Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | stimulated by 2 mM | Pseudomonas putida | |
2-mercaptoethanol | stimulated by 2 mM | Pseudomonas alcaligenes | |
Fe2+ | - |
Pseudomonas putida | |
Fe2+ | - |
Pseudomonas alcaligenes | |
glycerol | stimulated by 10% (v/v) | Pseudomonas putida | |
glycerol | stimulated by 10% (v/v) | Pseudomonas alcaligenes | |
L-cysteine | stimulated by 2 mM | Pseudomonas putida | |
L-cysteine | stimulated by 2 mM | Pseudomonas alcaligenes |
General Stability | Organism |
---|---|
highly unstable in 50 mM MOPS buffer with no additives | Pseudomonas putida |
highly unstable in 50 mM MOPS buffer with no additives | Pseudomonas alcaligenes |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | 5 mM complete inactivation; complete inhibition at 5 mM | Pseudomonas alcaligenes | |
Cu2+ | 5 mM complete inactivation; complete inhibition at 5 mM | Pseudomonas putida | |
EDTA | 60.24% residual activity at 5 mM | Pseudomonas alcaligenes | |
EDTA | 4.95% residual activity at 5 mM | Pseudomonas putida | |
Mg2+ | 66% residual activity at 5 mM | Pseudomonas putida | |
Mn2+ | 73.96% residual activity at 5 mM | Pseudomonas alcaligenes | |
Mn2+ | 22.36% residual activity at 5 mM | Pseudomonas putida | |
additional information | K+, Na+, Mg2+, and Ca2+ do not inhibit the activity | Pseudomonas alcaligenes | |
additional information | K+, Na+, and Ca2+, do not inhibit the activity | Pseudomonas putida | |
Zn2+ | 2.58% residual activity at 5 mM | Pseudomonas alcaligenes | |
Zn2+ | 4.69% residual activity at 5 mM | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0043 | - |
3-Isopropylgentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas putida | |
0.011 | - |
3-Bromogentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas putida | |
0.015 | - |
3-Bromogentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas alcaligenes | |
0.036 | - |
3-Methylgentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas putida | |
0.053 | - |
3-Isopropylgentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas alcaligenes | |
0.076 | - |
3-Methylgentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas alcaligenes | |
0.092 | - |
gentisate | - |
Pseudomonas alcaligenes | |
0.092 | - |
gentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas alcaligenes | |
0.143 | - |
gentisate | - |
Pseudomonas putida | |
0.143 | - |
gentisate | in 0.1 M phosphate buffer, pH 7.4, at 23°C | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | strongly stimulated by 0.1 mM Fe2+ | Pseudomonas putida | |
Fe2+ | strongly stimulated by 0.1 mM Fe2+ | Pseudomonas alcaligenes | |
additional information | K+, Na+, and Ca2+, do not stimulate the activity | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39000 | - |
4 * 39000, SDS-PAGE | Pseudomonas alcaligenes |
41000 | - |
2 * 41000 | Pseudomonas putida |
41000 | - |
2 * 41000, gel filtration | Pseudomonas putida |
82000 | - |
gel filtration | Pseudomonas putida |
154000 | - |
gel filtration | Pseudomonas alcaligenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas alcaligenes | - |
- |
- |
Pseudomonas alcaligenes NCIB 9867 | - |
- |
- |
Pseudomonas putida | - |
- |
- |
Pseudomonas putida NCIB 9869 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas putida |
- |
Pseudomonas alcaligenes |
ammonium sulfate fractionation, DEAE-Sepharose fast flow column chromatography | Pseudomonas putida |
ammonium sulfate fractionation, DEAE-Sepharose fast flow column chromatography | Pseudomonas alcaligenes |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.45 | - |
crude extract, at 23°C | Pseudomonas alcaligenes |
0.72 | - |
crude extract, at 23°C | Pseudomonas putida |
23.2 | - |
after 51.56fold purification, at 23°C | Pseudomonas alcaligenes |
43.14 | - |
after 59.92fold purification, at 23°C | Pseudomonas putida |
Storage Stability | Organism |
---|---|
-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 |
-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 |
-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 |
-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 |
-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 |
-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 |
4°C, MOPS buffer with no additives, 24 h, 22.5% residual activity | Pseudomonas alcaligenes |
4°C, MOPS buffer with no additives, 24 h, 33.9% residual activity | Pseudomonas putida |
4°C, MOPS buffer with no additives, 96 h, complete loss of activity | Pseudomonas putida |
4°C, MOPS buffer with no additives, 96 h, complete loss of activity | Pseudomonas alcaligenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,5-dihydroxybenzoate + O2 | - |
Pseudomonas putida | maleylpyruvate | - |
? | |
2,5-dihydroxybenzoate + O2 | - |
Pseudomonas alcaligenes | maleylpyruvate | - |
? | |
2,5-dihydroxybenzoate + O2 | - |
Pseudomonas alcaligenes NCIB 9867 | maleylpyruvate | - |
? | |
2,5-dihydroxybenzoate + O2 | - |
Pseudomonas putida NCIB 9869 | maleylpyruvate | - |
? | |
3-bromogentisate + O2 | - |
Pseudomonas putida | 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-bromogentisate + O2 | - |
Pseudomonas alcaligenes | 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-bromogentisate + O2 | - |
Pseudomonas alcaligenes NCIB 9867 | 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-bromogentisate + O2 | - |
Pseudomonas putida NCIB 9869 | 3-bromo-4-hydroxy-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-isopropyl-gentisate + O2 | - |
Pseudomonas putida | 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-isopropyl-gentisate + O2 | - |
Pseudomonas alcaligenes | 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-isopropyl-gentisate + O2 | - |
Pseudomonas alcaligenes NCIB 9867 | 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-isopropyl-gentisate + O2 | - |
Pseudomonas putida NCIB 9869 | 4-hydroxy-3-isopropyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-methylgentisate + O2 | - |
Pseudomonas putida | 4-hydroxy-3-methyl-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 NCIB 9867 | 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
3-methylgentisate + O2 | - |
Pseudomonas putida NCIB 9869 | 4-hydroxy-3-methyl-6-oxo-hepta-2,4-dienedioic acid | - |
? | |
gentisate + O2 | - |
Pseudomonas putida | maleylpyruvate | - |
? | |
gentisate + O2 | - |
Pseudomonas alcaligenes | maleylpyruvate | - |
? | |
gentisate + O2 | - |
Pseudomonas alcaligenes NCIB 9867 | maleylpyruvate | - |
? | |
gentisate + O2 | - |
Pseudomonas putida NCIB 9869 | maleylpyruvate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 41000 | Pseudomonas putida |
homodimer | 2 * 41000, gel filtration | Pseudomonas putida |
homotetramer | 4 * about 39000, gel filtration | Pseudomonas alcaligenes |
tetramer | 4 * 39000, SDS-PAGE | Pseudomonas alcaligenes |
Synonyms | Comment | Organism |
---|---|---|
P25X gentisate 1,2-dioxygenase | - |
Pseudomonas alcaligenes |
P35X gentisate 1,2-dioxygenase | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Pseudomonas putida |
50 | - |
- |
Pseudomonas alcaligenes |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
retains 35% of its activity when incubated at 50°C for 5 min | Pseudomonas putida |
50 | - |
retains 73% of its activity when incubated at 50°C for 5 min | Pseudomonas alcaligenes |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Pseudomonas putida |
8 | - |
- |
Pseudomonas alcaligenes |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 7.5 | - |
Pseudomonas alcaligenes |
7 | 9 | - |
Pseudomonas putida |
7 | 9 | - |
Pseudomonas alcaligenes |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudomonas putida | isoelectric focusing | 4.8 | 4.6 |
Pseudomonas alcaligenes | isoelectric focusing | - |
4.85 |