Enzymes and methods for cleaving n-glycans from glycoproteins
AssigneeThe Regents of the University of California
InventorsDaniel GARRIDO, J. Bruce GERMAN, Carlito B. LEBRILLA, David A. MILLS
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Abstract
Provided herein are deglycosylating enzymes that remove a broad range of N-glycans from N-glycosylated proteins. Further provided are methods of recombinantly producing and expressing the deglycosylating enzymes. The presently described deglycosylating enzymes can be used to produce free glycans for characterization, and for prebiotic and immunostimulatory uses. In addition, the presently described deglycosylating enzymes can be used to produce deglycosylated proteins for characterization, to improve digestion, and to reduce immunogenicity.
Claims21
1-19. (canceled)
A composition comprising released N-glycans, wherein the released N-glycans are generated by contacting a glycoprotein with an enzyme at least 80% identical to SEQ ID NO:4 or SEQ ID NO:5 that cleaves high mannose glycans, complex glycans, hybrid N-glycans, or a combination thereof from the glycoprotein, thereby generating the released N-glycans.
A method of making a food comprising prebiotics, the method comprising adding released N-glycans to a food, wherein the released N-glycans are generated by contacting a glycoprotein comprising high mannose glycans, complex glycans, hybrid N-glycans, or a combination thereof with an enzyme at least 80% identical to SEQ ID NO:4 or SEQ ID NO:5 that cleaves the high mannose glycans, complex glycans, hybrid N-glycans, or a combination thereof from the glycoprotein.
A method of releasing glycans from glycoproteins comprising high mannose glycans, complex glycans, hybrid N-glycans, or a combination thereof, the method comprising,
- contacting the glycoproteins with an enzyme at least 80% identical to SEQ ID NO:4 or SEQ ID NO:5 that cleaves the high mannose glycans, complex glycans, hybrid N-glycans, or a combination thereof from the glycoproteins, thereby releasing glycans from the glycoproteins and deglycosylating the glycoproteins.
The method of claim 39, wherein the enzyme lacks a transmembrane domain.
Dependencies are inferred from the claim text.