Open AccessAn oligosaccharyltransferase from Leishmania major increases the N‐glycan occupancy on recombinant glycoproteins produced in Nicotiana benthamiana
Author(s) -
Castilho Alexandra,
Beihammer Gernot,
Pfeiffer Christina,
Göritzer Kathrin,
MonteroMorales Laura,
Vavra Ulrike,
Maresch Daniel,
GrünwaldGruber Clemens,
Altmann Friedrich,
Steinkellner Herta,
Strasser Richard
Publication year - 2018
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12906
Subject(s) - nicotiana benthamiana , glycosylation , glycoprotein , biology , n linked glycosylation , glycan , recombinant dna , endoplasmic reticulum , biochemistry , microbiology and biotechnology , gene
Summary N‐glycosylation is critical for recombinant glycoprotein production as it influences the heterogeneity of products and affects their biological function. In most eukaryotes, the oligosaccharyltransferase is the central‐protein complex facilitating the N‐glycosylation of proteins in the lumen of the endoplasmic reticulum ( ER ). Not all potential N‐glycosylation sites are recognized in vivo and the site occupancy can vary in different expression systems, resulting in underglycosylation of recombinant glycoproteins. To overcome this limitation in plants, we expressed Lm STT 3D, a single‐subunit oligosaccharyltransferase from the protozoan Leishmania major transiently in Nicotiana benthamiana, a well‐established production platform for recombinant proteins. A fluorescent protein‐tagged Lm STT 3D variant was predominately found in the ER and co‐located with plant oligosaccharyltransferase subunits. Co‐expression of Lm STT 3D with immunoglobulins and other recombinant human glycoproteins resulted in a substantially increased N‐glycosylation site occupancy on all N‐glycosylation sites except those that were already more than 90% occupied. Our results show that the heterologous expression of Lm STT 3D is a versatile tool to increase N‐glycosylation efficiency in plants.