System and method for the production of recombinant glycosylated proteins in a prokaryotic host

Abstract

A system and a method for the production of recombinant N-glycosylated target proteins. The system comprises a prokaryotic organism (e.g. Escherichia coli ) into which is introduced a genetic information encoding for a metabolic apparatus capable of carrying out the requested N-glycosylation of the target protein. Said prokaryotic organism also contains the genetic information required for the expression of one or more recombinant target proteins. The metabolic apparatus preferably comprises specific glycosyltransferases for the assembly of the oligosaccharide on a lipid carrier and an OTase that covalently links this oligosaccharide to specific residues of the desired protein.

1 . A system for the production of recombinant N-glycosylated target proteins, the system comprising a prokaryotic organism into which is introduced a genetic information encoding for a metabolic apparatus capable of carrying out the requested N-glycosylation of the target protein, wherein said prokaryotic organism also contains the genetic information required for the expression of one or more recombinant target proteins.  
   
   
       2 . The system of  claim 1 , wherein the metabolic apparatus comprises specific glycosyltransferases for the assembly of the oligosaccharide on a lipid carrier and an OTase that covalently links this oligosaccharide to specific residues of the desired target protein.  
   
   
       3 . The system of  claim 1 , wherein the prokaryotic organism is  Escherichia coli.    
   
   
       4 . The system of  claim 2 , wherein the prokaryotic organism is producing N-glycanes with a specific structure which is defined by the type of the specific glycosyltransferases.  
   
   
       5 . A method of producing recombinant N-glycosylated target proteins, the method comprising the introduction of a genetic information encoding for a metabolic apparatus capable of carrying out the requested N-glycosylation of the target protein into a prokaryotic organism, wherein also the genetic information required for the expression of one or more recombinant target proteins is introduced into said prokaryotic organism.  
   
   
       6 . The method of  claim 5 , wherein the metabolic apparatus comprises specific glycosyltransferases for the assembly of the oligosaccharide on a lipid carrier and an OTase, the OTase covalently linking this oligosaccharide to specific residues of the desired target protein.  
   
   
       7 . The method of one of claims  5  or  6 , wherein the prokaryotic organism is  Escherichia coli.    
   
   
       8 . The method of  claim 6 , wherein—by selection of specific glycosyltranferases—the prokaryotic organism is producing N-glycanes with a specific structure which is defined by the type of the specific glycosyltransferases.  
   
   
       9 . Utilization of the system of  claim 1  for the production of target proteins for the development of medicaments or for the production of medicaments for the treatment of humans or animals or plant.  
   
   
       10 . Proteins for nutrition and/or pharmaceutical purposes, produced with the system of one of claims  1 .  
   
   
       11 . Vaccines, Cytokines and the like medicaments for human or animals or plant, produced with the system of  claim 1 .  
   
   
       12 . Industrial enzymes, functional food, cosmetics, packaging materials or textiles comprising proteins produced with the system of claim.  
   
   
       13 . Utilization of a medicament produced with the system of  claim 1  for the therapy of human or animal or plant diseases.  
   
   
       14 . The system of  claim 2 , wherein the prokaryotic organism is  Escherichia coli.    
   
   
       15 . The system of  claim 3 , wherein the prokaryotic organism is producing N-glycanes with a specific structure which is defined by the type of the specific glycosyltransferases.  
   
   
       16 . Utilization of the method of  claim 5  for the production of target proteins for the development of medicaments or for the production of medicaments for the treatment of humans or animals or plant.  
   
   
       17 . Proteins for nutrition and/or pharmaceutical purposes, produced with the method of  claim 5 .  
   
   
       18 . Vaccines, Cytokines and the like medicaments for human or animals or plant, produced with the method of  claim 5 .  
   
   
       19 . Industrial enzymes, functional food, cosmetics, packaging materials or textiles comprising proteins produced with the method of  claim 5 .  
   
   
       20 . Utilization of a medicament produced with the method of  claim 5  for the therapy of human or animal or plant diseases.