Erythrose reductase, its cDNA and cell which the cDNA express

Abstract

Protein having an erythrose reductase activity; a DNA encoding the protein; a cell to which a DNA has been transferred in a manner such that the DNA is capable of expressing an erythrose reductases type III, II or I the DNA encodes; and a method for producing erythritol, comprising acting the erythrose reductases type III, II or I or a cell to which erythrose reductases type III, II or I has been transferred in a manner capable of expressing the erythrose reductases type III, II or I on D-erythrose and harvesting the produced erythritol.

1 . A protein described in (A) or (B) below: 
 (A) a protein having an amino acid sequence of SEQ ID NO: 2 in the Sequence Listing;    (B) a protein having an amino acid sequence of SEQ ID NO: 2 in the Sequence Listing, wherein the amino acid sequence includes substitution, deletion, insertion, addition or inversion of one or several amino acids, and wherein the protein has an erythrose reductase activity.    
     
     
         2 . A DNA encoding a protein described in (A) or (B) below: 
 (A) a protein having an amino acid sequence of SEQ ID NO: 2 in the Sequence Listing;    (B) a protein having an amino acid sequence of SEQ ID NO: 2 in the Sequence Listing, wherein the amino acid sequence includes substitution, deletion, insertion, addition or inversion of one or several amino acids, and wherein the protein has an erythrose reductase activity.    
     
     
         3 . The DNA as claimed in  claim 2 , wherein the DNA comprises one described in (a) or (b) below: 
 (a) a DNA containing a base sequence comprising at least nucleotides Nos. 1 to 399 out of the nucleotide sequence described in SEQ ID NO: 1 in the Sequence Listing;    (b) a DNA hybridizing with a base sequence comprising at least nucleotides Nos. 1 to 399 out of the nucleotide sequence described in SEQ ID NO: 1 in the Sequence Listing or a probe prepared therefrom under a stringent conditions and encoding a protein having an erythrose reductase activity.    
     
     
         4 . The DNA as claimed in  claim 3 , wherein the stringent condition is a condition under which washing is performed at a salt concentration corresponding to 2×SSC containing 0.1% SDS at 60° C.  
     
     
         5 . The DNA as claimed in  claim 2 , wherein the DNA comprises a DNA described in (c) or (d) below: 
 (c) a DNA containing a base sequence comprising at least nucleotides Nos. 408 to 1119 out of the nucleotide sequence described in SEQ ID NO: 1 in the Sequence Listing;    (d) a DNA hybridizing with a base sequence comprising at least nucleotides Nos. 408 to 1119 out of the nucleotide sequence described in SEQ ID NO: 1 in the Sequence Listing or a probe prepared therefrom under a stringent condition, and encoding a protein having an erythrose reductase activity.    
     
     
         6 . The DNA as claimed in  claim 5 , wherein the stringent condition is a condition under which washing is performed at a salt concentration corresponding to 2×SSC containing 0.1% SDS at 60° C.  
     
     
         7 . A cell to which has been transferred, a DNA as claimed in  claim 2 , in a manner such that the DNA is capable of expressing an erythrose reductase type III that the DNA encodes.  
     
     
         8 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 7 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         9 . A protein described in (C) or (D) below: 
 (C) a protein having an amino acid sequence of SEQ ID NO: 4 in the Sequence Listing;    (D) a protein having an amino acid sequence of SEQ ID NO: 4 in the Sequence Listing, wherein the amino acid sequence includes substitution, deletion, insertion, addition or inversion of one or several amino acids, and wherein the protein has an erythrose reductase activity.    
     
     
         10 - 16 . (canceled)  
     
     
         17 . A protein described in (E) or (F) below: 
 (E) a protein having an amino acid sequence of SEQ ID NO: 6 in the Sequence Listing;    (F) a protein having an amino acid sequence of SEQ ID NO: 6 in the Sequence Listing, wherein the amino acid sequence includes substitution, deletion, insertion, addition or inversion of one or several amino acids, and wherein the protein has an erythrose reductase activity.    
     
     
         18 . A DNA encoding a protein described in (E) or (F) below: 
 (E) a protein having an amino acid sequence of SEQ ID NO: 6 in the Sequence Listing;    (F) a protein having an amino acid sequence of SEQ ID NO: 6 in the Sequence Listing, wherein the amino acid sequence includes substitution, deletion, insertion, addition or inversion of one or several amino acids, and wherein the protein has an erythrose reductase activity.    
     
     
         19 . The DNA as claimed in  claim 18 , wherein the DNA comprises one described in (i) or (j) below: 
 (i) a DNA containing a base sequence comprising at least nucleotides Nos. 1 to 399 out of the nucleotide sequence described in SEQ ID NO: 5 in the Sequence Listing;    (j) a DNA hybridizing with a base sequence comprising at least nucleotides Nos. 1 to 399 out of the nucleotide sequence described in SEQ ID NO: 5 in the Sequence Listing or a probe prepared therefrom under a stringent condition, and encoding a protein having an erythrose reductase activity.    
     
     
         20 . The DNA as claimed in  claim 19 , wherein the stringent condition is a condition under which washing is performed at a salt concentration corresponding to 2×SSC containing 0.1% SDS at 60° C.  
     
     
         21 . The DNA as claimed in  claim 18 , wherein the DNA comprises a DNA described in (k) or (l) below: 
 (k) a DNA containing a base sequence comprising at least nucleotides Nos. 408 to 1121 out of the nucleotide sequence described in SEQ ID NO: 5 in the Sequence Listing;    (l) a DNA hybridizing with a base sequence comprising at least nucleotides Nos. 408 to 1121 out of the nucleotide sequence described in SEQ ID NO: 5 in the Sequence Listing or a probe prepared therefrom under a stringent condition, and encoding a protein having an erythrose reductase activity.    
     
     
         22 . The DNA as claimed in  claim 21 , wherein the stringent condition is a condition under which washing is performed at a salt concentration corresponding to 2×SSC containing 0.1% SDS at 60° C.  
     
     
         23 . A cell to which has been transferred, a DNA as claimed in  claim 18 , in a manner such that the DNA is capable of expressing an erythrose reductase type I that the DNA encodes.  
     
     
         24 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 23 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         25 . A method for producing erythritol, comprising the steps of acting the protein, having an erythrose reductase activity as claimed in  claim 1 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         26 . A method for producing erythritol, comprising the steps of acting the cell, as claimed in  claim 7 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         27 . A cell to which has been transferred, a DNA as claimed in  claim 3 , in a manner such that the DNA is capable of expressing an erythrose reductase type III that the DNA encodes.  
     
     
         28 . A cell to which has been transferred, a DNA as claimed in  claim 4 , in a manner such that the DNA is capable of expressing an erythrose reductase type III that the DNA encodes.  
     
     
         29 . A cell to which has been transferred, a DNA as claimed in  claim 5 , in a manner such that the DNA is capable of expressing an erythrose reductase type III that the DNA encodes.  
     
     
         30 . A cell to which has been transferred, a DNA as claimed in  claim 6 , in a manner such that the DNA is capable of expressing an erythrose reductase type III that the DNA encodes.  
     
     
         31 . A cell to which has been transferred, a DNA as claimed in  claim 19 , in a manner such that the DNA is capable of expressing an erythrose reductase type I that the DNA encodes.  
     
     
         32 . A cell to which has been transferred, a DNA as claimed in  claim 20 , in a manner such that the DNA is capable of expressing an erythrose reductase type I that the DNA encodes.  
     
     
         33 . A cell to which has been transferred, a DNA as claimed in  claim 21 , in a manner such that the DNA is capable of expressing an erythrose reductase type I that the DNA encodes.  
     
     
         34 . A cell to which has been transferred, a DNA as claimed in  claim 22 , in a manner such that the DNA is capable of expressing an erythrose reductase type I that the DNA encodes.  
     
     
         35 . A method for producing erythritol, comprising the steps of acting the protein, having an erythrose reductase activity as claimed in  claim 9 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         36 . A method for producing erythritol, comprising the steps of acting the protein, having an erythrose reductase activity as claimed in  claim 17 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         37 . (canceled)  
     
     
         38 . A method for producing erythritol, comprising the steps of acting the cell, as claimed in  claim 23 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         39 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 27 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         40 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 28 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         41 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 29 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         42 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 30 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         43 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 31 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         44 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 32 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         45 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 33 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         46 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 34 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         47 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 27 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         48 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 28 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         49 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 29 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         50 . A method for producing erythrose reductase type III, comprising the steps of cultivating a cell, as claimed in  claim 30 , in a medium to produce and accumulate erythrose reductase type III in a culture liquid, and harvesting the erythrose reductase type III from the culture liquid.  
     
     
         51 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 31 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         52 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 32 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         53 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 33 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         54 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 34 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.  
     
     
         55 . A method for producing erythritol, comprising the steps of acting the protein, having an erythrose reductase activity as claimed in  claim 2 , on D-erythrose, and harvesting a produced erythritol.  
     
     
         56 . A method for producing erythrose reductase type I, comprising the steps of cultivating a cell, as claimed in  claim 35 , in a medium to produce and accumulate erythrose reductase type I in a culture liquid, and harvesting the erythrose reductase type I from the culture liquid.