A combined method for producing homogeneous glycoproteins with eukaryotic N-glycosylation.
We describe a new method for producing homogeneous eukaryotic N-glycoproteins. The method involves the engineering and functional transfer of the Campylobacter jejuni glycosylation machinery in Escherichia coli to express glycosylated proteins with the key GlcNAc-Asn linkage. The bacterial glycans were then trimmed and remodeled in vitro by enzymatic transglycosylation to fulfill a eukaryotic N-glycosylation. It provides a potentially general platform for producing eukaryotic N-glycoproteins.
Schwarz
Flavio
F
Institute of Microbiology, Department of Biology, Eidgenössische Technische Hochschule (ETH) Zurich, Zurich, Switzerland.
Huang
Wei
W
Li
Cishan
C
Schulz
Benjamin L
BL
Lizak
Christian
C
Palumbo
Alessandro
A
Numao
Shin
S
Neri
Dario
D
Aebi
Markus
M
Wang
Lai-Xi
LX
D016123:Campylobacter jejuni / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D056890:Eukaryota / Q000737:chemistry / Q000378:metabolism*; D005818:Genetic Engineering; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry; D006031:Glycosylation
D006023:Glycoproteins