title: An engineered eukaryotic protein glycosylation pathway in Escherichia coli.
abstract: We performed bottom-up engineering of a synthetic pathway in Escherichia coli for the production of eukaryotic trimannosyl chitobiose glycans and the transfer of these glycans to specific asparagine residues in target proteins. The glycan biosynthesis was enabled by four eukaryotic glycosyltransferases, including the yeast uridine diphosphate-N-acetylglucosamine transferases Alg13 and Alg14 and the mannosyltransferases Alg1 and Alg2. By including the bacterial oligosaccharyltransferase PglB from Campylobacter jejuni, we successfully transferred glycans to eukaryotic proteins.
authors.affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors.forename: Juan D
authors.initials: JD
authors.lastname: Valderrama-Rincon
authors.forename: Adam C
authors.initials: AC
authors.lastname: Fisher
authors.forename: Judith H
authors.initials: JH
authors.lastname: Merritt
authors.forename: Yao-Yun
authors.initials: YY
authors.lastname: Fan
authors.forename: Craig A
authors.initials: CA
authors.lastname: Reading
authors.forename: Krishan
authors.initials: K
authors.lastname: Chhiba
authors.forename: Christian
authors.initials: C
authors.lastname: Heiss
authors.forename: Parastoo
authors.initials: P
authors.lastname: Azadi
authors.forename: Markus
authors.initials: M
authors.lastname: Aebi
authors.forename: Matthew P
authors.initials: MP
authors.lastname: DeLisa
mesh_terms: D016123:Campylobacter jejuni / Q000201:enzymology; D004187:Disaccharides / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D006031:Glycosylation; D006602:Hexosyltransferases / Q000378:metabolism; D008364:Mannosyltransferases / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D011134:Polysaccharides / Q000096:biosynthesis*; D015202:Protein Engineering; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D004187:Disaccharides; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011134:Polysaccharides; D029701:Saccharomyces cerevisiae Proteins; C032438:chitobiose; C083733:ALG2 protein, S cerevisiae; C506082:Alg13 protein, S cerevisiae; C506083:Alg14 protein, S cerevisiae; D006602:Hexosyltransferases; D008364:Mannosyltransferases; D017351:N-Acetylglucosaminyltransferases; C035777:chitobiosyldiphosphodolichol beta-mannosyltransferase; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase