Institute of Bioscience and Biotechnology and Marine Center for Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 202, Taiwan.
J Agric Food Chem. 2010 Apr 28;58(8):4825-30. doi: 10.1021/jf100222b.
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 degrees C was 12.9 min, and its thermal inactivation rate constant K(d) was 5.37 x 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 degrees C with chymotrypsin. The Michaelis constant (K(m)) value for dithionitrobenzoate (DTNB) was 1.59 mM.
从药用蘑菇樟芝（T. camphorata）中鉴定出一种编码假定硫氧还蛋白还原酶（TR）的 cDNA。与来自其他生物体的 TR 的推导氨基酸序列进行比对显示出高度的同一性（59-74%）。为该 TR 创建了一个三维（3-D）同源结构。在酵母中过量表达并纯化了功能性樟芝 TR（TcTR）。纯化的酶在 10%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上显示出单聚体形式。该酶在 60°C 下失活的半衰期为 12.9 分钟，其热失活速率常数 K(d)为 5.37 x 10(-2) min(-1)。该酶的最适 pH 为 pH 8，在存在 0.1 M 咪唑的情况下保留约 76%的活性。该酶在 37°C 下与胰凝乳蛋白酶孵育 10 分钟后，活性为 50%。二硫代硝基苯甲酸（DTNB）的米氏常数（K(m)）值为 1.59 mM。
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