RRC ID |
29693
|
著者 |
Mitsudome T, Xu J, Nagata Y, Masuda A, Iiyama K, Morokuma D, Li Z, Mon H, Lee JM, Kusakabe T.
|
タイトル |
Expression, purification, and characterization of endo-β-N-acetylglucosaminidase H using baculovirus-mediated silkworm protein expression system.
|
ジャーナル |
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.
|
巻・号 |
172(8)
|
ページ |
3978-88
|
公開日 |
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
|
引用数 |
24
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
カイコ |
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 |