Gene Encoding Transcriptional Inducer for Maltase Gene and Maltose Transporter Gene and use Thereof
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.
1 . A polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1;
(b) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2;
(c) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2 in which one or more amino acids thereof are deleted, substituted, inserted and/or added, and having a transcriptional induction activity of maltase and maltose transporter gene;
(d) a polynucleotide comprising a polynucleotide encoding a protein having an amino acid sequence having 60% or higher identity with the amino acid sequence of SEQ ID NO: 2, and said protein having a transcriptional induction activity of maltase and maltose transporter gene;
(e) a polynucleotide comprising a polynucleotide which hybridizes to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1 under stringent conditions, and which encodes a protein having a transcriptional induction activity of maltase and maltose transporter gene; and
(f) a polynucleotide comprising a polynucleotide which hybridizes to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of the polynucleotide encoding the protein having the amino acid sequence of SEQ ID NO: 2 under stringent conditions, and which encodes a protein having a transcriptional induction activity of maltase and maltose transporter gene.
2 . The polynucleotide according to claim 1 selected from the group consisting of:
(g) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, or encoding the amino acid sequence of SEQ ID NO: 2 in which 1 to 10 amino acids thereof are deleted, substituted, inserted, and/or added, and wherein said protein has a transcriptional induction activity of maltase and maltose transporter gene;
(h) a polynucleotide comprising a polynucleotide encoding a protein having 90% or higher identity with the amino acid sequence of SEQ ID NO: 2, and having a transcriptional induction activity of maltase and maltose transporter gene; and
(i) a polynucleotide comprising a polynucleotide which hybridizes to a polynucleotide consisting of a nucleotide sequence of SEQ ID NO: 1 or which hybridizes to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1, under high stringent conditions, which encodes a protein having a transcriptional induction activity of maltase and maltose transporter gene.
3 . The polynucleotide according to claim 1 comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1.
4 . The polynucleotide according to claim 1 comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2.
5 . The polynucleotide according to claim 1 , wherein the polynucleotide is DNA.
6 . A protein encoded by the polynucleotide according to claim 1 .
7 . A vector containing the polynucleotide according to claim 1 .
8 . A yeast into which the vector according to claim 7 has been introduced.
9 . The yeast according to claim 8 , wherein maltose assimilation ability is increased by introducing the vector containing the polynucleotide.
10 . The yeast according to claim 8 , wherein maltose assimilation ability is increased by increasing an expression level of the protein encoded by the polynucleotide.
11 . A method for producing an alcoholic beverage by using the yeast according to claim 8 .
12 . The method according to claim 11 , wherein the brewed alcoholic beverage is a malt beverage.
13 . An alcoholic beverage produced by the method according to claim 11 .
14 . A method for assessing a test yeast for its maltose assimilation ability, comprising using a primer or probe designed based on the nucleotide sequence of a gene having the nucleotide sequence of SEQ ID NO: 1 and encoding a transcriptional inducer for maltase gene and maltose transporter gene.
15 . A method for assessing a test yeast for its maltose assimilation ability, comprising: culturing the test yeast; and measuring the expression level of the gene having the nucleotide sequence of SEQ ID NO: 1 and encoding a transcriptional inducer for maltase gene and maltose transporter gene.
16 . A method for selecting a yeast, comprising: culturing test yeasts; quantifying the protein of claim 6 or measuring the expression level of the gene having the nucleotide sequence of SEQ ID NO: 1 and encoding a transcriptional inducer for maltase gene and maltose transporter gene; and selecting a test yeast having an amount of the protein or the gene expression level according to desired maltose assimilation ability.
17 . The method for selecting a yeast according to claim 16 , comprising: culturing a reference yeast and test yeasts; measuring for each yeast the expression level of the gene having the nucleotide sequence of SEQ ID NO: 1 and encoding a transcriptional inducer for maltase gene and maltose transporter gene; and selecting a test yeast having gene expression level higher than that in the reference yeast.
18 . The method for selecting a yeast according to claim 16 , comprising: culturing a reference yeast and test yeasts; quantifying the protein encoded by the polynucleotide in each yeast; and selecting a test yeast having a larger amount of the protein than that in the reference yeast.
19 . A method for producing an alcoholic beverage comprising: conducting fermentation using the yeast according to claim 8 or a yeast selected by the method according to claim 16 .