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Literature summary for 5.2.1.4 extracted from

  • Fang, T.; Li, D.F.; Zhou, N.Y.
    Identification and clarification of the role of key active site residues in bacterial glutathione S-transferase zeta/maleylpyruvate isomerase (2011), Biochem. Biophys. Res. Commun., 410, 452-456.
    View publication on PubMed
Search for more literature for 5.2.1.4

Cloned(Commentary)

Cloned (Comment) Organism
N-terminal His-tagged wild type enzyme and mutants are expressed in Escherichia coli BL21(DE3) cells Ralstonia sp.

Protein Variants

Protein Variants Comment Organism
D102A completely inactive Ralstonia sp.
H104A the mutant shows 15.25% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
H38A completely inactive Ralstonia sp.
N108A the mutant shows 50.83% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
P53A completely inactive Ralstonia sp.
Q49A the mutant shows 1.15% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
Q64A completely inactive Ralstonia sp.
R109A the mutant shows 0.88% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
R110A completely inactive Ralstonia sp.
R176A the mutant shows 12.2% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
R8A the mutant shows 12.26% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
R8A/R176A the mutant shows 0.18% catalytic efficiency compared to the wild type enzyme Ralstonia sp.
S9A completely inactive Ralstonia sp.
T11A the mutant shows 97.81% catalytic efficiency compared to the wild type enzyme Ralstonia sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.344
-
 maleylpyruvate mutant enzyme R109A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
0.346
-
 maleylpyruvate mutant enzyme N108A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
0.407
-
 maleylpyruvate mutant enzyme H104A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
0.566
-
 maleylpyruvate wild type enzyme, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
0.69
-
 maleylpyruvate mutant enzyme T11A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
1.029
-
 maleylpyruvate mutant enzyme R8A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
2.355
-
 maleylpyruvate mutant enzyme R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
3.339
-
 maleylpyruvate mutant enzyme R8A/R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
6.517
-
 maleylpyruvate mutant enzyme Q49A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.

Organism

Organism UniProt Comment Textmining
Ralstonia sp.
-
-
-
Ralstonia sp. U2
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and gel filtration Ralstonia sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Maleylpyruvate
-
 Ralstonia sp. Fumarylpyruvate
-
 ?
Maleylpyruvate
-
 Ralstonia sp. U2 Fumarylpyruvate
-
 ?

Synonyms

Synonyms Comment Organism
NagL
-
 Ralstonia sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.54
-
 maleylpyruvate mutant enzyme R109A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
1.06
-
 maleylpyruvate mutant enzyme R8A/R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
11.11
-
 maleylpyruvate mutant enzyme H104A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
13.45
-
 maleylpyruvate mutant enzyme Q49A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
22.59
-
 maleylpyruvate mutant enzyme R8A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
31.47
-
 maleylpyruvate mutant enzyme N108A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
51.45
-
 maleylpyruvate mutant enzyme R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
101.3
-
 maleylpyruvate wild type enzyme, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
120.9
-
 maleylpyruvate mutant enzyme T11A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.

Cofactor

Cofactor Comment Organism Structure
glutathione dependent on reduced glutathione Ralstonia sp.

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.316
-
 maleylpyruvate mutant enzyme R8A/R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
1.579
-
 maleylpyruvate mutant enzyme R109A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
2.063
-
 maleylpyruvate mutant enzyme Q49A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
21.84
-
 maleylpyruvate mutant enzyme R176A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
21.96
-
 maleylpyruvate mutant enzyme R8A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
27.31
-
 maleylpyruvate mutant enzyme H104A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
91.31
-
 maleylpyruvate mutant enzyme N108A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
175.1
-
 maleylpyruvate mutant enzyme T11A, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.
179.1
-
 maleylpyruvate wild type enzyme, in 50 mM phosphate buffer, pH 8.0, at 22°C Ralstonia sp.