Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Synechocystis sp. |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion method, using 15% (w/v) PEG 5000MME, 1 mM dithiothreitol, 3% tascimate and 100 mM HEPES, pH 6.8 | Synechocystis sp. |
Protein Variants | Comment | Organism |
---|---|---|
C169A | inactive | Synechocystis sp. |
E142A | inactive | Synechocystis sp. |
E53A | inactive | Synechocystis sp. |
K135A | inactive | Synechocystis sp. |
N118A | inactive | Synechocystis sp. |
W146A | inactive | Synechocystis sp. |
W146C | inactive | Synechocystis sp. |
W146D | inactive | Synechocystis sp. |
W146E | inactive | Synechocystis sp. |
W146F | the mutant shows reduced activity compared to the wild type enzyme | Synechocystis sp. |
W146G | inactive | Synechocystis sp. |
W146H | the mutant shows reduced activity compared to the wild type enzyme | Synechocystis sp. |
W146I | inactive | Synechocystis sp. |
W146K | inactive | Synechocystis sp. |
W146L | inactive | Synechocystis sp. |
W146M | inactive | Synechocystis sp. |
W146N | inactive | Synechocystis sp. |
W146P | inactive | Synechocystis sp. |
W146Q | inactive | Synechocystis sp. |
W146R | inactive | Synechocystis sp. |
W146S | inactive | Synechocystis sp. |
W146T | inactive | Synechocystis sp. |
W146V | inactive | Synechocystis sp. |
W146Y | the mutant shows reduced activity compared to the wild type enzyme | Synechocystis sp. |
Y59A | inactive | Synechocystis sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.98 | - |
Benzonitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
1.13 | - |
2-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
1.21 | - |
Benzonitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
1.26 | - |
Benzonitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
1.26 | - |
2-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
1.3 | - |
2-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.33 | - |
Benzonitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.47 | - |
2-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
3.16 | - |
3-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
3.2 | - |
3-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
3.94 | - |
3-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
4.41 | - |
3-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
7.95 | - |
2-butenenitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
9.01 | - |
Fumaronitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
9.16 | - |
2-butenenitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
10.77 | - |
Fumaronitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
11.06 | - |
Fumaronitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
11.45 | - |
Fumaronitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Synechocystis sp. | Q55949 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni2+ affinity column chromatography and Superdex 75 gel filtration | Synechocystis sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.05 | - |
mutant enzyme W146F, with 2-butenenitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
0.29 | - |
wild type enzyme, with 2-butenenitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
0.68 | - |
wild type enzyme, with 3-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
0.97 | - |
wild type enzyme, with benzonitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1 | - |
mutant enzyme W146F, with benzonitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.18 | - |
mutant enzyme W146Y, with benzonitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.47 | - |
mutant enzyme W146H, with benzonitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.57 | - |
mutant enzyme W146Y, with fumaronitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.6 | - |
mutant enzyme W146F, with 3-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.69 | - |
mutant enzyme W146Y, with 3-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.82 | - |
mutant enzyme W146H, with 3-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.85 | - |
mutant enzyme W146F, with fumaronitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
1.95 | - |
mutant enzyme W146H, with fumaronitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
2.7 | - |
wild type enzyme, with 2-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
2.72 | - |
mutant enzyme W146H, with 2-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
2.93 | - |
mutant enzyme W146F, with 2-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
3.08 | - |
mutant enzyme W146Y, with 2-cyanopyridine as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
3.2 | - |
wild type enzyme, with fumaronitrile as substrate, at pH 7.0 and 30°C | Synechocystis sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-butenenitrile + H2O | worst substrate | Synechocystis sp. | 2-butenic acid + NH3 | - |
? | |
2-cyanopyridine + 2 H2O | second best substrate | Synechocystis sp. | pyridine 2-carboxylic acid + NH3 | - |
? | |
3-cyanopyridine + 2 H2O | - |
Synechocystis sp. | pyridine 3-carboxylic acid + NH3 | - |
? | |
benzonitrile + 2 H2O | - |
Synechocystis sp. | benzoate + NH3 | - |
? | |
fumaronitrile + H2O | best substrate | Synechocystis sp. | fumarate + NH3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NIT | - |
Synechocystis sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
2-butenenitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.19 | - |
2-butenenitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.45 | - |
3-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.65 | - |
Benzonitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.67 | - |
Benzonitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.79 | - |
Benzonitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
0.98 | - |
Benzonitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.07 | - |
3-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
1.13 | - |
3-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
1.21 | - |
3-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.8 | - |
2-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
1.81 | - |
2-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.95 | - |
2-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
2.05 | - |
2-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
7.42 | - |
Fumaronitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
8.75 | - |
Fumaronitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
9.22 | - |
Fumaronitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
15.13 | - |
Fumaronitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00328 | - |
2-butenenitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.0239 | - |
2-butenenitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.141 | - |
3-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.243 | - |
3-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.307 | - |
3-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
0.357 | - |
3-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
0.531 | - |
Benzonitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.537 | - |
Benzonitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
0.689 | - |
Fumaronitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
0.737 | - |
Benzonitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
0.764 | - |
Fumaronitrile | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
0.806 | - |
Benzonitrile | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. | |
0.834 | - |
Fumaronitrile | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.224 | - |
2-Cyanopyridine | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
1.392 | - |
2-Cyanopyridine | mutant enzyme W146H, at pH 7.0 and 30°C | Synechocystis sp. | |
1.548 | - |
2-Cyanopyridine | mutant enzyme W146F, at pH 7.0 and 30°C | Synechocystis sp. | |
1.679 | - |
Fumaronitrile | wild type enzyme, at pH 7.0 and 30°C | Synechocystis sp. | |
1.814 | - |
2-Cyanopyridine | mutant enzyme W146Y, at pH 7.0 and 30°C | Synechocystis sp. |