Expression of recombinant sea urchin cellulase SnEG54 using mammalian cell lines

Okumura, Fumihiko; Kameda, Hiroyuki; Ojima, Takao; Hatakeyama, Shigetsugu

Expression of recombinant sea urchin cellulase SnEG54 using mammalian cell lines

Okumura, Fumihiko; Kameda, Hiroyuki; Ojima, Takao et al.

Biochemical and Biophysical Research Communications, 2010, 395(3), 352-355

Permalink : https://hdl.handle.net/2115/43096

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アクセス権	open access
URI	https://hdl.handle.net/2115/43096
タイトル	Expression of recombinant sea urchin cellulase SnEG54 using mammalian cell lines
著者	Okumura, Fumihiko ; Kameda, Hiroyuki ; Ojima, Takao ; Hatakeyama, Shigetsugu
著者名 Okumura, Fumihiko
著者名 Kameda, Hiroyuki
e-Rad_Researcher 30160865 著者名 Ojima, Takao
e-Rad_Researcher 70294973 著者名 Hatakeyama, Shigetsugu
言語	英語
キーワード	Sea urchin
	Stronglyocentrotus nudus
	Cellulase
	HeLa
	HEK293T
発行日	2010-05-07
出版者	Elsevier
収録物名	Biochemical and Biophysical Research Communications
巻(号)	395(3)
ページ	352-355
抄録	We previously identified the cellulase SnEG54 from Japanese purple sea urchin Strongylocentrotus nudus, the molecular mass of which is about 54 kDa on SDS-PAGE. It is difficult to express and purify a recombinant cellulase protein using bacteria such as E. coil or yeast. In this study, we generated mammalian expression vectors encoding SnEG54 to transiently express SnEG54 in mammalian cells. Both SnEG54 expressed in mammalian cells and SnEG54 released into the culture supernatant showed hydrolytic activity toward carboxymethyl cellulose. By using a retroviral expression system, we also established a mammalian cell line that constitutively produces SnEG54. Unexpectedly, SnEG54 released into the culture medium was not stable, and the peak time showing the highest concentration was approximately 1 to 2 days after seeding into fresh culture media. These findings suggest that non-mammalian sea urchin cellulase can be generated in human cell lines but that recombinant SnEG54 is unstable in culture medium due to an unidentified mechanism.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0006-291X
関連情報	PMID 20381456
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.bbrc.2010.04.016