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Literature summary for 1.14.13.24 extracted from

  • Montersino, S.; van Berkel, W.J.
    Functional annotation and characterization of 3-hydroxybenzoate 6-hydroxylase from Rhodococcus jostii RHA1 (2012), Biochim. Biophys. Acta, 1824, 433-442.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
2,5-Dihydroxybenzoate i.e. gentisate, strongly stimulates the NADH oxidase activity of the enzyme Rhodococcus jostii
3-hydroxybenzoate strongly stimulates activity Rhodococcus jostii
additional information enzyme activity is not stimulated by salicylate Rhodococcus jostii

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Top10 cells Rhodococcus jostii

Inhibitors

Inhibitors Comment Organism Structure
Cl-
-
Rhodococcus jostii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.046
-
3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
0.048
-
NADH in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
0.051
-
2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
0.14
-
2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
0.25
-
3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
0.39
-
NADPH in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
47000
-
2 * 47000, SDS-PAGE Rhodococcus jostii
108000
-
gel filtration Rhodococcus jostii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-hydroxybenzoate + NADH + H+ + O2 Rhodococcus jostii the enzyme clearly prefers NADH as external electron donor over NADPH 2,5-dihydroxybenzoate + NAD+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Rhodococcus jostii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography and Q-Sepharose column chromatography Rhodococcus jostii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6
-
crude extract, in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
24
-
after purification, in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,3,4-trihydroxybenzoate + NADH + H+ + O2
-
Rhodococcus jostii 2,3,4,6-tetrahydroxybenzoate + NAD+ + H2O
-
?
2,3-dihydroxybenzoate + NADH + H+ + O2
-
Rhodococcus jostii 2,3,6-trihydroxybenzoate + NAD+ + H2O
-
?
3,4-dihydroxybenzoate + NADH + H+ + O2 very poor substrate Rhodococcus jostii ?
-
?
3,5-dihydroxybenzoate + NADH + H+ + O2
-
Rhodococcus jostii 2,3,5,6-tetrahydroxybenzoate + NAD+ + H2O
-
?
3-hydroxy-4-aminobenzoate + NADH + H+ + O2
-
Rhodococcus jostii 4-amino-2,5-dihydroxybenzoate + NAD+ + H2O
-
?
3-hydroxy-4-methoxybenzoate + NADH + H+ + O2
-
Rhodococcus jostii 4-methoxy-2,5-dihydroxybenzoate + NAD+ + H2O
-
?
3-hydroxy-4-methylbenzoate + NADH + H+ + O2
-
Rhodococcus jostii 4-methyl-2,5-dihydroxybenzoate + NAD+ + H2O
-
?
3-hydroxybenzoate + NADH + H+ + O2 the enzyme clearly prefers NADH as external electron donor over NADPH Rhodococcus jostii 2,5-dihydroxybenzoate + NAD+ + H2O
-
?
3-hydroxybenzoate + NADPH + H+ + O2 the enzyme clearly prefers NADH as external electron donor over NADPH Rhodococcus jostii 2,5-dihydroxybenzoate + NADP+ + H2O
-
?
additional information no activity with 2,5-dihydroxybenzoate Rhodococcus jostii ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 47000, SDS-PAGE Rhodococcus jostii

Synonyms

Synonyms Comment Organism
3-hydroxybenzoate 6-hydroxylase
-
Rhodococcus jostii
3HB6H
-
Rhodococcus jostii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7
-
2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
10
-
2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
13
-
3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
35
-
3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
around pH 8.0 Rhodococcus jostii

Cofactor

Cofactor Comment Organism Structure
FAD the enzyme contains a non-covalently bound FAD cofactor Rhodococcus jostii
NADH
-
Rhodococcus jostii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
50
-
2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
50
-
3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
200
-
2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
800
-
3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii