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Low-yield higher-alcohol saccharomyces cerevisiae engineering bacterium and construction method thereof

A technology of Saccharomyces cerevisiae and higher alcohols, which is applied in the field of bioengineering, can solve the problems of high content of higher alcohols, and achieve the effects of not being easy to reverse mutations, wide application prospects, and ensuring safety

Active Publication Date: 2010-12-15
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem of higher alcohol content in the process of fermentation and production of liquor, and to provide a low-yield higher alcohol Saccharomyces cerevisiae engineering bacteria and its construction method

Method used

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  • Low-yield higher-alcohol saccharomyces cerevisiae engineering bacterium and construction method thereof
  • Low-yield higher-alcohol saccharomyces cerevisiae engineering bacterium and construction method thereof
  • Low-yield higher-alcohol saccharomyces cerevisiae engineering bacterium and construction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1: Construction of Saccharomyces cerevisiae Engineering Bacteria with Low Yield and Higher Alcohol

[0023] (1) Construction of genetic engineering strains

[0024] The construction process of genetically engineered strains is as follows: figure 1 shown. Diploid 2.1190 was haploidized to generate haploid strains a-8 and α-22, and genetically engineered haploid strains were constructed by two homologous recombination methods, and the KanMX resistance gene was removed and the haploid strains were hybridized , to generate diploid genetically engineered strains.

[0025] The construction process of the recombinant plasmid pUC-BBAK is as follows: figure 2 shown. By PCR amplification technology, the sequence BA and BB fragments on both sides of the BAT2 gene for homologous recombination to knock out the gene and the KanMX resistance gene from the pUG6 plasmid were amplified, and the three fragments were amplified in accordance with the pUC19 plasmid as a vector. ...

Embodiment 2

[0034] Embodiment 2: Saccharomyces cerevisiae engineering strain fermentation experiment

[0035] (1) Fermentation experiments of transformants and haploid parents

[0036]Parents a-8 and α-22 and their corresponding BAT2 gene complete deletion transformants were fermented with alcohol thick mash at the same time. After the fermentation, the fermentation performance and higher alcohol production of the strains were measured. The results are shown in Table 1. The results showed that the isobutanol, isoamyl alcohol and higher alcohol contents of parent a-8 were 70.37 mg / L, 208.33 mg / L and 312.93 mg / L respectively, and the isobutanol, isoamyl alcohol and The total contents of higher alcohols were 35.37mg / L, 137.62mg / L and 204.13mg / L respectively, that is, the isobutanol, isoamyl alcohol and higher alcohols of the a-8 transformant were reduced by 49.74% and 33.94% respectively compared with the parent a-8 and 34.77%; the isobutanol, isoamyl alcohol and higher alcohol content of t...

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Abstract

The invention discloses low-yield high-alcohol saccharomyces cerevisiae engineering bacterium and a construction method thereof. The saccharomyces cerevisiae AY-20 engineering bacterium has the preservation number of CGMCC No.3930. Under the condition that other fermenting performances are not influenced, the content of the converted sub strain is respectively reduced by 55.19 percent and 34.43 percent compared with isobutanol and isoamyl alcohol of a parent strain, and the content of total higher alcohol is reduced by 35.01 percent. The invention can be realized by inserting a nucleotide sequence into start bacterium of saccharomyces cerevisiae 2.1190 or deleting partial or complete amino acid transaminase coding gene (BAT2). If the BAT2 is deleted by 100 percent, the back mutation is not easy to generate and a KanMX resistance gene in a recombinant strain is eliminated so as to ensure the safety of the strain and fermenting products. The screened engineering bacterium has no special requirements on the fermenting equipments and conditions and can be used by equipments and conditions in a general wine factory, so the invention has wide application potentials and can bring remarkable economic benefit for industrial production of a while spirit factory.

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 Saccharomyces cerevisiae engineering bacteria with low production of higher alcohols and a construction method thereof. 【Background technique】: [0002] Baijiu is a unique distilled liquor in China with a unique flavor and is deeply loved. Higher alcohol is one of the main components of liquor flavor substances, and it plays an important role in forming the flavor of liquor and making the liquor full and thick. Appropriate content of higher alcohols and the coordination of various higher alcohols can make the taste of liquor soft and coordinated, full and mellow, and unique in taste; but if the content of higher alcohols in liquor is low, the taste of liquor will become monotonous and weak; When the content is too high, it will be out of balance with the components such as acid and ester, which will cause the l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12N1/19C12N15/09C12N15/11C12R1/865
Inventor 肖冬光张翠英郝欣郭学武陈叶福杜丽平
Owner TIANJIN UNIV OF SCI & TECH
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