EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
1.13.11.8 | additional information | allosteric rate enhancement in the presence of non-substrate protocatechuate-like aldehydes such as vanillin | Sphingobium sp. | |
1.13.11.8 | Vanillin | heteroallosteric activation of LigAB by vanillin at an allosteric binding site in LigAB, regulatory feed-forward activation mechanism, kinetics, overview. Vanillin is the pre-pre-substrate of LigAB. Enhancement by vanillin is specific to protocatechuate. Glu86b is directly involved in a hydrogen bond network through the carboxylate with Ndelta of His127b, believed to be the catalytic base that performs the initial substrate deprotonation. The His127b methylene also contributes a small fraction to the identified allosteric binding pocket | Sphingobium sp. |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.13.11.8 | recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha | Sphingobium sp. |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.13.11.8 | A18W | site-directed mutagenesis, mutation in the betaa-subunit, residue Ala18b is located opening of the putative allosteric pocket, the mutant displays reduced activity and altered rate enhancement by vanillin compared to the wild-type enzyme | Sphingobium sp. |
1.13.11.8 | F103H | site-directed mutagenesis, mutation in the alpha-subunit, residue Phe103a is located in the putative allosteric pocket, the mutant displays reduced activity and altered rate enhancement by vanillin compared to the wild-type enzyme | Sphingobium sp. |
1.13.11.8 | F103L | site-directed mutagenesis, mutation in the alpha-subunit, residue Phe103a is located in the putative allosteric pocket, the mutant displays reduced activity and altered rate enhancement by vanillin compared to the wild-type enzyme | Sphingobium sp. |
1.13.11.8 | F103T | site-directed mutagenesis, mutation in the alpha-subunit, residue Phe103a is located in the putative allosteric pocket, the mutant displays reduced activity and altered rate enhancement by vanillin compared to the wild-type enzyme | Sphingobium sp. |
1.13.11.8 | F103V | site-directed mutagenesis, mutation in the alpha-subunit, residue Phe103a is located in the putative allosteric pocket, the mutant displays reduced activity and altered rate enhancement by vanillin compared to the wild-type enzyme | Sphingobium sp. |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.13.11.8 | protocatechuate | substrate inhibition | Sphingobium sp. |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.8 | additional information | - |
additional information | Michaelis-Menten steady-state kinetics measuring O2 consumption | Sphingobium sp. | |
1.13.11.8 | 0.066 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant A18W | Sphingobium sp. | |
1.13.11.8 | 0.081 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme | Sphingobium sp. | |
1.13.11.8 | 0.108 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103H | Sphingobium sp. | |
1.13.11.8 | 0.13 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme, with vanillin | Sphingobium sp. | |
1.13.11.8 | 0.21 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103T | Sphingobium sp. | |
1.13.11.8 | 0.26 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103V | Sphingobium sp. | |
1.13.11.8 | 1.6 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103L | Sphingobium sp. |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.8 | protocatechuate + O2 | Sphingobium sp. | - |
4-carboxy-2-hydroxymuconate semialdehyde | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.13.11.8 | Sphingobium sp. | G2IQQ4 AND G2IQQ3 | alpha- and beta-chain of LigAB; formerly Sphingomonas paucimobilis SYK-6 | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.13.11.8 | recombinant His6-tagged wild-type and mutant enzymes anaerobically from Escherichia coli strain DH5alpha by nickel affinity chromatography, tag cleavage anaerobically by thrombin cleavage | Sphingobium sp. |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.8 | additional information | LigAB has the broadest substrate utilization profile | Sphingobium sp. | ? | - |
? | |
1.13.11.8 | protocatechuate + O2 | - |
Sphingobium sp. | 4-carboxy-2-hydroxymuconate semialdehyde | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.13.11.8 | LigAB | - |
Sphingobium sp. |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.13.11.8 | 25 | - |
assay at | Sphingobium sp. |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.8 | 33 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103L | Sphingobium sp. | |
1.13.11.8 | 89 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103V | Sphingobium sp. | |
1.13.11.8 | 110 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103H | Sphingobium sp. | |
1.13.11.8 | 112 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103T | Sphingobium sp. | |
1.13.11.8 | 122 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant A18W | Sphingobium sp. | |
1.13.11.8 | 178 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme | Sphingobium sp. | |
1.13.11.8 | 230 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme, with vanillin | Sphingobium sp. |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.13.11.8 | 7.5 | - |
assay at | Sphingobium sp. |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.8 | 17 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme, with vanillin | Sphingobium sp. | |
1.13.11.8 | 18 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme | Sphingobium sp. |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.13.11.8 | evolution | protocatechuate 4,5-dioxygenase (LigAB) from Sphingobium sp. strain SYK-6 is the defining member of the type II extradiol dioxygenase superfamily, a.k.a. PCA dioxygenase superfamily or PCADSF, computational docking and sequence comparisons of the enzyme with PCA dioxygenase superfamily LigAB enzymes | Sphingobium sp. |
1.13.11.8 | physiological function | the enzyme plays a key aromatic ring-opening role in the metabolism of several lignin derived aromatic compounds | Sphingobium sp. |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.8 | 21 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103L | Sphingobium sp. | |
1.13.11.8 | 340 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103V | Sphingobium sp. | |
1.13.11.8 | 530 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103T | Sphingobium sp. | |
1.13.11.8 | 1100 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant F103H | Sphingobium sp. | |
1.13.11.8 | 1800 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme, with vanillin | Sphingobium sp. | |
1.13.11.8 | 1900 | - |
protocatechuate | pH 7.5, 25°C, recombinant mutant A18W | Sphingobium sp. | |
1.13.11.8 | 2200 | - |
protocatechuate | pH 7.5, 25°C, recombinant wild-type enzyme | Sphingobium sp. |