Gene encoding glycogen synthesis initiator and use thereof
The present invention relates to a gene encoding glycogen synthesis initiator 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 GLG1 or GLG2 gene encoding a Glg1p or Glg2p which is a glycogen synthesis initiator in brewer's yeast, especially non-ScGLG1 gene or non-ScGLG2 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 or SEQ ID NO: 3;
(b) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4;
(c) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4 in which one or more amino acids thereof are deleted, substituted, inserted and/or added, and having a glycogen synthesis initiation activity;
(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 or SEQ ID NO: 4, and said protein having a glycogen synthesis initiation activity;
(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 or SEQ ID NO: 3 under stringent conditions, and which encodes a protein having a glycogen synthesis initiation activity; 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 or SEQ ID NO: 4 under stringent conditions, and which encodes a protein having a glycogen synthesis initiation activity.
2 . The polynucleotide according to claim 1 selected from the group consisting of:
(a) a polynucleotide comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or encoding the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4 in which 1 to 10 amino acids thereof are deleted, substituted, inserted, and/or added, and wherein said protein has a glycogen synthesis initiation activity;
(b) a polynucleotide comprising a polynucleotide encoding a protein having 90% or higher identity with the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4, and having a glycogen synthesis initiation activity; and
(c) a polynucleotide comprising a polynucleotide which hybridizes to a polynucleotide consisting of a nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3 or which hybridizes to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3, under high stringent conditions, which encodes a protein having a glycogen synthesis initiation activity.
3 . The polynucleotide according to claim 1 comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3.
4 . The polynucleotide according to claim 1 comprising a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4.
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 drying-resistant property is increased.
10 . The yeast according to claim 8 , wherein low-temperature storage-resistant property is increased.
11 . A yeast having a drying-resistant property which is increased by increasing an expression level of the protein of claim 6 .
12 . A yeast having a low-temperature storage-resistant property which is increased by increasing an expression level of the protein of claim 6 .
13 . A method for producing an alcoholic beverage by using the yeast according to claim 8 .
14 . The method according to claim 13 , wherein the brewed alcoholic beverage is a malt beverage.
15 . The method according to claim 13 , wherein the brewed alcoholic beverage is wine.
16 . An alcoholic beverage produced by the method according to claim 13 .
17 . A method for assessing a test yeast for its drying-resistant property and/or low-temperature storage-resistant property, comprising using a primer or probe designed based on the nucleotide sequence of a gene having the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3 and encoding a glycogen synthesis initiator.
18 . A method for assessing a test yeast for its drying-resistant property and/or low-temperature storage-resistant property, comprising: culturing the test yeast; and measuring the expression level of the gene having the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3 and encoding a glycogen synthesis initiator.
19 . 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 or SEQ ID NO: 3 and encoding a glycogen synthesis initiator; and selecting a test yeast having an amount of the protein or the gene expression level according to favorable drying-resistant property and/or low-temperature storage-resistant property.
20 . The method for selecting a yeast according to claim 19 , 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 or SEQ ID NO: 3 and encoding a glycogen synthesis initiator; and selecting a test yeast having the gene expression higher than that in the reference yeast.
21 . The method for selecting a yeast according to claim 19 , 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.
22 . A method for producing an alcoholic beverage comprising: conducting fermentation using the yeast according to claim 8 or a yeast selected by a method comprising: culturing test yeasts; quantifying the protein encoded by the polynucleotide or measuring the expression level of the gene having the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 3 and encoding a glycogen synthesis initiator; and selecting a test yeast having an amount of the protein or the gene expression level according to favorable drying-resistant property and/or low-temperature storage-resistant property.