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Literature summary extracted from

  • Feng, Y.; Khoo, H.E.; Poh, C.L.
    Purification and characterization of gentisate 1,2-dioxygenases from Pseudomonas alcaligenes NCIB 9867 and Pseudomonas putida NCIB 9869 (1999), Appl. Environ. Microbiol., 65, 946-950.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.13.11.4 2-mercaptoethanol stimulated by 2 mM Pseudomonas putida
1.13.11.4 2-mercaptoethanol stimulated by 2 mM Pseudomonas alcaligenes
1.13.11.4 Fe2+
-
 Pseudomonas putida
1.13.11.4 Fe2+
-
 Pseudomonas alcaligenes
1.13.11.4 glycerol stimulated by 10% (v/v) Pseudomonas putida
1.13.11.4 glycerol stimulated by 10% (v/v) Pseudomonas alcaligenes
1.13.11.4 L-cysteine stimulated by 2 mM Pseudomonas putida
1.13.11.4 L-cysteine stimulated by 2 mM Pseudomonas alcaligenes

General Stability

EC Number General Stability Organism
1.13.11.4 highly unstable in 50 mM MOPS buffer with no additives Pseudomonas putida
1.13.11.4 highly unstable in 50 mM MOPS buffer with no additives Pseudomonas alcaligenes

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.13.11.4 Cu2+ 5 mM complete inactivation; complete inhibition at 5 mM Pseudomonas alcaligenes
1.13.11.4 Cu2+ 5 mM complete inactivation; complete inhibition at 5 mM Pseudomonas putida
1.13.11.4 EDTA 60.24% residual activity at 5 mM Pseudomonas alcaligenes
1.13.11.4 EDTA 4.95% residual activity at 5 mM Pseudomonas putida
1.13.11.4 Mg2+ 66% residual activity at 5 mM Pseudomonas putida
1.13.11.4 Mn2+ 73.96% residual activity at 5 mM Pseudomonas alcaligenes
1.13.11.4 Mn2+ 22.36% residual activity at 5 mM Pseudomonas putida
1.13.11.4 additional information K+, Na+, Mg2+, and Ca2+ do not inhibit the activity Pseudomonas alcaligenes
1.13.11.4 additional information K+, Na+, and Ca2+, do not inhibit the activity Pseudomonas putida
1.13.11.4 Zn2+ 2.58% residual activity at 5 mM Pseudomonas alcaligenes
1.13.11.4 Zn2+ 4.69% residual activity at 5 mM Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.11.4 0.0043
-
 3-Isopropylgentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas putida
1.13.11.4 0.011
-
 3-Bromogentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas putida
1.13.11.4 0.015
-
 3-Bromogentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas alcaligenes
1.13.11.4 0.036
-
 3-Methylgentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas putida
1.13.11.4 0.053
-
 3-Isopropylgentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas alcaligenes
1.13.11.4 0.076
-
 3-Methylgentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas alcaligenes
1.13.11.4 0.092
-
 gentisate
-
 Pseudomonas alcaligenes
1.13.11.4 0.092
-
 gentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas alcaligenes
1.13.11.4 0.143
-
 gentisate
-
 Pseudomonas putida
1.13.11.4 0.143
-
 gentisate in 0.1 M phosphate buffer, pH 7.4, at 23°C Pseudomonas putida

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.13.11.4 Fe2+ strongly stimulated by 0.1 mM Fe2+ Pseudomonas putida
1.13.11.4 Fe2+ strongly stimulated by 0.1 mM Fe2+ Pseudomonas alcaligenes
1.13.11.4 additional information K+, Na+, and Ca2+, do not stimulate the activity Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.13.11.4 39000
-
 4 * 39000, SDS-PAGE Pseudomonas alcaligenes
1.13.11.4 41000
-
 2 * 41000 Pseudomonas putida
1.13.11.4 41000
-
 2 * 41000, gel filtration Pseudomonas putida
1.13.11.4 82000
-
 gel filtration Pseudomonas putida
1.13.11.4 154000
-
 gel filtration Pseudomonas alcaligenes

Organism

EC Number Organism UniProt Comment Textmining
1.13.11.4 Pseudomonas alcaligenes
-
-
-
1.13.11.4 Pseudomonas alcaligenes NCIB 9867
-
-
-
1.13.11.4 Pseudomonas putida
-
-
-
1.13.11.4 Pseudomonas putida NCIB 9869
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.11.4
-
 Pseudomonas putida
1.13.11.4
-
 Pseudomonas alcaligenes
1.13.11.4 ammonium sulfate fractionation, DEAE-Sepharose fast flow column chromatography Pseudomonas putida
1.13.11.4 ammonium sulfate fractionation, DEAE-Sepharose fast flow column chromatography Pseudomonas alcaligenes

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.13.11.4 0.45
-
 crude extract, at 23°C Pseudomonas alcaligenes
1.13.11.4 0.72
-
 crude extract, at 23°C Pseudomonas putida
1.13.11.4 23.2
-
 after 51.56fold purification, at 23°C Pseudomonas alcaligenes
1.13.11.4 43.14
-
 after 59.92fold purification, at 23°C Pseudomonas putida

Storage Stability

EC Number Storage Stability Organism
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 12 months, 3.5% residual activity Pseudomonas putida
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 12 months, 42.9% residual activity Pseudomonas alcaligenes
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 3 months, 42.5% residual activity Pseudomonas putida
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 3 months, 86.5% residual activity Pseudomonas alcaligenes
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 6 months, 23.1% residual activity Pseudomonas putida
1.13.11.4 -20°C, MOPS buffer with 0.1 mM Fe2+, 2 mM L-cysteine, and 10% (v/v) glycerol, 6 months, 70.9% residual activity Pseudomonas alcaligenes
1.13.11.4 4°C, MOPS buffer with no additives, 24 h, 22.5% residual activity Pseudomonas alcaligenes
1.13.11.4 4°C, MOPS buffer with no additives, 24 h, 33.9% residual activity Pseudomonas putida
1.13.11.4 4°C, MOPS buffer with no additives, 96 h, complete loss of activity Pseudomonas putida
1.13.11.4 4°C, MOPS buffer with no additives, 96 h, complete loss of activity 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 putida maleylpyruvate
-
 ?
1.13.11.4 2,5-dihydroxybenzoate + O2
-
 Pseudomonas alcaligenes maleylpyruvate
-
 ?
1.13.11.4 2,5-dihydroxybenzoate + O2
-
 Pseudomonas alcaligenes NCIB 9867 maleylpyruvate
-
 ?
1.13.11.4 2,5-dihydroxybenzoate + O2
-
 Pseudomonas putida NCIB 9869 maleylpyruvate
-
 ?
1.13.11.4 3-bromogentisate + O2
-
 Pseudomonas putida 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
 ?
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-bromogentisate + O2
-
 Pseudomonas alcaligenes NCIB 9867 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-bromogentisate + O2
-
 Pseudomonas putida NCIB 9869 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-isopropyl-gentisate + O2
-
 Pseudomonas putida 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-isopropyl-gentisate + O2
-
 Pseudomonas alcaligenes 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-isopropyl-gentisate + O2
-
 Pseudomonas alcaligenes NCIB 9867 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-isopropyl-gentisate + O2
-
 Pseudomonas putida NCIB 9869 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-methylgentisate + O2
-
 Pseudomonas putida 4-hydroxy-3-methyl-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 NCIB 9867 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 3-methylgentisate + O2
-
 Pseudomonas putida NCIB 9869 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid
-
 ?
1.13.11.4 gentisate + O2
-
 Pseudomonas putida maleylpyruvate
-
 ?
1.13.11.4 gentisate + O2
-
 Pseudomonas alcaligenes maleylpyruvate
-
 ?
1.13.11.4 gentisate + O2
-
 Pseudomonas alcaligenes NCIB 9867 maleylpyruvate
-
 ?
1.13.11.4 gentisate + O2
-
 Pseudomonas putida NCIB 9869 maleylpyruvate
-
 ?

Subunits

EC Number Subunits Comment Organism
1.13.11.4 dimer 2 * 41000 Pseudomonas putida
1.13.11.4 homodimer 2 * 41000, gel filtration Pseudomonas putida
1.13.11.4 homotetramer 4 * about 39000, gel filtration Pseudomonas alcaligenes
1.13.11.4 tetramer 4 * 39000, SDS-PAGE Pseudomonas alcaligenes

Synonyms

EC Number Synonyms Comment Organism
1.13.11.4 P25X gentisate 1,2-dioxygenase
-
 Pseudomonas alcaligenes
1.13.11.4 P35X gentisate 1,2-dioxygenase
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.13.11.4 50
-
-
 Pseudomonas putida
1.13.11.4 50
-
-
 Pseudomonas alcaligenes

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.13.11.4 50
-
 retains 35% of its activity when incubated at 50°C for 5 min Pseudomonas putida
1.13.11.4 50
-
 retains 73% of its activity when incubated at 50°C for 5 min Pseudomonas alcaligenes

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.11.4 8
-
-
 Pseudomonas putida
1.13.11.4 8
-
-
 Pseudomonas alcaligenes

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.13.11.4 5 7.5
-
 Pseudomonas alcaligenes
1.13.11.4 7 9
-
 Pseudomonas putida
1.13.11.4 7 9
-
 Pseudomonas alcaligenes

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.13.11.4 Pseudomonas putida isoelectric focusing 4.8 4.6
1.13.11.4 Pseudomonas alcaligenes isoelectric focusing
-
 4.85