Beer yeast capable of producing beer with low acetaldehyde content, and domestication method of beer yeast

A brewer's yeast, low acetaldehyde technology, applied in the direction of using microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as poor acetaldehyde content reduction effect, poor strain stability, etc. The effect of enhancing the average activity and improving the metabolic capacity

Active Publication Date: 2020-09-15
TSINGTAO BREWERY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the technical problems existing in the prior art such as poor effect of reducing acetaldehyde content and poor strain stability caused by mutagenesis, the present invention proposes a beer strain capable of producing beer with low acetaldehyde content that does not require severe mutagenesis and has stable performance. Brewer's yeast and its domestication method

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  • Beer yeast capable of producing beer with low acetaldehyde content, and domestication method of beer yeast
  • Beer yeast capable of producing beer with low acetaldehyde content, and domestication method of beer yeast
  • Beer yeast capable of producing beer with low acetaldehyde content, and domestication method of beer yeast

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Embodiment 1

[0059] The present embodiment provides a brewer's yeast domestication method capable of producing beer with low acetaldehyde content, and its specific method is as follows:

[0060] ·Cultivation medium and its formula:

[0061] Ethanol-YNB liquid medium ( / L), the formula is: ethanol 10-20g, ammonium sulfate 5g, biotin 2μg, calcium pantothenate 400μg, folic acid 2μg, inositol 2000μg, nicotinic acid 400μg, p-aminobenzoic acid 200μg, pyridoxine hydrochloride Pyridoxine 400μg, riboflavin 200μg, thiamine hydrochloride 400μg, boric acid 500μg, copper sulfate 40μg, potassium iodide 100μg, ferric chloride 200μg, manganese sulfate 400μg, sodium molybdate 200μg, zinc sulfate 400μg, potassium dihydrogen phosphate 1g, sulfuric acid Magnesium 0.5g, sodium chloride 0.1g, calcium chloride 0.1g;

[0062] Disulfiram liquid medium ( / L), the formula is: ethanol 10-20g, 4-methylpyrazole 0.4g, disulfiram 0.2mg, ammonium sulfate 5g, biotin 2μg, calcium pantothenate 400μg, folic acid 2μg, muscle 2...

Embodiment 2

[0073] The present embodiment utilizes the bacterial strain fermentation experiment that the domestication method provided in Example 1 carries out, and its concrete steps are as follows:

[0074] (1) Pick multiple single colonies of starting strains and place them in YPD medium for static culture at 28°C for 48 hours, then centrifuge to obtain yeast sludge, wash them twice with deionized water, and inoculate them in ethanol-YNB liquid with 2% ethanol added In culture medium, OD after inoculation 600 = 0.5;

[0075] (2) Add the inoculated bacterial solution into the fully automatic high-throughput microbial droplet culture instrument, select the adaptive monitoring mode, and make the starting strain in the ethanol-YNB liquid medium with 2% ethanol without any repressor After culturing for 30 hours, droplets were generated;

[0076] (3) Transfer the generated droplets into the ethanol-YNB liquid medium supplemented with 0.1g / L 4-methylpyrazole, and automatically subculture fo...

Embodiment 3

[0088] The present embodiment carries out fermentation performance stability experiment to selected bacterial strain on the basis of embodiment 2, and its specific method is as follows:

[0089] The bacterial strains screened in Example 2 were used for 10 generations of subculture in YPD liquid medium. The specific method of subculture is: use an inoculation needle to inoculate in a small test tube containing 5 mL of YPD liquid medium. After the passaging was completed, the 1st generation strain, the 5th generation strain and the 10th generation strain were subjected to fermentation experiments. The fermentation results and flavor substances are shown in Table 3.

[0090] Table 3 Fermentation results of beer yeast 1st generation strain, 5th generation strain and 10th generation strain

[0091]

[0092] The content of acetaldehyde and flavor substances of the above strains were obtained through fermentation experiments on the screened 1st generation strains, 5th generation a...

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Abstract

The invention proposes beer yeast capable of producing beer with the low acetaldehyde content, and a domestication method of the beer yeast. The beer yeast is DY-01, a preservation number of the beeryeast is CGMCC No. 19840, and the beer yeast is preserved in the China General Microbiological Culture Collection Center on May 19, 2020. The domestication method mainly comprises the steps such as adaptive evolution treatment, domestication treatment, enzyme labeling determination, and fermentation result evaluation and screening. The domestication method of the invention does not perform drasticmutagenesis on the strain, and does not produce drastic changes to the strain's genome, but an adaptive evolution method is adopted, and an alcohol dehydrogenase repressor and an acetaldehyde dehydrogenase repressor are added into a culture medium, so that activities of both alcohol dehydrogenase (ADH2) and acetaldehyde dehydrogenase (ALDH) of the beer yeast are improved, and the ability that acetaldehyde is metabolized into ethanol and acetic acid is further improved. The beer yeast and domestication method have broad application prospects in the field of beer yeast and a domestication method thereof.

Description

technical field [0001] The invention belongs to the field of brewer's yeast and its domestication method, and in particular relates to a strain of brewer's yeast capable of producing beer with low acetaldehyde content and its domestication method. Background technique [0002] Acetaldehyde is the volatile aldehyde with the highest content in beer, accounting for 60% of the total aldehydes in beer. The level of acetaldehyde will affect the flavor and aging speed of beer. When the content is too high, the beer will produce unpleasant grass flavor or immature green apple flavor, while the content of acetaldehyde in mature high-quality beer is generally 3-8mg / L. Therefore, reducing the acetaldehyde content in beer is an important goal of countless beer practitioners. [0003] At present, many measures to reduce the content of acetaldehyde in beer have also been developed, mainly including the improvement of process parameters and the acquisition of engineering bacteria throug...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12N1/36C12R1/865
CPCC12N1/18C12N1/36
Inventor 余俊红尹花侯晓平陈璐贺扬赵玉祥万秀娟陈嵘张欣
Owner TSINGTAO BREWERY
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