Reduced paucimannosidic N‐glycan formation by suppression of a specific β‐hexosaminidase from Nicotiana benthamiana

• DOI: 10.1111/pbi.12602
• Publication Date: 2016-07-15T15:10:51Z
• Authors: Yun‐Ji Shin; Alexandra Castilho; Martina Dicker; Flavio Sádio; Ulrike Vavra; Clemens Grünwald‐...; Tae‐Ho Kwon; Friedrich Altmann; Herta Steinkellner; Richard Strasser
• Keywords: N-linked glycosylation

Plants are attractive hosts for the production of recombinant glycoproteins therapeutic use. Recent advances in glyco-engineering facilitate elimination nonmammalian-type glycosylation and introduction missing pathways customized N-glycan formation. However, some therapeutically relevant exhibit unwanted truncated (paucimannosidic) N-glycans that lack GlcNAc residues at nonreducing terminal end. These paucimannosidic increase product heterogeneity may affect biological function drugs. Here, we identified two enzymes, β-hexosaminidases (HEXOs) account formation Nicotiana benthamiana, a widely used expression host proteins. Subcellular localization studies showed HEXO1 is vacuolar protein HEXO3 mainly located plasma membrane N. benthamiana leaf epidermal cells. Both enzymes functional can complement corresponding HEXO-deficient Arabidopsis thaliana mutants. In planta demonstrated core α1,3-fucose enhances trimming from Fc domain human IgG. Finally, using RNA interference, show suppression be applied to amounts complex on plant-produced α1-antitrypsin.