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Literature summary extracted from

  • Huang, Y.; Zhao, K.X.; Shen, X.H.; Jiang, C.Y.; Liu, S.J.
    Genetic and biochemical characterization of a 4-hydroxybenzoate hydroxylase from Corynebacterium glutamicum (2008), Appl. Microbiol. Biotechnol., 78, 75-83.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.14.13.2 dithiothreitol 4 mM, slightly increases activity Corynebacterium glutamicum
1.14.13.2 EDTA 5 mM or 20 mM, slightly increases activity Corynebacterium glutamicum

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.2 D38A kcat/KM for 4-hydroxybenzoate is 16.8fold higher than wild-type value Corynebacterium glutamicum
1.14.13.2 D38Y kcat/KM for 4-hydroxybenzoate is 11.8fold higher than wild-type value Corynebacterium glutamicum
1.14.13.2 D38Y/T42R kcat/KM for 4-hydroxybenzoate is 32fold higher than wild-type value Corynebacterium glutamicum
1.14.13.2 T42R kcat/KM for 4-hydroxybenzoate is 7.2fold higher than wild-type value Corynebacterium glutamicum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.2 Cl- 50 mM, significant inhibition. No inhibition at 5 mM Corynebacterium glutamicum
1.14.13.2 Cu2+ 1 mM, moderate inhibition Corynebacterium glutamicum
1.14.13.2 Fe3+ 1 mM, moderate inhibition Corynebacterium glutamicum
1.14.13.2 Mn2+ 1 mM, moderate inhibition Corynebacterium glutamicum
1.14.13.2 NEM 0.1 mM, 59% inhibition Corynebacterium glutamicum
1.14.13.2 Zn2+ 1 mM, moderate inhibition Corynebacterium glutamicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.2 0.008
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38Y/T42R Corynebacterium glutamicum
1.14.13.2 0.012
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38A Corynebacterium glutamicum
1.14.13.2 0.014
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme T42R Corynebacterium glutamicum
1.14.13.2 0.026
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38Y Corynebacterium glutamicum
1.14.13.2 0.036
-
 4-hydroxybenzoate pH 8.0, 25°C, wild-type enzyme Corynebacterium glutamicum
1.14.13.2 0.04
-
 NADPH pH 8.0, 25°C, mutant enzyme D38A Corynebacterium glutamicum
1.14.13.2 0.045
-
 NADPH pH 8.0, 25°C, mutant enzyme T42R Corynebacterium glutamicum
1.14.13.2 0.047
-
 NADPH pH 8.0, 25°C, mutant enzyme D38Y Corynebacterium glutamicum
1.14.13.2 0.084
-
 NADPH pH 8.0, 25°C, mutant enzyme D38Y/T42R Corynebacterium glutamicum
1.14.13.2 0.195
-
 NADPH pH 8.0, 25°C, wild-type enzyme Corynebacterium glutamicum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.13.2 additional information metal ions (at 1 mM) Co2+, Ni2+, Mg2+, and Hg2+ shows little influence on hydroxylase activity Corynebacterium glutamicum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.2 45000
-
 2 * 45000, SDS-PAGE Corynebacterium glutamicum
1.14.13.2 80000
-
 gel filtration, non-denaturing PAGE Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.2 4-hydroxybenzoate + NADPH + O2 Corynebacterium glutamicum the enzyme prefers NADPH to NADH. 4HBA hydroxylase can be induced by 4HBA, 4-cresol, and 4-hydroxybenzaldehyde protocatechuate + NADP+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.2 Corynebacterium glutamicum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.2
-
 Corynebacterium glutamicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.2 4-hydroxybenzoate + NADH + O2 the enzyme prefers NADPH to NADH Corynebacterium glutamicum protocatechuate + NAD+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADPH + O2 the enzyme prefers NADPH to NADH. 4HBA hydroxylase can be induced by 4HBA, 4-cresol, and 4-hydroxybenzaldehyde Corynebacterium glutamicum protocatechuate + NADP+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADPH + O2 the enzyme prefers NADPH to NADH Corynebacterium glutamicum protocatechuate + NADP+ + H2O
-
 ?
1.14.13.2 additional information no hydroxylase activity is observed with 3-hydroxybenzoate, resorcinol, 3-nitrophenol, benzoate, 2,4-dihydroxybenzoate, 4-chlorobenzoate, 4-aminobenzoate, 4-aminophenol, 4-nitrophenol, and 4-dinitrobenzene Corynebacterium glutamicum ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.2 dimer 2 * 45000, SDS-PAGE Corynebacterium glutamicum

Synonyms

EC Number Synonyms Comment Organism
1.14.13.2 4-hydroxybenzoate hydroxylase
-
 Corynebacterium glutamicum
1.14.13.2 4HBA 3-hydroxylase
-
 Corynebacterium glutamicum
1.14.13.2 ncgl1032
-
 Corynebacterium glutamicum
1.14.13.2 pobACg
-
 Corynebacterium glutamicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.2 2.8
-
 4-hydroxybenzoate pH 8.0, 25°C, wild-type enzyme Corynebacterium glutamicum
1.14.13.2 7.5
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme T42R Corynebacterium glutamicum
1.14.13.2 7.8
-
 NADPH pH 8.0, 25°C, wild-type enzyme Corynebacterium glutamicum
1.14.13.2 12.4
-
 NADPH pH 8.0, 25°C, mutant enzyme T42R Corynebacterium glutamicum
1.14.13.2 15.2
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38A Corynebacterium glutamicum
1.14.13.2 18.8
-
 NADPH pH 8.0, 25°C, mutant enzyme D38A Corynebacterium glutamicum
1.14.13.2 19.6
-
 NADPH pH 8.0, 25°C, mutant enzyme D38Y Corynebacterium glutamicum
1.14.13.2 20.4
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38Y/T42R Corynebacterium glutamicum
1.14.13.2 24.3
-
 4-hydroxybenzoate pH 8.0, 25°C, mutant enzyme D38Y Corynebacterium glutamicum
1.14.13.2 46.4
-
 NADPH pH 8.0, 25°C, mutant enzyme D38Y/T42R Corynebacterium glutamicum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.2 8.5
-
-
 Corynebacterium glutamicum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.13.2 7.5 9
-
 Corynebacterium glutamicum

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.2 NADH the enzyme prefers NADPH to NADH Corynebacterium glutamicum
1.14.13.2 NADPH the enzyme prefers NADPH to NADH Corynebacterium glutamicum