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Literature summary 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

EC Number Activating Compound Comment Organism Structure
1.14.13.24 2,5-Dihydroxybenzoate i.e. gentisate, strongly stimulates the NADH oxidase activity of the enzyme Rhodococcus jostii
1.14.13.24 3-hydroxybenzoate strongly stimulates activity Rhodococcus jostii
1.14.13.24 additional information enzyme activity is not stimulated by salicylate Rhodococcus jostii

Cloned(Commentary)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.24 Cl-
-
 Rhodococcus jostii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.24 0.046
-
 3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 0.048
-
 NADH in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 0.051
-
 2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 0.14
-
 2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 0.25
-
 3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 0.39
-
 NADPH in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.24 47000
-
 2 * 47000, SDS-PAGE Rhodococcus jostii
1.14.13.24 108000
-
 gel filtration Rhodococcus jostii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 Rhodococcus jostii the enzyme clearly prefers NADH as external electron donor over NADPH 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.24 Rhodococcus jostii
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.24 6
-
 crude extract, in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 24
-
 after purification, in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.24 2,3,4-trihydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 2,3,4,6-tetrahydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 2,3-dihydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 2,3,6-trihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3,4-dihydroxybenzoate + NADH + H+ + O2 very poor substrate Rhodococcus jostii ?
-
 ?
1.14.13.24 3,5-dihydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 2,3,5,6-tetrahydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxy-4-aminobenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 4-amino-2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxy-4-methoxybenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 4-methoxy-2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxy-4-methylbenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 4-methyl-2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 the enzyme clearly prefers NADH as external electron donor over NADPH Rhodococcus jostii 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADPH + H+ + O2 the enzyme clearly prefers NADH as external electron donor over NADPH Rhodococcus jostii 2,5-dihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.24 additional information no activity with 2,5-dihydroxybenzoate Rhodococcus jostii ?
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.14.13.24 3-hydroxybenzoate 6-hydroxylase
-
 Rhodococcus jostii
1.14.13.24 3HB6H
-
 Rhodococcus jostii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.24 7
-
 2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 10
-
 2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 13
-
 3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 35
-
 3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.24 8
-
 around pH 8.0 Rhodococcus jostii

Cofactor

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.24 50
-
 2,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 50
-
 3,5-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 200
-
 2,3-Dihydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii
1.14.13.24 800
-
 3-hydroxybenzoate in 50 mM Tris-SO4 (pH 8.0), at 25°C Rhodococcus jostii