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A brewer's yeast strain with low-yielding higher alcohol performance and its construction method

A technology of brewer's yeast and higher alcohol, applied in the field of bioengineering, can solve the problems of unsatisfactory effect, lower content of higher alcohol, great difference in regulation effect, etc., achieve good fermentation performance and growth performance, increase content and reduce higher alcohol Effect

Active Publication Date: 2021-09-07
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In modern wheat beer fermentation, there are many technological methods to reduce the content of higher alcohols. For example, Yang Xiaolan reduced the content of higher alcohols by adjusting the way of adding syrup in beer fermentation. not ideal

Method used

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  • A brewer's yeast strain with low-yielding higher alcohol performance and its construction method
  • A brewer's yeast strain with low-yielding higher alcohol performance and its construction method
  • A brewer's yeast strain with low-yielding higher alcohol performance and its construction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Embodiment 1: Construction of the Saccharomyces cerevisiae strain of double knockout BIO3 gene

[0097] The starting strain S. cerevisiae S17 was constructed by homologous recombination to construct recombinant genetically engineered strains.

[0098] According to the yeast genome data in Genebank and the integrated plasmid sequence, the primers in the following Example 1 were designed.

[0099] Table 1 PCR primers required for double knockout of BIO3 gene

[0100]

[0101]

[0102] The main construction process of the recombinant strain is as follows (see the attached S17-DΔbio3-k-p homologous recombination strain construction flow chart figure 1 ):

[0103] 1) Amplification of the fragment required for one allele knockout of the BIO3 gene

[0104] Using the genome of Saccharomyces cerevisiae S17 as a template, BIO3A-F and BIO3A-R were used as primer pairs to PCR amplify the upstream homologous sequence BIO3A fragment required for BIO3 gene knockout, with a le...

Embodiment 2

[0123] Embodiment 2: Construction of the Saccharomyces cerevisiae strain of double knockout BIO5 gene

[0124] The starting strain S. cerevisiae S17 was constructed by homologous recombination to construct recombinant genetically engineered strains.

[0125] According to the yeast genome data in Genebank and the integrated plasmid sequence, the primers in the following Example 2 were designed.

[0126] Table 2 PCR primers required for double knockout of BIO5 gene

[0127]

[0128]

[0129] The main construction process of the recombinant strain is as follows (see the attached S17-DΔbio5-k-p homologous recombination strain construction flow chart Figure 10 shown):

[0130] 1) Amplification of the fragment required for one allele knockout of the BIO5 gene

[0131] Using the genome of Saccharomyces cerevisiae S17 as a template, BIO5A-F and BIO5A-R were used as primer pairs to PCR amplify the upstream homologous sequence BIO5A fragment required for BIO5 gene knockout, wi...

Embodiment 3

[0150] Example 3: Fermentation experiment of recombinant strains S17-DΔbio3-k-p, S17-DΔbio5-k-p wheat beer

[0151] 1) Fermentation process roadmap (refer to attached Figure 19 shown):

[0152] 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 ...

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Abstract

The invention discloses a brewer's yeast strain with low-yielding higher alcohol performance and a construction method thereof, which belongs to the technical field of bioengineering. The low-yielding higher-alcohol brewer's yeast strain of the invention starts from knocking out brewer's yeast by regulating the biotin synthesis pathway of brewer's yeast The complete sequence of the gene encoding the precursor substance of biotin synthesis that promotes the biotin synthesis function of brewer's yeast in the strain is realized, and the specific genes are the BIO3 gene encoding 7,8-diamino-nonanoic acid aminotransferase and the encoding 8 BIO5 gene for ‑amino‑7‑oxononanoate ethyl ester carrier protein. 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 respectively reduced by 16.85% and 23.55%. 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, relates to the breeding of industrial microorganisms, in particular to a brewer's yeast strain with low-yielding higher alcohol performance and a construction method thereof. Background technique [0002] Flavor substances in beer mainly include higher alcohols, esters, aldehydes, etc. Among them, higher alcohol is one of the important chemical substances forming beer flavor and mouthfeel. Appropriate higher alcohol content can make the taste and aroma of beer plump, soft and harmonious, but excessive higher alcohol is the main source of off-flavors in beer. Excessive n-propanol smells like ether and has a bitter taste; excessive butanol It will form the typical fusel oil odor of beer, and produce unpleasant bitterness at the same time; if amyl alcohol exceeds the standard, there will be putrefaction and sweat odor; when 2-phenylethanol is close to the threshold, there will be sour taste ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12N15/90C12N15/65C12R1/865
CPCC07K14/395C12N9/1096C12N15/65C12N15/905
Inventor 肖冬光王亚平孙中贯张翠英陈叶福杜丽平郭学武
Owner TIANJIN UNIV OF SCI & TECH
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