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

• Published in: Journal of Asia-Pacific entomology pp. 404 - 408
• Main Authors: 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, 이재만
• Format: Journal Article
• Language: English
• Published: 한국응용곤충학회 01.06.2019
• Subjects: 농수해양학
• ISSN: 1226-8615; 1876-7990
• DOI: 10.1016/j.aspen.2019.01.009

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 Oglycosidase. Thus, our study provides important clues to produce highly active rEndoEF O-glycosidases employing silkworm-BEVS as an alternative. KCI Citation Count: 0