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Gene for transcription induction factor of encoding maltase and maltose transfer protein gene and application thereof

A maltase and transporter technology, applied in genetic engineering, plant genetic improvement, enzymes, etc., can solve problems such as insufficiency and achieve the effect of improving fermentation speed

Inactive Publication Date: 2007-10-03
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to these problems, conventional techniques for high-concentration brewing, such as fermenting wort with twice the normal concentration, are insufficient

Method used

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  • Gene for transcription induction factor of encoding maltase and maltose transfer protein gene and application thereof
  • Gene for transcription induction factor of encoding maltase and maltose transfer protein gene and application thereof
  • Gene for transcription induction factor of encoding maltase and maltose transfer protein gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1: Cloning of a gene (nonScMALR) encoding a transcriptional inducer of maltase and maltose transporter genes

[0096] As a result of searching using the comparison database described in JP-A-2004-283169, the gene nonScMALR (SEQ ID NO: 1) encoding a transcription inducer of maltase and maltose transporter genes unique to Saccharomyces cerevisiae was found. Based on the base sequence information obtained, primers nonScMALR_F (sequence number: 3) / nonScMALR_R (sequence number: 4) for amplifying the full-length gene were designed respectively, and the strain Saccharomyces pastorianus Weihenstephan34 / 70 strain (sometimes It can be abbreviated as "W34 / 70 strain") chromosomal DNA as a template, and a DNA fragment containing the full-length nonScMALR gene was obtained by PCR.

[0097] The nonScMALR gene fragment obtained above was inserted into pCR2.1-TOPO vector (manufactured by Invitrogen) by TA cloning. The base sequence of the nonScMALR gene was analyzed by the Sang...

Embodiment 2

[0098] Example 2: Analysis of nonScMALR gene expression in beer brewing trials

[0099] Saccharomyces pastorianus W34 / 70 strain was used for beer brewing experiment, and the mRNA extracted from the fermenting Saccharomyces yeast cell was detected by Saccharomyces DNA microarray.

[0100] Wort extract concentration 12.69%

[0101] Wort capacity 70L

[0102] Wort dissolved oxygen concentration 8.6ppm

[0103] Fermentation temperature 15°C

[0104] Yeast input amount 12.8×10 6 cells / mL

[0105] The fermented liquid was sampled over time, and the changes over time in the amount of yeast proliferation (Fig. 1) and apparent extract concentration (Fig. 2) were observed. At the same time, the yeast cells were sampled, and the prepared mRNA was biotinylated to hybridize with the brewer's yeast DNA microarray described in Japanese Patent Laid-Open No. 2004-283169. Signal detection was performed using a gene chip operating system (GCOS; GeneChip Operating Software 1.0, manufactured...

Embodiment 3

[0106] Example 3: Preparation of nonScMALR high expression strain

[0107] The nonScMALR / pCR2.1-TOPO described in Example 1 was digested with restriction enzymes SacI and NotI to prepare a DNA fragment including the full length of the protein coding region. This fragment was ligated with pYCGPYNot treated with restriction enzymes SacI and NotI to construct a nonScMALR high expression vector nonScMALR / pYCGPYNot. pYCGPYNot is a YCp-type yeast expression vector, and the introduced gene is highly expressed through the promoter of pyruvate kinase gene PYK1. Selectable markers in yeast include the aminoglycoside antibiotic (Geneticin) resistance gene G418 r , the selectable marker in E. coli contains the ampicillin resistance gene Amp r .

[0108] Using the high expression vector prepared by the above method, the Saccharomyces pastorianus UPMT3 strain was transformed by the method described in Japanese Patent Laid-Open No. Hei 07-303475. The UPMT3 strain is a bacterial strain in...

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Abstract

The present invention relates to a gene encoding a transcriptional inducer for maltase gene and maltose transporter gene and use thereof, in particular, a brewer's yeast with high maltose assimilation ability, alcoholic beverages produced with said yeast, and a method for producing said beverages. More particularly, the present invention relates to a yeast, whose maltose assimilation is enhanced by amplifying expression level of MALR gene encoding MalRp, a maltase and maltose transporter transcription factor in brewer's yeast, especially non-ScMALR 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 maltase and a transcription inducing factor of a maltose transporter gene and uses thereof, and particularly relates to a brewer's yeast excellent in maltose fermentability, an alcoholic beverage produced using the yeast, a production method thereof, and the like. More specifically, the present invention relates to the gene MALR that encodes maltase and maltose transporter gene MalRp of Saccharomyces cerevisiae, and particularly improves the ability to ferment maltose by increasing the expression level of the characteristic nonScMALR gene in Saccharomyces cerevisiae Yeast of the present invention and a method for producing alcoholic beverages using the same. Background technique [0002] In beer production, the original wort with an extract concentration of about 11% is fermented to obtain beer with an alcohol concentration of about 4.5-5%. In order to improve the productivity of beer, a high-concentra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/24C12N15/63C12N1/19C12C11/02C12Q1/68C12Q1/04C12G3/00
CPCC07K14/395C12N9/2408
Inventor 中尾嘉宏儿玉由纪子下永朋子
Owner SUNTORY HLDG LTD
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