O-acetylhomoserinesulfhydorelace gene and use thereof
The present invention relates to a brewery yeast having controlled hydrogen sulfide-producing capability, a process for producing alcoholic beverages with controlled hydrogen sulfide amount. More particularly, the present invention relates to a yeast whose hydrogen sulfide-producing capability that increases the product flavor is controlled by enhancing the expression level of MET17 gene encoding brewery yeast O-acetylhomoserinesulfhydorelace Met17p, particularly non-ScMET17 gene specific to lager brewing yeast, and to a method for producing alcoholic beverages with said yeast.
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 with one or more amino acids thereof being deleted, substituted, inserted and/or added, and having an O-acetylhomoserinesulfhydorelace 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, and having an O-acetylhomoserinesulfhydorelace 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 under stringent conditions, and which encodes a protein having an O-acetylhomoserinesulfhydorelace 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 of the amino acid sequence of SEQ ID NO:2 under stringent conditions, and which encodes a protein having an O-acetylhomoserinesulfhydorelace activity.
2 . The polynucleotide of claim 1 selected from the group consisting of:
(a) a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, or encoding an amino acid sequence of SEQ ID NO: 2 wherein 1 to 10 amino acids thereof is deleted, substituted, inserted, and/or added, and wherein said protein has O-acetylhomoserinesulfhydorelace activity;
(b) a polynucleotide encoding a protein having 90% or higher identity with the amino acid sequence of SEQ ID NO: 2, and having O-acetylhomoserinesulfhydorelace activity; and
(c) a polynucleotide which hybridizes to SEQ ID NO: 1 or which hybridizes to a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1 under stringent conditions, and which encodes a protein having O-acetylhomoserinesulfhydorelace activity.
3 . The polynucleotide of claim 1 comprising a polynucleotide consisting of SEQ ID NO: 1.
4 . The polynucleotide of claim 1 comprising a polynucleotide encoding a protein consisting of SEQ ID NO: 2.
5 . The polynucleotide of claim 1 , wherein the polynucleotide is DNA.
6 . A protein encoded by the polynucleotide of claim 1 .
7 . A vector comprising the polynucleotide of claim 1 .
8 . A yeast comprising the vector of claim 7 .
9 . The yeast of claim 8 , wherein a hydrogen sulfide-producing ability is reduced by introducing the vector.
10 . The yeast of claim 9 , wherein a hydrogen sulfide-producing ability is reduced by increasing an expression level of the protein encoded by the polynucleotide.
11 . A method for producing an alcoholic beverage comprising culturing the yeast of claim 8 .
12 . The method for producing an alcoholic beverage of claim 11 , wherein the brewed alcoholic beverage is a malt beverage.
13 . The method for producing an alcoholic beverage of claim 11 , wherein the brewed alcoholic beverage is wine.
14 . An alcoholic beverage produced by the method of claim 11 .
15 . A method for assessing a test yeast for its hydrogen sulfide-producing capability, comprising using a primer or a probe designed based on a nucleotide sequence of an O-acetylhomoserinesulfhydorelace gene having the nucleotide sequence of SEQ ID NO: 1.
16 . A method for assessing a test yeast for its hydrogen sulfide-producing capability, comprising: culturing a test yeast; and measuring an expression level of an O-acetylhomoserinesulfhydorelace gene having the nucleotide sequence of SEQ ID NO: 1.
17 . A method for selecting a yeast, comprising: culturing test yeasts; quantifying the protein according to claim 6 or measuring an expression level of an O-acetylhomoserinesulfhydorelace gene having the nucleotide sequence of SEQ ID NO: 1; and selecting a test yeast having said protein amount or said gene expression level according to a target capability of producing hydrogen sulfide.
18 . The method for selecting a yeast according to claim 17 , comprising: culturing a reference yeast and test yeasts; measuring an expression level of an O-acetylhomoserinesulfhydorelace gene having the nucleotide sequence of SEQ ID NO: 1 in each yeast; and selecting a test yeast having the gene expressed higher than that in the reference yeast.
19 . The method for selecting a yeast according to claim 17 , comprising: culturing a reference yeast and test yeasts; quantifying the protein in each yeast; and selecting a test yeast having said protein for a larger amount than that in the reference yeast.
20 . A method for producing an alcoholic beverage comprising:
(a) conducting fermentation for producing an alcoholic beverage using the yeast according to claim 8 , or a yeast selected using a method for selecting a yeast, comprising: culturing test yeasts; quantifying the protein encoded by the polynucleotide or measuring an expression level of an O-acetylhomoserinesulfhydorelace gene having the nucleotide sequence of SEQ ID NO: 1; and selecting a test yeast having said protein amount or said gene expression level according to a target capability of producing hydrogen sulfide; and
(b) adjusting the production amount of hydrogen sulfide.