Patent ID: 7368108

Claim:
An in vitro method of glycosylating a polypeptide comprising an Asn or an Asp residue, the method comprising the steps of contacting the polypeptide with a glycosyl donor molecule having a GlcNAc residue and a PNGase-F amidase under conditions suitable for the linkage of the GlcNAc residue on the glycosyl donor molecule to the Asn or Asp residue on the polypeptide, wherein said PNGase-F amidase comprises at least one amino acid substitution of an amino acid residue for an active site acidic amino acid residue selected from the group consisting of Asp at position 60, Glu at position 206 and Glu at position 118 corresponding to a wild-type PNGase-F amidase sequence (SEQ ID NO:01).