Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system

Akihiro Morio, Jian Xu, Akitsu Masuda, Yurie Kinoshita, Masato Hino, Daisuke Morokuma, Hatsumi M. Goda, Nozomu Okino, Makoto Ito, Hiroaki Mon, Ryosuke Fujita, Takahiro Kusakabe, Man Lee

研究成果: ジャーナルへの寄稿記事

1 引用 (Scopus)

抄録

The O-glycosidase, endo-α-N-acetylgalactosaminidase from Enterococcus faecalis (endoEF) catalyzes the cleavage of core 1 and core 3 type O-linked disaccharides between GalNAc and serine or threonine residues from glycoproteins. The endoEF has broad substrate specificity and thus is extensively utilized for the structural and functional analysis of the O-linked glycans. In this study, we expressed and purified the recombinant endoEF (rEndoEF) by using the silkworm-baculovirus expression vector system (Silkworm-BEVS) and confirmed the deglycosylation activity of rEndoEF targeting reporter glycoproteins, which was equivalent to the commercial O-glycosidase. Thus, our study provides important clues to produce highly active rEndoEF O-glycosidases employing silkworm-BEVS as an alternative.

元の言語英語
ページ(範囲)404-408
ページ数5
ジャーナルJournal of Asia-Pacific Entomology
22
発行部数2
DOI
出版物ステータス出版済み - 6 1 2019

Fingerprint

glycosidases
Enterococcus faecalis
Baculoviridae
silkworms
glycoproteins
disaccharides
substrate specificity
threonine
serine
polysaccharides

All Science Journal Classification (ASJC) codes

  • Insect Science

これを引用

Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system. / Morio, Akihiro; Xu, Jian; Masuda, Akitsu; Kinoshita, Yurie; Hino, Masato; Morokuma, Daisuke; Goda, Hatsumi M.; Okino, Nozomu; Ito, Makoto; Mon, Hiroaki; Fujita, Ryosuke; Kusakabe, Takahiro; Lee, Man.

:: Journal of Asia-Pacific Entomology, 巻 22, 番号 2, 01.06.2019, p. 404-408.

研究成果: ジャーナルへの寄稿記事

Morio, Akihiro ; Xu, Jian ; Masuda, Akitsu ; Kinoshita, Yurie ; Hino, Masato ; Morokuma, Daisuke ; Goda, Hatsumi M. ; Okino, Nozomu ; Ito, Makoto ; Mon, Hiroaki ; Fujita, Ryosuke ; Kusakabe, Takahiro ; Lee, Man. / Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system. :: Journal of Asia-Pacific Entomology. 2019 ; 巻 22, 番号 2. pp. 404-408.
@article{02c9bf8b1ce84d4cbaffa0563232452c,
title = "Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system",
abstract = "The O-glycosidase, endo-α-N-acetylgalactosaminidase from Enterococcus faecalis (endoEF) catalyzes the cleavage of core 1 and core 3 type O-linked disaccharides between GalNAc and serine or threonine residues from glycoproteins. The endoEF has broad substrate specificity and thus is extensively utilized for the structural and functional analysis of the O-linked glycans. In this study, we expressed and purified the recombinant endoEF (rEndoEF) by using the silkworm-baculovirus expression vector system (Silkworm-BEVS) and confirmed the deglycosylation activity of rEndoEF targeting reporter glycoproteins, which was equivalent to the commercial O-glycosidase. Thus, our study provides important clues to produce highly active rEndoEF O-glycosidases employing silkworm-BEVS as an alternative.",
author = "Akihiro Morio and Jian Xu and Akitsu Masuda and Yurie Kinoshita and Masato Hino and Daisuke Morokuma and Goda, {Hatsumi M.} and Nozomu Okino and Makoto Ito and Hiroaki Mon and Ryosuke Fujita and Takahiro Kusakabe and Man Lee",
year = "2019",
month = "6",
day = "1",
doi = "10.1016/j.aspen.2019.01.009",
language = "English",
volume = "22",
pages = "404--408",
journal = "Journal of Asia-Pacific Entomology",
issn = "1226-8615",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system

AU - Morio, Akihiro

AU - Xu, Jian

AU - Masuda, Akitsu

AU - Kinoshita, Yurie

AU - Hino, Masato

AU - Morokuma, Daisuke

AU - Goda, Hatsumi M.

AU - Okino, Nozomu

AU - Ito, Makoto

AU - Mon, Hiroaki

AU - Fujita, Ryosuke

AU - Kusakabe, Takahiro

AU - Lee, Man

PY - 2019/6/1

Y1 - 2019/6/1

N2 - The O-glycosidase, endo-α-N-acetylgalactosaminidase from Enterococcus faecalis (endoEF) catalyzes the cleavage of core 1 and core 3 type O-linked disaccharides between GalNAc and serine or threonine residues from glycoproteins. The endoEF has broad substrate specificity and thus is extensively utilized for the structural and functional analysis of the O-linked glycans. In this study, we expressed and purified the recombinant endoEF (rEndoEF) by using the silkworm-baculovirus expression vector system (Silkworm-BEVS) and confirmed the deglycosylation activity of rEndoEF targeting reporter glycoproteins, which was equivalent to the commercial O-glycosidase. Thus, our study provides important clues to produce highly active rEndoEF O-glycosidases employing silkworm-BEVS as an alternative.

AB - The O-glycosidase, endo-α-N-acetylgalactosaminidase from Enterococcus faecalis (endoEF) catalyzes the cleavage of core 1 and core 3 type O-linked disaccharides between GalNAc and serine or threonine residues from glycoproteins. The endoEF has broad substrate specificity and thus is extensively utilized for the structural and functional analysis of the O-linked glycans. In this study, we expressed and purified the recombinant endoEF (rEndoEF) by using the silkworm-baculovirus expression vector system (Silkworm-BEVS) and confirmed the deglycosylation activity of rEndoEF targeting reporter glycoproteins, which was equivalent to the commercial O-glycosidase. Thus, our study provides important clues to produce highly active rEndoEF O-glycosidases employing silkworm-BEVS as an alternative.

UR - http://www.scopus.com/inward/record.url?scp=85062386153&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062386153&partnerID=8YFLogxK

U2 - 10.1016/j.aspen.2019.01.009

DO - 10.1016/j.aspen.2019.01.009

M3 - Article

VL - 22

SP - 404

EP - 408

JO - Journal of Asia-Pacific Entomology

JF - Journal of Asia-Pacific Entomology

SN - 1226-8615

IS - 2

ER -