Cloned (Comment) | Organism |
---|---|
His-tagged version expressed in Escherichia coli BL21(DE3) | Sphingomonas paucimobilis |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method | Sphingomonas paucimobilis |
Protein Variants | Comment | Organism |
---|---|---|
D248A | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
D248N | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
H180A | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
H180C | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
H31N | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
H33N | prototypical metal-coordinating residue in the amidohydrolase superfamily | Sphingomonas paucimobilis |
R124M | active site residue | Sphingomonas paucimobilis |
R130M | active site residue | Sphingomonas paucimobilis |
R183M | active site residue | Sphingomonas paucimobilis |
R217M | active site residue | Sphingomonas paucimobilis |
Y156F | active site residue | Sphingomonas paucimobilis |
Y49F | active site residue | Sphingomonas paucimobilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5-hydroxyisophthalic acid | - |
Sphingomonas paucimobilis | |
pyridine-2,4-dicarboxylic acid | - |
Sphingomonas paucimobilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.041 | - |
2-Pyrone-4,6-dicarboxylate | H180C mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.048 | - |
2-Pyrone-4,6-dicarboxylate | wild type protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.064 | - |
2-Pyrone-4,6-dicarboxylate | R183M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.066 | - |
2-Pyrone-4,6-dicarboxylate | D248N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.067 | - |
2-Pyrone-4,6-dicarboxylate | R217M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.089 | - |
2-Pyrone-4,6-dicarboxylate | H33N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.09 | - |
2-Pyrone-4,6-dicarboxylate | R124M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.116 | - |
2-Pyrone-4,6-dicarboxylate | D248A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.214 | - |
2-Pyrone-4,6-dicarboxylate | H180A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.217 | - |
2-Pyrone-4,6-dicarboxylate | Y49F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.238 | - |
2-Pyrone-4,6-dicarboxylate | Y156F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.465 | - |
2-Pyrone-4,6-dicarboxylate | R130M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.72 | - |
2-Pyrone-4,6-dicarboxylate | H31N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sphingomonas paucimobilis | O87170 | - |
- |
Purification (Comment) | Organism |
---|---|
immobilized metal ion affinity chromatography (Ni2+), gel filtration | Sphingomonas paucimobilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-pyrone-4,6-dicarboxylate + H2O | - |
Sphingomonas paucimobilis | 4-oxalomesaconate | the product occurs in two tautomeric forms. 4-oxalomesaconate is predominant at pH 10, while at pH 6 4-carboxy-2-hydroxymuconate is observed | r |
Synonyms | Comment | Organism |
---|---|---|
2-pyrone-4,6-dicarboxylate lactonase | - |
Sphingomonas paucimobilis |
LigI | - |
Sphingomonas paucimobilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0013 | - |
2-Pyrone-4,6-dicarboxylate | D248A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.02 | - |
2-Pyrone-4,6-dicarboxylate | R124M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.023 | - |
2-Pyrone-4,6-dicarboxylate | D248N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.4 | - |
2-Pyrone-4,6-dicarboxylate | H31N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.42 | - |
2-Pyrone-4,6-dicarboxylate | R217M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
3.6 | - |
2-Pyrone-4,6-dicarboxylate | H33N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
5 | - |
2-Pyrone-4,6-dicarboxylate | R130M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
8 | - |
2-Pyrone-4,6-dicarboxylate | H180A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
8.7 | - |
2-Pyrone-4,6-dicarboxylate | Y156F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
26 | - |
2-Pyrone-4,6-dicarboxylate | Y49F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
32 | - |
2-Pyrone-4,6-dicarboxylate | H180C mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
196 | - |
2-Pyrone-4,6-dicarboxylate | R183M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
342 | - |
2-Pyrone-4,6-dicarboxylate | wild type protein, pH 8.25, 30°C | Sphingomonas paucimobilis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
5-hydroxyisophthalic acid | pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.075 | - |
pyridine-2,4-dicarboxylic acid | pH 8.25, 30°C | Sphingomonas paucimobilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.016 | - |
2-Pyrone-4,6-dicarboxylate | D248A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.2 | - |
2-Pyrone-4,6-dicarboxylate | D248N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.2 | - |
2-Pyrone-4,6-dicarboxylate | R124M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
0.5 | - |
2-Pyrone-4,6-dicarboxylate | H31N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
6.3 | - |
2-Pyrone-4,6-dicarboxylate | R217M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
11 | - |
2-Pyrone-4,6-dicarboxylate | R130M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
37 | - |
2-Pyrone-4,6-dicarboxylate | H180A mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
37 | - |
2-Pyrone-4,6-dicarboxylate | Y156F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
41 | - |
2-Pyrone-4,6-dicarboxylate | H33N mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
120 | - |
2-Pyrone-4,6-dicarboxylate | Y49F mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
780 | - |
2-Pyrone-4,6-dicarboxylate | H180C mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
3000 | - |
2-Pyrone-4,6-dicarboxylate | R183M mutant protein, pH 8.25, 30°C | Sphingomonas paucimobilis | |
7100 | - |
2-Pyrone-4,6-dicarboxylate | wild type protein, pH 8.25, 30°C | Sphingomonas paucimobilis |