CN111154662A - High-concentration beer strain and screening method thereof - Google Patents
High-concentration beer strain and screening method thereof Download PDFInfo
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- CN111154662A CN111154662A CN202010014507.8A CN202010014507A CN111154662A CN 111154662 A CN111154662 A CN 111154662A CN 202010014507 A CN202010014507 A CN 202010014507A CN 111154662 A CN111154662 A CN 111154662A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C12/00—Processes specially adapted for making special kinds of beer
- C12C12/002—Processes specially adapted for making special kinds of beer using special microorganisms
- C12C12/006—Yeasts
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
The invention relates to a high-concentration beer strain and a screening method thereof, belonging to the technical field of beer brewing. The screening method comprises the following steps: domestication: inoculating the starting strain into a compound culture medium containing predetermined amino acid for acclimatization for 3-7 times, wherein the sugar degree of the compound culture medium is 25-30 DEG P; separation: carrying out fermentation culture on the culture solution obtained by the last domestication, and selecting yeast meeting a preset standard as preset separation yeast; coating the colony of the selected predetermined separated yeast on a composite culture medium plate with the sugar degree of 25-30 DEG P, streaking, culturing, separating, activating, and separating by a gradient dilution method to obtain a single colony; screening: inoculating the single colony obtained by separation into 18 ° P wort, culturing at 12 deg.C, adding 8% alcohol when the sugar degree of wort is reduced to 12 ° P, culturing at 16 deg.C, and selecting high alcohol concentration resistant strain. The obtained yeast can resist 30-degree P ultrahigh-concentration wort for fermentation and produce beer with 13.8% alcoholic strength.
Description
Technical Field
The invention relates to the technical field of beer brewing, in particular to a high-concentration beer strain and a screening method thereof.
Background
Along with the development of the refined beer in the world, various beers with various flavors and bright characteristics emerge. In the aspect of high-alcohol beer, the alcohol content is generally improved by adding the separated alcohol back through beer distillation, but the method has high energy consumption and complex process, is not applicable today when energy conservation and consumption reduction are advocated and environmental protection and high efficiency are achieved, so the purpose of high alcohol content is achieved by screening ultrahigh-concentration yeast to be matched with a high-concentration brewing method.
For brewing with high alcohol content, the conventional technology relates to 20 percent tolerance°The fermentation research of the yeast with high concentration of the P wort is limited to a laboratory stage and is not applied to large-scale production. The high-gravity brewing of the modern beer products is 16-18°P wort brewing, rarely over 18°P wheat juice is brewed to produce beer. The main reason is that the fermentation performance and quality stability of high-concentration yeast are difficult to control, the higher the wort concentration is, the worse the living environment of yeast is, and the higher the osmotic pressure and alcoholic strength which the yeast needs to bear, so that the common beer yeast is difficult to be competent for the work, and must be bred to resist the workHigh-concentration wort and has good fermentation performance and genetic stability.
Disclosure of Invention
In view of the above, it is desirable to provide a beer strain with high concentration and a method for screening the same, by which a strain obtained by screening can be fermented while enduring wort and alcohol with high concentration.
A method for screening high-concentration beer strains comprises the following steps:
domestication: inoculating the starting strain into a compound culture medium containing predetermined amino acid for acclimatization for 3-7 times, wherein the sugar degree of the compound culture medium is 25-30 DEG P;
separation: carrying out fermentation culture on the culture solution obtained by the last domestication, detecting the mortality rate and the alcoholic strength of the yeast, and selecting the yeast meeting a predetermined standard as a predetermined separation yeast; coating the colony of the selected predetermined isolated yeast on a composite culture medium plate with the sugar degree of 25-30 DEG P, streaking, culturing and separating, finding out a single colony, carrying out single colony separation, selecting a colony which grows well, inoculating the colony into wort liquid, activating, and separating the single colony by adopting a gradient dilution method;
screening: inoculating the single colony obtained by separation into 18 ° P wort, culturing at 12 deg.C, adding 8% alcohol when sugar degree is reduced to 12 ° P, culturing at 15 deg.C, and selecting high alcohol concentration resistant strain.
The inventor finds in earlier studies that free amino acid is one of the most important factors for yeast growth, so that the content of amino acid in wort directly affects each biosynthesis pathway, and the method for screening high-concentration beer strains is characterized in that the inventor effectively shortens the number of experiments through skillful experimental design (such as DOE design) and calculation test steps, considers the most effective parameter information affecting the yeast ultra-high concentration fermentation performance, grasps the type and concentration of main amino acid affecting yeast biological metabolism in the fermented ultra-high concentration wort, and utilizes the affected amino acid as a sieve to prepare a culture medium, thereby screening the yeast capable of resisting the fermentation of the ultra-high concentration wort.
In the screening step, the bacterial colony is inoculated in an environment with a lower wort content than that in the acclimatization step, and is cultured at a temperature lower than the acclimatization temperature, and after the wort content is reduced to a predetermined concentration, alcohol is added and the temperature is raised for culture, so that yeast with high-concentration resistance and alcohol content can be screened.
In one embodiment, in the acclimating step, the predetermined amino acids are: 40-120mg/L aspartic acid, 25-75mg/L glutamic acid, 55-165mg/L asparagine, 40-120mg/L serine, 5-15mg/L glutamine, 30-90mg/L histidine, 20-60mg/L glycine, 35-105mg/L threonine, 60-180mg/L alanine, 75-225mg/L arginine, 55-165mg/L tyrosine, 70-210mg/L valine, 20-60mg/L methionine, 40-120mg/L isoleucine, 35-105mg/L tryptophan, 65-195mg/L phenylalanine, 85-255mg/L leucine, 55-165mg/L lysine, 5-15mg/L cysteine, 180-540mg/L proline. The said amino acid has excellent domesticating effect.
In one embodiment, the predetermined amino acid is: 80mg/L aspartic acid, 50mg/L glutamic acid, 110mg/L asparagine, 80mg/L serine, 10mg/L glutamine, 90mg/L histidine, 60mg/L glycine, 70mg/L threonine, 120mg/L alanine, 150mg/L arginine, 110mg/L tyrosine, 140mg/L valine, 40mg/L methionine, 80mg/L isoleucine, 70mg/L tryptophan, 195mg/L phenylalanine, 170mg/L leucine, 110mg/L lysine, 10mg/L cysteine, 360mg/L proline. The strain domestication is carried out by adopting the predetermined amino acid, and the optimal domestication effect is achieved.
In one implementation, in the acclimatization step, the complex culture medium further includes complex sugar, where the complex sugar is: 21.5-25.6g/L glucose, 2.9-3.8g/L fructose, 12.1-14.5g/L sucrose, 172.7-180.5g/L maltose, 41.8-46.9g/L maltotriose, 52.3-56.7g/L maltotetraose. The compound sugar is a carbon source formula required by yeast in a 30-DEG P compound culture medium used for domestication.
In one embodiment, the acclimatization step is carried out at a temperature of 15 ℃.
In one embodiment, in the acclimation step, the starting bacterial species is selected from the group consisting of: laggera yeast. The strain is used as a starting strain, and high-concentration beer strains with good effects can be obtained.
In one embodiment, the isolated culture is a fermentation culture comprising: inoculating the culture solution obtained after domestication into 18-degree P wort, culturing at 25 deg.C for 24h for activation, streaking the activated bacteria solution on compound culture medium without alcohol addition, and fermenting and culturing at 15 deg.C. Individual strains were isolated by streaking.
In one implementation, the domestication step further comprises an activation step, wherein in the activation step, a starting strain is picked and inoculated into 18-DEG P wort to be cultured for 24 hours at 25 ℃.
In one embodiment, the screening step is followed by an evaluation step in which the streaked individual strains are subjected to a pilot test to evaluate their suitability for production.
Specifically, the above-mentioned pilot test may be performed by the following method: selecting the screened strains for one circle; inoculating into 10mL of wort, and culturing at 28 deg.C for 24 hr; and then, completely filling the bacteria liquid in the activated test tube into a 30-DEG P wheat juice triangular flask added with 30mg/L histidine, 20mg/L glycine and 65mg/L phenylalanine, sealing by using a fermentation plug, culturing and fermenting for 8 days at 12 ℃, weighing every day, recording the weight loss amount every day, and indicating that the fermentation is finished when the weight loss amount every day is within 0.2 g. By this evaluation step, a strain capable of being propagated and rapidly fermented with 30 ° P wort was selected.
The invention also discloses a high-concentration beer strain obtained by screening the high-concentration beer strain screening method.
Compared with the prior art, the invention has the following beneficial effects:
the screening method of the high-concentration beer strains effectively shortens the number of experiments through ingenious experimental design and calculation test steps, simultaneously grasps the types and the concentrations of main amino acids influencing the biological metabolism of yeast in the fermented ultra-high concentration wort after considering the most effective parameter information influencing the fermentation performance of the yeast ultra-high concentration, and screens out the yeast capable of resisting the fermentation of the ultra-high concentration wort by using the influenced amino acids as a screen to prepare a culture medium. The obtained yeast can resist 30-degree P ultrahigh-concentration wort for fermentation and produce beer with 13.8% alcoholic strength.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following examples. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The starting strains used in the following examples are all conventional commercially available Lagrangian yeasts.
Example 1
A method for screening high-concentration beer strains, wherein the starting strain is Lagranzyme, comprises the following steps:
1. designing the concentration of amino acid in the culture medium.
1) The amino acid content of the 18 ℃ P wort and the 30 ℃ P wort were measured, and the results are shown in the following table.
TABLE 1 amino acid content of wort
The amino acid concentrations were calculated using the DOE design based on the above table of amino acid concentrations and are shown in the following table.
TABLE 2 amino acid concentrations
2) The compound sugar is prepared according to sugar components required by yeast, and specifically comprises the following components: 21.5-25.6g/L glucose, 2.9-3.8g/L fructose, 12.1-14.5g/L sucrose, 172.7-180.5g/L maltose, 41.8-46.9g/L maltotriose, 52.3-56.7g/L maltotetraose.
3) The complex medium was prepared from 25g/L agar according to the amino acid concentration and complex sugar in Table 2 above.
2. And (5) domesticating.
Inoculating the starting strain into the composite culture medium for domestication for 5 times, wherein the sugar degree of the composite culture medium is 30 DEG P; the method comprises the following specific steps:
and (3) pouring all the bacteria liquid in the activated test tube into a 500mL triangular flask, adding 300mL of the compound culture medium into each flask, and performing acclimatization at 15 ℃ for 5 times. And (4) carrying out fermentation culture on the culture solution obtained by the last domestication, and detecting the mortality rate and the alcoholic strength of the yeast.
3. And (5) separating.
The method comprises the following specific steps: the original strain is picked up, inoculated into 10mL of 18 DEG P wheat juice and cultured for 24h at 25 ℃ for activation. Then the activated bacteria liquid is streaked on the compound culture medium without alcohol addition, and cultured at 15 ℃. By streaking, individual strains were isolated and stored.
4. And (4) screening.
Selecting the separated single strains, inoculating the strains into 10mL of 18 DEG P wort, culturing at 25 ℃ for 24h, activating, pouring all the bacteria liquid in the activated test tube into a 500mL triangular flask, adding 300mL of the compound culture medium without adding alcohol into each flask, and fermenting at 15 ℃ until the content of diacetyl is detected to be reduced to below 0.25 ppm. After the fermentation is finished, the fermentation conditions of the fermentation broth yeast including the yeast mortality and the ethanol yield are detected, and the results are shown in the following table.
TABLE 3 fermentation assay results
Number of rounds | Mortality of Yeast (Single) | Alcohol content (Single) |
1 | 21.9 | 8.5 |
2 | 23.5 | 9.2 |
3 | 26.7 | 9.7 |
4 | 19.4 | 7.6 |
5 | 18.8 | 9.9 |
6 | 20.1 | 7.3 |
7 | 25.6 | 8.1 |
8 | 22.0 | 9.4 |
9 | 21.3 | 8.7 |
10 | 19.9 | 10.8 |
11 | 15.3 | 13.8 |
12 | 18.4 | 9.2 |
13 | 17.6 | 10.9 |
14 | 24.6 | 11.2 |
15 | 22.8 | 8.4 |
16 | 16.2 | 9.2 |
17 | 23.3 | 8.3 |
18 | 26.1 | 10.6 |
19 | 21.5 | 7.9 |
20 | 20.5 | 9.6 |
21 | 27.3 | 8.8 |
22 | 19.6 | 9.4 |
From the above test results, the 11 th round amino acid formulation enables the yeast to tolerate sugar levels of 30 ° P and produce higher alcohol levels.
Using the yeast of the 11 th round as a predetermined isolated yeast; and (3) coating the colony of the selected predetermined isolated yeast in a composite culture medium plate with the 11 th round of amino acid proportion and the sugar degree of 30 DEG P for streak culture and separation, finding out a single colony for single colony separation, selecting a colony with good growth, inoculating the colony into 10mL of wort liquid for activation, and separating by adopting a gradient dilution method to obtain the single colony.
Inoculating the single colony obtained by the separation into 18-degree P wort, culturing at 12 ℃, adding 8% alcohol when the wort degree is reduced to 12-degree P, culturing at 15 ℃, and selecting a strain capable of resisting high alcohol concentration according to the yeast death rate after the sugar degree is reduced to 5-degree P.
5. And (6) evaluating.
And (3) carrying out fermentation culture on the newly obtained strain: selecting the screened strains for one circle; inoculating into 10mL of wort, and culturing at 28 deg.C for 24 hr; and then, completely filling the bacteria liquid in the activated test tube into a 30-DEG P wheat juice triangular flask added with 30mg/L histidine, 20mg/L glycine and 65mg/L phenylalanine, sealing by using a fermentation plug, culturing and fermenting for 8 days at 12 ℃, weighing every day, recording the weight loss amount every day, and indicating that the fermentation is finished when the weight loss amount every day is within 0.2 g. After the fermentation was completed, the fermentation index was measured and the results are shown in the following table.
TABLE 4 fermentation index
As can be seen from the indexes, the fermentation performance indexes of the strains bred by the culture medium using the screening method are greatly improved compared with those of the original strains.
Example 2
The selected strain obtained by selection in example 1 above was fermented by the following fermentation steps:
1) saccharifying, preparing raw wort with concentration of 16 ° P from 65-74% malt and 25-36% rice, adding syrup into boiling pan to adjust wort concentration to 30 ° P, cooling, and adding 30mg/L histidine, 20mg/L glycine and 65mg/L phenylalanine;
2) fermenting, cooling wort, charging oxygen at 10-12ppm, inoculating strain, and controlling inoculum size at 17-21 × 106The fermentation temperature is 15 ℃, and the pressure-increasing sugar degree is controlled to be 11-13 ℃ P;
3) the performance index of the fermented product was measured, and the results are shown in the following table.
TABLE 5 fermentation Performance index
From the above results, it can be seen that the above breeding yielded a high-gravity beer strain that could be fermented at 30P ultra-high gravity wort and 14% alcohol.
Example 3
A method for screening high-concentration beer strains comprises the following steps of selecting Laggera yeast as an initial strain, screening according to the method in example 1 to obtain a breeding strain, and performing fermentation culture on the breeding strain:
selecting the screened strains for one circle; inoculating into 10mL of wort, and culturing at 28 deg.C for 24 hr; and then, completely filling the bacteria liquid in the activated test tube into a 300mL 30-degree P wheat juice triangular flask added with 30mg/L histidine, 20mg/L glycine and 65mg/L phenylalanine, sealing by using a fermentation plug, culturing and fermenting for 8 days at 12 ℃, weighing every day, recording the weight loss amount every day, and indicating that the fermentation is finished when the weight loss amount per day is within 0.2 g. After the fermentation was completed, the fermentation index was measured and the results are shown in the following table.
TABLE 6 fermentation index
As can be seen from the indexes, the fermentation performance indexes of the strains bred by the culture medium using the screening method are greatly improved compared with those of the original strains.
Example 4
A method for screening high-concentration beer strains comprises the following steps of selecting Laggera yeast as an initial strain, screening according to the method in example 1 to obtain a breeding strain, and performing fermentation culture on the breeding strain:
selecting the screened strains for one circle; inoculating into 10mL of wort, and culturing at 28 deg.C for 24 hr; and then, completely filling the bacteria liquid in the activated test tube into a 30-DEG P wheat juice triangular flask added with 30mg/L histidine, 20mg/L glycine and 65mg/L phenylalanine, sealing by using a fermentation plug, culturing and fermenting for 8 days at 12 ℃, weighing every day, recording the weight loss amount every day, and indicating that the fermentation is finished when the weight loss amount every day is within 0.2 g. After the fermentation was completed, the fermentation index was measured and the results are shown in the following table.
TABLE 7 fermentation indices
As can be seen from the indexes, the fermentation performance indexes of the strains bred by the culture medium using the screening method are greatly improved compared with those of the original strains.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A method for screening high-concentration beer strains is characterized by comprising the following steps:
domestication: inoculating the starting strain into a compound culture medium containing predetermined amino acid for acclimatization for 3-7 times, wherein the sugar degree of the compound culture medium is 25-30 DEG P;
separation: carrying out fermentation culture on the culture solution obtained by the last domestication, detecting the mortality rate and the alcoholic strength of the yeast, and selecting the yeast meeting a predetermined standard as a predetermined separation yeast; coating the colony of the selected predetermined isolated yeast on a composite culture medium plate with the sugar degree of 25-30 DEG P, carrying out streak culture and separation, finding out a single colony, carrying out single colony separation, selecting a colony which grows well, inoculating the colony into wort liquid for activation, and separating by adopting a gradient dilution method to obtain a single colony;
screening: inoculating the single colony obtained by separation into 18 ° P wort, culturing at 12 deg.C, adding 8% alcohol when the sugar degree of wort is reduced to 12 ° P, culturing at 16 deg.C, and selecting high alcohol concentration resistant strain.
2. The method for screening high gravity beer yeast according to claim 1, wherein in said acclimatization step, the predetermined amino acids are: 40-120mg/L aspartic acid, 25-75mg/L glutamic acid, 55-165mg/L asparagine, 40-120mg/L serine, 5-15mg/L glutamine, 30-90mg/L histidine, 20-60mg/L glycine, 35-105mg/L threonine, 60-180mg/L alanine, 75-225mg/L arginine, 55-165mg/L tyrosine, 70-210mg/L valine, 20-60mg/L methionine, 40-120mg/L isoleucine, 35-105mg/L tryptophan, 65-195mg/L phenylalanine, 85-255mg/L leucine, 55-165mg/L lysine, 5-15mg/L cysteine, 180-540mg/L proline.
3. The method for screening high gravity beer bacterial species according to claim 2, wherein said predetermined amino acids are: 80mg/L aspartic acid, 50mg/L glutamic acid, 110mg/L asparagine, 80mg/L serine, 10mg/L glutamine, 90mg/L histidine, 60mg/L glycine, 70mg/L threonine, 120mg/L alanine, 150mg/L arginine, 110mg/L tyrosine, 140mg/L valine, 40mg/L methionine, 80mg/L isoleucine, 70mg/L tryptophan, 195mg/L phenylalanine, 170mg/L leucine, 110mg/L lysine, 10mg/L cysteine, 360mg/L proline.
4. The method for screening high-concentration beer strains according to claim 1, wherein in the acclimation step, the compound culture medium further comprises compound sugar, and the compound sugar comprises: 21.5-25.6g/L glucose, 2.9-3.8g/L fructose, 12.1-14.5g/L sucrose, 172.7-180.5g/L maltose, 41.8-46.9g/L maltotriose, 52.3-56.7g/L maltotetraose.
5. The method for screening a high gravity beer yeast strain according to claim 1, wherein the temperature is maintained at 15 ℃ in the acclimatization step.
6. The method for screening high gravity beer yeast according to claim 1, wherein in the acclimatization step, the starting strain is selected from the group consisting of: laggera yeast.
7. The method for screening high gravity beer yeast according to claim 1, wherein the fermentation culture in the isolation culture specifically comprises: inoculating the culture solution obtained after domestication into 18-degree P wheat juice, culturing at 25 deg.C for 24 hr for activation, pouring the activated bacteria solution into compound culture medium without alcohol, and fermenting and culturing at 15 deg.C.
8. The method for screening high-gravity beer strains according to claim 1, wherein the acclimatization step further comprises an activation step, wherein in the activation step, starting strains are picked and inoculated into 18-degree P wort for culturing at 25 ℃ for 24 h.
9. The method for screening a high-gravity beer yeast strain according to claim 1, further comprising an evaluation step of evaluating the suitability of the screened high-gravity tolerant strain for production by a pilot test after the screening step.
10. A high gravity beer yeast strain obtained by the screening method for a high gravity beer yeast strain according to any one of claims 1 to 9.
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