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Literature summary extracted from

  • Seibold, B.; Matthes, M.; Eppink, M.H.M.; Lingens, F.; Van Berkel, W.J.H.; Mueller, R.
    4-Hydroxybenzoate hydroxylase from Pseudomonas sp. CBS3. Purification, characterization, gene cloning, sequence analysis and assignment of structural features determining the coenzyme specificity (1996), Eur. J. Biochem., 239, 469-478.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.2
-
 Pseudomonas sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.2 Cl- competitive with respect to NADH Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.2 0.075
-
 4-hydroxybenzoate 35°C Pseudomonas sp.
1.14.13.2 0.08
-
 NADH 25°C Pseudomonas sp.
1.14.13.2 0.12
-
 NADH 35°C Pseudomonas sp.
1.14.13.2 0.14
-
 NADPH 25°C Pseudomonas sp.
1.14.13.2 0.16
-
 NADPH 35°C Pseudomonas sp.
1.14.13.2 0.18
-
 NADH 45°C Pseudomonas sp.
1.14.13.2 0.2
-
 NADPH at 45°C Pseudomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.2 43632
-
 2 * 43632, calculation from nucleotide sequence Pseudomonas sp.
1.14.13.2 44000
-
 2 * 44000, SDS-PAGE Pseudomonas sp.
1.14.13.2 90000
-
 gel filtration Pseudomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.2 Pseudomonas sp.
-
 CBS3
-
1.14.13.2 Pseudomonas sp. CBS3
-
 CBS3
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.2
-
 Pseudomonas sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.2 additional information
-
-
 Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.2 2,4-dihydroxybenzoate + NADPH + O2 8% of the activity with 4-hydroxybenzoate Pseudomonas sp. 2,3,4-trihydroxybenzoate + 2,4,5-trihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.2 2,4-dihydroxybenzoate + NADPH + O2 8% of the activity with 4-hydroxybenzoate Pseudomonas sp. CBS3 2,3,4-trihydroxybenzoate + 2,4,5-trihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.2 2-chloro-4-hydroxybenzoate + NADH + O2 40% of the activity with 4-hydroxybenzoate Pseudomonas sp. ?
-
 ?
1.14.13.2 2-chloro-4-hydroxybenzoate + NADH + O2 40% of the activity with 4-hydroxybenzoate Pseudomonas sp. CBS3 ?
-
 ?
1.14.13.2 2-fluoro-4-hydroxybenzoate + NADH + O2 50% of the activity with 4-hydroxybenzoate Pseudomonas sp. ?
-
 ?
1.14.13.2 2-fluoro-4-hydroxybenzoate + NADH + O2 50% of the activity with 4-hydroxybenzoate Pseudomonas sp. CBS3 ?
-
 ?
1.14.13.2 4-hydroxybenzoate + NADH + O2
-
 Pseudomonas sp. protocatechuate + NAD+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADH + O2
-
 Pseudomonas sp. CBS3 protocatechuate + NAD+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADPH + O2
-
 Pseudomonas sp. protocatechuate + NADP+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADPH + O2
-
 Pseudomonas sp. CBS3 protocatechuate + NADP+ + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.2 dimer 2 * 44000, SDS-PAGE Pseudomonas sp.
1.14.13.2 dimer 2 * 43632, calculation from nucleotide sequence Pseudomonas sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.2 45
-
-
 Pseudomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.2 6.6
-
 NADPH 25°C Pseudomonas sp.
1.14.13.2 6.8
-
 NADH 25°C Pseudomonas sp.
1.14.13.2 7.4
-
 4-hydroxybenzoate 35°C Pseudomonas sp.
1.14.13.2 8.6
-
 NADPH 35°C Pseudomonas sp.
1.14.13.2 11.6
-
 NADH 35°C Pseudomonas sp.
1.14.13.2 13.3
-
 NADPH 45°C Pseudomonas sp.
1.14.13.2 16.9
-
 NADH 45°C Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.2 8
-
-
 Pseudomonas sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.2 FAD Km: 150 nM Pseudomonas sp.
1.14.13.2 additional information helix H2 is involved in determining the coenzyme specificity Pseudomonas sp.
1.14.13.2 NADH activity with NADH is higher than activity with NADPH Pseudomonas sp.

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.13.2 50
-
 Cl-
-
 Pseudomonas sp.