論文 - 詳細
| RRC ID | 42497 |
|---|---|
| 著者 | Okumura S, Saitoh H, Wasano N, Katayama H, Higuchi K, Mizuki E, Inouye K. |
| タイトル | Efficient solubilization, activation, and purification of recombinant Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli. |
| ジャーナル | Protein Expr Purif |
| Abstract |
A cytotoxic protein Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli was solubilized in 10 mM HCl. Protein concentration of saturated solution of the recombinant Cry45Aa in 10 mM HCl was about 25 times higher than that in the buffer of previous method (in 50 mM sodium carbonate buffer, pH 10.5, containing 1 mM EDTA, and 10 mM dithiothreitol). The Cry45Aa solubilized in the acidic solution was activated by pepsin as an alternative to proteinase K in the previous method. Cytotoxic activity against CACO-2 cells of the pepsin-treated Cry45Aa was almost identical to the proteinase K-treated protein. The pepsin-treated Cry45Aa was purified by cation-exchange chromatography. The concentration of the purified protein was 539 microg/ml, which was 27-fold higher than that of the activated Cry45Aa by the previously method. The cytotoxic activity of the purified protein was stable in broad pH region (pH 2.0-11.0) for 3 days, and 97% cytotoxic activity remained after incubation at 30 degrees C for 360 min. |
| 巻・号 | 47(1) |
| ページ | 144-51 |
| 公開日 | 2006-5-1 |
| DOI | 10.1016/j.pep.2005.10.011 |
| PII | S1046-5928(05)00356-6 |
| PMID | 16307894 |
| MeSH | Bacillus thuringiensis Toxins Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / isolation & purification* Bacterial Proteins / toxicity Bacterial Toxins / chemistry Bacterial Toxins / genetics Bacterial Toxins / isolation & purification* Bacterial Toxins / toxicity Caco-2 Cells Chromatography, Ion Exchange Cloning, Molecular Endotoxins / chemistry Endotoxins / genetics Endotoxins / isolation & purification* Endotoxins / toxicity Escherichia coli / genetics* Hemolysin Proteins / chemistry Hemolysin Proteins / genetics Hemolysin Proteins / isolation & purification* Hemolysin Proteins / toxicity Humans Inclusion Bodies / chemistry* Inclusion Bodies / genetics* Inclusion Bodies / metabolism Inclusion Bodies / physiology Molecular Sequence Data Recombinant Proteins / chemistry Recombinant Proteins / genetics* Recombinant Proteins / isolation & purification* Recombinant Proteins / toxicity Solubility |
| IF | 1.513 |
| 引用数 | 14 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | CACO-2(RCB0988) |