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Literature summary extracted from
- Molecular and biochemical characterization of the xlnD-encoded 3-hydroxybenzoate 6-hydroxylase involved in the degradation of 2,5-xylenol via the gentisate pathway in Pseudomonas alcaligenes NCIMB 9867 (2005), J. Bacteriol., 187, 7696-7702.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.24 | additional information | isozyme II is strictly inducible by specific aromatic substrates | Pseudomonas alcaligenes |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.13.24 | gene xlnD, DNA and amino acid sequence determination and analysis, subcloning and overexpression of the His-tagged enzyme in Escherichia coli strain DH5alpha, S17-1, and BL21(DE3) | Pseudomonas alcaligenes |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.13.24 | additional information | construction of a P25X xlnD knockout mutant strain G50, no induction after growth on lactate and 3-hydroxy-4-methylbenzoate in contrast to the wild-type enzyme, slightly reduced activity with 3-hydroxybenzoate and gentisate | Pseudomonas alcaligenes |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.24 | Ca2+ | - |
Pseudomonas alcaligenes |  |
| 1.14.13.24 | Cu2+ | complete inhibition at 0.05 mM | Pseudomonas alcaligenes | |
| 1.14.13.24 | Fe2+ | complete inhibition at 5 mM | Pseudomonas alcaligenes | |
| 1.14.13.24 | Hg2+ | complete inhibition at 0.05 mM | Pseudomonas alcaligenes | |
| 1.14.13.24 | Mn2+ | complete inhibition at 5 mM | Pseudomonas alcaligenes | |
| 1.14.13.24 | Zn2+ | - |
Pseudomonas alcaligenes | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.24 | 0.071 | - |
3-hydroxy-4-methylbenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADPH | Pseudomonas alcaligenes | |
| 1.14.13.24 | 0.079 | - |
3-hydroxybenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADH | Pseudomonas alcaligenes | |
| 1.14.13.24 | 0.095 | - |
3-hydroxy-4-methylbenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADH | Pseudomonas alcaligenes | |
| 1.14.13.24 | 0.108 | - |
3-hydroxybenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADPH | Pseudomonas alcaligenes | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.24 | K+ | slight activation of 13% at 5 mM | Pseudomonas alcaligenes | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 43000 | - |
3 * 43000, His-tagged recombinant enzyme, SDS-PAGE | Pseudomonas alcaligenes |
| 1.14.13.24 | 130000 | - |
His-tagged recombinant enzyme, native PAGE and gel filtration | Pseudomonas alcaligenes |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | Pseudomonas alcaligenes | the enzyme is involved in the degradation of 2,5-xylenol and 3,5-xylenol via the gentisate pathway, overview | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | Pseudomonas alcaligenes P25X | the enzyme is involved in the degradation of 2,5-xylenol and 3,5-xylenol via the gentisate pathway, overview | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | additional information | Pseudomonas alcaligenes | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | ? | - |
? | |
| 1.14.13.24 | additional information | Pseudomonas alcaligenes P25X | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.13.24 | Pseudomonas alcaligenes | Q9F131 | gene xlnD, two isozymes I and II | - |
| 1.14.13.24 | Pseudomonas alcaligenes P25X | Q9F131 | gene xlnD, two isozymes I and II | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.13.24 | recombinnat His-tagged enzyme from Escherichia coli by metal chelating chromatography | Pseudomonas alcaligenes |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 1.14.13.24 | additional information | effects of carbon sources for growth on enzyme substrate specificity, overview | Pseudomonas alcaligenes | - |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 1.63 | - |
recombinant His-tagged enzyme, substrate 3-hydroxy-5-methylbenzoate | Pseudomonas alcaligenes |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.13.24 | -80°C, purified recombinant enzyme, 50 mM MOPS or 100 mM potassium phosphate, 10% v/v glycerol, 0.2 mM FAD, 60% activity remaining after 3 months | Pseudomonas alcaligenes |
| 1.14.13.24 | 22°C, rom temperature, purified recombinant enzyme, 50 mM MOPS or 100 mM potassium phosphate, 10% v/v glycerol, 0.2 mM FAD, complete loss of activity after 12 h | Pseudomonas alcaligenes |
| 1.14.13.24 | 4°C, purified recombinant enzyme, 50 mM MOPS or 100 mM potassium phosphate, 10% v/v glycerol, 0.2 mM FAD, complete loss of activity after 3 days | Pseudomonas alcaligenes |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.24 | 3-aminobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | 3-aminobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | ? | - |
? | |
| 1.14.13.24 | 3-bromobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | 3-bromobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | ? | - |
? | |
| 1.14.13.24 | 3-chlorobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | 3-fluorobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | 3-hydroxy-3-methylbenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | 2,5-dihydroxy-3-methylbenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-hydroxy-4-methylbenzoate + NAD(P)H + O2 | the substrate is an intermediate in the degradation of 2,5-xylenol, higher activity with the recombinant His-tagged enzyme compared to the native enzyme | Pseudomonas alcaligenes | 2,5-dihydroxy-4-methylbenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-hydroxy-5-methylbenzoate + NAD(P)H + O2 | the substrate is an intermediate in the degradation of 3,5-xylenol, higher activity with the recombinant His-tagged enzyme compared to the native enzyme, preferred substrate | Pseudomonas alcaligenes | 2,5-dihydroxy-3-methylbenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | i.e. gentisate | ? | |
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | the enzyme is involved in the degradation of 2,5-xylenol and 3,5-xylenol via the gentisate pathway, overview | Pseudomonas alcaligenes | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | i.e. gentisate | ? | |
| 1.14.13.24 | 3-hydroxybenzoate + NAD(P)H + O2 | the enzyme is involved in the degradation of 2,5-xylenol and 3,5-xylenol via the gentisate pathway, overview | Pseudomonas alcaligenes P25X | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | - |
? | |
| 1.14.13.24 | 3-methylthiobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | additional information | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | Pseudomonas alcaligenes | ? | - |
? | |
| 1.14.13.24 | additional information | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | Pseudomonas alcaligenes P25X | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.13.24 | trimer | 3 * 43000, His-tagged recombinant enzyme, SDS-PAGE | Pseudomonas alcaligenes |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.13.24 | 3-hydroxybenzoate-6-hydroxylase | - |
Pseudomonas alcaligenes |
| 1.14.13.24 | XlnD | - |
Pseudomonas alcaligenes |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 25 | - |
- |
Pseudomonas alcaligenes |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 15 | 50 | no activity below 15°C and above 50°C | Pseudomonas alcaligenes |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 7.5 | - |
- |
Pseudomonas alcaligenes |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.24 | 6 | 9 | - |
Pseudomonas alcaligenes |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.24 | NADH | XlnD can utilize either NADH or NADPH as the electron donor, NADH is preferred | Pseudomonas alcaligenes | |
| 1.14.13.24 | NADPH | XlnD can utilize either NADH or NADPH as the electron donor, NADH is preferred | Pseudomonas alcaligenes | |
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