Fusion Enzymes

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.

1 - 69 . (canceled) 
     
     
         70 . A fungal host cell comprising an expression vector comprising 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 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, wherein the acceptor glycan is attached to a heterologous polypeptide. 
     
     
         71 . The fungal host cell of claim  1 , wherein the host cell is selected from the group consisting of  Trichoderma, Aspergillus, Fusarium, Chrysosporium, Magnaporthe, Mycellopthora, Neurospora,  or  Penicillium.