Fig. 2
From: Homogeneous production and characterization of recombinant N-GlcNAc-protein in Pichia pastoris
Endo-T expressed in Golgi or ER of P. pastoris to produce N-GlcNAc modified proteins. a Schematic presentation of the glycoengineering process of P. pastoris to produce N-GlcNAc modified proteins. b P. pastoris strains were detected using Western Blot with anti-Flag antibody. Lane 1: P. pastoris Pir-Endo-T; Lane 2: P. pastoris MNS1-EndoT; Lane 3: P. pastoris MNN9-EndoT; c human GalNAc-T1 secreted in different P. pastoris strains and detected using Western Blot with anti-His antibody. Lane 1: P. pastoris WT; Lanes 2–3: P. pastoris MNS1-EndoT; Lanes 4–5: P. pastoris MNN9-EndoT. G0–2 stands for the protein with 0–2 glycans. d Purified human GalNAc-T1 treated with PNGase F and analyzed by SDS-PAGE. Lane 1: before PNGase F treatment; Lane 2: treated with inactivated (boiled) PNGase F; Lane 3: treated with PNGase F. M stands for the protein marker