Abstract
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 (K m) value for dithionitrobenzoate (DTNB) was 1.59 mM.
| Original language | English |
|---|---|
| Pages (from-to) | 4825-4830 |
| Number of pages | 6 |
| Journal | Journal of Agricultural and Food Chemistry |
| Volume | 58 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2010 Apr 28 |
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All Science Journal Classification (ASJC) codes
- Chemistry(all)
- Agricultural and Biological Sciences(all)
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Cloning, expression, and characterization of a thioredoxin reductase cDNA from Taiwanofungus camphorata. / Huang, Chih Yu; Ken, Chuian Fu; Chi, Hsiang Hui; Wen, Lisa; Lin, Chi Tsai.
In: Journal of Agricultural and Food Chemistry, Vol. 58, No. 8, 28.04.2010, p. 4825-4830.Research output: Contribution to journal › Article
TY - JOUR
T1 - Cloning, expression, and characterization of a thioredoxin reductase cDNA from Taiwanofungus camphorata
AU - Huang, Chih Yu
AU - Ken, Chuian Fu
AU - Chi, Hsiang Hui
AU - Wen, Lisa
AU - Lin, Chi Tsai
PY - 2010/4/28
Y1 - 2010/4/28
N2 - 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 (K m) value for dithionitrobenzoate (DTNB) was 1.59 mM.
AB - 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 (K m) value for dithionitrobenzoate (DTNB) was 1.59 mM.
UR - http://www.scopus.com/inward/record.url?scp=77951273199&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77951273199&partnerID=8YFLogxK
U2 - 10.1021/jf100222b
DO - 10.1021/jf100222b
M3 - Article
C2 - 20307095
AN - SCOPUS:77951273199
VL - 58
SP - 4825
EP - 4830
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
SN - 0021-8561
IS - 8
ER -