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Gene encoding glycogen synthase and use thereof

A technique of glycogen synthase and coding, which is applied in the field of alcoholic beverages and the manufacture of alcoholic beverages, can solve the problems of poor low-temperature preservation of baker's yeast, and achieve the effect of eliminating difficult problems and reducing weight

Inactive Publication Date: 2007-09-26
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is because baker's yeast Saccharomyces cerevisiae has poor low-temperature preservation compared to brewing yeast such as beer and sake fermented at low temperature.

Method used

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  • Gene encoding glycogen synthase and use thereof
  • Gene encoding glycogen synthase and use thereof
  • Gene encoding glycogen synthase and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1 Cloning of the gene (nonScGSY1) encoding glycogen synthase

[0104] As a result of searching using the comparative database described in JP-A-2004-283169, the nonScGSY1 gene (SEQ ID NO: 1) encoding glycogen synthase of Saccharomyces cerevisiae was found. According to the obtained nucleotide sequence information, the primers nonScGSY1_for (sequence number 5) / nonScGSY1_rv (sequence number 6) used to amplify the full-length gene were respectively designed, and the strain Saccharomyces pastorianus Weihenstepan34 / 70 strain (referred to as "W34 / 70" Strain") chromosomal DNA as a template for PCR to obtain a DNA fragment including the full-length gene of nonScGSY1.

[0105] The nonScGSY1 gene fragment obtained above was inserted into pCR2.1-TOPO vector (manufactured by Invitrogen) by TA cloning. The nucleotide sequence of the nonScGSY1 gene was analyzed and determined by the Sanger method (F. Sanger, Science, 214, 1215, 1981).

Embodiment 2

[0106] Example 2 Expression analysis of nonScGSY1 gene in beer trial brewing

[0107] Saccharomyces pastorianus W34 / 70 strain was used for beer trial brewing, and the mRNA extracted from the fermenting Saccharomyces pastorianus was detected by Saccharomyces DNA microarray.

[0108] Wort extract concentration 12.69%

[0109] Wort volume 70L

[0110] Dissolved oxygen concentration in wort 8.6ppm

[0111] Fermentation temperature 15°C

[0112] The amount of yeast added 12.8×10 6 cells / mL

[0113] The fermented liquid was sampled over time to observe the changes in the amount of yeast proliferation (Figure 1) and the apparent concentration of the extract (Figure 2). At the same time, the yeast cells were sampled to prepare mRNA, and the prepared mRNA was labeled with biotin to be hybridized with the brewer's yeast DNA microarray. Signal detection was performed using the GeneChip Operating System (GCOS; GeneChip Operating Software 1.0, manufactured by Affymetrix Corporation),...

Embodiment 3

[0114] Example 3 Construction of nonScGSY1 high expression strain

[0115] The nonScGSY1 / pCR2.1-TOPO obtained by the method described in Example 1 was digested with restriction enzymes SacI and NotI to prepare a DNA fragment containing the nonScGSY1 gene. The fragment was connected to pYCGPYNot treated with restriction enzymes SacI and NotI to construct nonScGSY1 high expression vector nonScGSY1 / pYCGPYNot. pYCGPYNot is a YCp-type yeast expression vector, and the introduced gene is highly expressed through the promoter of pyruvate kinase gene PYK1. Yeast selectable markers include the aminoglycoside antibiotic (Geneticin) resistance gene G418 r , selectable markers for E. coli include the ampicillin resistance gene Amp r .

[0116] Using the high expression vector obtained by the above method, adopt the method described in JP 07-303475 to transform AJL4004 strain, adopt YPD plate medium (1% yeast extract, 2% yeast extract) containing aminoglycoside antibiotics (Geneticin) 30...

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Abstract

The present invention relates to a gene encoding glycogen synthase and use thereof, in particular, a yeast for practical use with superior resistance property to dryness and / or low-temperature storage, alcoholic beverages produced with said yeast, and a method for producing said beverages. More particularly, the present invention relates to a yeast, whose resistance property to dryness and / or resistance property to low-temperature storage is enhanced by amplifying expression level of GSY1 or GSY2 gene encoding a Gsy1p or Gsy2p which is a glycogen synthase in brewer's yeast, especially non-ScGSY1 gene or non-ScGSY2 gene specific to a lager brewing yeast and to a method for producing alcoholic beverages with said yeast, etc.

Description

technical field [0001] The present invention relates to a gene encoding glycogen synthase and its use, and in particular to a practical yeast with good desiccation resistance and / or low temperature storage resistance, an alcoholic beverage produced using the yeast, and a production method thereof. More specifically, the present invention relates to the expression of nonScGSY1 or nonScGSY2 gene of Saccharomyces cerevisiae by increasing the expression of GSY1 or GSY2 gene of glycogen synthase Gsy1p or Gsy2p of Saccharomyces cerevisiae so as to make desiccation resistance and / or low temperature preservation Yeast with improved performance, a method for producing alcoholic beverages using the yeast, and the like. In addition, the yeast of the present invention can also be used as baker's yeast or industrial yeast. Background technique [0002] The beer brewing process is characterized in that the yeast after fermentation is recovered and used in the next fermentation (called co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/63C12N1/19C12C11/02C12G1/00C12Q1/68C12Q1/04
CPCC12G1/0203C12C12/004C12C12/006C12N1/04C12N9/1051
Inventor 中尾嘉宏儿玉由纪子下永朋子
Owner SUNTORY HLDG LTD
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