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题名: Cloning, Expression, and Characterization of a Thioredoxin Reductase cDNA from Taiwanofungus Camphorata
作者: Huang, Chih-Yu;Ken, Chuian-Fu;Chi, Hsiang-Hui;Wen, Lisa;Lin, Chi-Tsai
贡献者: 生物學系
关键词: Taiwanofungus camphorata;Three-dimensional (3-D) homology structure;Thioredoxin reductase (TR);Dithionitrobenzoate [DTNB, 5-5-dithiobis(2-nitrobenzoic acid)];Thioredoxin (Trx)
日期: 2010-04
上传时间: 2013-06-05T08:48:24Z
出版者: American Chemical Society
摘要: A cDNA encoding putative thioredoxin reductase (TR) was identified from a medicinal mushroom, Taiwanofungus camphorata (T. camphorata). Alignment of the deduced amino acid sequence with TRs from other organisms showed high levels of identity (59−74%). A three-dimensional (3-D) homology structure was created for this TR. Functional T. camphorata TR (TcTR) was overexpressed in yeast and purified. The purified enzyme showed a monomic form on a 10% sodium dodecyl sulfate−polyacrylamide gel electrophoresis (SDS−PAGE). The enzyme’s half-life of deactivation at 60 °C was 12.9 min, and its thermal inactivation rate constant Kd was 5.37 × 10−2 min−1. The optimal pH for the enzyme was pH 8 and retained about 76% activity in the presence of 0.1 M imidazole. The enzyme showed 50% activity after 10 min of incubation at 37 °C with chymotrypsin. The Michaelis constant (Km) value for dithionitrobenzoate (DTNB) was 1.59 mM.
關聯: Journal of Agricultural and Food Chemistry, 58(8): 4825-4830
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