Cloned (Comment) | Organism |
---|---|
gene ncgl2923, sequence determination, analysis, and comparisons, functional overexpression in of N-terminally His-tagged Ncgl2923 in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Corynebacterium glutamicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0534 | - |
3-hydroxybenzoate | recombinant enzyme, with NADH, pH 7.5, 25°C | Corynebacterium glutamicum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
49000 | - |
3 * 49000, recombinant Ncgl2923, SDS-PAGE | Corynebacterium glutamicum |
125000 | - |
recombinant Ncgl2923, gel filtration | Corynebacterium glutamicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxybenzoate + NADH + H+ + O2 | Corynebacterium glutamicum | - |
2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADH + H+ + O2 | Corynebacterium glutamicum ATCC 13032 | - |
2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADPH + H+ + O2 | Corynebacterium glutamicum | - |
2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADPH + H+ + O2 | Corynebacterium glutamicum ATCC 13032 | - |
2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | Q8NLB6 | gene ncgl2923 | - |
Corynebacterium glutamicum ATCC 13032 | Q8NLB6 | gene ncgl2923 | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged Ncgl2923 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity | Corynebacterium glutamicum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
3-hydroxybenzoate + NADH + H+ + O2 = 2,5-dihydroxybenzoate + NAD+ + H2O | catalytic mechanism involving FAD, overview | Corynebacterium glutamicum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
6.92 | - |
purified recombinant enzyme, with NADH, pH 7.5, 25°C | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxybenzoate + NADH + H+ + O2 | - |
Corynebacterium glutamicum | 2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADH + H+ + O2 | - |
Corynebacterium glutamicum ATCC 13032 | 2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADPH + H+ + O2 | - |
Corynebacterium glutamicum | 2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
3-hydroxybenzoate + NADPH + H+ + O2 | - |
Corynebacterium glutamicum ATCC 13032 | 2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? |
Subunits | Comment | Organism |
---|---|---|
trimer | 3 * 49000, recombinant Ncgl2923, SDS-PAGE | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxybenzoate 6-hydroxylase | - |
Corynebacterium glutamicum |
Ncgl2923 | - |
Corynebacterium glutamicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Corynebacterium glutamicum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | high activity range | Corynebacterium glutamicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | 3fold activation at 0.015 mM. Sequence analysis of FAD/NADPH-binding domain of Ncgl2923, overview | Corynebacterium glutamicum | |
NADH | dependent on, preferred cofactor | Corynebacterium glutamicum | |
NADPH | less active than NADH | Corynebacterium glutamicum |
General Information | Comment | Organism |
---|---|---|
metabolism | strain ATCC 13032 metabolizes 3-hydroxybenzoate, the 3-hydroxybenzoate 6-hydroxylase is involved in the initial step of 3-hydroxybenzoate catabolismate via gentisate | Corynebacterium glutamicum |
physiological function | the enzyme catalyzes the NADH-dependent conversion of 3-hydroxybenzoate to gentisate | Corynebacterium glutamicum |