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Literature summary for 3.1.1.57 extracted from

  • Hobbs, M.E.; Malashkevich, V.; Williams, H.J.; Xu, C.; Sauder, J.M.; Burley, S.K.; Almo, S.C.; Raushel, F.M.
    Structure and catalytic mechanism of LigI: Insight into the amidohydrolase enzymes of cog3618 and lignin degradation (2012), Biochemistry, 51, 3497-3507.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
His-tagged version expressed in Escherichia coli BL21(DE3) Sphingomonas paucimobilis

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method Sphingomonas paucimobilis

Protein Variants

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

Inhibitors Comment Organism Structure
5-hydroxyisophthalic acid
-
Sphingomonas paucimobilis
pyridine-2,4-dicarboxylic acid
-
Sphingomonas paucimobilis

KM Value [mM]

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

Organism UniProt Comment Textmining
Sphingomonas paucimobilis O87170
-
-

Purification (Commentary)

Purification (Comment) Organism
immobilized metal ion affinity chromatography (Ni2+), gel filtration Sphingomonas paucimobilis

Substrates and Products (Substrate)

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

Synonyms Comment Organism
2-pyrone-4,6-dicarboxylate lactonase
-
Sphingomonas paucimobilis
LigI
-
Sphingomonas paucimobilis

Turnover Number [1/s]

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 [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 [mM/s]

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