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; Jae Man Lee

doi:10.1016/j.aspen.2019.01.009

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, Jae Man Lee

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

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.

Original language	English
Pages (from-to)	404-408
Number of pages	5
Journal	Journal of Asia-Pacific Entomology
Volume	22
Issue number	2
DOIs	https://doi.org/10.1016/j.aspen.2019.01.009
Publication status	Published - Jun 2019

All Science Journal Classification (ASJC) codes

Insect Science

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Morio, A., Xu, J., Masuda, A., Kinoshita, Y., Hino, M., Morokuma, D., Goda, H. M., Okino, N., Ito, M., Mon, H., Fujita, R., Kusakabe, T., & Lee, J. M. (2019). Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system. Journal of Asia-Pacific Entomology, 22(2), 404-408. https://doi.org/10.1016/j.aspen.2019.01.009

Morio, A, Xu, J, Masuda, A, Kinoshita, Y, Hino, M, Morokuma, D, Goda, HM, Okino, N, Ito, M, Mon, H , Fujita, R , Kusakabe, T & Lee, JM 2019, 'Expression, purification, and characterization of highly active endo-α-N-acetylgalactosaminidases expressed by silkworm-baculovirus expression system', Journal of Asia-Pacific Entomology, vol. 22, no. 2, pp. 404-408. https://doi.org/10.1016/j.aspen.2019.01.009

@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 Lee, {Jae Man}",

note = "Funding Information: The NIAS-Bm-oyanagi2 (BmO2) cell line for propagation of recombinant BmNPVs was kindly provided by Dr. Imanishi (National Institute of Agrobiological Sciences, Japan). This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number JP15H04612 . ",

year = "2019",

month = jun,

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",

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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, Jae Man

N1 - Funding Information: The NIAS-Bm-oyanagi2 (BmO2) cell line for propagation of recombinant BmNPVs was kindly provided by Dr. Imanishi (National Institute of Agrobiological Sciences, Japan). This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number JP15H04612 .

PY - 2019/6

Y1 - 2019/6

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.

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M3 - Article

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ER -

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

Abstract

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