A low-yielding high-alcohol brewer's yeast strain and its construction method

A technology of brewer's yeast and higher alcohol, which is applied in the field of bioengineering, can solve the problems of lowering the content of higher alcohol, large differences in regulation effects, unsatisfactory application, etc., and achieve the effect of reducing higher alcohol, fermentation performance and growth performance

Active Publication Date: 2021-06-25
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these reports are about the construction of beer yeast and lower fermenting yeast, and there are few reports on the transformation and breeding of higher alcohol metabolism-related genes of upper fermenting yeast used in wheat beer
In addition, there are many technological methods in the fermentation of modern wheat beer to reduce the content of higher alcohols. For example, Hanlong reduced the content of higher alcohols by changing the process parameters such as yeast inoculation amount, fermentation temperature, and fermentation pressure in beer fermentation. However, in actual production, The control effect varies greatly between different batches, and the application is not ideal

Method used

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  • A low-yielding high-alcohol brewer's yeast strain and its construction method
  • A low-yielding high-alcohol brewer's yeast strain and its construction method
  • A low-yielding high-alcohol brewer's yeast strain and its construction method

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1: Construction of low-yield higher alcohol brewer's yeast engineering strains

[0056] The starting strain S. cerevisiae S17 was constructed by homologous recombination to construct recombinant genetically engineered strains.

[0057] According to the yeast genome data in Genebank and the integrated plasmid sequence, the primers in the following examples were designed.

[0058] The PCR primers used in the present embodiment of table 1

[0059]

[0060] The main construction process of the strain is as follows (homologous recombination strain construction flow chart is attached figure 1 shown):

[0061] 1) Amplification of the fragment required for one allelic knockout of the GAT1 gene

[0062] Using the genome of Saccharomyces cerevisiae S17 as a template, using GAT1A-F and GAT1A-R as primers to PCR amplify an allelic knockout upstream homologous sequence GAT1A fragment of the GAT1 gene, the length is 564bp; using the genome of Saccharomyces cerevisiae S1...

Embodiment 2

[0081] Example 2: Fermentation experiment of recombinant strain S17-DΔgat1-k-p wheat beer

[0082] 1) Fermentation process roadmap: refer to the attached Figure 10 .

[0083] 2) Process conditions: crushing conditions: the degree of crushing is suitable for wheat malt without whole grains, and the degree of crushing is not too fine, so as not to cause excessive filtration pressure; liquefaction and saccharification conditions: the crushed wheat malt is mixed with a material-to-water ratio of 1:4 Proportionally add warm water at 30°C, stir well, place in a constant temperature water bath, keep at 30°C for 30 minutes, raise the temperature to 65°C at 2.0°C / min, keep for 90 minutes, rapidly raise the temperature to 78°C, and keep for 10 minutes. Fully stir once every 5 minutes during the saccharification process; filter conditions: filter the wheat wort after saccharification while it is hot, and wash the lees with hot water at 75°C for 3 times; ‰ of bitter hops (based on malt...

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Abstract

The invention discloses a beer yeast strain with low yield of higher alcohol and a construction method thereof, belonging to the technical field of bioengineering. The brewer's yeast strain with low production of higher alcohol of the present invention is obtained by knocking out the key regulatory factor GAT1 gene sequence of the nitrogen catabolism inhibition gene transcription of brewer's yeast. After the brewer's yeast strain with low production of higher alcohol is fermented in the fermentation medium with wheat as raw material, the total amount of higher alcohol of the parent strain is reduced by 21.42%. The fermentation performance and growth performance of the bacterial strain of the present invention are good, and there is no influence on the growth performance of the recombinant bacterial strain or other conditions.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to the breeding of industrial microorganisms, in particular to a beer yeast strain with low yield and higher alcohol and a construction method thereof. Background technique [0002] Flavor substances in beer mainly include higher alcohols, esters, aldehydes, phenols, acids, diketones, etc. Among them, higher alcohol is one of the important chemical substances that form beer flavor and mouthfeel. The literature "Formation of Higher Alcohol in Beer Production and Breeding of Low-yield Higher Alcohol Yeast Strains" and "Research Progress in Controlling Higher Alcohol Content in Above-fermented Wheat Beer" pointed out that appropriate higher alcohol content can make beer taste and aroma full , the role of soft and harmonious wine body, but the high content of higher alcohol will lead to the formation of off-flavors in beer, which not only affects the drinking taste and flavor quali...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/90C12C12/04C12R1/865
CPCC12C12/04C12N9/00C12N15/905
Inventor 肖冬光孙中贯王亚平张翠英陈叶福杜丽平郭学武
Owner TIANJIN UNIV OF SCI & TECH
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