Alkaliphilic and thermophilic microorganisms and enzymes obtained therefrom

Abstract

The present invention provides thermophilic alkaliphilic bacteria and thermostable alkaline polypeptides obtainable therefrom. It also provides a method for producing polypeptides according to the invention, nucleic acids encoding such polypeptides and compositions comprising such polypeptides. It also relates to the use of enzymes obtainable from these novel organisms in the detergent industry, the paper and pulp industry and the textile industry.

1 . A pure culture of thermophilic alkaliphilic bacteria wherein the bacteria are members of the genus Thermopallium.  
     
     
         2 . A pure culture of thermophilic alkaliphilic bacteria according to  claim 1 , wherein the bacteria are members of the species  Thermopallium natronophilum.    
     
     
         3 . A pure culture of thermophilic bacteria according to  claim 1 , wherein the bacteria are members of the strain  Thermopallium Natronophilum  Tg9a, as deposited under number DSM 9460.  
     
     
         4 . A pure culture of thermophilic bacteria, wherein the nucleotide sequence of the 16S rRNA of said bacteria show from 92 to 100% identity to the 16S rRNA gene sequence of SEQ ID No. 1.  
     
     
         5 . A pure culture of bacteria, wherein the nucleotide sequence of the 16S rRNA of the bacteria shows 95-100% identity to the 16S rRNA gene sequence of SEQ ID No. 1.  
     
     
         6 . A pure culture of bacteria according to  claim 5 , wherein the bacteria are alkaliphilic or thermophilic.  
     
     
         7 . A polypeptide obtainable from bacteria as defined in any one of the preceding claims by culturing bacteria capable of producing the polypeptide under conditions conducive to the production of the polypeptide and recovering said polypeptide.  
     
     
         8 . A polypeptide according to  claim 7 , wherein the polypeptide is an enzyme.  
     
     
         9 . A polypeptide according to  claim 8 , wherein the enzyme is an amylase or a pullulanase.  
     
     
         10 . An isolated alkaline pullulanase having (a) an N-terminal amino acid sequence shown in SEQ. ID. No. 2, or an internal amino acid sequence as shown in SEQ. ID. 3; or (b) a variant thereof differing by deletion, substitution or insertion of one or more amino acids, and having pullulanase activity.  
     
     
         11 . An isolated alkaline amylase having (a) an internal amino acid sequence as shown in SEQ. ID. 4; or (b) a variant thereof differing by deletion, substitution or insertion of one or more amino acids, and having amylase activity.  
     
     
         12 . A composition comprising a polypeptide as defined in claims  7 - 9 , a pullulanase as defined in  claim 10  or an amylase as defined in  claim 11 .  
     
     
         13 . A composition according to  claim 12 , wherein the composition is a detergent composition.  
     
     
         14 . A method for obtaining a polypeptide as defined in claims  7 - 9 , a pullulanase as defined in  claim 10  or an amylase as defined in  claim 11 , comprising (a) culturing bacteria according to claims  1 - 6 , (b) recovering fractions comprising a polypeptide as defined in claims  7 - 9 , a pullulanase as defined in  claim 10  or an amylase as defined in  claim 11 .  
     
     
         15 . A method for producing a composition according to  claim 11  or  12  comprising (a) culturing bacteria according to claims  1 - 6 , (b) recovering fractions comprising a polypeptide as defined in claims  7 - 9 , a pullulanase as defined in  claim 10  or an amylase as defined in  claim 11 .  
     
     
         16 . An isolated nucleic acid encoding a polypeptide according to claims  7 - 9  or an enzyme having an amino acid sequence as shown in SEQ. ID. No. 2, 3 or 4.  
     
     
         17 . A recombinant nucleic acid comprising a nucleic acid according to  claim 16 .  
     
     
         18 . A host cell comprising recombinant nucleic acid according to  claim 17 .  
     
     
         19 . A method for producing a polypeptide according to claims  7 - 9 , a pullulanase as defined in  claim 10  or an amylase as defined in  claim 11  comprising (a) culturing a host cell according to  claim 18  so that it expresses the polypeptide, pullulanase or amylase, and (b) recovering the resulting polypeptide, pullulanase or amylase.  
     
     
         20 . The use of a polypeptide according to claims  7 - 9 , a pullulanase as defined in  claim 9  or an amylase as defined in  claim 11  in the manufacture of paper or pulp, in the manufacture of a textile, or in the formulation of a detergent composition.