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Literature summary extracted from

  • Tan, C.L.; Yeo, C.C.; Khoo, H.E.; Poh, C.L.
    Replacement of tyrosine 181 by phenylalanine in gentisate 1,2-dioxygenase I from Pseudomonas alcaligenes NCIMB 9867 enhances catalytic activities (2005), J. Bacteriol., 187, 7543-7545.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.11.4 expressed in Escherichia coli BL21 cells Pseudomonas alcaligenes

Protein Variants

EC Number Protein Variants Comment Organism
1.13.11.4 D120K complete loss of activity Pseudomonas alcaligenes
1.13.11.4 E223A 102% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 G123N 75% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 G164T complete loss of activity Pseudomonas alcaligenes
1.13.11.4 K338Y 156% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 M146T complete loss of activity Pseudomonas alcaligenes
1.13.11.4 M169T complete loss of activity Pseudomonas alcaligenes
1.13.11.4 N153H 183% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 N43T 317% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 S113P complete loss of activity Pseudomonas alcaligenes
1.13.11.4 T260C complete loss of activity Pseudomonas alcaligenes
1.13.11.4 V326Q complete loss of activity Pseudomonas alcaligenes
1.13.11.4 V36A complete loss of activity Pseudomonas alcaligenes
1.13.11.4 Y17C 161% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 Y181D 68% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 Y181F mutant demonstrates 4-, 3-, 6-, and 16fold increase in relative activity towards 2,5-dihydroxybenzoate, 3-fluoro-, 4-methyl-, and 3-methylgentisate, respectively, and shows 464% relative activity compared to the wild type enzyme for 2,5-dihydroxybenzoate Pseudomonas alcaligenes
1.13.11.4 Y181H 98% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
1.13.11.4 Y284I 260% relative activity compared to the wild type enzyme Pseudomonas alcaligenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.11.4 0.02326
-
4-Chlorogentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.04591
-
3-Bromogentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.06069
-
4-Chlorogentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.06234
-
3-Methylgentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.06769
-
3-Isopropylgentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.06843
-
2,5-Dihydroxybenzoate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.07258
-
4-Methylgentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.07426
-
4-Methylgentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.07669
-
3-Fluorogentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.07826
-
3-Bromogentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.07937
-
3-Isopropylgentisate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.0862
-
2,5-Dihydroxybenzoate wild type enzyme, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.08734
-
3-Methylgentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes
1.13.11.4 0.08962
-
3-Fluorogentisate mutant enzyme Y181F, in 0.1 M KH2PO4 buffer (pH 7.4) at 23°C Pseudomonas alcaligenes

Organism

EC Number Organism UniProt Comment Textmining
1.13.11.4 Pseudomonas alcaligenes
-
-
-
1.13.11.4 Pseudomonas alcaligenes NCIMB 9867
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.11.4 GSTrap fast flow column chromatography Pseudomonas alcaligenes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.11.4 2,5-dihydroxybenzoate + O2
-
Pseudomonas alcaligenes maleylpyruvate
-
?
1.13.11.4 2,5-dihydroxybenzoate + O2
-
Pseudomonas alcaligenes NCIMB 9867 maleylpyruvate
-
?
1.13.11.4 3-bromogentisate + O2
-
Pseudomonas alcaligenes 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 3-fluorogentisate + O2
-
Pseudomonas alcaligenes 3-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 3-fluorogentisate + O2
-
Pseudomonas alcaligenes NCIMB 9867 3-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 3-isopropylgentisate + O2
-
Pseudomonas alcaligenes 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 3-methylgentisate + O2
-
Pseudomonas alcaligenes 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 3-methylgentisate + O2
-
Pseudomonas alcaligenes NCIMB 9867 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 4-chlorogentisate + O2
-
Pseudomonas alcaligenes 2-chloro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 4-chlorogentisate + O2
-
Pseudomonas alcaligenes NCIMB 9867 2-chloro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 4-methylgentisate + O2
-
Pseudomonas alcaligenes 4-hydroxy-2-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
1.13.11.4 4-methylgentisate + O2
-
Pseudomonas alcaligenes NCIMB 9867 4-hydroxy-2-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?

Synonyms

EC Number Synonyms Comment Organism
1.13.11.4 gentisate dioxygenase
-
Pseudomonas alcaligenes