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Literature summary for 1.14.13.2 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
Search for more literature for 1.14.13.2

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
-
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
p-hydroxybenzoate hydroxylase
-
 Pseudomonas aeruginosa
PobA
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Pseudomonas aeruginosa
NADPH
-
 Pseudomonas aeruginosa

kcat/KM [mM/s]

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