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Saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as building and application of saccharomyces cerevisiae strain

The technology of Saccharomyces cerevisiae strain and yeast strain is applied in the field of bioengineering, which can solve the problem of not simultaneously regulating key genes of higher alcohol metabolism, and achieve the effects of broad market prospects, improved expression, and overcoming inconsistency in flavor.

Pending Publication Date: 2016-03-09
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the related research on the breeding of yeast strains with high ester yield and low yield of higher alcohol in liquor and rice wine, there is no report on the simultaneous regulation of key genes related to higher alcohol metabolism and ester metabolism

Method used

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  • Saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as building and application of saccharomyces cerevisiae strain
  • Saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as building and application of saccharomyces cerevisiae strain
  • Saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as building and application of saccharomyces cerevisiae strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Construction of Saccharomyces cerevisiae with high ester yield and low yield of higher alcohol

[0049] The starting strain CICC32315 used in this example. The Escherichia coli DH5a was purchased from Takara Company. The YPD medium is a universal complete medium, and the solid medium contains 2% imported agar powder.

[0050] Based on the yeast genome data in Genebank and the integrated plasmid sequence, the following primers were designed.

[0051] The primers used in the present embodiment of table 1

[0052]

[0053]

[0054] Note: The underlined part is the restriction site.

[0055] The PCR amplification system used in the present embodiment of table 2

[0056]

[0057] (1) Construction of recombinant plasmid pUC-PAK

[0058] The construction process of the recombinant plasmid pUC-PAK is as follows: figure 1 shown;

[0059] Plasmid pPGK1 and carrier plasmid pUC19 were digested with restriction endonuclease HindIII respectively, and the di...

Embodiment 2

[0075] Example 2: Fermentation experiment of Saccharomyces cerevisiae strain with high ester yield and low yield of higher alcohol

[0076] (1) Corn thick mash fermentation experiments of recombinant strains and starting strains

[0077] The recombinant strain and the starting strain were subjected to corn mash fermentation experiments at the same time, the fermentation process roadmap: corn flour→soaking→liquefaction→saccharification→cooling→inoculation→fermentation→steaming wine→measurement indicators;

[0078] Process conditions:

[0079] Soaking conditions: 60670°C, soaking for 20 minutes; liquefaction conditions: 85690°C, adding high-temperature-resistant α-amylase, liquefying for 90 minutes; saccharification conditions: 55660°C, adding glucoamylase, saccharifying for 20 minutes;

[0080] Ingredients: corn flour 60g, water 130mL, high temperature resistant α-amylase 2×10 4 U / mL, 30μL, glucoamylase 1×10 5 U / mL, 90μL, 7.5×10 2 U / mL acid protease 1.2mL; nutrient salt 1mL...

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Abstract

The invention discloses a saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as a building method of the saccharomyces cerevisiae strain, and belongs to the technical field of bioengineering. According to the building method provided by the invention, through completely knocking out an amino acid transaminase gene BAT2 and an ester hydrolase gene IAH1 in an original strain, and selecting a strong promoter PGK1 over-expression alcohol acetyltransferase I gene ATF1 at the same time, the saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol is obtained. Compared with a parent strain, other fermentation performances of built recombinant bacteria are not affected, the total quantity of acetic acid ester is obviously increased and reaches 1303.6mg / L, wherein the content of ethyl acetate is 52 times that of the original strain, isoamyl acetate is increased to 73.7mg / L, the content of main higher alcohol is 151.8mg / L and is reduced by 61.4 percent in comparison with that of the original strain. By using the saccharomyces cerevisiae, ester yield is significantly increased while the higher alcohol yield is reduced, the higher requirements of white spirit related fields on yeast are met and the application prospect is wide.

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 strain with high yield of ester and low yield of higher alcohol, its construction method and application. Background technique: [0002] Baijiu is a unique distilled liquor in China, with unique flavor and much love. Higher alcohols and esters are very important components in liquor. They coexist in a certain proportion with other flavor substances in the liquor body, cooperate, supplement, set off and restrict each other, and give full play to their respective characteristics to form liquor with different aromas and styles. There are many types of esters, most of which exist in the form of ethyl esters in liquor, which have a fruity aroma and taste, and are the main carrier of the aroma of wine, making people feel joyful. Most of the esters in liquor have a positive effect on the flavor ...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12G3/02C12R1/865
CPCC12N9/1096C12G3/02C12N9/1029C12N9/18C12Y203/01C12Y206/01C12Y301/01
Inventor 张翠英肖冬光李维陈叶福郭学武杜丽平董健马立娟王建辉
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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