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Literature summary for 1.14.13.2 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

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxybenzoate + NADH + O2 the enzyme prefers NADPH to NADH Corynebacterium glutamicum protocatechuate + NAD+ + H2O
-
?
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
-
?
4-hydroxybenzoate + NADPH + O2 the enzyme prefers NADPH to NADH Corynebacterium glutamicum protocatechuate + NADP+ + H2O
-
?
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

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

Synonyms

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
Corynebacterium glutamicum

pH Range

pH Minimum pH Maximum Comment Organism
7.5 9
-
Corynebacterium glutamicum

Cofactor

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