RRC ID 29693
Author Mitsudome T, Xu J, Nagata Y, Masuda A, Iiyama K, Morokuma D, Li Z, Mon H, Lee JM, Kusakabe T.
Title Expression, purification, and characterization of endo-β-N-acetylglucosaminidase H using baculovirus-mediated silkworm protein expression system.
Journal Appl Biochem Biotechnol
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.
Volume 172(8)
Pages 3978-88
Published 2014-4-1
DOI 10.1007/s12010-014-0814-5
PMID 24599668
MeSH Animals Baculoviridae / genetics* Bombyx / cytology Bombyx / genetics* Cell Line Gene Expression Genetic Engineering / methods* Glycosylation Larva / genetics Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / biosynthesis Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / genetics* Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / isolation & purification* Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / metabolism Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism
IF 2.277
Times Cited 24
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Silkworms b31 c11 d011 d17 d33 e10 e80 g05 g30 g35 l90 l311 m90 m421 n17 n21 n22 n41 n70 o321 p53 r211 t90 u125 xu33