Screening method for genes of brewing yeast

Abstract

The present invention provides (A) a method for the selection of genes participating in the desired brewing character, which comprises preparing a database compiling the data of the whole genome sequence of industrial yeast, particularly a brewing yeast used for alcoholic beverages; selecting gene participating in a desired brewing character that the brewing yeast specifically possesses; and carrying out functional analysis of the gene by disruption or overexpression; (B) a DNA array in which oligonucleotide(s) selected based on the data base compiling the data of the whole genome sequences of an industrial yeast, (C) a breeding method for constructing improved cultures achieving the desired brewing character, (D) a method for producing an alcohol or an alcoholic beverage in which productivity and quality are improved using the yeast, (E) a gene which is specific to the improved brewing yeast, and (F) a peptide encoded by the gene.

1 . A screening method for genes participating in increase in productivity and/or improvement in flavor in the production of an alcohol or an alcoholic beverage, characterized in that, (a) the whole genome sequence of industrial yeast is analyzed, (b) the sequence is compared with that of  Saccharomyces cerevisiae , (c) a gene of the industrial yeast encoding an amino acid sequence having 70 to 97% identity to an amino acid sequence encoded by the gene of  Saccharomyces cerevisiae  is selected, and (d) functional analysis of the selected gene is carried out, whereby the character given to the yeast by the gene is identified.  
     
     
         2 . The method according to  claim 1 , wherein a DNA array is used for the functional analysis in (d).  
     
     
         3 . The method according to  claim 2 , wherein said DNA array comprises one or more of oligonucleotides adhered to a solid support; 
 said one or more oligonucleotides comprise a DNA sequence having 10 to 30 nucleotides existing in an open reading frame of the whole genome sequence of an industrial yeast and not existing in the region other than the region of said 10 to 30 nucleotides sequence in the whole genome sequence, or its complementary DNA sequence.    
     
     
         4 . The method according to  claim 3 , wherein said one or more of oligonucleotides are hybrized under a stringent condition.  
     
     
         5 . The method according to  claim 2 , wherein said DNA array, comprises one or more oligonucleotides adhered to a solid support; 
 said one or more oligonucleotides comprise a DNA sequence having 10 to 30 nucleotides existing in a non-coding region of the whole genome sequence of an industrial yeast and not existing in the region other than the region of said 10 to 30 nucleotides sequence in the whole genome sequence, or its complementary DNA sequence.    
     
     
         6 . The method according to  claim 5 , wherein said one or more of oligonucleotides are hybridized under a stringent condition.  
     
     
         7 . The method according to  claim 3 , wherein said DNA array comprises two or more oligonucleotides.  
     
     
         8 . The method according to  claim 1 , wherein the industrial yeast is brewing yeast.  
     
     
         9 . The method according to  claim 1 , wherein the brewing yeast is beer yeast.  
     
     
         10 . A gene which is obtained by the screening method according to  claim 1 .  
     
     
         11 . The gene according to  claim 10 , which is characterized by being able to increase the concentration of sulfite in a culture medium of yeast when said gene is expressed in yeast.  
     
     
         12 . DNA which comprises a DNA sequence represented by SEQ ID NO: 1 or 2, and DNA which hybridizes to said DNA under stringent condition.  
     
     
         13 . DNA which encodes a polypeptide having an amino acid sequence represented by SEQ ID NO: 3 or 4, and DNA which encodes polypeptide having an amino acid sequence in which one to several amino acid residues are deficient, substituted, added or a combination thereof in an amino acid sequence represented by SEQ ID NO: 3 or 4.  
     
     
         14 . A recombinant vector containing the gene of  claim 10 .  
     
     
         15 . The recombinant vector according to  claim 14 , wherein a promoter, a terminator, or both are placed adjacent to said gene.  
     
     
         16 . The recombinant vector according to  claim 15 , wherein said promoter shows constitutive expression.  
     
     
         17 . The recombinant vector according to  claim 15 , wherein said promoter is a promoter of glyceraldehyde-3-phosphate dehydrogenase gene.  
     
     
         18 . A transformant containing the gene according to  claim 10 .  
     
     
         19 . The transformant according to  claim 18 , wherein said transformant belongs to yeast of genus  Saccharomyces.    
     
     
         20 . A polypeptide encoded by the gene according to  claim 10  or a polypeptide having an amino acid sequence in which one to several amino acid residues are deficient, substituted, added, or a combination thereof in an amino acid sequence in the said polypeptide.  
     
     
         21 . A polypeptide having an amino acid sequence represented by SEQ ID NO: 3 or 4 or a polypeptide having an amino acid sequence in which one to several amino acid residues are deficient, substituted, added, or a combination thereof in the amino acid sequence represented by SEQ ID NO: 3 or 4.  
     
     
         22 . A method for the production of an alcohol or an alcoholic beverage comprising subjecting the transformant according to  claim 18  to fermentation in a sugar-containing medium selected from the group consisting of wort, grape juice, rice juice and glucose syrup.  
     
     
         23 . A breeding method of yeast which is suitable for the production of an alcohol or an alcoholic beverage comprising controlling expression of the gene according to  claim 10 .  
     
     
         24 . The breeding method according to  claim 23 , wherein the yeast belongs to the genus  Saccharomyces.    
     
     
         25 . Yeast obtained by the breeding method according to  claim 23 .  
     
     
         26 . A method for the production of an alcohol or an alcoholic beverage comprising using the yeast according to  claim 25 .  
     
     
         27 . An alcohol or an alcoholic beverage which is produced using the method for the production of an alcohol or an alcoholic beverage according to  claim 26 .  
     
     
         28 . A DNA array comprising one or more oligonucleotides adhered to a solid support: 
 said one or more oligonucleotides comprise a DNA sequence having 10 to 30 nucleotides existing in an open reading frame of the whole genome sequence of an industrial yeast and not existing in the region other than the region of said 10 to 30 nucleotides sequence in the whole genome sequence, or its complementary DNA sequence.    
     
     
         29 . The DNA array according to  claim 28 , wherein said one or more of oligonucleotides are hybridized under a stringent condition.  
     
     
         30 . A DNA array comprising one or more oligonuclaeotides adhered to a solid support: 
 said one or more oligonucleotides comprise a DNA sequence having 10 to 30 nucleotides existing in a non-coding region of the whole genome sequence of an industrial yeast and not existing in the region other than the region of said 10 to 30 nucleotides sequence in the whole genome sequence, or its complementarty DNA sequence.    
     
     
         31 . The DNA array according to  claim 30 , wherein said one or more of oligonucleotides are hybridized under a stringent condition.  
     
     
         32 . The DNA array according to  claim 28 , wherein said DNA array comprises two or more oligonucleotides.  
     
     
         33 . The method according to  claim 5 , wherein said DNA array comprises two or more oligonucleotides.  
     
     
         34 . A recombinant vector containing the DNA of  claim 11 .  
     
     
         35 . The recombinant vector according to  claim 34 , wherein a promoter, a terminator, or both are placed adjacent to said DNA.  
     
     
         36 . The recombinant vector according to  claim 35 , wherein said promoter shows constitutive expression.  
     
     
         37 . The recombinant vector according to  claim 35 , wherein said promoter is a promoter of glyceraldehyde-3-phosphate dehydrogenase gene.  
     
     
         38 . A recombinant vector containing the DNA of  claim 12 .  
     
     
         39 . The recombinant vector according to  claim 38 , wherein a promoter, a terminator, or both are placed adjacent to said DNA.  
     
     
         40 . The recombinant vector according to  claim 39 , wherein said promoter shows constitutive expression.  
     
     
         41 . The recombinant vector according to  claim 39 , wherein said promoter is a promoter of glyceraldehyde-3-phosphate dehydrogenase gene.  
     
     
         42 . A transformant containing the DNA according to  claim 12 .  
     
     
         43 . The transformant according to  claim 42 , wherein said transformant belongs to yeast of genus  Saccharomyces.    
     
     
         44 . A transformant containing the DNA according to  claim 13 .  
     
     
         45 . The transformant according to  claim 44 , wherein said transformant belongs to yeast of genus  Saccharomyces.    
     
     
         46 . The DNA array according to  claim 30 , wherein said DNA array comprises two or more oligonucleotides.