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US 9,399,764 B2Granted
Fusion enzymes
AssigneesNovartis International Pharmaceutical Ltd., Glykos Finland Oy
InventorsJari Natunen, Anne Kanerva, Jukka Hiltunen, Markku Saloheimo, Heli Viskari, Anne Huuskonen
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Abstract
The present disclosure relates to recombinant proteins having N-acetylglucosaminyltransferase activity. The present disclosure further relates to methods for producing complex N-glycans including the steps of providing host cells containing such recombinant proteins and culturing the host cells such that the recombinant proteins are expressed.
Claims21
A method of producing a complex N-glycan, comprising:
- (1) providing a filamentous fungal or yeast host cell,
- wherein the host cell comprises a polynucleotide encoding a fusion protein comprising an N-acetylglucosaminyltransferase I catalytic domain and an N-acetylglucosaminyltransferase II catalytic domain,
- wherein the N-acetylglucosaminyltransferase II catalytic domain is positioned N-terminal to the N-acetylglucosaminyltransferase I catalytic domain, and
- wherein the alg3 gene is deleted from the host cell;
- and
- (2) culturing the filamentous fungal or yeast host cell such that the fusion protein is expressed, wherein the fusion protein catalyzes the transfer of N-acetylglucosamine to a terminal Manα3 residue and N-acetylglucosamine to a terminal Manα6 residue of an acceptor glycan to produce a complex N-glycan.
3 more elements
Dependencies are inferred from the claim text.