Sulfate ion transporter gene and use thereof

A sulfate ion and transporter technology, applied in the manufacture of alcoholic beverages and in the field of alcoholic beverages, to achieve the effects of long shelf life and excellent flavor stability

Inactive Publication Date: 2008-05-28
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The easiest way to increase the sulfite content in the product is to add sulfite, but at this time sulfite will be treated as a food additive, in terms of the freedom of product development and the negative impression of consumers on food additives wait will be a problem

Method used

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  • Sulfate ion transporter gene and use thereof
  • Sulfate ion transporter gene and use thereof
  • Sulfate ion transporter gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Example 1: Cloning of the SUL2 gene

[0111] Using the results of the comparative database search described in Japanese Patent Laid-Open No. 2004-283169, two novel SUL2 genes in the S. cerevisiae genome were found. Based on the homology with the nucleotide sequence of the Saccharomyces cerevisiae SUL2 gene published in SGD (Yeast Genome Database; http: / / www.yeastgenome.org / ), the novel high homology (99%) The found gene was defined as ScSUL2, and the gene with low homology (80%) was defined as non-ScSUL2. Based on the obtained nucleotide sequence information, primers for amplifying the respective full-length genes were designed, and the chromosomal DNA of the genome reading strain Saccharomyces pastorianus Weihenstephan 34 / 70 strain was used as a template to obtain the full-length DNA containing ScSUL2 or non-ScSUL2 by PCR. DNA fragments of the genes (both about 2.7 kb). ScSUL2 gene amplification uses primers ScSUL2_for (SEQ ID NO: 3) / ScSUL2_rv (SEQ ID NO: 4), non-S...

Embodiment 2

[0113] Example 2: Constitutive expression of the SUL2 gene

[0114] A DNA fragment of about 2.7 kb containing the ScSUL2 gene was prepared by treating the plasmid TOPO / ScSUL2 described in Example 1 with restriction enzymes SacI and NotI. This was ligated with pUP3GLP2 treated with restriction enzymes SacI and NotI to construct a ScSUL2 constitutive expression vector pUP-ScSUL2. And use the same method to construct non-ScSUL2 constitutive expression vector pUP-non-ScSUL2. The yeast expression vector pUP3GLP2 is a YIp (chromosomally integrated) vector containing the orotidine-5-phosphate decarboxylase gene URA3 as a homologous recombination site, and the introduced gene is passed through the glyceraldehyde-3-phosphate dehydrogenase gene TDH3 promoter / terminator and constitutive expression. As a selectable marker in yeast, the drug resistance gene YAP1 was introduced under the control of the promoter / terminator of the galactokinase gene GAL1, and its expression was induced in...

Embodiment 3

[0117] Example 3: Analysis of the amount of sulfite produced in beer test brewing

[0118] Using the parent strain TF010, ScSUL2 high-expression strains (2 strains) and non-ScSUL2 high-expression strains (2 strains) obtained in Example 2, the fermentation test was carried out under the following conditions.

[0119] Wort Extract Concentration 13%

[0120] Wort capacity 800mL

[0121] Wort dissolved oxygen concentration about 8ppm

[0122] Fermentation temperature was constant at 15°C

[0123] Yeast input amount 5g wet yeast cell / 800mL wort extract

[0124] Sampling of fermented mash over time, analysis of yeast proliferation (OD660) and sugar consumption over time, using figure 1 , figure 2 express. The quantification of sulfite concentration at the end of fermentation is to collect sulfite in hydrogen peroxide solution by distillation under acidic conditions, and then perform alkali titration [(Finance) Japan Brewing Association revised BCOJ beer analysis method], u...

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Abstract

The present invention relates to a sulfate ion transporter gene and uses thereof, and particularly relates to a brewer's yeast for producing an alcoholic beverage excellent in flavor stability, an alcoholic beverage produced using the yeast, a method for producing the same, and the like. More specifically, the present invention relates to increasing the expression level of the gene SUL2 encoding the brewer's yeast sulfate ion transporter Sul2p, especially the characteristic non-ScSUL2 gene in brewer's yeast, so as to improve the subunits that play a role in product aroma stabilization. Yeast capable of producing sulfate, method for producing alcoholic beverages using the yeast, and the like.

Description

technical field [0001] The present invention relates to a sulfate ion transporter gene and uses thereof, and particularly relates to a brewer's yeast for producing an alcoholic beverage excellent in flavor stability, an alcoholic beverage produced using the yeast, a method for producing the same, and the like. More specifically, the present invention relates to controlling the expression level of the gene SUL2 encoding the sulfate ion transporter Sul2p of Saccharomyces cerevisiae, especially the characteristic non-ScSUL2 gene in S. Yeast capable of producing sulfite, method for producing alcoholic beverages using the yeast, and the like. Background technique [0002] Sulfite is known as a compound with strong antioxidant activity, and is widely used as an antioxidant in the fields of food, beverages, pharmaceuticals, etc. ). Sulfites in alcoholic beverages are also used as antioxidants. For example, since they play an important role in ensuring the quality of wine that nee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/39C12N15/63C12N1/19C12C11/02C12G1/00C12Q1/68G01N33/68G01N33/569
CPCC07K14/395C12P11/00C12N1/18C12N15/81
Inventor 中尾嘉宏儿玉由纪子下永朋子
Owner SUNTORY HLDG LTD
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