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Literature summary for 1.13.11.2 extracted from

  • Viggiani, A.; Siani, L.; Notomista, E.; Birolo, L.; Pucci, P.; Di Donato, A.
    The role of the conserved residues His-246, His-199, and Tyr-255 in the catalysis of catechol 2,3-dioxygenase from Pseudomonas stutzeri OX1 (2004), J. Biol. Chem., 279, 48630-48639.
    View publication on PubMed
Search for more literature for 1.13.11.2

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) Pseudomonas stutzeri

Protein Variants

Protein Variants Comment Organism
H199N strongly reduced activity, significant changes in pH profile Pseudomonas stutzeri
H246A strongly reduced activity, significant changes in pH profile Pseudomonas stutzeri
H246N strongly reduced activity, significant changes in pH profile Pseudomonas stutzeri
Y255F strongly reduced activity Pseudomonas stutzeri

Inhibitors

Inhibitors Comment Organism Structure
H2O2 partially reversible by addition of Fe(NH4)2(SO4)2 Pseudomonas stutzeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0015
-
 catechol wild type enzyme, pH 7.5, 25°C Pseudomonas stutzeri
0.005
-
 catechol Y255F mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri
0.434
-
 catechol H199N mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri
0.714
-
 catechol H246N mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
 SDS-PAGE Pseudomonas stutzeri
35100
-
 calculated from deduced amino acid sequence Pseudomonas stutzeri
35110
-
 mass sprectrometry Pseudomonas stutzeri
117000
-
 gel filtration Pseudomonas stutzeri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
catechol + O2 Pseudomonas stutzeri
-
 2-hydroxymuconate semialdehyde
-
 ?
catechol + O2 Pseudomonas stutzeri OX1
-
 2-hydroxymuconate semialdehyde
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas stutzeri Q8KLV4
-
-
Pseudomonas stutzeri OX1 Q8KLV4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Pseudomonas stutzeri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-methoxycatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri 2-hydroxy-3-methoxymuconate semialdehyde
-
 ?
3-methoxycatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri OX1 2-hydroxy-3-methoxymuconate semialdehyde
-
 ?
4-ethylcatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri 4-ethyl-2-hydroxymuconate semialdehyde
-
 ?
4-ethylcatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri OX1 4-ethyl-2-hydroxymuconate semialdehyde
-
 ?
4-formylcatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri 4-formyl-2-hydroxymuconate semialdehyde
-
 ?
4-formylcatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri OX1 4-formyl-2-hydroxymuconate semialdehyde
-
 ?
4-methylcatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri 2-hydroxy-4-methylmuconate semialdehyde
-
 ?
4-nitrocatechol + O2 rapid inactivation of enzyme during turnover Pseudomonas stutzeri 2-hydroxy-4-nitromuconate semialdehyde
-
 ?
catechol + O2
-
 Pseudomonas stutzeri 2-hydroxymuconate semialdehyde
-
 ?
catechol + O2 most effective substrate Pseudomonas stutzeri 2-hydroxymuconate semialdehyde
-
 ?
catechol + O2
-
 Pseudomonas stutzeri OX1 2-hydroxymuconate semialdehyde
-
 ?
catechol + O2 most effective substrate Pseudomonas stutzeri OX1 2-hydroxymuconate semialdehyde
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 35000, SDS-PAGE, mass spectrometry Pseudomonas stutzeri

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.46
-
 catechol H199N mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri
1.6
-
 catechol H246N mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri
8.8
-
 catechol Y255F mutant enzyme, pH 7.5, 25°C Pseudomonas stutzeri
200
-
 catechol wild type enzyme, pH 7.5, 25°C Pseudomonas stutzeri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 rapid loss of activity below pH 6, very slow decrease of activity above pH 6 Pseudomonas stutzeri

pH Stability

pH Stability pH Stability Maximum Comment Organism
6
-
 rapid loss of activity below pH 6 Pseudomonas stutzeri