Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta-gami2(DE3)pLysS cells | Nostoc punctiforme |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 0.1 M HEPES-NaOH (pH 7.5), 14% (w/v) polyethylene glycol 8000, and 8% (v/v) ethylene glycol | Nostoc punctiforme |
Protein Variants | Comment | Organism |
---|---|---|
A69L | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
A69M | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
I296A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
L288M | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
L59F/D168G/A234D/I296N | the mutant exhibits higher stability and specific activity than the wild type enzyme | Nostoc punctiforme |
M295A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
M40L | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
M65A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
V132A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
V133A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
V291A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
Y292F | the mutant shows increased catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
Y292H | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
Y292W | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Nostoc punctiforme |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
L-tryptophan | mutant enzyme V132A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.019 | - |
L-tryptophan | mutant enzyme V133A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.021 | - |
L-tryptophan | mutant enzyme Y292H, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.03 | - |
L-tryptophan | mutant enzyme Y292W, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.039 | - |
L-tryptophan | mutant enzyme Y292F, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.045 | - |
L-tryptophan | wild type enzyme, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.047 | - |
L-tryptophan | mutant enzyme M295A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.047 | - |
L-tryptophan | mutant enzyme M40L, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.047 | - |
L-tryptophan | mutant enzyme V291A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.059 | - |
L-tryptophan | mutant enzyme I296A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.2 | - |
L-tryptophan | mutant enzyme M65A, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.52 | - |
L-tryptophan | mutant enzyme L288M, at pH 11.0 and 25°C | Nostoc punctiforme | |
0.82 | - |
L-tryptophan | mutant enzyme A69M, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.1 | - |
L-tryptophan | mutant enzyme A69L, at pH 11.0 and 25°C | Nostoc punctiforme |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-tryptophan + NAD+ + H2O | Nostoc punctiforme | - |
(indol-3-yl)pyruvate + NH3 + NADH + H+ | - |
r | |
L-tryptophan + NAD+ + H2O | Nostoc punctiforme NIES-2108 | - |
(indol-3-yl)pyruvate + NH3 + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Nostoc punctiforme | W8CV45 | - |
- |
Nostoc punctiforme NIES-2108 | W8CV45 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Nostoc punctiforme |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-tryptophan + NAD+ + H2O | - |
Nostoc punctiforme | (indol-3-yl)pyruvate + NH3 + NADH + H+ | - |
r | |
L-tryptophan + NAD+ + H2O | - |
Nostoc punctiforme NIES-2108 | (indol-3-yl)pyruvate + NH3 + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 40000, SDS-PAGE | Nostoc punctiforme |
Synonyms | Comment | Organism |
---|---|---|
L-Trp dehydrogenase | - |
Nostoc punctiforme |
TrpDH | - |
Nostoc punctiforme |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.39 | - |
L-tryptophan | mutant enzyme L288M, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.1 | - |
L-tryptophan | mutant enzyme M65A, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.2 | - |
L-tryptophan | mutant enzyme A69L, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.2 | - |
L-tryptophan | mutant enzyme A69M, at pH 11.0 and 25°C | Nostoc punctiforme | |
2.6 | - |
L-tryptophan | mutant enzyme V133A, at pH 11.0 and 25°C | Nostoc punctiforme | |
2.9 | - |
L-tryptophan | mutant enzyme V291A, at pH 11.0 and 25°C | Nostoc punctiforme | |
3.5 | - |
L-tryptophan | mutant enzyme Y292H, at pH 11.0 and 25°C | Nostoc punctiforme | |
4.6 | - |
L-tryptophan | mutant enzyme V132A, at pH 11.0 and 25°C | Nostoc punctiforme | |
8 | - |
L-tryptophan | mutant enzyme M295A, at pH 11.0 and 25°C | Nostoc punctiforme | |
8.5 | - |
L-tryptophan | mutant enzyme Y292W, at pH 11.0 and 25°C | Nostoc punctiforme | |
10 | - |
L-tryptophan | mutant enzyme I296A, at pH 11.0 and 25°C | Nostoc punctiforme | |
12 | - |
L-tryptophan | mutant enzyme M40L, at pH 11.0 and 25°C | Nostoc punctiforme | |
20 | - |
L-tryptophan | wild type enzyme, at pH 11.0 and 25°C | Nostoc punctiforme | |
21 | - |
L-tryptophan | mutant enzyme Y292F, at pH 11.0 and 25°C | Nostoc punctiforme |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Nostoc punctiforme |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.75 | - |
L-tryptophan | mutant enzyme L288M, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.1 | - |
L-tryptophan | mutant enzyme A69L, at pH 11.0 and 25°C | Nostoc punctiforme | |
1.5 | - |
L-tryptophan | mutant enzyme A69M, at pH 11.0 and 25°C | Nostoc punctiforme | |
5.5 | - |
L-tryptophan | mutant enzyme M65A, at pH 11.0 and 25°C | Nostoc punctiforme | |
62 | - |
L-tryptophan | mutant enzyme V291A, at pH 11.0 and 25°C | Nostoc punctiforme | |
140 | - |
L-tryptophan | mutant enzyme V133A, at pH 11.0 and 25°C | Nostoc punctiforme | |
170 | - |
L-tryptophan | mutant enzyme I296A, at pH 11.0 and 25°C | Nostoc punctiforme | |
170 | - |
L-tryptophan | mutant enzyme M295A, at pH 11.0 and 25°C | Nostoc punctiforme | |
170 | - |
L-tryptophan | mutant enzyme Y292H, at pH 11.0 and 25°C | Nostoc punctiforme | |
260 | - |
L-tryptophan | mutant enzyme M40L, at pH 11.0 and 25°C | Nostoc punctiforme | |
270 | - |
L-tryptophan | mutant enzyme V132A, at pH 11.0 and 25°C | Nostoc punctiforme | |
280 | - |
L-tryptophan | mutant enzyme Y292W, at pH 11.0 and 25°C | Nostoc punctiforme | |
440 | - |
L-tryptophan | wild type enzyme, at pH 11.0 and 25°C | Nostoc punctiforme | |
540 | - |
L-tryptophan | mutant enzyme Y292F, at pH 11.0 and 25°C | Nostoc punctiforme |