English  |  正體中文  |  简体中文  |  Items with full text/Total items : 6480/11652
Visitors : 20237571      Online Users : 280
RC Version 3.2 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
Scope Adv. Search
LoginUploadHelpAboutAdminister

Please use this identifier to cite or link to this item: http://ir.ncue.edu.tw/ir/handle/987654321/16769

Title: Characterization of Copper/Zinc-Superoxide Dismutase from Pagrus major cDNA and Enzyme Stability
Authors: Ken, Chuian-Fu;Weng, De-Feng;Duan, Kow-Jen;Lin, Chi-Tsai
Contributors: 生物學系
Keywords: Pagrus major;Expression;Escherichia coli;PCR;pET-20b(+)
Date: 2002-02
Issue Date: 2013-06-05T08:48:08Z
Publisher: American Chemical Society
Abstract: 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.
Relation: Journal of Agricultural and Food Chemistry, 50(4): 784-789
Appears in Collections:[生物學系] 期刊論文

Files in This Item:

File SizeFormat
index.html0KbHTML397View/Open


All items in NCUEIR are protected by copyright, with all rights reserved.

 


DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - Feedback