Gene encoding protein having trehalose synthesis-promoting activity and use thereof

A protein and trehalose technology, applied in genetic engineering, plant genetic improvement, application, etc., can solve the problems of poor low-temperature preservation of baker's yeast, and achieve the effect of eliminating thorny 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 protein having trehalose synthesis-promoting activity and use thereof
  • Gene encoding protein having trehalose synthesis-promoting activity and use thereof
  • Gene encoding protein having trehalose synthesis-promoting activity and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1 Cloning of a gene (nonScTSL1) encoding a protein having trehalose synthesis-promoting activity

[0102] As a result of searching using the comparison database described in Japanese Patent Application Laid-Open No. 2004-283169, the nonScTSL1 gene (SEQ ID NO: 1) encoding a protein having trehalose synthesis-promoting activity of S. cerevisiae was found. According to the obtained nucleotide sequence information, the primers nonScTSL1_for (sequence number 3) / nonScTSL1_rv (sequence number 4) used to amplify the full-length gene were respectively designed, and the strain Saccharomyces pastorianus Weihenstepan34 / 70 strain (referred to as "W34 / The chromosomal DNA of 70 strains") was used as a template for PCR to obtain a DNA fragment including the full-length gene of nonScTSL1.

[0103] The nonScTSL1 gene fragment obtained above was inserted into pCR2.1-TOPO vector (manufactured by Invitrogen) by TA cloning. The nucleotide sequence of the nonScTSL1 gene was analyzed...

Embodiment 2

[0104] Example 2 Expression analysis of nonScTSL1 gene in beer trial brewing

[0105] Brewery yeast Saccharomyces pastorianus W34 / 70 strain was used for trial brewing, and the mRNA extracted from the fermenting yeast cell was detected by Saccharomyces DNA microarray.

[0106] Wort extract concentration 12.69%

[0107] Wort volume 70L

[0108] Dissolved oxygen concentration in wort 8.6ppm

[0109] Fermentation temperature 15°C

[0110] The amount of yeast added 12.8×10 6 cells / mL

[0111] 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), and t...

Embodiment 3

[0112] Example 3 Construction of nonScTSL1 high expression strain

[0113] The nonScTSL1 / 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 nonScTSL1 gene. The fragment was connected to pYCGPYNot treated with restriction enzymes SacI and NotI to construct nonScTSL1 high expression vector nonScTSL1 / 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 .

[0114] Using the high expression vector obtained by the above method, adopt the method described in Japanese Patent Application Laid-Open 07-303475 to transform the AJL4004 strain, adopt the YPD plate medium (1% yeast extract, 2 % polypeptone, 2% glucos...

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Abstract

The present invention relates to a gene encoding a protein having a trehalose synthesis-promoting activity 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 TSL1 gene encoding a protein Tsl1p having a trehalose synthesis-promoting activity in brewer's yeast, especially non-ScTSL1 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 a protein with trehalose synthesis-promoting activity and uses thereof, and in particular to a practical yeast with good desiccation resistance and / or low-temperature storage resistance, an alcoholic beverage produced using the yeast, a method for producing the beverage, and the like. More specifically, the present invention relates to improving desiccation resistance and / or low-temperature storage resistance by increasing the expression level of the TSL1 gene encoding the protein Tsl1p of Saccharomyces cerevisiae, particularly the characteristic gene nonScTSL1 of Saccharomyces cerevisiae. yeast, a method for producing an alcoholic beverage using the yeast, etc. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/39C12N15/63C12N1/19C12C11/02C12G1/00C12Q1/68C12Q1/04
CPCC12C12/006C12N9/90C12G1/02C12C12/004
Inventor 中尾嘉宏儿玉由纪子下永朋子
Owner SUNTORY HLDG LTD
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