Polypeptides Having Glucoamylase Activity and Polynucleotides Encoding Same

Abstract

The present invention relates to isolated polypeptides having glucoamylase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

1 . An isolated polypeptide having glucoamylase activity, selected from the group consisting of:
 (a) a polypeptide comprising an amino acid sequence having preferably at least 72%, more preferably at least 73%, more preferably at least 74%, more preferably at least 75%, more preferably at least 76%, more preferably at least 77%, more preferably at least 78%, more preferably at least 79%, more preferably at least 80%, more preferably at least 81%, more preferably at least 82%, more preferably at least 83%, more preferably at least 84%, more preferably at least 85%, more preferably at least 86%, more preferably at least 87%, more preferably at least 88%, more preferably at least 89%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, most preferably at least 94%, and even most preferably at least 95%, such as at least 95%, at least 97%, at least 98%, at least 99% or even 100% identity to the mature polypeptide of SEQ ID NO: 2;   (b) a polypeptide comprising an amino acid sequence having preferably at least 72%, more preferably at least 73%, more preferably at least 74%, more preferably at least 75%, more preferably at least 76%, more preferably at least 77%, more preferably at least 78%, more preferably at least 79%, more preferably at least 80%, more preferably at least 81%, more preferably at least 82%, more preferably at least 83%, more preferably at least 84%, more preferably at least 85%, more preferably at least 86%, more preferably at least 87%, more preferably at least 88%, more preferably at least 89%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, most preferably at least 94%, and even most preferably at least 95%, such as at least 95%, at least 97%, at least 98%, at least 99% or even 100% identity to the catalytic domain shown as amino acids 21 to 472 of SEQ ID NO: 2;   (c) a polypeptide encoded by a polynucleotide that hybridizes under preferably at least medium-high stringency conditions, and most preferably at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence contained in SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii);   (d) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having preferably at least 72%, more preferably at least 73%, more preferably at least 74%, more preferably at least 75%, more preferably at least 76%, more preferably at least 77%, more preferably at least 78%, more preferably at least 79%, more preferably at least 80%, more preferably at least 81%, more preferably at least 82%, more preferably at least 83%, more preferably at least 84%, more preferably at least 85%, more preferably at least 86%, more preferably at least 87%, more preferably at least 88%, more preferably at least 89%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, most preferably at least 94%, and even most preferably at least 95%, such as at least 95%, at least 97%, at least 98%, at least 99% or even 100% identity to the mature polypeptide coding sequence of SEQ ID NO: 1; and   (e) a variant comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of the mature polypeptide of SEQ ID NO: 2.   
     
     
         2 . The polypeptide of  claim 1 , which is encoded by a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 1; or a subsequence thereof encoding a fragment having glucoamylase activity. 
     
     
         3 . The polypeptide of  claim 1 , wherein the mature polypeptide is amino acids 20 to 573 of SEQ ID NO: 2. 
     
     
         4 . An isolated polypeptide having carbohydrate-binding activity, comprising a carbohydrate binding module having at least 80%, more preferably at least 81%, more preferably at least 82%, more preferably at least 83%, more preferably at least 84%, more preferably at least 85%, more preferably at least 86%, more preferably at least 87%, more preferably at least 88%, more preferably at least 89%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, most preferably at least 94%, and even most preferably at least 95%, such as at least 95%, at least 97%, at least 98%, at least 99% or even 100% identity to the carbohydrate-binding domain shown as amino acids 480 to 573 of SEQ ID NO: 2. 
     
     
         5 . A hybrid enzyme which comprises a catalytic domain and a carbohydrate-binding domain of  claim 4 . 
     
     
         6 . The hybrid enzyme of  claim 5 , wherein the catalytic domain has an enzyme activity selected from the group consisting of alpha-amylase, amylopullulanase, beta-amylase, CGTase, glucoamylase, isoamylase, maltogenic amylase, and pullulanase. 
     
     
         7 . An isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of  claim 1 . 
     
     
         8 . A nucleic acid construct comprising the polynucleotide of  claim 7  operably linked to one or more (several) control sequences that direct the production of the polypeptide in an expression host. 
     
     
         9 . A recombinant expression vector comprising the nucleic acid construct of  claim 8 . 
     
     
         10 . A recombinant host cell comprising the nucleic acid construct of  claim 8 . 
     
     
         11 . A method of producing the polypeptide of  claim 1 , comprising: (a) cultivating a host cell comprising a nucleic acid construct comprising a nucleotide sequence encoding the polypeptide under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide. 
     
     
         12 . A method of producing the polypeptide of  claim 1 , comprising: (a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding the polypeptide under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide. 
     
     
         13 . A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         14 . A composition comprising an alpha-amylase and a polypeptide of  claim 1 . 
     
     
         15 . A process of producing a fermentation product from starch-containing material comprising the steps of:
 (a) liquefying starch-containing material;   (b) saccharifying the liquefied material; and   (c) fermenting with a fermenting organism;   
       wherein step (b) is carried out using at least a glucoamylase according to  claim 1 . 
     
     
         16 . A process of producing a fermentation product from starch-containing material, comprising the steps of:
 (a) saccharifying starch-containing material at a temperature below the initial gelatinization temperature of said starch-containing material; and   (b) fermenting with a fermenting organism,   
       wherein step (a) is carried out using at least a glucoamylase according to  claim 1 .