Cloned (Comment) | Organism |
---|---|
mutant enzymes are expressed in Escherichia coli BL21(DE3)(pLysS) cells | Ensifer sp. AS08 |
Protein Variants | Comment | Organism |
---|---|---|
H465A | the mutant shows no significant change in affinity toward substrates, but the apparent kcat values decrease to approximately 1/20 of the apparent kcat value of the wild type enzyme | Ensifer sp. AS08 |
H465A/N507A | inactive | Ensifer sp. AS08 |
N507A | inactive | Ensifer sp. AS08 |
N509A | the mutant shows the less kcat values toward NPEOav10 than toward PEG-1000 | Ensifer sp. AS08 |
N90A | the mutant shows the less kcat values toward NPEOav10 than toward PEG-1000 | Ensifer sp. AS08 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.1 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
2.4 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
2.5 | - |
PEG-1000 | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
3.3 | - |
PEG-1000 | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
4.1 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
4.5 | - |
PEG-1000 | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ensifer sp. AS08 | Q2HXX0 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA affinity chromatography | Ensifer sp. AS08 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
PEG-1000 + 2,6-dichloroindophenol | 520% activity compared to PEG-mono-4-nonylphenyl ether, averaging 2 ethoxy units | Ensifer sp. AS08 | ? | - |
? | |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units + 2,6-dichloroindophenol | 93% activity compared to PEG-mono-4-nonylphenyl ether, averaging 2 ethoxy units | Ensifer sp. AS08 | ? | - |
? | |
PEG-mono-4-nonylphenyl ether, averaging 2 ethoxy units + 2,6-dichloroindophenol | 100% activity | Ensifer sp. AS08 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ethoxy chain nonylphenol dehydrogenase | - |
Ensifer sp. AS08 |
NPEO-DH | - |
Ensifer sp. AS08 |
polyethylene glycol dehydrogenase | - |
Ensifer sp. AS08 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.63 | - |
PEG-1000 | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
1.3 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
5.5 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
8.4 | - |
PEG-1000 | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
11 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
12 | - |
PEG-1000 | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Ensifer sp. AS08 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
PEG-1000 | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
0.54 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme H465A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
1.3 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
1.9 | - |
PEG-1000 | mutant enzyme N90A, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
3.6 | - |
PEG-1000 | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 | |
5.2 | - |
PEG-mono-4-nonylphenyl ether, averaging 10 ethoxy units | wild type enzyme, in 0.1 M Tris-HCl buffer (pH 8.0), at 25°C | Ensifer sp. AS08 |