Lookup NU author(s): Professor Phillip Wright
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© 2017 Elsevier Inc. Although Escherichia coli has been engineered to perform N-glycosylation of recombinant proteins, an optimal glycosylating strain has not been created. By inserting a codon optimised Campylobacter oligosaccharyltransferase onto the E. coli chromosome, we created a glycoprotein platform strain, where the target glycoprotein, sugar synthesis and glycosyltransferase enzymes, can be inserted using expression vectors to produce the desired homogenous glycoform. To assess the functionality and glycoprotein producing capacity of the chromosomally based OST, a combined Western blot and parallel reaction monitoring mass spectrometry approach was applied, with absolute quantification of glycoprotein. We demonstrated that chromosomal oligosaccharyltransferase remained functional and facilitated N-glycosylation. Although the engineered strain produced less total recombinant protein, the glycosylation efficiency increased by 85%, and total glycoprotein production was enhanced by 17%.
Author(s): Strutton B, Jaffe SRP, Pandhal J, Wright PC
Publication type: Article
Publication status: Published
Journal: Biochemical and Biophysical Research Communications
Print publication date: 01/01/2018
Online publication date: 04/11/2017
Acceptance date: 03/11/2017
ISSN (print): 0006-291X
ISSN (electronic): 1090-2104
Publisher: Elsevier BV
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