Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | kcat and Km values are increased by the elevation of temperature, whereas the kcat/Km values are not largely changed | Thermus thermophilus HB8 |
Cloned (Comment) | Organism |
---|---|
into vector pET-11a and overexpressed in Escherichia coli BL21 (DE3) cells | Thermus thermophilus HB8 |
Crystallization (Comment) | Organism |
---|---|
by oil-microbatch method, at 1.65 A resolution. TAD follows an ordered sequential mechanism in which the coenzyme binds first | Thermus thermophilus HB8 |
Protein Variants | Comment | Organism |
---|---|---|
D242N | mutation results in a large decrease in thermostability of TAD without dissociation into dimer or monomer | Thermus thermophilus HB8 |
F141A | mutation results in decrease in thermostability of TAD without dissociation into dimer or monomer | Thermus thermophilus HB8 |
F249A | mutation results in decrease in thermostability of TAD without dissociation into dimer or monomer | Thermus thermophilus HB8 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | uncompetitive inhibition with respect to NAD+ and competitive inhibition with respect to D-galactose | Thermus thermophilus HB8 | |
2,3-benzofuran | uncompetitive inhibition with respect to NAD+ and competitive inhibition with respect to D-galactose | Thermus thermophilus HB8 | |
catechol | - |
Thermus thermophilus HB8 | |
Hexestrol | uncompetitive inhibition with respect to NAD+ and competitive inhibition with respect to D-galactose | Thermus thermophilus HB8 | |
indazole | - |
Thermus thermophilus HB8 | |
indole | uncompetitive inhibition with respect to NAD+ and competitive inhibition with respect to D-galactose | Thermus thermophilus HB8 | |
additional information | EDTA (1 mM) and p-chloromercuriphenylsulfonate and p-chloromercuribenzoate (0.1 mM) have no effect on the activity | Thermus thermophilus HB8 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
isatin | - |
Thermus thermophilus HB8 | |
0.14 | - |
1-Acenaphthenol | - |
Thermus thermophilus HB8 | |
0.17 | - |
(S)-tetralol | - |
Thermus thermophilus HB8 | |
0.25 | - |
4-chromanol | - |
Thermus thermophilus HB8 | |
0.65 | - |
(S)-indanol | - |
Thermus thermophilus HB8 | |
1.3 | - |
(R)-indanol | - |
Thermus thermophilus HB8 | |
1.9 | - |
alpha-tetralone | - |
Thermus thermophilus HB8 | |
2 | - |
phenyl-1,2-propanedione | - |
Thermus thermophilus HB8 | |
2 | 3 | D-galactosamine | - |
Thermus thermophilus HB8 | |
2.5 | - |
(R)-tetralol | - |
Thermus thermophilus HB8 | |
3.7 | - |
2-Cyclohexen-1-ol | - |
Thermus thermophilus HB8 | |
5.5 | - |
D-fucose | - |
Thermus thermophilus HB8 | |
27 | - |
cis-benzene dihydrodiol | - |
Thermus thermophilus HB8 | |
33 | - |
2-deoxy-D-galactose | - |
Thermus thermophilus HB8 | |
78 | - |
D-galactose | - |
Thermus thermophilus HB8 | |
131 | - |
D-talose | - |
Thermus thermophilus HB8 | |
139 | - |
D-glucosamine | - |
Thermus thermophilus HB8 | |
162 | - |
L-fucose | - |
Thermus thermophilus HB8 | |
178 | - |
6-deoxy-D-glucose | - |
Thermus thermophilus HB8 | |
250 | - |
cellobiose | - |
Thermus thermophilus HB8 | |
262 | - |
L-arabinose | - |
Thermus thermophilus HB8 | |
300 | - |
2-deoxy-D-glucose | - |
Thermus thermophilus HB8 | |
580 | - |
D-glucose | - |
Thermus thermophilus HB8 | |
781 | - |
lactose | - |
Thermus thermophilus HB8 | |
841 | - |
D-xylose | - |
Thermus thermophilus HB8 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
27000 | - |
4 * 27000, SDS-PAGE | Thermus thermophilus HB8 |
100000 | - |
gel filtration | Thermus thermophilus HB8 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus HB8 | Q5SLC4 | - |
- |
Purification (Comment) | Organism |
---|---|
by sonication, heating at 70°C for 10 min, ion-exchange chromatography, gel filtration and ultrafiltration | Thermus thermophilus HB8 |
Storage Stability | Organism |
---|---|
4°C | Thermus thermophilus HB8 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-indanol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
(R)-tetralol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
(S)-(+)-1,2,3,4-tetrahydro-1-naphthol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
(S)-(+)-1-indanol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
(S)-indanol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
(S)-tetralol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
1-acenaphthenol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
2-cyclohexen-1-ol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
2-deoxy-D-galactose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
2-deoxy-D-glucose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
4-chromanol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
6-deoxy-D-glucose | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
alpha-tetralone + NADH + H+ | - |
Thermus thermophilus HB8 | ? + NAD+ | - |
? | |
cellobiose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
cis-benzene dihydrodiol + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
D-fucose + NAD+ | - |
Thermus thermophilus HB8 | D-fucono-delta-lactone + NADH + H+ | - |
? | |
D-galactosamine + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
D-galactose + NAD+ | - |
Thermus thermophilus HB8 | D-galactono-gamma-lactone + NADH + H+ | - |
? | |
D-glucosamine + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
D-glucose + NAD+ | - |
Thermus thermophilus HB8 | D-glucono-delta-lactone + NADH + H+ | - |
? | |
D-talose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
D-xylose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
glucono-delta-lactone + NADH + H+ | - |
Thermus thermophilus HB8 | ? + NAD+ | - |
? | |
isatin + NADH + H+ | - |
Thermus thermophilus HB8 | ? + NAD+ | - |
? | |
L-arabinose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
L-fucose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
lactose + NAD+ | - |
Thermus thermophilus HB8 | ? + NADH + H+ | - |
? | |
additional information | TAD possesses the catalytic triad Ser136, Tyr149, and Lys153. No activity with L-xylose, D-ribose and 2-deoxy-D-ribose. TAD does not oxidize ketoses (D-fructose and D-sorbose), sugar alcohols (D-sorbitol, galactitol, inositol, L-arabitol and xylitol), uronic acids (D-galacturonic acid and D-glucuronic acid), aldonic acids (L-gulonic acid and 2-keto-L-gulonic acid), aromatic alcohols (benzyl alcohol and catechol), aliphatic alcohols (1-nonanol, 1-decanol, 3-hydroxybutyric acid and 2,3-butanediol), and hydroxysteroids (androsterone, testosterone, 20alpha/beta-hydroxyprogesterone, cholic acid, cortisol and corticosterone). No significant activity ketones (4-nitroacetophenone, camphorquinone, 2,3-hexanedione and diacetyl) and aldehydes (4-nitrobenzaldehyde, pyridine-3-aldehyde, 1-nonanal and citral) | Thermus thermophilus HB8 | ? | - |
? | |
phenyl-1,2-propanedione + NADH + H+ | - |
Thermus thermophilus HB8 | ? + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 27000, SDS-PAGE | Thermus thermophilus HB8 |
Synonyms | Comment | Organism |
---|---|---|
TAD | - |
Thermus thermophilus HB8 |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | 85 | - |
Thermus thermophilus HB8 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
85 | - |
approximately 40% of the activity is inactivated by the incubation of the enzyme solution (0.3 mg/ml in 10 mM potassium phosphate, pH 7.0) for 15 min at 95°C | Thermus thermophilus HB8 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.075 | - |
(R)-tetralol | - |
Thermus thermophilus HB8 | |
0.1067 | - |
L-fucose | - |
Thermus thermophilus HB8 | |
0.167 | - |
(R)-indanol | - |
Thermus thermophilus HB8 | |
0.3117 | - |
D-galactosamine | - |
Thermus thermophilus HB8 | |
0.423 | - |
D-xylose | - |
Thermus thermophilus HB8 | |
0.6217 | - |
D-glucosamine | - |
Thermus thermophilus HB8 | |
3.017 | - |
D-talose | - |
Thermus thermophilus HB8 | |
3.5 | - |
2-Cyclohexen-1-ol | - |
Thermus thermophilus HB8 | |
3.85 | - |
D-glucose | - |
Thermus thermophilus HB8 | |
3.9 | - |
alpha-tetralone | - |
Thermus thermophilus HB8 | |
4.55 | - |
L-arabinose | - |
Thermus thermophilus HB8 | |
5.767 | - |
cellobiose | - |
Thermus thermophilus HB8 | |
6.35 | - |
6-deoxy-D-glucose | - |
Thermus thermophilus HB8 | |
6.93 | - |
2-deoxy-D-glucose | - |
Thermus thermophilus HB8 | |
7.383 | - |
cis-benzene dihydrodiol | - |
Thermus thermophilus HB8 | |
8.25 | - |
D-galactose | - |
Thermus thermophilus HB8 | |
8.73 | - |
D-fucose | - |
Thermus thermophilus HB8 | |
9.23 | - |
lactose | - |
Thermus thermophilus HB8 | |
12.8 | - |
2-deoxy-D-galactose | - |
Thermus thermophilus HB8 | |
16.83 | - |
phenyl-1,2-propanedione | - |
Thermus thermophilus HB8 | |
40.3 | - |
4-chromanol | - |
Thermus thermophilus HB8 | |
138.7 | - |
(S)-indanol | - |
Thermus thermophilus HB8 | |
161.7 | - |
(S)-tetralol | - |
Thermus thermophilus HB8 | |
433.3 | - |
1-Acenaphthenol | - |
Thermus thermophilus HB8 | |
981.7 | - |
isatin | - |
Thermus thermophilus HB8 |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
2 | 13 | - |
Thermus thermophilus HB8 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | TAD does not exhibit dehydrogenase activity with NADP+ as the coenzyme | Thermus thermophilus HB8 | |
NAD+ | - |
Thermus thermophilus HB8 |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
Hexestrol | - |
Thermus thermophilus HB8 | |
0.025 | - |
1,10-phenanthroline | - |
Thermus thermophilus HB8 | |
0.042 | - |
2,3-benzofuran | - |
Thermus thermophilus HB8 | |
0.48 | - |
indole | - |
Thermus thermophilus HB8 |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.063 | - |
- |
Thermus thermophilus HB8 | Hexestrol | |
0.18 | - |
- |
Thermus thermophilus HB8 | 1,10-phenanthroline | |
0.2 | - |
- |
Thermus thermophilus HB8 | 2,3-benzofuran | |
0.48 | - |
- |
Thermus thermophilus HB8 | indole | |
1.2 | - |
- |
Thermus thermophilus HB8 | catechol | |
1.9 | - |
- |
Thermus thermophilus HB8 | indazole |