Peptidase and its uses

Abstract

The invention relates to the uses of a new characterized TET protein showed restricted to N-terminus glycine residues exopeptidase. The invention also relates to a method comprising said use of said new characterized TET protein as a N-terminus glycine residues specific exopeptidase. The invention further relates to a support wherein it is immobilized on said new characterized TET protein as a N-terminus glycine residues specific exopeptidase.

1 . A method for providing a N-terminus glycine residues specific exopeptidase, wherein said N-terminus glycine residues specific exopeptidase is provided by a TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity. 
     
     
         2 . A method for the modification of all or part of the polypeptide content of a substrate comprising peptides, polypeptides and/or proteins harbouring a N-terminus glycine residue, wherein said modification is performed by at least a TET protein harbouring at least a N-terminus glycine residues specific exopeptidase activity, said at least TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1,
 or any homologous protein derived from said at least TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity.   
     
     
         3 . The method according to  claim 1 , wherein said TET protein or said derived protein originates from an extremophilic microorganism belonging to the Thermococcales order. 
     
     
         4 . The method according to  claim 3 , wherein said extremophilic microorganism is  Pyrococcus horikoshii.    
     
     
         5 . The method according to  claim 2 , wherein peptides, polypeptides and/or proteins of said substrate are obtained from food industry or health industry or chemical industry, preferably proteins from fermented products or soya products or sea food products or cheese products. 
     
     
         6 . Method for degrading, from the N-terminus part, a polypeptide harbouring a glycine residue at its N-terminal part, said method comprising a step of contacting said polypeptide harbouring a glycine residue at its N-terminal part with
 at least a TET protein harbouring at least a N-terminus glycine residues specific exopeptidase activity, said TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or   at least one homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity,   
       and possibly a step of recovering the resulting N-terminal glycine free peptides. 
     
     
         7 . Method for modifying all or part of the polypeptide content of a substrate comprising peptides, polypeptides and/or proteins, wherein at least one of the peptides, polypeptides and/or proteins of said substrate harbours a N-terminus glycine residue, said method comprising a step of contacting said substrate with
 at least a TET protein harbouring at least a N-terminus glycine residues specific exopeptidase activity, said TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or   at least one homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity,   
       and possibly a step of recovering the modified polypeptide content of said substrate. 
     
     
         8 . The method according to  claim 6 , wherein said step of contacting comprises the activation of the said TET protein or derived protein using as enzyme cofactor at least one of the metal ions of the group comprising: Ni 2+ , Co 2+  and Mn 2+ , preferably using Ni 2+  as enzyme cofactor. 
     
     
         9 . The method according to  claim 6 , wherein said step of contacting is carried out from pH 9 to pH 10, preferably at pH 9.5. 
     
     
         10 . The method according to  claim 6 , wherein said step of contacting is carried out from 80° C. to 100° C., preferably at 85° C. 
     
     
         11 . The method according to  claim 6 , wherein said at least TET protein or said derived protein is immobilized on a solid support, preferably on a filter cartridge or on silica beads or on magnetic beads or on organic polymeric materials, or on inorganic polymeric materials or on membrane devices or in microcapsules. 
     
     
         12 . A method for the modification of all or part of the polypeptide content of a substrate comprising peptides, polypeptides and/or proteins, wherein at least one of the peptides, polypeptides and/or proteins of said substrate harbours a N-terminus glycine residue, said method comprises the contact of said polypeptide content with a solid support, and wherein is immobilized on said solid support
 at least a TET protein harbouring at least a N-terminus glycine residues specific exopeptidase activity, said TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or   at least one homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity.   
     
     
         13 . The method according to  claim 12 , wherein said solid support is a filter cartridge or silica beads or magnetic beads or on organic polymeric materials, or on inorganic polymeric materials or on membrane devices or in microcapsules. 
     
     
         14 . The method according to  claim 12 , wherein peptides, polypeptides and/or proteins of said substrate are obtained from food, chemical and health industries, or from biomass.