CN117866770A - Method for separating yeast for fermenting tartary buckwheat yellow rice wine - Google Patents
Method for separating yeast for fermenting tartary buckwheat yellow rice wine Download PDFInfo
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- CN117866770A CN117866770A CN202311830951.7A CN202311830951A CN117866770A CN 117866770 A CN117866770 A CN 117866770A CN 202311830951 A CN202311830951 A CN 202311830951A CN 117866770 A CN117866770 A CN 117866770A
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 49
- 244000130270 Fagopyrum tataricum Species 0.000 title claims abstract description 33
- 235000014693 Fagopyrum tataricum Nutrition 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 26
- 235000019991 rice wine Nutrition 0.000 title claims description 11
- 239000001963 growth medium Substances 0.000 claims abstract description 46
- 241000235342 Saccharomycetes Species 0.000 claims abstract description 34
- 235000014101 wine Nutrition 0.000 claims abstract description 33
- 239000007788 liquid Substances 0.000 claims abstract description 17
- 230000001954 sterilising effect Effects 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000012216 screening Methods 0.000 claims abstract description 7
- 239000008223 sterile water Substances 0.000 claims abstract description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 7
- 238000005303 weighing Methods 0.000 claims abstract description 7
- 238000004140 cleaning Methods 0.000 claims abstract description 4
- 235000013339 cereals Nutrition 0.000 claims description 60
- 241000894006 Bacteria Species 0.000 claims description 14
- 238000010790 dilution Methods 0.000 claims description 10
- 239000012895 dilution Substances 0.000 claims description 10
- 238000002386 leaching Methods 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- 238000012258 culturing Methods 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 7
- 238000009835 boiling Methods 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 239000003085 diluting agent Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 238000007865 diluting Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 abstract description 37
- 238000000926 separation method Methods 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 2
- 238000000855 fermentation Methods 0.000 description 14
- 230000004151 fermentation Effects 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000010025 steaming Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
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- 230000032683 aging Effects 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000009982 effect on human Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- -1 esters and aldehydes Chemical class 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
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- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for separating yeast used for fermenting tartary buckwheat yellow wine, belonging to the technical field of separating yellow wine yeast. The separation method comprises the following steps: cleaning and sterilizing the operation table top to enable the operation table top to reach a preliminary sterile state; further sterilization was performed on the counter top using an ultraviolet lamp at an irradiation operation of 10 minutes per square meter; weighing 10g of tartary buckwheat yellow wine fermented grains, adding 100mL of sterile water, and shaking in a shaking table at a constant temperature of 28 ℃ for 20min to obtain a yeast enrichment liquid; the invention takes the yellow wine fermented grains as the extraction raw material, adopts the WL-Daqu distiller's grains flat-plate culture medium to separate and extract the saccharomycetes for fermenting the tartary buckwheat yellow wine, and has the advantages of more targeted screening of the required strain, time saving, efficiency improvement, strong pertinence and wide screening range.
Description
Technical Field
The invention relates to the technical field of yellow wine saccharomycetes separation, in particular to a method for separating a yellow wine saccharomycetes used for fermenting tartary buckwheat.
Background
Yeast widely exists in starter propagation, accumulation, cellar, air and airing hall, is an important product in the production process of yellow wine, and the growth metabolism condition of the yeast has important influence on the yield and flavor of the yellow wine. Yeast converts starch in the wine raw material into sugar under the action of unique enzymes, and then ferments the sugar into alcohol and carbon dioxide.
The yeast is the subtle operation, so that the alcohol degree and the aroma of the wine are improved. In addition, some organic compounds, such as esters and aldehydes, are produced by metabolic processes of yeasts. These substances have important taste characteristics in wine, and directly affect the taste of wine. Thus, yeasts play an important "flavourist" role in the fermentation of wine.
Disclosure of Invention
The invention aims to solve the problem that the metabolism of saccharomycetes affects the taste quality of wine in the prior art, and provides a method for separating saccharomycetes used for fermenting tartary buckwheat yellow wine.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a method for separating yeast used for fermenting tartary buckwheat yellow rice wine, which comprises the following steps:
s1, cleaning and sterilizing an operation table top to enable the operation table top to reach a preliminary sterile state;
s2, further sterilizing the operation table by using ultraviolet lamps at an irradiation operation of 10 minutes per square meter;
s3, weighing 10g of tartary buckwheat yellow wine fermented grains, adding 100mL of sterile water, and shaking for 20min in a shaking table at a constant temperature of 28 ℃ to obtain a yeast enrichment liquid;
s4, taking out the culture medium, adding 200mL/L Daqu distiller ' S grain leaching solution, adding 100mL of sterile water into the rest, placing in an autoclave for sterilization at 121 ℃ for 30min, taking out the culture medium after the sterilization is completed, cooling at normal temperature to form a WL-Daqu distiller ' S grain flat-plate culture medium, and placing the WL-Daqu distiller ' S grain flat-plate culture medium aside for later use;
s5, dividing the saccharomycete enrichment liquid into three parts, and diluting according to gradient;
s6, absorbing 100 mu L of diluent from three yeast enrichment liquids respectively, coating the diluted liquid on a WL-Daqu distiller 'S grains flat culture medium, and placing the WL-Daqu distiller' S grains flat culture medium into an incubator at 28 ℃ for culturing for 48-72 hours;
s7, taking out the WL-Daqu distiller 'S grains flat culture medium, selecting suspicious colonies, scribing on the surface of the WL-Daqu distiller' S grains flat culture medium, and culturing for 48-72 hours at the temperature of 28 ℃ in an incubator again;
s8, taking out the WL-Daqu distiller' S grains flat-plate culture medium, selecting single yeast colony, streaking and storing;
s9, periodically observing and recording the growth condition of the saccharomycetes, and avoiding introducing mixed bacteria.
Preferably, the concentration gradient of the yeast after dilution is 10 respectively -3 、10 -4 、10 -5 Three gradients.
Preferably, the WL-Daqu distiller's grains plate medium is placed in an incubator and inverted.
Preferably, the preparation method of the WL-Daqu distiller's grains leaching solution comprises the following steps:
s1, weighing vinasse and Daqu, wherein the mass ratio of the vinasse to the Daqu is 1:1;
s2, mixing the weighed distilled grain with Daqu, adding distilled pure water with the mass five times of that of the distilled grain, boiling, and continuing for 30-45 seconds after boiling;
s3, cooling the completed WL-Daqu distiller' S grains leaching liquor to room temperature.
Preferably, the steps S6-S9 are performed using WL-Daqu distillers' grains plate medium.
Preferably, after yeast is selected by dilution plating, single colonies are selected by secondary streaking.
Compared with the prior art, the invention provides a method for separating the saccharomycetes used for fermenting the tartary buckwheat yellow rice wine, which has the following beneficial effects:
1. the invention takes the yellow wine fermented grains as the extraction raw material, adopts the WL-Daqu distiller's grains flat-plate culture medium to separate and extract the saccharomycetes for fermenting the tartary buckwheat yellow wine, and has the advantages of more targeted screening of the required strain, time saving, efficiency improvement, strong pertinence and wide screening range.
2. The invention combines the traditional method for separating saccharomycetes, adds the distillers 'grains extract liquid of the distiller's yeast to better simulate the environment of fermenting yellow wine, is favorable for separating the proper saccharomycetes in the distilled grains, and selectively selects the saccharomycetes with good fermentation performance in a targeted way, thereby achieving the purpose of separation and having wide screening range.
3. The invention can effectively separate high-purity saccharomycetes from the fermented grains of the tartary buckwheat yellow wine, and provides excellent strain resources for further fermentation production of the tartary buckwheat yellow wine. Meanwhile, the WL-Daqu distiller's grains flat culture medium is used for culturing and screening, so that the survival rate and purity of the saccharomycetes can be improved, and better quality saccharomycetes can be obtained. On the WL-Daqu distiller's grains flat-plate culture medium, bacterial colony is in cream (pale yellow) to green, the surface is in spherical protrusion, and the smooth, opaque and creamy bacterial strain is Saccharomyces cerevisiae, so that the WL-Daqu distiller's grains flat-plate culture medium is used for culturing compared with a common culture medium, and a better quality saccharomycete strain can be obtained.
4. According to the invention, the WL-Daqu distiller's grains flat-plate culture medium is placed in the incubator for inversion, so that the uniform distribution of the water and oxygen of the culture medium can be better maintained, and the growth and propagation of saccharomycetes are facilitated. Meanwhile, the problems of mixed bacteria pollution and the like can be found in time by regularly observing and recording the growth condition of the saccharomycetes, and the separation purity and the culture effect are ensured.
In conclusion, the separation method has the advantages of simple operation, high separation purity, good culture effect and the like, can effectively separate high-purity saccharomycetes from the fermented grains of the tartary buckwheat yellow wine, and provides excellent strain resources for further fermentation production of the tartary buckwheat yellow wine.
Drawings
Fig. 1 is a simplified flow chart of the operation of the method in this implementation.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments.
Example 1
A method for separating yeast used for fermenting tartary buckwheat yellow rice wine, which comprises the following steps:
s1, cleaning and sterilizing an operation table top to enable the operation table top to reach a preliminary sterile state;
s2, further sterilizing the operation table by using ultraviolet lamps at an irradiation operation of 10 minutes per square meter;
s3, weighing 10g of tartary buckwheat yellow wine fermented grains, adding 100mL of sterile water, and shaking for 20min in a shaking table at a constant temperature of 28 ℃ to obtain a yeast enrichment liquid;
s4, taking out the culture medium, adding 200mL/L Daqu distiller ' S grain leaching solution, adding 100mL of sterile water into the rest, placing in an autoclave for sterilization at 121 ℃ for 30min, taking out the culture medium after the sterilization is completed, cooling at normal temperature to form a WL-Daqu distiller ' S grain flat-plate culture medium, and placing the WL-Daqu distiller ' S grain flat-plate culture medium aside for later use;
s5, dividing the saccharomycete enrichment liquid into three parts, and diluting according to gradient;
s6, absorbing 100 mu L of diluent from three yeast enrichment liquids respectively, coating the diluted liquid on a WL-Daqu distiller 'S grains flat culture medium, and placing the WL-Daqu distiller' S grains flat culture medium into an incubator at 28 ℃ for culturing for 48-72 hours;
s7, taking out the WL-Daqu distiller 'S grains flat culture medium, selecting suspicious colonies, streaking on the surface of the WL-Daqu distiller' S grains flat culture medium, and culturing for 48-72 hours at the temperature of 28 ℃ in an incubator again.
S8, taking out the WL-Daqu distiller' S grains flat-plate culture medium, selecting single yeast colony, streaking and preserving.
S9, periodically observing and recording the growth condition of the saccharomycetes, and avoiding introducing mixed bacteria.
In this example, the concentration gradients of the yeast after dilution were 10 -3 、10 -4 、10 -5 Three gradients.
In this example, the WL-Daqu stillage plate medium is placed in an incubator with inversion.
In this example, steps S6-S9 were performed using WL-Daqu distillers' grains plate medium.
In this example, after yeast was screened by the dilution plating method, single colonies were screened by secondary streaking.
Example 2 further improvement based on the method of example 1 for separating yeasts used to ferment tartary buckwheat yellow rice wine: i.e.
The preparation method of the WL-Daqu distiller's grains leaching solution comprises the following steps:
s1, weighing vinasse and Daqu, wherein the mass ratio of the vinasse to the Daqu is 1:1;
s2, mixing the weighed distilled grain with Daqu, adding distilled pure water with the mass five times of that of the distilled grain, boiling, and continuing for 30-45 seconds after boiling;
s3, cooling the completed WL-Daqu distiller' S grains leaching liquor to room temperature.
Example 3 the dilution flask method of the present protocol screens for yeasts as follows:
s1, preparing a culture medium and a culture dish, and selecting a culture medium suitable for yeast growth, wherein the culture medium is a Daqu distiller 'S grains culture dish and a WL-Daqu distiller' S grains flat-plate culture medium;
s2, taking out a certain amount of bacterial colonies from the saccharomycete culture medium or saccharomycete suspension in the liquid culture medium as a test sample;
s3, absorbing 100 mu L of diluent from three yeast enrichment liquids respectively, and coating the diluted liquid on the WL-Daqu distiller' S grains flat-plate culture medium.
S4, taking out the WL-Daqu vinasse plate culture medium, and selecting suspicious colonies to score on the surface of the WL-Daqu vinasse plate culture medium;
s5, taking out the WL-Daqu distiller' S grains flat-plate culture medium, selecting single yeast colony, streaking and preserving.
Example 4 dilution to different concentration split for yeast;
the dilution plate method is a commonly used microorganism culture method for isolating and identifying microorganisms. In the dilution plate method, the yeast culture is diluted according to a certain proportion to prepare bacterial solutions with different concentrations. The bacterial solutions in each tube were then inoculated onto a plate of medium, and bacterial solutions of each concentration formed a colony on the medium. By observing the characteristics and the number of colonies, the type and the number of yeasts can be initially identified, and before the experiment of the invention, various concentration gradients have been tried in advance, as detailed in the following table:
from the above table it can be derived from 10 -3 、10 -4 、10 -5 In the yeast culture experiment, the culture experiment time is relatively central, and the visibility of the yeast is relatively good when the yeast is streaked after the yeast is cultured, so that 10 is selected in the method -3 、10 -4 、10 -5 Three gradients were used as the basis for concentration culture.
Example 5 in S9 of example 1, the method of avoiding the introduction of the mixed bacteria is further refined:
firstly, it is to be understood that in the operation of the process of the present invention, the types of the miscellaneous bacteria are generally bacteria, mold and other yeasts, which greatly affect the quality of the finished product, and therefore, need to be avoided, the following operations (the following operations need to be carried out in synchronization with the operation method in example 1)
S1, before, during and after the experiment, the saccharomycete is bred and fermented by utilizing surrounding nutrient substances, and if other bacteria, viruses and the like exist in the surrounding environment, the nutrient substances are preempted, so that the saccharomycete cannot grow normally. Therefore, a clean environment is maintained during fermentation, avoiding contamination by other bacteria and impurities.
S2, in the experiment, the materials used in the fermentation process are kept fresh and clean and have no germ and other bacterial pollution so as to avoid the generation of mixed bacteria and heterogeneous bacteria.
S3, in the experiment, proper temperature and humidity are needed in the fermentation process of the saccharomycetes, and if the temperature is too high or too low, or the humidity is too high or too low, the propagation and the fermentation of the saccharomycetes are not facilitated.
S4, during experimental culture, a proper starter can promote the propagation and fermentation of saccharomycetes, and reduce the influence of mixed bacteria and heterogeneous bacteria.
S5, in the fermentation process, time and fermentation conditions are strictly controlled, so that the influence of external factor interference in the fermentation process on the normal growth of saccharomycetes is avoided.
The operation is realized, and the main culture thalli can be further prevented from being corroded by mixed bacteria during the growth of the saccharomycetes.
Further, the fermented grains of the tartary buckwheat yellow wine can be selected at any period in the production process, and the fermented grains of the tartary buckwheat yellow wine refer to the tartary buckwheat yellow wine after fermentation, and are wine prepared by taking tartary buckwheat as a main raw material. The preparation process comprises the steps of soaking, steaming, inoculating saccharomycetes, fermenting, filtering, ageing and the like of the tartary buckwheat, wherein when the tartary buckwheat yellow wine unstrained is prepared, the tartary buckwheat with high quality is selected, and is fermented through the steps of soaking, steaming, inoculating saccharomycetes and the like, after the alcoholic fermentation is completed, the wine liquid is filtered, and the aged and post-treated are carried out, so that the tartary buckwheat yellow wine unstrained is finally obtained, and has unique taste and nutritional value, such as rich amino acid, vitamin, mineral substance and the like, and has a certain health care effect on human bodies.
The present invention is not limited to the above-mentioned embodiments, and any person skilled in the art, based on the technical solution of the present invention and the inventive concept thereof, can be replaced or changed within the scope of the present invention.
Claims (6)
1. The method for separating the yeast for fermenting the tartary buckwheat yellow rice wine is characterized by comprising the following steps of:
s1, cleaning and sterilizing an operation table top to enable the operation table top to reach a preliminary sterile state;
s2, further sterilizing the operation table by using ultraviolet lamps at an irradiation operation of 10 minutes per square meter;
s3, weighing 10g of tartary buckwheat yellow wine fermented grains, adding 100mL of sterile water, and shaking for 20min in a shaking table at a constant temperature of 28 ℃ to obtain a yeast enrichment liquid;
s4, taking out the culture medium, adding 200mL/L Daqu distiller ' S grains leaching solution, adding 100mL of sterile water into the rest, placing in an autoclave for sterilization at 121 ℃ for 30min, taking out the culture medium after the sterilization is finished, cooling at normal temperature to form a WL-Daqu distiller ' S grains flat-plate culture medium, and placing the WL-Daqu distiller ' S grains flat-plate culture medium aside for later use;
s5, dividing the saccharomycete enrichment liquid into three parts, and diluting according to gradient;
s6, absorbing 100 mu L of diluent from three yeast enrichment liquids respectively, coating the diluted liquid on a WL-Daqu distiller 'S grains flat culture medium, and placing the WL-Daqu distiller' S grains flat culture medium into an incubator at 28 ℃ for culturing for 48-72 hours;
s7, taking out the WL-Daqu distiller 'S grains flat culture medium, selecting suspicious colonies, scribing on the surface of the WL-Daqu distiller' S grains flat culture medium, and culturing for 48-72 hours at the temperature of 28 ℃ in an incubator again;
s8, taking out the WL-Daqu distiller' S grains flat-plate culture medium, selecting single yeast colony, streaking and storing;
s9, periodically observing and recording the growth condition of the saccharomycetes, and avoiding introducing mixed bacteria.
2. The method for separating yeast used for fermenting tartary buckwheat yellow rice wine according to claim 1, wherein the concentration gradient of the yeast after dilution is 10 respectively -3 、10 -4 、10 -5 Three gradients.
3. The method for separating yeast for fermenting tartary buckwheat yellow rice wine according to claim 2, wherein the WL-daqu distiller's grains plate culture medium is placed in an incubator to be inverted.
4. The method for separating yeast for fermenting tartary buckwheat yellow wine according to claim 1, wherein the preparation method of the WL-Daqu distiller's grains leaching liquor is as follows:
s1, weighing vinasse and Daqu, wherein the mass ratio of the vinasse to the Daqu is 1:1;
s2, mixing the weighed distilled grain with Daqu, adding distilled pure water with the mass five times of that of the distilled grain, boiling, and continuing for 30-45 seconds after boiling;
s3, cooling the completed WL-Daqu distiller' S grains leaching liquor to room temperature.
5. The method for separating yeast for fermenting tartary buckwheat yellow rice wine according to claim 1, wherein the step S6-S9 is performed by using WL-Daqu distiller' S grains flat medium.
6. The method for separating yeast for fermenting tartary buckwheat yellow rice wine according to claim 1, wherein after screening the yeast by a dilution plate method, single colonies are screened by secondary streaking.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311830951.7A CN117866770A (en) | 2023-12-27 | 2023-12-27 | Method for separating yeast for fermenting tartary buckwheat yellow rice wine |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311830951.7A CN117866770A (en) | 2023-12-27 | 2023-12-27 | Method for separating yeast for fermenting tartary buckwheat yellow rice wine |
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