title: Expression, purification, and characterization of endo-β-N-acetylglucosaminidase H using baculovirus-mediated silkworm protein expression system.
abstract: Endo-β-N-acetylglucosaminidase (Endo H) from Streptomyces plicatus hydrolyzes the core di-GlcNAc units of asparagine-linked oligosaccharides and is regarded as an important tool for glycobiology research. In the present study, we established a large-scale system to produce secreted Endo H using a silkworm-baculovirus expression system (silkworm-BES). The recombinant Endo H purified from silkworm hemolymph had activity comparable to that from recombinant Escherichia coli. As well as its well-characterized substrate RNase B, the Endo H from silkworm-BES was able to deglycosylate the high-mannose glycoproteins from silkworm hemolymph. Interestingly, the secretion amount of recombinant Endo H was significantly varied among the different silkworm strains, which could provide valuable information for larger-scale protein productions from silkworm-BES.
authors.affiliation: Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki 6-10-1, Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Mitsudome
authors.forename: Jian
authors.initials: J
authors.lastname: Xu
authors.forename: Yudai
authors.initials: Y
authors.lastname: Nagata
authors.forename: Atsushi
authors.initials: A
authors.lastname: Masuda
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iiyama
authors.forename: Daisuke
authors.initials: D
authors.lastname: Morokuma
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Li
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mon
authors.forename: Jae Man
authors.initials: JM
authors.lastname: Lee
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kusakabe
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D012831:Bombyx / Q000166:cytology / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D006031:Glycosylation; D007814:Larva / Q000235:genetics; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase