN102154080B discloses a preparation method of strong aromatic white spirit pit mud added with esterified liquid, which comprises the steps of adding 5kg of esterified liquid into a mixed infiltration body mixed and infiltrated by 35 kg of yellow clay with the fineness of 10 meshes and the pH value of 5.5-6.0, 3 kg of old pit mud, 20 kg of peat, 10kg of yeast powder, 5kg of bean cake powder, 5kg of double-wheel bottom aromatic fermented grains, 0.35 kg of silkworm chrysalis powder, 2kg of yellow water, 10kg of hot wine grains, 10kg of 20-22 DEG tail wine and 5kg of nutrient solution, inoculating 10kg of compound caproic acid bacteria enriched liquid, stirring and mixing to prepare pit mud fermented grains, sealing the peripheries of the pit mud fermented grains by using plastic films, carrying out heat preservation culture at 33-35 ℃ for 30-35 days under anaerobic conditions, and detecting the components and the number of beneficial viable microorganisms to obtain a strong aromatic white spirit pit mud product added with esterified liquid. The method is used for culturing pit mud in the process of the strong aromatic Chinese spirits. Nitrogen source and trace elements are highlighted, and the esterification liquid influences the generation components of the flavor and taste substances of the wine product. However, the culture period of the pit mud is too short, the number and the types of microorganisms in the pit mud cannot be guaranteed to reach a certain order of magnitude, and the beneficial microorganisms in the pit mud cannot be guaranteed to form a stable community result, so that the yield and the quality of the raw wine cannot be guaranteed to be stable after wine brewing.
The Chinese patent application with publication number CN106893657A discloses a preparation method of pit mud, which comprises the following steps: (1) selecting sun-dried and crushed loess, old pit mud and pit sealing mud, uniformly mixing, and paving on a cement ground; (2) selecting Daqu powder, aroma-producing yeast, fresh distiller's grains, bean cake powder, compound fertilizer and esterified red yeast rice, scattering into the mixture of loess, old pit mud and pit sealing mud, and stirring uniformly; (3) pouring the tail wine, the yellow serofluid and the purified water into the mixture, and naturally absorbing to form a mud blank; (4) mixing caproic acid bacteria expanding culture solution and pit mud culture solution into the embryo, and treading the mixture to be soft or stirring the mixture by using a mud mixer; (5) piling the uniformly stirred mud embryo and the cement road surface facing the sun, tightly covering the mud embryo and the cement road surface with a plastic film, sealing and fermenting, and keeping the temperature and the rain. The method adopts manual culture of the aged pit mud, the fermentation temperature rise is stable, the gas production is rich, the basic maturity can be achieved after one or two months of accumulation fermentation culture, all indexes can meet the requirements of the aged pit mud, and a good effect is achieved after the new workshop is put into operation. However, the addition of the chemical fertilizer in the patent can cause the pit mud to be hardened and easily aged, and the adverse phenomena can bring different foreign flavors to the base wine, which is not beneficial to the improvement of the quality of the base wine.
Disclosure of Invention
In order to overcome the defects, the invention aims to provide white spirit pit mud containing esterified liquid and a preparation process and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a white spirit pit mud containing esterified liquid is prepared from the following raw materials:
10-15 kg of Daqu powder, 10-15 kg of bean cake powder, 55-65 kg of wet distiller's grains and 0.1-0.15 m of pit skin mud30.03-0.08 m of aged cellar mud31-1.6 kg of peat, 0.2-0.8 kg of micro-fertilizer, 0.2-0.8 kg of organic bone, 0.4-0.9kg of monopotassium phosphate, 15-25 kg of alcohol, 130-150L of caproic acid bacteria tertiary seed liquid, 90-110 kg of esterification liquid, 200-300 kg of cool boiled water and 0.8-1.2 m of loess3。
Preferably, the water content of the yeast powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) x 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
Preferably, the loess is loess with surface area below 50-100cm, and is obtained by drying in the sun, removing impurities, and pulverizing.
Preferably, the preparation method of the third-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 0.8-1.2 g old cellar mud in a large test tube containing enrichment culture medium, treating in 75-85 deg.C water for 10-15min, cooling, placing in an anaerobic incubator, and anaerobically culturing at 33-36 deg.C for 6-8 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on a separation culture medium under the aseptic condition, and carrying out anaerobic culture for 5-7 days in a culture dish at the temperature of 35-37 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 5-7 days at 35-37 deg.C in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium to perform amplification culture in a triangular flask, and performing anaerobic culture at 35-37 ℃ for 5-7 days;
(4) preparing a seed solution: a. first-stage seed liquid, namely inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 25-35%, and culturing for 6-8 d at 35-37 ℃ by adopting a screening culture medium;
b. inoculating mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 25-35%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 35-37 deg.C for 6-8 days;
c. and (3) third-stage seed liquid, inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 20-25%, covering the inner cover and the outer cover of the plastic pot with a screening culture medium, and culturing for 6-8 days at the temperature of 35-37 ℃.
Preferably, the anaerobic atmosphere in step (1) or (2) is 90% N2、5% CO2And 5% of H2。
Preferably, the formula of the separation medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
Preferably, the formulation of the screening medium in step (3) or step (4) is: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
Preferably, the preparation method of the esterification solution comprises the following steps:
170-190 kg of warm boiled water with the temperature of 40-45 ℃, 25-35 kg of pot bottom water, 100-110 kg of 15-20% vol tail water, 22-25 kg of red yeast rice, 40-50kg of yellow water, 85-100 kg of 90-95% vol ethanol and 3-8 kg of caproic acid are sequentially added into the sterilized pottery jar, the mixture is uniformly stirred, the mouth of the pottery jar is completely sealed by plastic paper, the environmental temperature is kept at 33-37 ℃, and the pottery jar is cultured for 55-65 days to obtain the pottery jar.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, sealing with plastic film, and culturing at 20-30 deg.C for 180-200 days.
An application of pit mud containing esterified liquid in brewing wine.
The invention has the following positive beneficial effects:
1. the monascus is rich in functional bacteria with esterase production capacity such as monascus and the like, the esterification capacity can reach the maximum value at a proper temperature, and ethanol in the raw materials and caproic acid are subjected to acid-alcohol esterification reaction through esterification to produce a large amount of ethyl caproate; the bottom boiler water is water at the bottom of a retort in the distillation process of the white spirit, the pH value is 3.5-4.0, the tail water is the part which flows out finally in the distillation process of the white spirit, the yellow water is brown yellow liquid which is infiltrated at the bottom of a cellar during fermentation of fermented grains, and the bottom boiler water, the tail water and the yellow water have certain nutrient components besides various acid, alcohol and ester components, can provide a carbon source and growth factors for the propagation and metabolism of functional bacteria in red yeast rice, and are favorable for the esterification reaction; the ethyl caproate is produced by the esterification reaction of ethanol and caproic acid under the action of esterifying enzyme, and the fragrance is increased. The esterified liquid obtained by the invention has high content of fragrant esters, and when the esterified liquid is used for culturing pit mud, the aging period of the pit mud can be obviously shortened, the content of brewing functional microorganisms in the pit mud and the pit bottom fragrance are increased, and the quality of white spirit is improved.
The inoculation amount of each grade of the caproic acid bacteria seed liquid is larger, so that the culture period can be shortened to the maximum extent, and the purity of the inoculated bacterial liquid can be fully ensured, so that the functional strains can always keep absolute advantages, and the obtained three-grade caproic acid bacteria seed liquid has high concentration, high purity and strong caproic acid production capacity.
2. The loess is used as the main body of the pit mud, and the yeast powder contains functional microorganisms and provides fragrant components and nutrient substances; the wet vinasse is the wine steamed after the wine making, so that the porosity of pit mud is improved, part of wine making functional microorganisms are provided, and the weight and the content of flavor components in the pit mud are increased; the addition of the esterifying solution increases the weight and content of flavor substances in the manmade pit mud and the types of related functional bacteria, and shortens the period that the pit mud can generate the peculiar smell of the aged pit mud after long-term use; the addition of the caproic acid bacterium three-level seed liquid increases the quantity of clostridium in pit mud, and is beneficial to the generation of corresponding ester substances in the brewing process of white spirit; the pit skin mud is pit mud covering the covered grains at least in the pit, the aged pit mud is pit mud with the pit age of 10-20 years, the type and the number of brewing functional bacteria in the artificial pit mud are greatly enriched, and the flavor components in the white spirit brewing process are not too single; organic bones, trace elements, monopotassium phosphate, peat and bean cake powder can increase the content of growth factors necessary for the growth of microorganisms such as a carbon source, a nitrogen source and phosphorus in pit mud, are beneficial to the reproduction and metabolism of the microorganisms in the pit mud, and further can promote the aging of the pit mud and improve the quality of base liquor; the method is characterized in that the pit mud environment is formed in the fermented grain fermentation process of the alcohol, pit mud microorganisms adaptive to the brewing environment are domesticated directionally, and meanwhile, the pit mud microorganisms react with acid substances produced by the microorganisms in the pit mud, so that the content of ester substances in the pit mud is increased, and the fragrance of the pit mud is improved. The pit mud obtained by the method is grey brown, pure in fragrance, strong in old pit mud smell, slightly fragrant in wine, lasting in fragrance and free of other foreign flavors; soft and fine hand feeling, no thorn hand feeling, bubbling section, uniform texture, no impurity and obvious sticky feeling, pH is 5.8-6.4, water content is 38-42%, humus is 18-22%, ammoniacal nitrogen is 351-423 mg/Kg, available phosphorus is 288-336 mg/Kg, available potassium is 395-437mg/Kg, total bacterial acid is 3.4-8.1 × 109CFU/g, total spore acid 2.8-7.2X 105CFU/g, which reaches the quality of the common pit mud aged for 10 years; the pit mud prepared by the method is used for brewing wine for the first time, and the obtained wine baseThe conventional physicochemical indexes of the total acid, the total ester and the like in the white spirit are close to the results of the wine produced by a 10-year cellar pit, even exceed the results of the 10-year cellar pit, and the sensory evaluation of the white spirit obtained by the invention is equivalent to that of the 10-year cellar pit. Therefore, the use of the pit mud shortens the aging period of the pit mud, is beneficial to improving the quality of white spirit and the production efficiency, and can be widely applied to the enlargement of the pit pool of the wine enterprise and the improvement of the pit mud.
Detailed Description
The invention will be further illustrated with reference to some specific embodiments.
Example 1
A white spirit pit mud containing esterified liquid is prepared from the following raw materials:
10kg of Daqu powder, 10kg of bean cake powder, 60 kg of wet distiller's grains and 0.1 m of pit skin mud30.05 m of aged pit mud31.5kg of peat, 0.4 kg of micro-fertilizer, 0.3 kg of organic bone, 0.9kg of monopotassium phosphate, 15 kg of alcohol, 130L of caproic acid bacteria tertiary seed liquid, 100 kg of esterification liquid, 200 kg of cool boiled water and 1 m of loess3。
The water content of the Daqu powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) is multiplied by 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
The wet vinasse is vinasse steamed after brewing, and the water content is 68-70%.
The loess is loess with surface area below 50-100cm, and is prepared by drying in the sun, removing impurities, and pulverizing.
The preparation method of the three-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 1g of old cellar mud in a large test tube containing enrichment medium, treating in 75 deg.C water for 15min, cooling, placing in an anaerobic incubator, and performing anaerobic culture at 33 deg.C for 7 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on an isolation culture medium under the aseptic condition, and carrying out anaerobic culture for 5 days in a culture dish at the temperature of 35 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 6 days at 35 ℃ in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium, performing amplification culture in a triangular flask, and performing anaerobic culture at 35 ℃ for 7 days;
(4) preparing a seed solution: a. first-stage seed liquid, namely inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 25%, and culturing for 7 d at 35 ℃ by adopting a screening culture medium;
b. inoculating the mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 30%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 35 deg.C for 7 d;
c. and (3) third-stage seed liquid, namely inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 20%, adopting a screening culture medium, covering the inner cover and the outer cover of the plastic pot, and culturing for 7 d at the temperature of 35 ℃.
The anaerobic atmosphere in the step (1) or (2) is 90 percent of N2、5% CO2And 5% of H2。
The formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
The formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
The preparation method of the esterification liquid comprises the following steps:
and (3) sequentially adding 170 kg of warm boiled water at 40 ℃, 25 kg of bottom pot water, 100 kg of tail water at 15-20% vol, 24 kg of Zhonghui red yeast rice, 40 kg of yellow water, 90 kg of ethanol at 90-95% vol and 5kg of caproic acid into the sterilized pottery jar, uniformly stirring, completely sealing the jar opening by using plastic paper, keeping the environmental temperature at 33-37 ℃, and culturing for 55 days to obtain the product.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria tertiary seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, covering with plastic film, sealing, and culturing at 20-30 deg.C for 180 days.
Example 2
A white spirit pit mud containing esterified liquid is prepared from the following raw materials:
12 kg of Daqu powder, 15 kg of bean cake powder, 55 kg of wet distiller's grains and 0.15 m of pit skin mud30.06 m of aged pit mud31 kg of peat, 0.2 kg of micro-fertilizer, 0.6 kg of organic bone, 0.5 kg of monopotassium phosphate, 20 kg of alcohol, 140L of caproic acid bacteria tertiary seed liquid, 90 kg of esterification liquid, 220 kg of cool boiled water and 0.8 m of loess3。
The water content of the Daqu powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) is multiplied by 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
The wet vinasse is vinasse steamed after brewing, and the water content is 68-70%.
The loess is loess with surface area below 50-100cm, and is prepared by drying in the sun, removing impurities, and pulverizing.
The preparation method of the three-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 0.8 g old cellar mud in a large test tube containing enrichment medium, treating in water at 80 deg.C for 10min, cooling, placing in an anaerobic incubator, and performing anaerobic culture at 35 deg.C for 6 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on an isolation culture medium under the aseptic condition, and carrying out anaerobic culture for 7 days in a culture dish at the temperature of 36 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 7 days at 36 ℃ in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium, performing amplification culture in a triangular flask, and performing anaerobic culture at 36 ℃ for 6 days;
(4) preparing a seed solution: a. inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 30%, and culturing for 7 d at 36 ℃ by adopting a screening culture medium;
b. inoculating the mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 30%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 36 deg.C for 7 d;
c. and (3) third-stage seed liquid, namely inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 25%, covering the inner cover and the outer cover of the plastic pot by adopting a screening culture medium, and culturing for 7 d at 37 ℃.
The anaerobic atmosphere in the step (1) or (2) is 90 percent of N2、5% CO2And 5% of H2。
The formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
The formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
The preparation method of the esterification liquid comprises the following steps:
adding 180 kg of warm boiled water at 41 ℃, 30 kg of bottom pot water, 105 kg of tail water at 15-20% vol, 22 kg of Zhonghui red yeast rice, 45kg of yellow water, 85 kg of ethanol at 90-95% vol and 8 kg of caproic acid into the sterilized pottery jar in sequence, stirring uniformly, completely sealing the jar mouth with plastic paper, keeping the environmental temperature at 33-37 ℃, and culturing for 60 days to obtain the product.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, covering with plastic film, sealing, and culturing at 20-30 deg.C for 200 days.
Example 3
A white spirit pit mud containing esterified liquid is prepared from the following raw materials:
15 kg of Daqu powder, 15 kg of bean cake powder, 65 kg of wet distiller's grains and 0.12m of pit skin mud30.03 m of aged pit mud31.2kg of peat, 0.5 kg of micro-fertilizer, 0.2 kg of organic bone, 0.4 kg of monopotassium phosphate, 20 kg of alcohol, 150L of caproic acid bacteria tertiary seed liquid, 100 kg of esterification liquid, 250 kg of cool boiled water and 1 m of loess3。
The water content of the Daqu powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) is multiplied by 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
The wet vinasse is vinasse steamed after brewing, and the water content is 68-70%.
The loess is loess with surface area below 50-100cm, and is prepared by drying in the sun, removing impurities, and pulverizing.
The preparation method of the three-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 1.2 g old cellar mud in a large test tube containing enrichment medium, treating in water at 85 deg.C for 12min, cooling, placing in an anaerobic incubator, and performing anaerobic culture at 36 deg.C for 7 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on a separation culture medium under the aseptic condition, and carrying out anaerobic culture for 6 days in a culture dish at the temperature of 37 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 6 days at 37 ℃ in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium, performing amplification culture in a triangular flask, and performing anaerobic culture at 37 ℃ for 5 days;
(4) preparing a seed solution: a. inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 35%, and culturing for 6d at 37 ℃ by adopting a screening culture medium;
b. inoculating the mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 30%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 37 ℃ for 6 d;
c. and (3) third-stage seed liquid, namely inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 25%, adopting a screening culture medium, covering the inner cover and the outer cover of the plastic pot, and culturing for 6d at the temperature of 36 ℃.
The anaerobic atmosphere in the step (1) or (2) is 90 percent of N2、5% CO2And 5% of H2。
The formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
The formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
The preparation method of the esterification liquid comprises the following steps:
190 kg of warm boiled water with the temperature of 42 ℃, 35 kg of bottom boiler water, 110kg of tail water with the concentration of 15-20% vol, 25 kg of Zhonghui red yeast rice, 45kg of yellow water, 100 kg of ethanol with the concentration of 90-95% vol and 6kg of caproic acid are sequentially added into the sterilized pottery jar, the mixture is uniformly stirred, the mouth of the jar is completely sealed by plastic paper, the environmental temperature is kept at 33-37 ℃, and the mixture is cultured for 60 days, thus obtaining the product.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, covering with plastic film, sealing, and culturing at 20-30 deg.C for 190 days.
Example 4
A white spirit pit mud containing esterified liquid is prepared from the following raw materials:
13 kg of Daqu powder, 12 kg of bean cake powder, 55 kg of wet distiller's grains and 0.1 m of pit skin mud30.08 m of aged cellar mud31.6kg of peat, 0.8 kg of micro-fertilizer, 0.7 kg of organic bone, 0.7 kg of monopotassium phosphate, 25 kg of alcohol, 140L of caproic acid bacteria tertiary seed liquid, 110kg of esterification liquid, 300 kg of cool boiled water and 1.2m of loess3。
The water content of the Daqu powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) is multiplied by 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
The wet vinasse is vinasse steamed after brewing, and the water content is 68-70%.
The loess is loess with surface area below 50-100cm, and is prepared by drying in the sun, removing impurities, and pulverizing.
The preparation method of the three-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 1g of old cellar mud in a large test tube containing enrichment medium, treating in 75 deg.C water for 12min, cooling, placing in an anaerobic incubator, and performing anaerobic culture at 35 deg.C for 8 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on an isolation culture medium under the aseptic condition, and carrying out anaerobic culture for 6 days in a culture dish at the temperature of 35 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 5 days at 36 ℃ in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium, performing amplification culture in a triangular flask, and performing anaerobic culture at 36 ℃ for 7 days;
(4) preparing a seed solution: a. first-stage seed liquid, namely inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 30%, and culturing for 8 d at 36 ℃ by adopting a screening culture medium;
b. inoculating the mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 25%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 36 deg.C for 7 d;
c. and (3) third-stage seed liquid, namely inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 20%, covering an inner cover and an outer cover of the plastic pot by adopting a screening culture medium, and culturing for 6d at 37 ℃.
The anaerobic atmosphere in the step (1) or (2) is 90 percent of N2、5% CO2And 5% of H2。
The formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
The formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
The preparation method of the esterification liquid comprises the following steps:
adding 180 kg of 45 ℃ warm boiled water, 30 kg of bottom pot water, 110kg of 15-20% vol tail water, 24 kg of Zhonghui red yeast rice, 50kg of yellow water, 95 kg of 90-95% vol ethanol and 8 kg of caproic acid into the sterilized pottery jar in sequence, uniformly stirring, completely sealing the jar mouth with plastic paper, keeping the environmental temperature at 33-37 ℃, and culturing for 65 days to obtain the product.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, covering with plastic film, sealing, and culturing at 20-30 deg.C for 190 days.
Example 5
A white spirit pit mud containing esterified liquid is prepared from the following raw materials:
15 kg of Daqu powder, 12 kg of bean cake powder, 60 kg of wet distiller's grains and 0.15 m of pit skin mud30.05 m of aged pit mud31 kg of peat, 0.6 kg of micro-fertilizer, 0.5 kg of organic bone, 0.8 kg of monopotassium phosphate, 20 kg of alcohol, 140L of caproic acid bacteria tertiary seed liquid, 90 kg of esterification liquid, 280 kg of cool boiled water and 1 m of loess3。
The moisture of the yeast powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, and the esterifying power is 20-30U/g, 30-70U/g of protease activity, 1.0-9.0 times 10 of bacteria7CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
The wet vinasse is vinasse steamed after brewing, and the water content is 68-70%.
The loess is loess with surface area below 50-100cm, and is prepared by drying in the sun, removing impurities, and pulverizing.
The preparation method of the three-level seed liquid of the caproic acid bacteria comprises the following steps:
(1) strain enrichment: placing 1.2 g old cellar mud in a large test tube containing enrichment culture medium, treating in water at 80 deg.C for 13min, cooling, placing in an anaerobic incubator, and performing anaerobic culture at 35 deg.C for 7 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on an isolation culture medium under the aseptic condition, and carrying out anaerobic culture for 5 days in a culture dish at the temperature of 36 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 5 days at 36 ℃ in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium, performing amplification culture in a triangular flask, and performing anaerobic culture at 35 ℃ for 6 days;
(4) preparing a seed solution: a. first-stage seed liquid, namely inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 30%, and culturing for 6-8 d at 36 ℃ by adopting a screening culture medium;
b. inoculating mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 30%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 36 deg.C for 8 d;
c. and (3) third-stage seed liquid, namely inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 25%, adopting a screening culture medium, covering the inner cover and the outer cover of the plastic pot, and culturing for 8 d at the temperature of 36 ℃.
The anaerobic atmosphere in the step (1) or (2) is 90 percent of N2、5% CO2And 5% of H2。
The formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
The formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
The preparation method of the esterification liquid comprises the following steps:
170 kg of warm boiled water with the temperature of 40 ℃, 30 kg of bottom pot water, 105 kg of tail water with the concentration of 15-20% vol, 23 kg of Zhonghui red yeast rice, 45kg of yellow water, 90 kg of ethanol with the concentration of 90-95% vol and 5kg of caproic acid are sequentially added into the sterilized pottery jar, the mixture is uniformly stirred, the mouth of the jar is completely sealed by plastic paper, the environmental temperature is kept at 33-37 ℃, and the mixture is cultured for 60 days, thus obtaining the product.
A preparation process of pit mud containing esterified liquid comprises the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, covering with plastic film, sealing, and culturing at 20-30 deg.C for 200 days.
The performance test results of the esterified liquid and pit mud in the examples 1-5 of the invention are shown in tables 1 and 2 respectively, and meanwhile, aged pit mud with the age of 10 years is used as comparison.
TABLE 1 data of the test results of the esterified liquid of the present invention
As can be seen from Table 1, the esterified liquid of the present invention has ethyl caproate content of 3.07-3.58 g/L, ethyl lactate content of 0.77-1.08 g/L, and ethyl butyrate content of 0.02-0.03 g/L, and has high ester yield.
Table 2 results of property measurements of pit mud of the present invention
As can be seen from Table 2, the pit mud of the invention is grey brown, has pure fragrance, has strong old pit mud smell, slightly has wine fragrance, has lasting fragrance and no other foreign flavor; the hand feeling is soft, mature, fine and smooth, no thorn hand feeling exists, the section is foamed, the texture is uniform, no impurities exist, and the feeling of viscosity is obvious; pH5.8-6.4, water 38-42%, humus 18-22%, ammoniacal nitrogen 351-423 mg/Kg, available phosphorus 288-336 mg/Kg, available potassium 395-437mg/Kg, and total bacterial acid 3.4-8.1 × 109CFU/g, total spore acid 2.8-7.2X 105CFU/g. The aged cellar mud of the cellar aged 10 years in comparative example 1 is dark gray, has pure cellar aroma, has wine aroma, is rich and lasting in aroma, has no foreign flavor, is uniform and sticky in hand feeling and texture, is soft and fine, and has no impurities or thorny hand feeling; pH 6.1, water content 41%, humus 17%, ammoniacal nitrogen 320 mg/Kg, available phosphorus 272 mg/Kg, available potassium 312 mg/Kg, total bacterial acid 8.5X 108CFU/g, total acid of spore 2.3X 105CFU/g. Therefore, the physical and chemical indexes, the microbial indexes and the sensory indexes of the artificial pit mud prepared by the method reach the quality of the common pit mud aged for 10 years, even are higher than the quality of the high-quality pit mud aged for 10 years.
The application of pit mud containing esterified liquid in wine brewing comprises the following steps:
(1) cleaning pit mud with broom, and driving bamboo sticks with length of 20-25 cm and width of 2-3 cm into the pit wall, wherein the distance between the bamboo sticks is 15-25 cm, and the upper part and the lower part are in a shape of Chinese character pin;
(2) the pit mud is smeared on the pit wall and the pit bottom, the pit wall mud is 5-8 cm in thickness and the pit bottom mud is 10-15 cm in thickness, the pit bottom and the pit wall are smoothed by mud boards, Daqu powder is scattered, sorghum can be put for brewing, the conventional brewing process is adopted, the performance detection results of the obtained white spirit are shown in tables 3 and 4, and the brewing experiment is carried out by taking the aged pit mud of 10 years of pit age produced by Yangshao wine industry Limited in Henan as comparison.
TABLE 3 test results of the Chinese liquor obtained by the present invention
TABLE 4 sensory evaluation of the spirit of the invention
As can be seen from tables 3 and 4, the pit mud prepared by the method is used for brewing for the first time, the conventional physicochemical indexes of the white spirit such as total acid, total ester and the like in the obtained raw wine are close to the results of the wine produced by the pit with the age of 10 years, even exceed the results of the pit with the age of 10 years, and the sensory evaluation of the white spirit obtained by the method is equivalent to that of the pit mud with the age of 10 years, so that the use of the pit mud disclosed by the invention shortens the aging period of the pit mud, is beneficial to improving the quality of the white spirit and improving the production efficiency.
Claims (10)
1. A white spirit pit mud containing esterified liquid is characterized by being prepared from the following raw materials:
10-15 kg of Daqu powder, 10-15 kg of bean cake powder, 55-65 kg of wet distiller's grains and 0.1-0.15 m of pit skin mud30.03-0.08 m of aged cellar mud31-1.6 kg of peat, 0.2-0.8 kg of micro-fertilizer, 0.2-0.8 kg of organic bone, 0.4-0.9kg of monopotassium phosphate, 15-25 kg of alcohol, 130-150L of caproic acid bacteria tertiary seed liquid, 90-110 kg of esterification liquid, 200-300 kg of cool boiled water and 0.8-1.2 m of loess3。
2. The white spirit pit mud containing the esterified liquid as claimed in claim 1, wherein the moisture content of the Daqu powder is 9.0-11.0%, the acidity is 0.4-1.2mmol/10g, the starch is 38-40%, the saccharifying power is 700-1000U/g, the liquefying power is 0.5-1.2U/g, the fermenting power is 0.5-2.0U/g, the esterifying power is 20-30U/g, the protease activity is 30-70U/g, and the bacteria (1.0-9.0) x 107CFU/g, Yeast (1.0-9.0). times.105CFU/g, mold (1.0-9.0). times.105CFU/g, spore (1.0-9.0). times.107CFU/g。
3. The white spirit cellar mud containing the esterified liquid as claimed in claim 1, wherein the loess is loess with a surface area of 50-100cm below, and is obtained by drying in the sun, removing impurities and crushing.
4. The white spirit pit mud containing the esterified liquid as claimed in claim 1, wherein the preparation method of the caproic acid bacteria tertiary seed liquid comprises the following steps:
(1) strain enrichment: placing 0.8-1.2 g old cellar mud in a large test tube containing enrichment culture medium, treating in 75-85 deg.C water for 10-15min, cooling, placing in an anaerobic incubator, and anaerobically culturing at 33-36 deg.C for 6-8 days;
(2) and (3) strain purification: selecting the enrichment culture solution which generates the gas turbidity and caproic acid in the step (1), coating the enrichment culture solution on a separation culture medium under the aseptic condition, and carrying out anaerobic culture for 5-7 days in a culture dish at the temperature of 35-37 ℃; selecting single colony in the isolated culture medium, streaking and inoculating on the inclined plane of the isolated culture medium, and anaerobically culturing for 5-7 days at 35-37 deg.C in a test tube;
(3) screening of excellent strains: inoculating the single colony obtained in the step (2) into a test tube, adopting a screening culture medium to perform caproic acid production experiments, selecting a strain with prominent caproic acid production as a target strain, adopting the screening culture medium to perform amplification culture in a triangular flask, and performing anaerobic culture at 35-37 ℃ for 5-7 days;
(4) preparing a seed solution: a. first-stage seed liquid, namely inoculating the seeds subjected to the amplification culture in the step (3) into a 1L triangular flask according to the inoculation amount of 25-35%, and culturing for 6-8 d at 35-37 ℃ by adopting a screening culture medium;
b. inoculating mature primary seed liquid into a 5L plastic pot according to the inoculation amount of 25-35%, covering the inner and outer covers of the plastic pot with a screening culture medium, and culturing at 35-37 deg.C for 6-8 days;
c. and (3) third-stage seed liquid, inoculating the mature second-stage seed liquid into a 25L plastic pot according to the inoculation amount of 20-25%, covering the inner cover and the outer cover of the plastic pot with a screening culture medium, and culturing for 6-8 days at the temperature of 35-37 ℃.
5. The white spirit pit mud containing the esterified liquid according to claim 4, characterized in that the anaerobic atmosphere in the step (1) or (2) is 90% N2、5% CO2And 5% of H2。
6. The white spirit pit mud containing the esterified liquid according to claim 4, wherein the formula of the separation culture medium in the step (2) is as follows: 10g of peptone, 10g of beef extract, 3 g of yeast extract, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3 g of sodium acetate, 20 g of agar powder and 1000 mL of deionized water, and sterilizing at 121 ℃ for 20 min.
7. The white spirit pit mud containing the esterified liquid according to claim 4, wherein the formula of the screening culture medium in the step (3) or the step (4) is as follows: 5g of sodium acetate, 0.4 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate, 0.5 g of ammonium sulfate, 1g of yeast extract, 10g of calcium carbonate and 1000 mL of deionized water, wherein the pH value is 7.0, the mixture is sterilized at 121 ℃ for 20min, 20mL of 95% vol ethanol is added after sterilization, and the components are uniformly mixed to obtain the yeast extract.
8. The white spirit pit mud containing the esterified liquid as claimed in claim 1, wherein the preparation method of the esterified liquid comprises the following steps:
170-190 kg of warm boiled water with the temperature of 40-45 ℃, 25-35 kg of pot bottom water, 100-110 kg of 15-20% vol tail water, 22-25 kg of red yeast rice, 40-50kg of yellow water, 85-100 kg of 90-95% vol ethanol and 3-8 kg of caproic acid are sequentially added into the sterilized pottery jar, the mixture is uniformly stirred, the mouth of the pottery jar is completely sealed by plastic paper, the environmental temperature is kept at 33-37 ℃, and the pottery jar is cultured for 55-65 days to obtain the pottery jar.
9. A preparation process of the white spirit pit mud containing the esterified liquid as described in any one of claims 1 to 8, which is characterized by comprising the following steps:
mixing Daqu powder, bean cake powder, wet distiller's grains, pit skin mud, old pit mud, peat, micro-fertilizer, organic bone, and potassium dihydrogen phosphate, adding alcohol, caproic acid bacteria three-stage seed liquid, esterification liquid, and cool boiled water, mixing, adding loess, stirring with a mud mixing machine, stacking on flat ground, sealing with plastic film, and culturing at 20-30 deg.C for 180-200 days.
10. The application of the white spirit pit mud containing the esterified liquid as described in any one of claims 1 to 8 in brewing wine.
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| CN111500507A (en) * | 2020-05-28 | 2020-08-07 | 江南大学 | Pit mud flora enrichment method based on oligoculture and application thereof |
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