Sweet factory: an O-glycosylation competent Escherichia coli strain for the recombinant expression of complex biopharmaceuticals

Abstract

Glycosylation represents an important modification influencing the quality of recombinant expressed biopharmaceuticals by modulating the stability and activity of commercially relevant therapeutics. As a result, the importance of E. coli as a manufacturing platform has continuously decreased due to the lack of post-translational modifications during protein expression. The presented E. coli-based expression system was established to achieve mucin-type O-glycosylation in vivo combining the expression of the human glycosyltransferase GalNAc-T2 and the uridine 5’diphospho-N-acetylglucosamine (UDP-GlcNAc) 4-epimerase derived from Plesiomonas shigelloides (WbgU). T7Muc10, a synthetic protein with multiple glycosylation sites, and the granulocyte-colony stimulating factor G-CSF, a pharmaceutically relevant product, were included as potential target proteins.