Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | isozyme II is strictly inducible by specific aromatic substrates | Pseudomonas alcaligenes |
Cloned (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Pseudomonas alcaligenes | |
Cu2+ | complete inhibition at 0.05 mM | Pseudomonas alcaligenes | |
Fe2+ | complete inhibition at 5 mM | Pseudomonas alcaligenes | |
Hg2+ | complete inhibition at 0.05 mM | Pseudomonas alcaligenes | |
Mn2+ | complete inhibition at 5 mM | Pseudomonas alcaligenes | |
Zn2+ | - |
Pseudomonas alcaligenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.071 | - |
3-hydroxy-4-methylbenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADPH | Pseudomonas alcaligenes | |
0.079 | - |
3-hydroxybenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADH | Pseudomonas alcaligenes | |
0.095 | - |
3-hydroxy-4-methylbenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADH | Pseudomonas alcaligenes | |
0.108 | - |
3-hydroxybenzoate | pH 7.5, 23°C, recombinant enzyme, cofactor NADPH | Pseudomonas alcaligenes |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | slight activation of 13% at 5 mM | Pseudomonas alcaligenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
43000 | - |
3 * 43000, His-tagged recombinant enzyme, SDS-PAGE | Pseudomonas alcaligenes |
130000 | - |
His-tagged recombinant enzyme, native PAGE and gel filtration | Pseudomonas alcaligenes |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | - |
? | |
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 | - |
? | |
additional information | Pseudomonas alcaligenes | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | ? | - |
? | |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas alcaligenes | Q9F131 | gene xlnD, two isozymes I and II | - |
Pseudomonas alcaligenes P25X | Q9F131 | gene xlnD, two isozymes I and II | - |
Purification (Comment) | Organism |
---|---|
recombinnat His-tagged enzyme from Escherichia coli by metal chelating chromatography | Pseudomonas alcaligenes |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | effects of carbon sources for growth on enzyme substrate specificity, overview | Pseudomonas alcaligenes | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.63 | - |
recombinant His-tagged enzyme, substrate 3-hydroxy-5-methylbenzoate | Pseudomonas alcaligenes |
Storage Stability | Organism |
---|---|
-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 |
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 |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-aminobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
3-aminobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | ? | - |
? | |
3-bromobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
3-bromobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | ? | - |
? | |
3-chlorobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
3-fluorobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
3-hydroxy-3-methylbenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | 2,5-dihydroxy-3-methylbenzoate + NAD(P)+ + H2O | - |
? | |
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 | - |
? | |
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 | - |
? | |
3-hydroxybenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | i.e. gentisate | ? | |
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 | - |
? | |
3-hydroxybenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes P25X | 2,5-dihydroxybenzoate + NAD(P)+ + H2O | i.e. gentisate | ? | |
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 | - |
? | |
3-methylthiobenzoate + NAD(P)H + O2 | - |
Pseudomonas alcaligenes | ? | - |
? | |
additional information | degradation of 2,5-xylenol and 3,5-xylenol in strain P25X depends on isozyme I, not isozyme II | Pseudomonas alcaligenes | ? | - |
? | |
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 | Comment | Organism |
---|---|---|
trimer | 3 * 43000, His-tagged recombinant enzyme, SDS-PAGE | Pseudomonas alcaligenes |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxybenzoate-6-hydroxylase | - |
Pseudomonas alcaligenes |
XlnD | - |
Pseudomonas alcaligenes |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
- |
Pseudomonas alcaligenes |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 50 | no activity below 15°C and above 50°C | Pseudomonas alcaligenes |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Pseudomonas alcaligenes |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | - |
Pseudomonas alcaligenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | XlnD can utilize either NADH or NADPH as the electron donor, NADH is preferred | Pseudomonas alcaligenes | |
NADPH | XlnD can utilize either NADH or NADPH as the electron donor, NADH is preferred | Pseudomonas alcaligenes |