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Literature summary extracted from

  • Chen, Z.; Shen, X.; Wang, J.; Wang, J.; Yuan, Q.; Yan, Y.
    Rational engineering of p-hydroxybenzoate hydroxylase to enable efficient gallic acid synthesis via a novel artificial biosynthetic pathway (2017), Biotechnol. Bioeng., 114, 2571-2580 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.2 expressed in Escherichia coli BL21 Star (DE3) cells Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.2 Y385F the mutant displays higher activity toward 3,4-dihydroxybenzoic acid than the wild type enzyme Pseudomonas aeruginosa
1.14.13.2 Y385F/T294A the mutant displays much higher activity toward 3,4-dihydroxybenzoic acid than the wild type enzyme Pseudomonas aeruginosa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.2 0.01955
-
 4-hydroxybenzoate mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.03467
-
 4-hydroxybenzoate wild type enzyme, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.04838
-
 4-hydroxybenzoate mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.15702
-
 3,4-dihydroxybenzoic acid mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.22802
-
 3,4-dihydroxybenzoic acid mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.2 4-hydroxybenzoate + NADPH + H+ + O2 Pseudomonas aeruginosa
-
 protocatechuate + NADP+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.2 Pseudomonas aeruginosa
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.2 0.26
-
 mutant enzyme Y385F, with 4-hydroxybenzoate as substrate, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.45
-
 mutant enzyme Y385F, with 3,4-dihydroxybenzoic acid as substrate, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 1.04
-
 mutant enzyme Y385F/T294A, with 4-hydroxybenzoate as substrate, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 1.86
-
 mutant enzyme Y385F/T294A, with 3,4-dihydroxybenzoic acid as substrate, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 16.85
-
 wild type enzyme, with 4-hydroxybenzoate as substrate, at pH 8.0 and 30°C Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.2 3,4-dihydroxybenzoic acid + NADPH + H+ + O2 weak activity Pseudomonas aeruginosa gallic acid + NADP+ + H2O
-
 ?
1.14.13.2 4-hydroxybenzoate + NADPH + H+ + O2
-
 Pseudomonas aeruginosa protocatechuate + NADP+ + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.13.2 p-hydroxybenzoate hydroxylase
-
 Pseudomonas aeruginosa
1.14.13.2 PobA
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.2 0.2
-
 4-hydroxybenzoate mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.39
-
 3,4-dihydroxybenzoic acid mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 0.9
-
 4-hydroxybenzoate mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 1.69
-
 3,4-dihydroxybenzoic acid mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 14.12
-
 4-hydroxybenzoate wild type enzyme, at pH 8.0 and 30°C Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.2 FAD
-
 Pseudomonas aeruginosa
1.14.13.2 NADPH
-
 Pseudomonas aeruginosa

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.2 2
-
 3,4-dihydroxybenzoic acid mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 10
-
 4-hydroxybenzoate mutant Y385F, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 12
-
 3,4-dihydroxybenzoic acid mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 20
-
 4-hydroxybenzoate mutant Y385F/T294A, at pH 8.0 and 30°C Pseudomonas aeruginosa
1.14.13.2 410
-
 4-hydroxybenzoate wild type enzyme, at pH 8.0 and 30°C Pseudomonas aeruginosa