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Literature summary for 1.13.11.4 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
Search for more literature for 1.13.11.4

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
D120K complete loss of activity Pseudomonas alcaligenes
E223A 102% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
G123N 75% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
G164T complete loss of activity Pseudomonas alcaligenes
K338Y 156% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
M146T complete loss of activity Pseudomonas alcaligenes
M169T complete loss of activity Pseudomonas alcaligenes
N153H 183% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
N43T 317% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
S113P complete loss of activity Pseudomonas alcaligenes
T260C complete loss of activity Pseudomonas alcaligenes
V326Q complete loss of activity Pseudomonas alcaligenes
V36A complete loss of activity Pseudomonas alcaligenes
Y17C 161% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
Y181D 68% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
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
Y181H 98% relative activity compared to the wild type enzyme Pseudomonas alcaligenes
Y284I 260% relative activity compared to the wild type enzyme Pseudomonas alcaligenes

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Pseudomonas alcaligenes
-
-
-
Pseudomonas alcaligenes NCIMB 9867
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
gentisate dioxygenase
-
 Pseudomonas alcaligenes