title: The combination of plant translational enhancers and terminator increase the expression of human glucocerebrosidase in Nicotiana benthamiana plants.
abstract: Gaucher's disease is a lysosomal storage disorder caused by mutations in the gene encoding glucocerebrosidase (GCase). It is currently treated by enzyme replacement therapy using recombinant GCase expressed in mammalian cells. Plant production systems are among the most attractive alternatives for pharmaceutical protein production due to such advantages as low-cost, high-scalability, and safety from human pathogen contamination. Because of its high biomass yield, Nicotiana benthamiana could be an economical recombinant GCase production system. In this study, a translational enhancer and suitable terminator were utilized to obtain a powerful expression system for GCase production in N. benthamiana plants. Six plasmid constructs were used. The highest activity of 44.5units/mg protein (after subtraction of endogenous glucosidase activity of the wild-type plant) was observed in transgenic plants transformed with pAt-GC-HSP combined with a 5' untranslated region of the Arabidopsis alcohol dehydrogenase gene with the Arabidopsis heat shock protein terminator. These transgenic plant lines could pave the way to a stable plant-production system for low-cost, high-yield human GCase production.
authors.affiliation: International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan.
authors.forename: Juthamard
authors.initials: J
authors.lastname: Limkul
authors.affiliation: International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Misaki
authors.affiliation: Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), 8916-5 Takayama-cho, Ikoma, Nara 630-0192, Japan.
authors.forename: Ko
authors.initials: K
authors.lastname: Kato
authors.affiliation: International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan. Electronic address: fujiyama@icb.osaka-u.ac.jp.
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Fujiyama
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005962:Glucosylceramidase / Q000235:genetics* / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D013728:Terminator Regions, Genetic; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; D000426:Alcohol Dehydrogenase; D005962:Glucosylceramidase
keywords: Gaucher’s disease; Human glucocerebrosidase; Nicotiana benthamiana; Plant-made pharmaceuticals