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US 8,969,034 B2Granted
Endoglycosidases that cleave O-linked glycans
AssigneeNew England Biolabs, Inc.
InventorsDimitris Koutsioulis, Ellen Guthrie
Life of this patent
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Abstract
Methods and compositions have been described that relate to a newly identified polypeptide family wherein each member has O-glycosidase activity and specified sequence characteristics. This family of enzymes can be used for example for cleaving O-linked glycans and for synthesis of neoglycopeptides or neoglycoproteins.
Claims9
A preparation in a reaction vessel, comprising a recombinant polypeptide and glycerol, the recombinant polypeptide comprising amino acids 1-1155 of SEQ ID NO:
- 4, wherein the recombinant polypeptide consists of less than 1400 amino acids, and lacks a carbohydrate-binding domain at the C-terminal end.
A method, comprising:
- a) combining in a reaction vessel a eukaryotic glycoprotein or glycopeptide with a purified prokaryotic glycosidase having an amino acid sequence comprising:
- i) an amino acid sequence at least 35.2% identical to SEQ ID NO:
- 2;
- ii) SEQ ID NO:
- 1;
- and
- iii) an FDY amino acid sequence in a central conserved domain wherein the number of amino acids between the FDY and a C-terminal end of the polypeptide is no more than 200 amino acids,
- wherein the glycosidase polypeptide consists of less than 1400 amino acids and lacks a carbohydrate-binding domain at the C-terminal end;
- b) cleaving a Core 3 (GIcNAcβ1,3GaINAc) O-linked glycan in the glycoprotein or glycopeptide;
- and
- c) separating the glycan products cleaved from the glycoprotein or glycopeptide.
8 more elements
Dependencies are inferred from the claim text.