Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 cells | Pseudomonas alcaligenes |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas alcaligenes | - |
- |
- |
Pseudomonas alcaligenes NCIMB 9867 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
GSTrap fast flow column chromatography | Pseudomonas alcaligenes |
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 | Comment | Organism |
---|---|---|
gentisate dioxygenase | - |
Pseudomonas alcaligenes |