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Literature summary for 1.1.99.20 extracted from

  • Liu, X.; Ohta, T.; Kawabata, T.; Kawai, F.
    Catalytic mechanism of short ethoxy chain nonylphenol dehydrogenase belonging to a polyethylene glycol dehydrogenase group in the GMC oxidoreductase family (2013), Int. J. Mol. Sci., 14, 1218-1231.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.99.20

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzymes are expressed in Escherichia coli BL21(DE3)(pLysS) cells Ensifer sp. AS08

Protein Variants

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 [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

Organism UniProt Comment Textmining
Ensifer sp. AS08 Q2HXX0
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA affinity chromatography Ensifer sp. AS08

Substrates and Products (Substrate)

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

Synonyms Comment Organism
ethoxy chain nonylphenol dehydrogenase
-
 Ensifer sp. AS08
NPEO-DH
-
 Ensifer sp. AS08
polyethylene glycol dehydrogenase
-
 Ensifer sp. AS08

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
FAD
-
 Ensifer sp. AS08

kcat/KM [mM/s]

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