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Literature summary extracted from

  • Chang, H.K.; Zylstra, G.J.
    Examination and expansion of the substrate range of m-hydroxybenzoate hydroxylase (2008), Biochem. Biophys. Res. Commun., 371, 149-153.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.2 mobA gene from pSEB2 cloned into pZR80. Random mutations introduced into the mobA gene and cloned into pCR2.1-TOPO and transformed into Escherichia coli Top10 Comamonas testosteroni
1.14.13.23 gene mobA, DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Escherichia coli Comamonas testosteroni

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.2 A16T/S394P/D416A low ability to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 A400G transforms 3-aminophenol with efficiency almost like mutant A400G/K429R Comamonas testosteroni
1.14.13.2 A400G/K429R among mutants, highest enzymatic activity to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 H135P alters the enzyme's substrate specificity Comamonas testosteroni
1.14.13.2 H135P low ability to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 H135P/I217L/Y304H low ability to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 K326I lacks the ability to transform phenol to catechol as the wild-type Comamonas testosteroni
1.14.13.2 K429R can not transform 3-aminophenol at all Comamonas testosteroni
1.14.13.2 N102T/I259S/V399M low ability to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 N227H is not able to transform 3-aminophenol Comamonas testosteroni
1.14.13.2 N227H/D416A almost has the same transformation efficiency as mutant N227H/Q292R/D416A Comamonas testosteroni
1.14.13.2 N227H/Q292R/D416A among mutants, highest enzymatic activity to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 Q292R is not able to transform 3-aminophenol Comamonas testosteroni
1.14.13.2 R152L/F364V low ability to hydroxylate 3-aminophenol Comamonas testosteroni
1.14.13.2 V257A mutation enables the mutant to transform phenol to catechol, also has enhanced ability to transform resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3-chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol, thus broadens the substrate range. Is not capable of hydroxylating benzoate, o-hydroxybenzoate (salicylate), 2,4-dihydroxybenzoate, 2,6-dihydroxybenzoate, 2-chlorophenol, 3-aminophenol, 4-methoxybenzoate, 3-toluate, o-cresol, m-cresol, or p-cresol as the wild-type Comamonas testosteroni
1.14.13.23 A400G random mutagenesis, the substitution confers the ability to the mutant enzyme to transform 3-aminophenol to a related substituted catechol Comamonas testosteroni
1.14.13.23 D416A random mutagenesis, the substitution confers the ability to the mutant enzyme to transform 3-aminophenol to a related substituted catechol Comamonas testosteroni
1.14.13.23 H135P random mutagenesis, the substitution confers the ability to the mutant enzyme to transform 3-aminophenol to a related substituted catechol Comamonas testosteroni
1.14.13.23 K326I random mutagenesis, the substitution confers the ability to the mutant enzyme to transform phenol to catechol Comamonas testosteroni
1.14.13.23 additional information screening of random mutants from a cosmid library for altered substrate specificities Comamonas testosteroni
1.14.13.23 V257A random mutagenesis, the substitution confers the ability to the mutant enzyme to transform phenol to catechol, the mutant is also active with resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3-chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol Comamonas testosteroni

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.23 3-hydroxybenzoate + NADPH + H+ + O2 Comamonas testosteroni
-
 3,4-dihydroxybenzoate + NADP+ + H2O i.e. procatechuate ?
1.14.13.23 4-hydroxybenzoate + NADPH + H+ + O2 Comamonas testosteroni low activity 3,4-dihydroxybenzoate + NADP+ + H2O i.e. procatechuate ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.2 Comamonas testosteroni Q6SSJ6
-
-
1.14.13.2 Comamonas testosteroni GZ39 Q6SSJ6
-
-
1.14.13.23 Comamonas testosteroni Q6SSJ6 gene mobA
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.2 2,3-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 2,3-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni GZ39 ? + NADP+ + H2O
-
 ?
1.14.13.2 2,5-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 2,5-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni GZ39 ? + NADP+ + H2O
-
 ?
1.14.13.2 3,4-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 3,4-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni GZ39 ? + NADP+ + H2O
-
 ?
1.14.13.2 3,5-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 3,5-dihydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni GZ39 ? + NADP+ + H2O
-
 ?
1.14.13.2 3-chlorophenol + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 3-hydroxyanthranilate + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 4-chlorophenol + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 4-chlororesorcinol + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 4-nitrophenol + NADPH + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 hydroquinone + NADPH + H+ + O2
-
 Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.2 m-hydroxybenzoate + NADPH + O2
-
 Comamonas testosteroni protocatechuate + NADP+ + H2O
-
 ?
1.14.13.2 additional information is not capable of hydroxylating benzoate, o-hydroxybenzoate (salicylate), 2,4-dihydroxybenzoate, 2,6-dihydroxybenzoate, 2-chlorophenol, 3-aminophenol, 4-methoxybenzoate, 3-toluate, o-cresol, m-cresol, or p-cresol Comamonas testosteroni ?
-
 ?
1.14.13.2 additional information is not capable of hydroxylating benzoate, o-hydroxybenzoate (salicylate), 2,4-dihydroxybenzoate, 2,6-dihydroxybenzoate, 2-chlorophenol, 3-aminophenol, 4-methoxybenzoate, 3-toluate, o-cresol, m-cresol, or p-cresol Comamonas testosteroni GZ39 ?
-
 ?
1.14.13.2 p-hydroxybenzoate + NADPH + O2 is transformed to a lesser extent than m-hydroxybenzoate Comamonas testosteroni ? + NADP+ + H2O
-
 ?
1.14.13.23 3-hydroxybenzoate + NADPH + H+ + O2
-
 Comamonas testosteroni 3,4-dihydroxybenzoate + NADP+ + H2O i.e. procatechuate ?
1.14.13.23 4-hydroxybenzoate + NADPH + H+ + O2 low activity Comamonas testosteroni 3,4-dihydroxybenzoate + NADP+ + H2O i.e. procatechuate ?

Synonyms

EC Number Synonyms Comment Organism
1.14.13.2 m-hydroxybenzoate hydroxylase
-
 Comamonas testosteroni
1.14.13.2 MobA
-
 Comamonas testosteroni
1.14.13.23 m-hydroxybenzoate hydroxylase
-
 Comamonas testosteroni
1.14.13.23 MobA
-
 Comamonas testosteroni

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.2 NADPH
-
 Comamonas testosteroni