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Expression of recombinant sea urchin cellulase SnEG54 using mammalian cell lines

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Title: Expression of recombinant sea urchin cellulase SnEG54 using mammalian cell lines
Authors: Okumura, Fumihiko Browse this author
Kameda, Hiroyuki Browse this author
Ojima, Takao Browse this author →KAKEN DB
Hatakeyama, Shigetsugu Browse this author →KAKEN DB
Keywords: Sea urchin
Stronglyocentrotus nudus
Cellulase
HeLa
HEK293T
Issue Date: 7-May-2010
Publisher: Elsevier
Journal Title: Biochemical and Biophysical Research Communications
Volume: 395
Issue: 3
Start Page: 352
End Page: 355
Publisher DOI: 10.1016/j.bbrc.2010.04.016
PMID: 20381456
Abstract: 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.
Type: article (author version)
URI: http://hdl.handle.net/2115/43096
Appears in Collections:医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 畠山 鎮次

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