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Literature summary for 1.13.11.4 extracted from

  • Harpel, M.R.; Lipscomb, J.D.
    Gentisate 1,2-dioxygenase from pseudomonas. Purification, characterization, and comparison of the enzymes from Pseudomonas testosteroni and Pseudomonas acidovorans (1990), J. Biol. Chem., 265, 6301-6311.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
ascorbate
-
Comamonas testosteroni
ascorbate
-
Delftia acidovorans
dithiothreitol
-
Comamonas testosteroni
dithiothreitol
-
Delftia acidovorans

General Stability

General Stability Organism
cysteine stabilizes Comamonas testosteroni
cysteine stabilizes Delftia acidovorans
Fe2+ stabilizes Comamonas testosteroni
Fe2+ stabilizes Delftia acidovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.015
-
4-Fluorogentisate
-
Delftia acidovorans
0.018
-
3-Bromogentisate
-
Delftia acidovorans
0.025
-
4-Chlorogentisate
-
Delftia acidovorans
0.027
-
3-Methylgentisate
-
Delftia acidovorans
0.035
-
3-Fluorogentisate
-
Delftia acidovorans
0.044
-
4-Fluorogentisate
-
Comamonas testosteroni
0.045
-
1,4-dihydroxy-2-naphthoate
-
Delftia acidovorans
0.048
-
3-Isopropylgentisate
-
Delftia acidovorans
0.05
-
3-Fluorogentisate
-
Comamonas testosteroni
0.057
-
4-Methylgentisate
-
Delftia acidovorans
0.074
-
gentisate
-
Delftia acidovorans
0.085
-
gentisate
-
Comamonas testosteroni
0.096
-
O2
-
Comamonas testosteroni
0.14
-
3,4-Dimethylgentisate
-
Delftia acidovorans
0.24
-
4-Chlorogentisate
-
Comamonas testosteroni
0.29
-
1,4-dihydroxy-2-naphthoate
-
Comamonas testosteroni
0.33
-
3-Bromogentisate
-
Comamonas testosteroni
0.45
-
4-O-Methylgentisate
-
Comamonas testosteroni
0.52
-
4-Methylgentisate
-
Comamonas testosteroni
1.18
-
3-O-Methylgentisate
-
Comamonas testosteroni
1.2
-
4-Isopropylgentisate
-
Comamonas testosteroni
1.23
-
4-Isopropylgentisate
-
Comamonas testosteroni
1.75
-
3-Methylgentisate
-
Comamonas testosteroni
2.75
-
3-Isopropylgentisate
-
Comamonas testosteroni
3.65
-
3-Ethylgentisate
-
Comamonas testosteroni
7.7
-
3,4-Dimethylgentisate
-
Comamonas testosteroni

Metals/Ions

Metals/Ions Comment Organism Structure
Iron 1.9-3.9 mol Fe2+ per subunit Comamonas testosteroni
Iron 1.3-3.2 mol Fe2+ per subunit Delftia acidovorans
Iron (alphaFe)4 structure Comamonas testosteroni
Iron (alphaFe)4 structure Delftia acidovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40800
-
4 * 40800, SDS-PAGE Comamonas testosteroni
158000
-
gel filtration Comamonas testosteroni
164000
-
gel filtration Delftia acidovorans

Organism

Organism UniProt Comment Textmining
Comamonas testosteroni
-
-
-
Delftia acidovorans
-
-
-

Oxidation Stability

Oxidation Stability Organism
exposure of enzyme to oxidants, e.g. H2O2 or K3Fe(CN)6, at 1 mM concentrations results in complete loss of activity, even under anaerobic conditions Comamonas testosteroni
exposure of enzyme to oxidants, e.g. H2O2 or K3Fe(CN)6, at 1 mM concentrations results in complete loss of activity, even under anaerobic conditions Delftia acidovorans

Purification (Commentary)

Purification (Comment) Organism
-
Comamonas testosteroni
-
Delftia acidovorans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
484
-
-
Delftia acidovorans
498
-
-
Comamonas testosteroni

Storage Stability

Storage Stability Organism
-80°C, storage following rapid freezing in liquid N2, protein concentration above 10 mg/ml, about 15% loss of activity after 1 year Comamonas testosteroni
-80°C, storage following rapid freezing in liquid N2, protein concentration above 10 mg/ml, about 15% loss of activity after 1 year Delftia acidovorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,4-dihydroxy-2-naphthoate + O2
-
Comamonas testosteroni ?
-
?
1,4-dihydroxy-2-naphthoate + O2
-
Delftia acidovorans ?
-
?
2,5-dihydroxybenzoate + O2
-
Comamonas testosteroni maleylpyruvate
-
?
2,5-dihydroxybenzoate + O2
-
Delftia acidovorans maleylpyruvate
-
?
3,4-dimethylgentisate + O2
-
Comamonas testosteroni ?
-
?
3,4-dimethylgentisate + O2
-
Delftia acidovorans ?
-
?
3-bromogentisate + O2
-
Comamonas testosteroni 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-bromogentisate + O2
-
Delftia acidovorans 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-ethylgentisate + O2
-
Comamonas testosteroni 3-ethyl-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-ethylgentisate + O2
-
Delftia acidovorans 3-ethyl-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-fluorogentisate + O2
-
Comamonas testosteroni 3-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-fluorogentisate + O2
-
Delftia acidovorans 3-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-hydroxybenzoate + O2 minor pathway Comamonas testosteroni 3,4-dihydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-hydroxybenzoate + O2 main pathway Delftia acidovorans 3,4-dihydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
3-isopropylgentisate + O2
-
Comamonas testosteroni 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
?
3-isopropylgentisate + O2
-
Delftia acidovorans 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
?
3-methylgentisate + O2
-
Comamonas testosteroni 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
3-methylgentisate + O2
-
Delftia acidovorans 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
3-O-methylgentisate + O2
-
Comamonas testosteroni ?
-
?
3-O-methylgentisate + O2
-
Delftia acidovorans ?
-
?
4-chlorogentisate + O2
-
Comamonas testosteroni 2-chloro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
4-chlorogentisate + O2
-
Delftia acidovorans 2-chloro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
4-fluorogentisate + O2
-
Comamonas testosteroni 2-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
4-fluorogentisate + O2
-
Delftia acidovorans 2-fluoro-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
?
4-isopropylgentisate + O2
-
Comamonas testosteroni 4-hydroxy-2-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
?
4-isopropylgentisate + O2
-
Delftia acidovorans 4-hydroxy-2-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
?
4-methylgentisate + O2
-
Comamonas testosteroni 4-hydroxy-2-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
4-methylgentisate + O2
-
Delftia acidovorans 4-hydroxy-2-methyl-6-oxo-hepta-2,4-dienedioic acid
-
?
4-O-methylgentisate + O2
-
Comamonas testosteroni ?
-
?
4-O-methylgentisate + O2
-
Delftia acidovorans ?
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 37200-39800, SDS-PAGE Delftia acidovorans
tetramer 4 * 40800, SDS-PAGE Comamonas testosteroni

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
322 328 gentisate
-
Delftia acidovorans
367 642 gentisate
-
Comamonas testosteroni

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 9
-
Comamonas testosteroni
7 9
-
Delftia acidovorans

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 8 unstable below and above Comamonas testosteroni
6 8 unstable below and above Delftia acidovorans
7.4
-
highest stability Comamonas testosteroni
7.4
-
highest stability Delftia acidovorans