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Literature summary extracted from

  • Reddy, C.C.; Vaidyanathan, C.S.
    Purification and properties of benzoate-4-hydroxylase from a soil pseudomonad (1976), Arch. Biochem. Biophys., 177, 488-498.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.14.14.92 2-mercaptoethanol hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine Pseudomonas sp.
1.14.14.92 ascorbic acid hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine Pseudomonas sp.
1.14.14.92 dithiothreitol hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine Pseudomonas sp.

General Stability

EC Number General Stability Organism
1.14.14.92 freezing and thawing has no effect Pseudomonas sp.
1.14.14.92 quite stable in presence of benzoate Pseudomonas sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.14.92 4-Chlorobenzoate
-
Pseudomonas sp.
1.14.14.92 4-Methylbenzoate
-
Pseudomonas sp.
1.14.14.92 4-nitrobenzoate 38% inhibition Pseudomonas sp.
1.14.14.92 benzaldehyde
-
Pseudomonas sp.
1.14.14.92 benzoate methyl ester slight inhibition Pseudomonas sp.
1.14.14.92 Cu2+ 0.005 mM, 50% inhibition Pseudomonas sp.
1.14.14.92 Hg2+ 0.005 mM, 70% inhibition Pseudomonas sp.
1.14.14.92 m-Hydroxybenzoate 12% inhibition Pseudomonas sp.
1.14.14.92 salicylate 20% inhibition Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.14.92 0.045
-
NADH
-
Pseudomonas sp.
1.14.14.92 0.045
-
6,7-dimethyltetrahydropterin
-
Pseudomonas sp.
1.14.14.92 0.3
-
benzoate
-
Pseudomonas sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.14.92 Fe2+ preincubation for 10 min in the presence of benzoate results in maximal activity Pseudomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.14.92 120000
-
gel filtration Pseudomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
1.14.14.92 Pseudomonas sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.92 protamine sulfate, heat treatment, calcium phosphate gel, ammonium sulfate, DEAE-Sephadex, Sephadex G-150 Pseudomonas sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.14.92 33.92
-
-
Pseudomonas sp.

Storage Stability

EC Number Storage Stability Organism
1.14.14.92 0-4°C, enzyme concentration 0.050 mg/ml, pH 6.0-7.5, 15 days, no loss of activity Pseudomonas sp.
1.14.14.92 0-4°C, pH 7.2-8.0, Tris-HCl buffer, 24 h, 50% loss of activity Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.92 4-chlorobenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
Pseudomonas sp. ?
-
?
1.14.14.92 4-methylbenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
Pseudomonas sp. ?
-
?
1.14.14.92 benzoate + 6,7-dimethyltetrahydropterin + O2
-
Pseudomonas sp. 4-hydroxybenzoate + oxidized 6,7-dimethyltetrahydropterin + H2O
-
r
1.14.14.92 benzoate + NADH + H+ + O2
-
Pseudomonas sp. 4-hydroxybenzoate + NAD+ + H2O
-
r
1.14.14.92 benzoate + [reduced NADPH-hemoprotein reductase] + O2
-
Pseudomonas sp. 4-hydroxybenzoate + [oxidized NADPH-hemoprotein reductase] + H2O
-
?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.92 34
-
-
Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.92 7.2
-
-
Pseudomonas sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.14.92 5.5 8.2 approx. 30% of maximal activity at pH 5.5, approx. 35% of maximal activity at pH 8.2 Pseudomonas sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.92 tetrahydropteridine essential for activity, hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine Pseudomonas sp.

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.14.92 0.75
-
4-nitrobenzoate
-
Pseudomonas sp.