Characterization of Cryptopygus antarcticus Endo-beta-1,4-Glucanase from Bombyx mori Expression Systems SCIE SCOPUS

Cited 10 time in WEB OF SCIENCE Cited 10 time in Scopus
Title
Characterization of Cryptopygus antarcticus Endo-beta-1,4-Glucanase from Bombyx mori Expression Systems
Author(s)
Hong, Sun Mee; Sung, Ho Sun; Kang, Mee Hye; Kim, Choong-Gon; Lee, Youn-Ho; Kim, Dae-Jung; Lee, Jae Man; Kusakabe, Takahiro
KIOST Author(s)
Kim, Choong Gon(김충곤)Lee, Youn Ho(이윤호)
Alternative Author(s)
강미혜; 김충곤; 이윤호
Publication Year
2014-10
Abstract
Endo-beta-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10 A degrees C, but the expression yield was low. Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90 % purity by Ni-affinity chromatography. The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50 A degrees C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30 % of their maximal activity at 0 A degrees C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. Thermostability of Bm-CaCel from B. mori at 60 A degrees C was higher than that from E. coli, probably due to glycosylation.
ISSN
1073-6085
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/40412
DOI
10.1007/s12033-014-9767-8
Bibliographic Citation
MOLECULAR BIOTECHNOLOGY, v.56, no.10, pp.878 - 889, 2014
Publisher
HUMANA PRESS INC
Keywords
Baculovirus; Silkworm; Bacteria; Glycosyl hydrolase; Antarctic springtail; N-glycosylation
Type
Article
Language
English
Document Type
Article
Publisher
HUMANA PRESS INC
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