Fusion enzymes

摘要

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.

主权项

1. 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, andwherein 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.