資料載入中.....
|
請使用永久網址來引用或連結此文件:
http://ir.ncue.edu.tw/ir/handle/987654321/16769
|
| 題名: | Characterization of Copper/Zinc-Superoxide Dismutase from Pagrus major cDNA and Enzyme Stability |
| 作者: | Ken, Chuian-Fu;Weng, De-Feng;Duan, Kow-Jen;Lin, Chi-Tsai |
| 貢獻者: | 生物學系 |
| 關鍵詞: | Pagrus major;Expression;Escherichia coli;PCR;pET-20b(+) |
| 日期: | 2002-02
|
| 上傳時間: | 2013-06-05T08:48:08Z
|
| 出版者: | American Chemical Society |
| 摘要: | A full-length cDNA of 794 bp encoding a putative copper/zinc-superoxide dismutase (Cu/Zn-SOD) from Pagrus major was cloned by the PCR approach. Nucleotide sequence analysis of this cDNA clone revealed that it comprises a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed high similarity (53−91%) with the sequences of Cu/Zn-SOD from other species. Computer analysis of the residues required for coordinating copper (His-47, 49, 64, and 121) and zinc (His-64, 72, 81, and Asp-84), as well as the two cysteines (58 and 147) that form a single disulfide bond, were well conserved among all reported Cu/Zn-SOD sequences. To further characterize the Pagrus major Cu/Zn-SOD, the coding region was subcloned into an expression vector, pET-20b(+), and transformed into Escherichia coli BL21(DE3). The expression of the Cu/Zn-SOD was confirmed by enzyme activity stained on a native-gel and purified by Ni2+-nitrilotriacetic acid Sepharose superflow. Dimer was the major form of the enzyme in equilibrium. The dimerization of the enzyme was inhibited under acidic pH (below 4.0 or higher than 10.0). The half-life was 8.6 min and the inactivation rate constant (kd) was 9.69 × 10-2 min-1 at 70 °C. The enzyme activity was not significantly affected under 4% SDS or 0.5 M imidazole. The enzyme was resistant to proteolysis by both trypsin and chymotrypsin. |
| 關聯: | Journal of Agricultural and Food Chemistry, 50(4): 784-789 |
| 顯示於類別: | [生物學系] 期刊論文
|
文件中的檔案:
| 檔案 |
大小 | 格式 | 瀏覽次數 |
|---|
| index.html | 0Kb | HTML | 638 | 檢視/開啟 |
|
在NCUEIR中所有的資料項目都受到原著作權保護.
|
