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Literature summary extracted from

  • Sucharitakul, J.; Medhanavyn, D.; Pakotiprapha, D.; van Berkel, W.J.; Chaiyen, P.
    Tyr217 and His213 are important for substrate binding and hydroxylation of 3-hydroxybenzoate 6-hydroxylase from Rhodococcus jostii RHA1 (2016), FEBS J., 283, 860-881 .
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.24 H213A the mutant enzyme cannot catalyze hydroxylation Rhodococcus jostii
1.14.13.24 H213D the mutant enzyme can bind to 3-hydroxybenzoate with similar affinity as the wild-type enzyme and form C4a-hydroperoxy intermediate. It produces 2,5-dihydroxybenzoate with yields of 52% Rhodococcus jostii
1.14.13.24 H213E the mutant enzyme can bind to 3-hydroxybenzoate with similar affinity as the wild-type enzyme and form C4a-hydroperoxy intermediate. It produces 2,5-dihydroxybenzoate with yields of 92%. The hydroxylation rate constant of the mutant enzyme (35/s) is similar to that of wild-type enzyme (36/s) and this variant has an efficiency of hydroxylation (92%) similar to the wild-type enzyme (86%) Rhodococcus jostii
1.14.13.24 H213S the mutant enzyme can bind to 3-hydroxybenzoate with similar affinity as the wild-type enzyme and form C4a-hydroperoxy intermediate. It produces 2,5-dihydroxybenzoate with yields of 28% Rhodococcus jostii
1.14.13.24 Y217A the mutant enzyme does not show any perturbation of flavin absorption upon addition of 3-hydroxybenzoate Rhodococcus jostii
1.14.13.24 Y217F the mutant enzyme has a Kd value for 3-hydroxybenzoate binding of 7.5 mM, which is about 50fold larger than that found for wild-type enzyme. The results indicate that Tyr217 is necessary for substrate binding Rhodococcus jostii
1.14.13.24 Y217S the mutant enzyme does not show any perturbation of flavin absorption upon addition of 3-hydroxybenzoate Rhodococcus jostii

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.24 Rhodococcus jostii Q0SFK6
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Substrates and Products (Substrate)

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

Synonyms

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

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.24 FAD flavoprotein Rhodococcus jostii