AUTHOR=Uthailak Naphatsamon , Kajiura Hiroyuki , Misaki Ryo , Fujiyama Kazuhito 

TITLE=Transient Production of Human β-Glucocerebrosidase With Mannosidic-Type N-Glycan Structure in Glycoengineered Nicotiana benthamiana Plants

JOURNAL=Frontiers in Plant Science

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.683762

DOI=10.3389/fpls.2021.683762

ISSN=1664-462X

ABSTRACT=<p>Gaucher disease is an inherited lysosomal storage disorder caused by a deficiency of functional enzyme β-glucocerebrosidase (GCase). Recombinant GCase has been used in enzyme replacement therapy to treat Gaucher disease. Importantly, the terminal mannose <italic>N</italic>-glycan structure is essential for the uptake of recombinant GCase into macrophages via the mannose receptor. In this research, recombinant GCase was produced using <italic>Agrobacterium</italic>-mediated transient expression in both wild-type (WT) and <italic>N</italic>-acetylglucosaminyltransferase I (GnTI) downregulated <italic>Nicotiana benthamiana</italic> (ΔgntI) plants, the latter of which accumulates mannosidic-type <italic>N</italic>-glycan structures. The successfully produced functional GCase exhibited GCase enzyme activity. The enzyme activity was the same as that of the conventional mammalian-derived GCase. Notably, <italic>N</italic>-glycan analysis revealed that a mannosidic-type <italic>N</italic>-glycan structure lacking plant-specific <italic>N</italic>-glycans (β1,2-xylose and α1,3-fucose residues) was predominant in all glycosylation sites of purified GCase produced from ΔgntI plants. Our research provides a promising alternative plant line as a host for the production of recombinant GCase with a mannosidic-type <italic>N</italic>-glycan structure. This glycoengineered plant might be applicable to the production of other pharmaceutical proteins, especially mannose receptor targeted protein, for therapeutic uses.</p>