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Literature summary extracted from

  • Montersino, S.; Orru, R.; Barendregt, A.; Westphal, A.H.; van Duijn, E.; Mattevi, A.; van Berkel, W.J.
    Crystal structure of 3-hydroxybenzoate 6-hydroxylase uncovers lipid-assisted flavoprotein strategy for regioselective aromatic hydroxylation (2013), J. Biol. Chem., 288, 26235-26245.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.13.24 sitting drop vapor diffusion method, using 28% (w/v) PEG 4000, 0.2 M sodium acetate, and 0.1 M Tris-HCl (pH 8.5) Rhodococcus jostii

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.24 H213A inactive, but the strength of substrate binding is not affected by the mutation Rhodococcus jostii
1.14.13.24 H213S inactive Rhodococcus jostii
1.14.13.24 Q301E inactive Rhodococcus jostii
1.14.13.24 Y105F the mutant shows less affinity for the aromatic substrate and lower catalytic efficiency compared to the wild type enzyme Rhodococcus jostii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.24 0.027
-
 3-hydroxybenzoate wild type enzyme, at pH 8.0 and 25°C Rhodococcus jostii
1.14.13.24 0.186
-
 3-hydroxybenzoate mutant enzyme Y105F, at pH 8.0 and 25°C 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
-
 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.24 Rhodococcus jostii Q0SFK6
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus jostii 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.24 homodimer x-ray crystallography 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 3 6 3-hydroxybenzoate wild type enzyme, at pH 8.0 and 25°C Rhodococcus jostii
1.14.13.24 41
-
 3-hydroxybenzoate mutant enzyme Y105F, at pH 8.0 and 25°C Rhodococcus jostii

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.24 FAD each subunit contains a tightly but non-covalently bound FAD Rhodococcus jostii
1.14.13.24 NADH
-
 Rhodococcus jostii