論文 - 詳細
| RRC ID | 33143 |
|---|---|
| 著者 | Hong SM, Sung HS, Kang MH, Kim CG, Lee YH, Kim DJ, Lee JM, Kusakabe T. |
| タイトル | Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. |
| ジャーナル | Mol Biotechnol |
| Abstract |
Endo-β-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°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°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°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°C was higher than that from E. coli, probably due to glycosylation. |
| 巻・号 | 56(10) |
| ページ | 878-89 |
| 公開日 | 2014-10-1 |
| DOI | 10.1007/s12033-014-9767-8 |
| PMID | 24848382 |
| MeSH | Amino Acid Sequence Animals Arthropods / enzymology* Bombyx / metabolism* Cellulase / chemistry Cellulase / metabolism* Escherichia coli / metabolism Gene Expression* Glycosylation Hydrogen-Ion Concentration Molecular Sequence Data Plant Lectins / metabolism Polysaccharides / chemistry Pupa / metabolism Recombinant Proteins / chemistry Recombinant Proteins / isolation & purification Temperature |
| IF | 2.022 |
| 引用数 | 10 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
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